Author Report for: Wu JContact information:Wu JieDivisions of Neurology, Barrow Neurological Institute, St. Joseph's Hospital and Medical Center, Phoenix, ArizonaUSAPhone : Fax : Send E-Mail to Wu Jie
Chen R, Gao X, Nie T, Wu J, Wang L, Osman A, Feng Y, Li X, Zhang Y Ref: Acta Biochim Biophys Sin (Shanghai), :, 2023 : PubMed ESTHER: Chen_2023_Acta.Biochim.Biophys.Sin.(Shanghai)__ PubMedSearch: Chen 2023 Acta.Biochim.Biophys.Sin.(Shanghai) PubMedID: 37705347 Abstract Esterases/lipases from the GDSL family have potential applications in the hydrolysis and synthesis of important esters of pharmaceutical, food, and biotechnical interests. However, the structural and functional understanding of GDSL enzymes is still limited. Here, we report the crystal structure of the GDSL family esterase EstL5 complexed with PMSF at 2.34 A resolution. Intriguingly, the PMSF binding site is not located at the active site pocket but is situated in a surface cavity. At the active site, we note that there is a trapped crystallization solvent 1,6-hexanediol, which mimics the bound ester chain, allowing for further definition of the active site pocket of EstL5. The most striking structural feature of EstL5 is the presence of a unique channel, which extends approximately 18.9 A, with a bottleneck radius of 6.8 A, connecting the active-site pocket and the surface cavity. Replacement of Ser205 with the bulk aromatic residue Trp or Phe could partially block the channel at one end and perturb its access. Reduced enzymatic activity is found in the EstL5 S205W and EstL5 S205F mutants, suggesting the functional relevance of the channel to enzyme catalysis. Our study provides valuable information regarding the properties of the GDSL-family enzymes for designing more efficient and robust biocatalysts. Title: Genetic analysis and management of a familial hypercholesterolemia pedigree with polygenic variants: Case report Han Y, Zhang L, Tao H, Wu J, Zhai J Ref: Medicine (Baltimore), 102:e34534, 2023 : PubMed ESTHER: Han_2023_Medicine.(Baltimore)_102_e34534 PubMedSearch: Han 2023 Medicine.(Baltimore) 102 e34534 PubMedID: 37565868 Abstract RATIONALE: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disorder typically caused by low density lipoprotein receptor (LDLR) gene mutation. Herein, we reported a FH pedigree with polygenic variants: LDLR, apolipoprotein B (APOB), and epoxide hydrolase 2 (EPHX2). PATIENT CONCERNS: A 10-year-old boy mainly presented multiple skin xanthomas and hypercholesterolemia. His family visited our hospital and was performed with pedigree whole exome sequencing (WES) at 20 + 3 weeks gestation of the mother's second pregnancy. DIAGNOSES: Based on the clinical features and genetic analysis, the pedigree was diagnosed with familial hypercholesterolemia. INTERVENTIONS: After genetic counseling, the couple opted to continue the pregnancy. Treatment advice and follow-up were offered to them. OUTCOMES: A novel compound heterozygous LDLR mutation: c.1009G>T and c.68-2A>G, derived from his parents respectively was revealed through pedigree WES, meanwhile, a maternal APOB gene variant: c.1670A>G and a paternal EPHX2 gene variant: c.548 dup of the proband were found together. Furthermore, the same compound heterozygous LDLR mutation as his was confirmed in his sister without APOB and EPHX2 variants in her fetal stage. LESSONS: WES combined with clinical features is essential for the diagnosis of FH, however, prenatal genetic testing results might bring more challenges to prenatal genetic counseling. Furthermore, it is more important to provide the guidance and early intervention for such families in the long run. Title: The FoxO1-ATGL axis alters milk lipolysis homeostasis through PI3K/AKT signaling pathway in dairy goat mammary epithelial cells He Q, GaoLiang J, Zhang F, Yao W, Wu J, Song N, Luo J, Zhang Y Ref: J Anim Sci, :, 2023 : PubMed ESTHER: He_2023_J.Anim.Sci__ PubMedSearch: He 2023 J.Anim.Sci PubMedID: 37638641 Abstract Goat milk is enriched in fatty acids which beneficial to human health. Previous research has revealed that 98% of milk fat is composed of triglycerides. However, the mechanisms regulating of milk fat composition remain unclear. Forkhead box protein O1 (FoxO1) is a crucial regulatory factor involved in lipid metabolism across various cell types. Chromatin immunoprecipitation sequencing (ChIP)--seq data) and RNA sequencing (RNA-seq) data revealed thathave indicated a close association between FoxO1 was closed related toand lipid metabolism during lactation in dairy goats. The objective of this study was to investigate the mechanisms by which FoxO1 regulates lipid metabolism in goat mammary epithelial cells (GMECs). FoxO1 knockdown significantly downregulated the expression of adipose triglyceride lipase (ATGL) and suppressed the activity of the ATGL promoter. Consistently, the number of lipid droplets decreased significantly in FoxO1-overexpressing cells, and increased in ATGL-knockdown cells. To further verify the effect of FoxO1 on ATGL promoter activity, cells were transfected with four promoter fragments of different lengths. We found that the core region of the ATGL promoter was located between -882 bp and -524 bp, encompassing two FoxO1 binding sites (FKH1 and FKH2). Mutations in the FoxO1 binding sites significantly downregulated ATGL promoter activity in GMECs. Luciferase reporter assays demonstrated that FoxO1 overexpression markedly enhanced ATGL promoter activity. Furthermore, site-directed mutation confirmed that FKH1 and FKH2 sites were simultaneously mutated significantly attenuated the stimulatory effect of FoxO1 on ATGL promoter activities simultaneous mutation of FKH1 and FKH2 sites significantly attenuated the stimulatory effect of FoxO1 on ATGL promoter activity. ChIP assays showed that FoxO1 directly binds to the FKH2 element located in the ATGL promoter in vivo. Finally, immunofluorescence staining revealed that insulin promotes the translocation of FoxO1 from the nucleus to the cytoplasm, thereby attenuating the FoxO1-induced activation of the ATGL promoter. Collectively, these findings uncover a novel pathway where by FoxO1 may regulate lipid metabolism in GMECs specifically by modulating the transcriptional activity of ATGL. Title: Maize resistance to witchweed through changes in strigolactone biosynthesis Li C, Dong L, Durairaj J, Guan JC, Yoshimura M, Quinodoz P, Horber R, Gaus K, Li J and Bouwmeester HJ <21 more author(s)> Li C, Dong L, Durairaj J, Guan JC, Yoshimura M, Quinodoz P, Horber R, Gaus K, Li J, Setotaw YB, Qi J, De Groote H, Wang Y, Thiombiano B, Flokova K, Walmsley A, Charnikhova TV, Chojnacka A, Correia de Lemos S, Ding Y, Skibbe D, Hermann K, Screpanti C, De Mesmaeker A, Schmelz EA, Menkir A, Medema M, van Dijk ADJ, Wu J, Koch KE, Bouwmeester HJ (- 21) Ref: Science, 379:94, 2023 : PubMed ESTHER: Li_2023_Science_379_94 PubMedSearch: Li 2023 Science 379 94 PubMedID: 36603079 Abstract Maize (Zea mays) is a major staple crop in Africa, where its yield and the livelihood of millions are compromised by the parasitic witchweed Striga. Germination of Striga is induced by strigolactones exuded from maize roots into the rhizosphere. In a maize germplasm collection, we identified two strigolactones, zealactol and zealactonoic acid, which stimulate less Striga germination than the major maize strigolactone, zealactone. We then showed that a single cytochrome P450, ZmCYP706C37, catalyzes a series of oxidative steps in the maize-strigolactone biosynthetic pathway. Reduction in activity of this enzyme and two others involved in the pathway, ZmMAX1b and ZmCLAMT1, can change strigolactone composition and reduce Striga germination and infection. These results offer prospects for breeding Striga-resistant maize. Title: Carboxy-Functionalized Covalent Organic Framework as a Carrier for Lipase Immobilization and Its Application in Inhibitors Screening Liu X, Wu J, Yang S, Li L, Ji Y Ref: Appl Biochem Biotechnol, :, 2023 : PubMed ESTHER: Liu_2023_Appl.Biochem.Biotechnol__ PubMedSearch: Liu 2023 Appl.Biochem.Biotechnol PubMedID: 37819460 Abstract Covalent organic frameworks (COFs) with large specific surface areas, high porosity, good stability, and designable structure are promising carriers for immobilized enzymes. It is important to explore lipase inhibitors from natural foods as lipase inhibitors are closely related to the treatment of obesity. In this work, a carboxyl functionalized covalent organic framework (TpBD-3COOH) was prepared by solvothermal method for covalent immobilization of porcine pancreatic lipase (PPL) and obtained the enzyme-decorated COF (PPL@COF). The immobilized lipase showed wider pH and temperature tolerance with the same optimal pH and temperature of 7.5 and 50 degC compared to free lipase. After 6 successive reuses, the PPL@COF maintained 53.0% of its original activity. Immobilized lipase also displayed enhanced storage stability (55.4% after 14 days at 4 degC). When p-nitrophenyl acetate was applied as the substrate, the calculated Michaelis constant was 3.57 mM and the half maximal inhibitory concentration of orlistat was 3.20 microM. Finally, the PPL@COF was used for enzyme inhibitors screening from natural foods combined with UV spectrophotometry, and Hawthorn was screened for excellent lipase inhibitory activity. Title: Ultra-small magnetic Candida antarctica lipase B nanoreactors for enzyme synthesis of bixin-maltitol ester Lv D, Wang M, He W, Wu J, Liu X, Guan Y Ref: Food Chem, 421:136132, 2023 : PubMed ESTHER: Lv_2023_Food.Chem_421_136132 PubMedSearch: Lv 2023 Food.Chem 421 136132 PubMedID: 37094396 Abstract Bixin has desirable bioactivities but poor water solubility, which limits its practical applications. Enzymatic transesterification of methyl to alditol groups in bixin by Candida antarctica lipase B (CALB) improves bixin water solubility. Herein, magnetic CALB nanoreactors with diameter of 11.7 nm and CALB layer thickness of 3.5 nm were developed by covalently linking CALB onto silicon covered Fe(3)O(4) nanoparticles. The CALB loading capacity in nanoreactors achieved 30%. The Michaelis constant (Km) and maximum reaction rate of magnetic CALB nanoreactors were 56.1 mmol/L and 0.2 mmol/(L.min). Magnetic CALB nanoreactors could circularly catalyze bixin-maltitol ester synthesis and keep catalytic efficiency of 62.6% after eight repetitive enzymatic reactions. Additionally, the optimal bixin-maltitol ester synthesis procedure was heating bixin-maltitol mixture at molar ratio of 1:7 in anhydrous 2-methyl-2-butanol-dimethylsulfoxide (8:2, v/v) at 50 degreesC for 24 h. Bixin-maltitol ester showed improved water solubility at pH 5.5 and 7.0. Title: Design, synthesis, and biological evaluation of novel N-Benzyl piperidine derivatives as potent HDAC/AChE inhibitors for Alzheimer's disease Qin P, Ran Y, Xie F, Liu Y, Wei C, Luan X, Wu J Ref: Bioorganic & Medicinal Chemistry, 80:117178, 2023 : PubMed ESTHER: Qin_2023_Bioorg.Med.Chem_80_117178 PubMedSearch: Qin 2023 Bioorg.Med.Chem 80 117178 PubMedID: 36706609 Abstract The multitarget-directed ligands approach represents a potential strategy to provide effective treatments for Alzheimer's disease (AD) given its multifactorial pathology. Herein, a series of N-benzyl piperidine derivatives were designed, synthesized, and biologically characterized for dual inhibitions of histone deacetylase (HDAC) and acetylcholinesterase (AChE). Among the compounds tested, d5 and d10 exhibited dual enzyme inhibitions (d5: HDAC(IC50) = 0.17 microM, AChE(IC50) = 6.89 microM, d10: HDAC(IC50) = 0.45 microM, AChE(IC50) = 3.22 microM), and both compounds showed activities on scavenging free radical, metal chelating, and inhibiting Abeta aggregations. More importantly, both compounds exhibited promising neuroprotective activities in PC-12 cells and good AChE selectivity. Collectively, the multifunctional profiles of compound d5 and d10 encourage further optimization and exploration to develop more potent analogues as potential treatments for AD. Title: A new terrein dimer and a new meroterpene from the mangrove endophytic fungus Lichtheimia sp Ren JL, Li CM, Shen L, Wu J Ref: J Asian Nat Prod Res, :1, 2023 : PubMed ESTHER: Ren_2023_J.Asian.Nat.Prod.Res__1 PubMedSearch: Ren 2023 J.Asian.Nat.Prod.Res 1 PubMedID: 37042744 Abstract A new terrein dimer named lichtheicol A (1) and a new meroterpene named lichtheiterpene A (2), were isolated from the mangrove endophytic fungus Lichtheimia sp. J2B1, together with 10 known compounds (3-12). The planar structures and absolute configurations of 1 and 2 were established by a combination of extensive spectroscopic data analyses and electronic circular dichroism (ECD) calculations. Compounds 4 and 5 exhibited marked inhibitory effects against butyrylcholinesterase (BuChE) with IC(50) values of 0.71 and 0.53 microM, respectively. Title: Clinical and genetic characteristics of CEL-MODY (MODY8): a literature review and screening in Chinese individuals diagnosed with early-onset type 2 diabetes Sun S, Gong S, Li M, Wang X, Wang F, Cai X, Liu W, Luo Y, Zhang S and Ji L <14 more author(s)> Sun S, Gong S, Li M, Wang X, Wang F, Cai X, Liu W, Luo Y, Zhang S, Zhang R, Zhou L, Zhu Y, Ma Y, Ren Q, Zhang X, Chen J, Chen L, Wu J, Gao L, Zhou X, Li Y, Zhong L, Han X, Ji L (- 14) Ref: Endocrine, :, 2023 : PubMed ESTHER: Sun_2023_Endocrine__ PubMedSearch: Sun 2023 Endocrine PubMedID: 37726640 Abstract OBJECTIVE: CEL-related maturity-onset diabetes of the young (CEL-MODY, MODY8) is a special type of monogenetic diabetes caused by mutations in the carboxyl-ester lipase (CEL) gene. This study aimed to summarize the genetic and clinical characteristics of CEL-MODY patients and to determine the prevalence of the disease among Chinese patients with early-onset type 2 diabetes (EOD). METHODS: We systematically reviewed the literature associated with CEL-MODY in PubMed, Embase, Web of Science, China National Knowledge Infrastructure and Wanfang Data to analyze the features of patients with CEL-MODY. We screened and evaluated rare variants of the CEL gene in a cohort of 679 Chinese patients with EOD to estimate the prevalence of CEL-MODY in China. RESULTS: In total, 21 individuals reported in previous studies were diagnosed with CEL-MODY based on the combination of diabetes and pancreatic exocrine dysfunction as well as frameshift mutations in exon 11 of the CEL gene. CEL-MODY patients were nonobese and presented with exocrine pancreatic affection (e.g., chronic pancreatitis, low fecal elastase levels, pancreas atrophy and lipomatosis) followed by insulin-dependent diabetes. No carriers of CEL missense mutations were reported with exocrine pancreatic dysfunction. Sequencing of CEL in Chinese EOD patients led to the identification of the variant p.Val736Cysfs*22 in two patients. However, these patients could not be diagnosed with CEL-MODY because there were no signs that the exocrine pancreas was afflicted. CONCLUSION: CEL-MODY is a very rare disease caused by frameshift mutations affecting the proximal VNTR segments of the CEL gene. Signs of exocrine pancreatic dysfunction provide diagnostic clues for CEL-MODY, and genetic testing is vital for proper diagnosis. Further research in larger cohorts is needed to investigate the characteristics and prevalence of CEL-MODY in the Chinese population. Title: The effect of double filtration plasmapheresis and corticosteroids on patients with anti-dipeptidyl-peptidase-like protein 6 encephalitis Wan W, Pan Y, Chen Y, Bai S, Yao X, Lin Y, Wu J, Ni L, Mei Y and Guan Y <3 more author(s)> Wan W, Pan Y, Chen Y, Bai S, Yao X, Lin Y, Wu J, Ni L, Mei Y, Qiu H, Zhou Y, Hao Y, Guan Y (- 3) Ref: Ther Apher Dial, :, 2023 : PubMed ESTHER: Wan_2023_Ther.Apher.Dial__ PubMedSearch: Wan 2023 Ther.Apher.Dial PubMedID: 37461148 Gene_locus related to this paper: human-DPP6 Abstract INTRODUCTION: Anti-dipeptidyl-peptidase-like protein 6 (DPPX) encephalitis is a rare condition with varied symptoms including gastrointestinal issues, weight loss, cognitive and mental dysfunction, and hyperexcitability of the central nervous system. METHODS: We studied five patients with anti-DPPX encephalitis who received immunotherapy, specifically DFPP, at our hospital. We analyzed their clinical symptoms, lab results, electrophysiological and imaging findings, and outcomes with immunotherapy. RESULTS: Patients presented with cognitive dysfunction, tremor, seizures, psychiatric disturbances, and cerebellar and brainstem dysfunction. Magnetic resonance imaging (MRI) showed brain abnormalities in one patient and elevated cerebrospinal fluid (CSF) protein levels in two patients. Antibodies against DPPX were detected in all patients and in CSF in two patients. One patient had antibodies against anti-CV2/contactin response mediator protein 5 (CRMP5). All patients responded well to DFPP and corticosteroids. CONCLUSION: DFPP may be an effective treatment for anti-DPPX encephalitis. Further research is needed to understand disease progression and evaluate immunotherapy efficacy. Title: Repetitive transcranial magnetic stimulation may be superior to drug therapy in the treatment of Alzheimer's disease: A systematic review and Bayesian network meta-analysis Wei N, Liu H, Ye W, Xu S, Lu C, Dai A, Hou T, Zeng X, Wu J, Chen J Ref: CNS Neurosci Ther, :, 2023 : PubMed ESTHER: Wei_2023_CNS.Neurosci.Ther__ PubMedSearch: Wei 2023 CNS.Neurosci.Ther PubMedID: 37088953 Abstract BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive brain stimulation therapy that is primarily used to treat a variety of neuropsychiatric conditions. Recently, previous research reports stated that rTMS have the characteristics of neurorestorative in Alzheimer's disease (AD). However, the relevant clinical research evidence has not been fully summarized. METHODS: This article performed a network meta-analysis of individual participant data from eligible studies searched in PubMed, Embase, and the Cochrane Library from inception to March 31, 2022. The drug treatments involved were acetylcholinesterase inhibitors (AChEIs), N-methyl-d-aspartate (NMDA), anti-amyloid-beta (Abeta), and some new targeted therapeutic drugs. RESULTS: A total of 15, 548 individuals with AD disease in 57 randomized clinical trials (RCTs) were included in this meta-analysis. The results indicated that the patients who received rTMS treatment (standard mean difference [SMD]: 0.65; 95% confidence interval [CI]: 0.22-1.07) had a better MMSE score than placebo. Treatment outcome analysis showed that, compared with multiple pharmacological interventions, rTMS acquired the greatest probability rank with the best cognitive improvement in MMSE score [the surface under the cumulative ranking curve (SUCRA) 93.3%] and ADAS-cog score (SUCRA 86.7%). At the same time, rTMS treatment had the lowest rank in the adverse events (SUCRA 24.1%) except for the placebo group (SUCRA 19.1%). CONCLUSION: Compared with the current clinical drug treatment, rTMS demonstrated better cognitive function improvement and fewer adverse events in AD patients. Therefore, rTMS shows broad prospects in the treatment of Alzheimer's disease, and it is worth being widely popularized in clinic. Title: Discovery of two ent-atisane diterpenoid lactones with AChE inhibitory activity from the roots of Euphorbia fischeriana Wei J, Li Z, Shan M, Wu F, Li L, Ma Y, Wu J, Li X, Liu Y and Wu Z <2 more author(s)> Wei J, Li Z, Shan M, Wu F, Li L, Ma Y, Wu J, Li X, Liu Y, Hu Z, Zhang Y, Wu Z (- 2) Ref: Org Biomol Chem, :, 2023 : PubMed ESTHER: Wei_2023_Org.Biomol.Chem__ PubMedSearch: Wei 2023 Org.Biomol.Chem PubMedID: 37581482 Abstract Euphorlactone A (1), a rare rearranged ent-atisane norditerpenoid with an undescribed 3-nor-2,4-olide-ent-atisane scaffold, and euphorlactone B (2), a new ent-atisane diterpenoid with an unprecedented seven-membered lactone ring C, were isolated from the roots of Euphorbia fischeriana. Their planar structures with absolute configurations were extensively elucidated by analysis of 1D and 2D NMR data, electronic circular dichroism (ECD) calculations, Rh(2)(OCOCF(3))(4)-induced ECD curves, and single-crystal X-ray diffraction. Euphorlactone A (ELA) showed a remarkable AChE (acetylcholinesterase) inhibitory activity (IC(50) = 2.13 +/- 0.06 microM and K(i) = 0.058 microM), which was five times stronger than that of the positive control (rivastigmine, IC(50) = 12.46 +/- 0.82 microM), and further in vitro enzyme inhibition kinetic analysis and molecular docking studies were performed to investigate the AChE inhibitory mechanism. Title: Novel miR-108 and miR-234 target juvenile hormone esterase to regulate the response of Plutella xylostella to Cry1Ac protoxin Yang J, Chen S, Xu X, Lin S, Wu J, Lin G, Bai J, Song Q, You M, Xie M Ref: Ecotoxicology & Environmental Safety, 254:114761, 2023 : PubMed ESTHER: Yang_2023_Ecotoxicol.Environ.Saf_254_114761 PubMedSearch: Yang 2023 Ecotoxicol.Environ.Saf 254 114761 PubMedID: 36907089 Abstract Insect hormones, such as juvenile hormone (JH), precisely regulate insect life-history traits. The regulation of JH is tightly associated with the tolerance or resistance to Bacillus thuringiensis (Bt). JH esterase (JHE) is a primary JH-specific metabolic enzyme which plays a key role in regulating JH titer. Here, we characterized a JHE gene from Plutella xylostella (PxJHE), and found it was differentially expressed in the Bt Cry1Ac resistant and susceptible strains. Suppression of PxJHE expression with RNAi increased the tolerance of P. xylostella to Cry1Ac protoxin. To investigate the regulatory mechanism of PxJHE, two target site prediction algorithms were applied to predict the putative miRNAs targeting PxJHE, and the resulting putative miRNAs were subsequently verified for their function targeting PxJHE using luciferase reporter assay and RNA immunoprecipitation. MiR-108 or miR-234 agomir delivery dramatically reduced PxJHE expression in vivo, whilst only miR-108 overexpression consequently increased the tolerance of P. xylostella larvae to Cry1Ac protoxin. By contrast, reduction of miR-108 or miR-234 dramatically increased PxJHE expression, accompanied by the decreased tolerance to Cry1Ac protoxin. Furthermore, injection of miR-108 or miR-234 led to developmental defects in P. xylostella, whilst injection of antagomir did not cause any obvious abnormal phenotypes. Our results indicated that miR-108 or miR-234 can be applied as potential molecular targets to combat P. xylostella and perhaps other lepidopteran pests, providing novel insights into miRNA-based integrated pest management. Title: Frequency and polymorphism of acetylcholinesterase gene involved in the organophosphate resistance of Musca domestica in Guizhou Province, China Yang X, Mou R, Liang Q, Cheng J, Wu Y, Tan W, Wu J Ref: Archives of Insect Biochemistry & Physiology, :e22045, 2023 : PubMed ESTHER: Yang_2023_Arch.Insect.Biochem.Physiol__e22045 PubMedSearch: Yang 2023 Arch.Insect.Biochem.Physiol e22045 PubMedID: 37602787 Abstract Organophosphate (OP) resistance has been prevalent in Musca domestica populations worldwide since 1960s. Previous studies have demonstrated that point mutations of the acetylcholinesterase gene (Ace) are one of the important molecular mechanisms underlying OP resistance. However, few studies have investigated the molecular mechanisms of OP resistance in the past 10 years in China. In this study, we investigated the status of OP resistance and genetic diversity of Ace in the field populations of houseflies in Guizhou Province of China. The bioassays showed that the houseflies had 142-304-fold resistance to dichlorvos (DDVP) and 122-364-fold resistance to temephos, compared to the susceptible houseflies. Five nonsynonymous mutations (Y226F, V260L, G342A/V, F407Y) in Ace were detected among the 7 field populations, with an average frequency of 5.4%, 55%, 68%, 32%, and 94%, respectively, of which the Y226F mutation had not been reported previously. Eleven combinations of triple mutations (at positions 260, 342, and 407) were observed, of which the combination 260L/V+342A/V+407Y was predominant. The ZY and AS populations showed greatest diversity of allelic combination and the other five populations showed different distributions among different regions. These results indicate that the resistance to OPs is prevalent among the housefly populations and target-site insensitivity is the main cause of resistance in Guizhou Province. The difference in distribution and the allelic diversity of Ace in field populations may be due to the complexity and variability of insecticide application. It is necessary to monitor resistance to insecticides and conduct management of houseflies in Guizhou Province. Title: A multibiomarker approach to assess the ecotoxicological effects of diclofenac on Asian clam Corbicula fluminea (O. F. Muller, 1774) Yuan N, Ding J, Wu J, Bao E, Chu Y, Hu F Ref: Environ Sci Pollut Res Int, :, 2023 : PubMed ESTHER: Yuan_2023_Environ.Sci.Pollut.Res.Int__ PubMedSearch: Yuan 2023 Environ.Sci.Pollut.Res.Int PubMedID: 37438503 Abstract Diclofenac (DCF), one of the most current and widely used nonsteroidal anti-inflammatory drugs (NSAIDs), has been frequently detected in aquatic environments worldwide. However, the ecotoxicological effects of DCF on freshwater invertebrates remain largely unknown. In the present study, Corbicula fluminea were exposed to environmentally relevant concentrations of DCF (0, 2, 20, and 200 microg/L) for 28 days, and the potential adverse effects of DCF on siphoning behavior, antioxidant responses, and apoptosis were investigated. Our results showed that the siphon efficiencies of clams were significantly suppressed under DCF stress. DCF exerted neurotoxicity via reducing the activity of acetylcholinesterase (AChE) in gills and digestive gland of C. fluminea. Exposure to DCF induced antioxidant stress and increased malondialdehyde (MDA) levels in both gills and digestive gland of C. fluminea. Transcriptional alterations of apoptosis-related genes indicated that DCF might induce apoptosis by triggering mitochondrial apoptotic pathway. These findings can improve our understanding of the ecological risk of DCF in freshwater ecosystems. Title: Glimepiride, a novel soluble epoxide hydrolase inhibitor, protects against heart failure via increasing epoxyeicosatrienoic acids Zhao C, Jiang X, Peng L, Zhang Y, Li H, Zhang Q, Wang Y, Yang F, Wu J and Wang DW <4 more author(s)> Zhao C, Jiang X, Peng L, Zhang Y, Li H, Zhang Q, Wang Y, Yang F, Wu J, Wen Z, He Z, Shen J, Chen C, Wang DW (- 4) Ref: Journal of Molecular & Cellular Cardiology, 185:13, 2023 : PubMed ESTHER: Zhao_2023_J.Mol.Cell.Cardiol_185_13 PubMedSearch: Zhao 2023 J.Mol.Cell.Cardiol 185 13 PubMedID: 37871528 Inhibitor(s) related to this paper: Glimepiride Abstract BACKGROUND: Epoxyeicosatrienoic acids (EETs), which exert multiple endogenous protective effects, are hydrolyzed into less active dihydroxyeicosatrienoic acids (DHETs) by soluble epoxide hydrolase (sEH). However, commercial drugs related to EETs or sEH are not yet in clinical use. METHODS: Firstly, the plasma concentration of EETs and DHETs of 316 patients with heart failure (HF) were detected and quantitated by liquid chromatography-tandem mass spectrometry. Then, transverse aortic constriction (TAC)-induced HF was introduced in cardiomyocyte-specific Ephx2(-/-) mice. Moreover, Western blot, real-time PCR, luciferase reporter, ChIP assays were employed to explore the underlying mechanism. Finally, multiple sEH inhibitors were designed, synthesized, and validated in vitro and in vivo. RESULTS: The ratios of DHETs/EETs were increased in the plasma from patients with HF. Meanwhile, the expression of sEH was upregulated in the heart of patients and mice with HF, especially in cardiomyocytes. Cardiomyocyte-specific Ephx2(-/-) mice ameliorated cardiac dysfunction induced by TAC. Consistently, Ephx2 knockdown protected Angiotensin II (AngII)-treated cardiomyocytes via increasing EETs in vitro. Mechanistically, AngII could enhance the expression of transcript factor Krppel-like factor 15 (KLF15), which in turn upregulated sEH. Importantly, glimepiride was identified as a novel sEH inhibitor, which benefited from the elevated EETs during HF. CONCLUSIONS: Glimepiride attenuates HF in mice in part by increasing EETs. CLINICAL TRIAL IDENTIFIER: NCT03461107 (https://clinicaltrials.gov). Title: Landscape of the gut archaeome in association with geography, ethnicity, urbanization, and diet in the Chinese population Bai X, Sun Y, Li Y, Li M, Cao Z, Huang Z, Zhang F, Yan P, Wang L and Zuo T <9 more author(s)> Bai X, Sun Y, Li Y, Li M, Cao Z, Huang Z, Zhang F, Yan P, Wang L, Luo J, Wu J, Fan D, Chen H, Zhi M, Lan P, Zeng Z, Wu X, Miao Y, Zuo T (- 9) Ref: Microbiome, 10:147, 2022 : PubMed ESTHER: Bai_2022_Microbiome_10_147 PubMedSearch: Bai 2022 Microbiome 10 147 PubMedID: 36100953 Abstract BACKGROUND AND AIMS: The human gut is home to a largely underexplored microbiome component, the archaeome. Little is known of the impact of geography, urbanization, ethnicity, and diet on the gut archaeome in association with host health. We aim to delineate the variation of the human gut archaeome in healthy individuals and its association with environmental factors and host homeostasis. METHODS: Using metagenomic sequencing, we characterized the fecal archaeomes of 792 healthy adult subjects from 5 regions in China, spanning 6 ethnicities (Han, Zang, Miao, Bai, Dai, and Hani), consisting of both urban and rural residents for each ethnicity. In addition, we sampled 119 host variables (including lifestyle, diet, and blood parameters) and interrogated the influences of those factors, individually and combined, on gut archaeome variations. RESULTS: Population geography had the strongest impact on the gut archaeome composition, followed by urbanization, dietary habit, and ethnicity. Overall, the metadata had a cumulative effect size of 11.0% on gut archaeome variation. Urbanization decreased both the alpha-diversity (intrinsic microbial diversity) and the beta-diversity (inter-individual dissimilarities) of the gut archaeome, and the archaea-to-bacteria ratios in feces, whereas rural residents were enriched for Methanobrevibacter smithii in feces. Consumption of buttered milk tea (a characteristic diet of the rural Zang population) was associated with increased abundance of M. smithii. M. smithii was at the central hub of archaeal-bacterial interactions in the gut microecology, where it was positively correlated with the abundances of a multitude of short chain fatty acid (SCFA)-producing bacteria (including Roseburia faecis, Collinsella aerofaciens, and Prevotella copri). Moreover, a decreased abundance of M. smithii was associated with increased human blood levels of cholinesterase in the urban population, coinciding with the increasing prevalence of noncommunicable diseases (such as dementia) during urbanization. CONCLUSIONS: Our data highlight marked contributions of environmental and host factors (geography, urbanization, ethnicity, and habitual diets) to gut archaeome variations across healthy individuals, and underscore the impact of urbanization on the gut archaeome in association with host health in modern society. Video Abstract. Title: Iboga-type alkaloids with Indolizidino[8,7-b]Indole scaffold and bisindole alkaloids from Tabernaemontana bufalina Lour Chen SQ, Jia J, Hu JY, Wu J, Sun WT, Zheng M, Wang X, Zhu KK, Jiang CS and Cai YS <3 more author(s)> Chen SQ, Jia J, Hu JY, Wu J, Sun WT, Zheng M, Wang X, Zhu KK, Jiang CS, Yang SP, Zhang J, Wang SB, Cai YS (- 3) Ref: Phytochemistry, 196:113089, 2022 : PubMed ESTHER: Chen_2022_Phytochemistry_196_113089 PubMedSearch: Chen 2022 Phytochemistry 196 113089 PubMedID: 35074605 Abstract Phytochemical investigation on the aerial parts of Tabernaemontana bufalina Lour. (Apocynaceae) led to the identification of four undescribed monoterpenoid indole alkaloids named taberbufamines A-D, an undescribed natural product, and fourteen known indole alkaloids. The structures of the undescribed alkaloids were established by spectroscopic and computational methods, and their absolute configurations were further determined by quantum chemical TDDFT calculations and the experimental ECD spectra. Taberbufamines A and B possessed an uncommon skeleton incorporating an indolizidino [8,7-b]indole motif with a 2-hydroxymethyl-butyl group attached at the pyrrolidine ring. Biosynthetically, Taberbufamines A and B might be derived from iboga-type alkaloid through rearrangement. Vobatensine C showed significant bioactivity against A-549, Bel-7402, and HCT-116 cells with IC(50) values of 2.61, 1.19, and 1.74 microM, respectively. Ervahanine A showed antimicrobial activity against Bacillus subtilis, Mycobacterium smegmatis, and Helicobacter pylori with MIC values of 4, 8, and 16 microg/mL, respectively. 19(S)-hydroxyibogamine was shown as butyrylcholinesterase inhibitor (IC(50) of 20.06 microM) and alpha-glycosidase inhibitor (IC(50) of 17.18 microM), while tabernamine, ervahanine B, and ervadivaricatine B only showed alpha-glycosidase inhibitory activities with IC(50) values in the range of 0.95-4.61 microM. Title: Directional-path modification strategy enhances PET hydrolase catalysis of plastic degradation Chen XQ, Guo ZY, Wang L, Yan ZF, Jin CX, Huang QS, Kong DM, Rao DM, Wu J Ref: J Hazard Mater, 433:128816, 2022 : PubMed ESTHER: Chen_2022_J.Hazard.Mater_433_128816 PubMedSearch: Chen 2022 J.Hazard.Mater 433 128816 PubMedID: 35390614 Gene_locus related to this paper: thefu-q6a0i3 Abstract Poly (ethylene terephthalate) (PET) is a widely used type of general plastic that produces a significant amount of waste due to its non-degradable properties. We propose a novel directional-path modification (DPM) strategy, involving positive charge amino acid introduction and binding groove remodeling, and apply it to Thermobifida fusca cutinase to enhance PET degradation. The highest value of PET degradation (90%) was achieved in variant 4Mz (H184S/Q92G/F209I/I213K), exhibiting values almost 30-fold that of the wild-type. We employed molecular docking, molecular dynamics simulations, and QM/MM MD for the degradation process of PET, accompanied by acylation and deacylation. We found that the distance of nucleophilic attack was reduced from about 4.6 A in the wild type to 3.8 A in 4Mz, and the free energy barrier of 4Mz dropped from 14.3 kcal/mol to 7.1 kcal/mol at the acylation which was the rate-limiting step. Subsequently, the high efficiency and universality of the DPM strategy were successfully demonstrated in LCC, Est119, and BhrPETase enhancing the degradation activity of PET. Finally, the highest degradation rate of the pretreated commercial plastic bottles had reached to 73%. The present study provides insight into the molecular binding mechanism of PET into the PET hydrolases structure and proposes a novel DPM strategy that will be useful for the engineering of more efficient enzymes for PET degradation. Title: N-acylhomoserine lactonase-based hybrid nanoflowers: a novel and practical strategy to control plant bacterial diseases Chen Y, Liu P, Wu J, Yan W, Xie S, Sun X, Ye BC, Chu X Ref: J Nanobiotechnology, 20:347, 2022 : PubMed ESTHER: Chen_2022_J.Nanobiotechnology_20_347 PubMedSearch: Chen 2022 J.Nanobiotechnology 20 347 PubMedID: 35883097 Abstract BACKGROUND: The disease caused by plant pathogenic bacteria in the production, transportation, and storage of many crops has brought huge losses to agricultural production. N-acylhomoserine lactonases (AHLases) can quench quorum-sensing (QS) by hydrolyzing acylhomoserine lactones (AHLs), which makes them the promising candidates for controlling infections of QS-dependent pathogenic bacteria. Although many AHLases have been isolated and considered as a potentially effective preventive and therapeutic agents for bacterial diseases, the intrinsically poor ambient stability has seriously restricted its application. RESULTS: Herein, we showed that a spheroid enzyme-based hybrid nanoflower (EHNF), AhlX@Ni(3)(PO(4))(2), can be easily synthesized, and it exhibited 10 times AHL (3OC8-HSL) degradation activity than that with free AhlX (a thermostable AHL lactonase). In addition, it showed intriguing stability even at the working concentration, and retained ~ 100% activity after incubation at room temperature (25 degreesC) for 40 days and approximately 80% activity after incubation at 60 degreesC for 48 h. Furthermore, it exhibited better organic solvent tolerance and long-term stability in a complicated ecological environment than that of AhlX. To reduce the cost and streamline production processes, CSA@Ni(3)(PO(4))(2), which was assembled from the crude supernatants of AhlX and Ni(3)(PO(4))(2), was synthesized. Both AhlX@Ni(3)(PO(4))(2) and CSA@Ni(3)(PO(4))(2) efficiently attenuated pathogenic bacterial infection. CONCLUSIONS: In this study, we have developed N-acylhomoserine lactonase-based hybrid nanoflowers as a novel and efficient biocontrol reagent with significant control effect, outstanding environmental adaptability and tolerance. It was expected to overcome the bottlenecks of poor stability and limited environmental tolerance that have existed for over two decades and pioneered the practical application of EHNFs in the field of biological control. Title: A Simple Aptamer SERS Sensor Based on Mesoporous Silica for the Detection of Chlorpyrifos Dong S, Shi Q, He K, Wu J, Zhu Z, Feng J Ref: Foods, 11:, 2022 : PubMed ESTHER: Dong_2022_Foods_11_ PubMedSearch: Dong 2022 Foods 11 PubMedID: 36359944 Abstract Chlorpyrifos is an organophosphorus insecticide, which can be used to control a variety of chewing and piercing mouthparts pests in agricultural production. It can destroy the normal nerve impulse conduction by inhibiting the activity of acetylcholinesterase or cholinesterase in the nerves, causing a series of poisoning symptoms. In order to achieve the quantitative analysis of chlorpyrifos residues in agricultural products, an aptamer-controlled signal molecule release method was developed in this study. The signal molecule 4-ATP of surface-enhanced Raman spectroscopy (SERS) was loaded into aminated mesoporous silica nanoparticles (MSNs-NH(2)) prepared by the one pot method, and then coated with an aptamer of chlorpyrifos through electrostatic interaction. The specific binding of the aptamer and chlorpyrifos led to the release of 4-ATP, and the amount of 4-ATP released was positively correlated with the amount of chlorpyrifos. Finally, the standard curve of chlorpyrifos quantitative detection based on SERS was established. Meanwhile, Ag-carrying mesoporous silica (Ag@MSNs) was prepared as the reinforcement substrate for SERS detection. The results showed that there was a good linear correlation between the Raman intensity and the concentration of chlorpyrifos at 25-250 ng/mL, and the limit of detection (LOD) was 19.87 ng/mL. The recoveries of chlorpyrifos in the apple and tomato samples were 90.08-102.2%, with RSD < 3.32%. This method has high sensitivity, specificity, reproducibility and stability, and can be used for the quantitative detection of chlorpyrifos in the environment and agricultural products. Title: FASN promotes lymph node metastasis in cervical cancer via cholesterol reprogramming and lymphangiogenesis Du Q, Liu P, Zhang C, Liu T, Wang W, Shang C, Wu J, Liao Y, Chen Y and Yao S <5 more author(s)> Du Q, Liu P, Zhang C, Liu T, Wang W, Shang C, Wu J, Liao Y, Chen Y, Huang J, Tan H, Zhao Y, Xia M, Liu J, Yao S (- 5) Ref: Cell Death Dis, 13:488, 2022 : PubMed ESTHER: Du_2022_Cell.Death.Dis_13_488 PubMedSearch: Du 2022 Cell.Death.Dis 13 488 PubMedID: 35597782 Abstract Cervical cancer (CC) patients with lymph node metastasis (LNM) have a poor prognosis. Clarification of the detailed mechanisms underlying LNM may provide potential clinical therapeutic targets for CC patients with LNM. However, the molecular mechanism of LNM in CC is unclear. In the present study, we demonstrated that fatty acid synthase (FASN), one of the key enzymes in lipid metabolism, had upregulated expression in the CC samples and was correlated with LNM. Moreover, multivariate Cox proportional hazards analysis identified FASN as an independent prognostic factor of CC patients. Furthermore, gain-of-function and loss-of-function approaches showed that FASN promoted CC cell migration, invasion, and lymphangiogenesis. Mechanistically, on the one hand, FASN could regulate cholesterol reprogramming and then activate the lipid raft-related c-Src/AKT/FAK signaling pathway, leading to enhanced cell migration and invasion. On the other hand, FASN induced lymphangiogenesis by secreting PDGF-AA/IGFBP3. More importantly, knockdown of FASN with FASN shRNA or the inhibitors C75 and Cerulenin dramatically diminished LNM in vivo, suggesting that FASN plays an essential role in LNM of CC and the clinical application potential of FASN inhibitors. Taken together, our findings uncover a novel molecular mechanism in LNM of CC and identify FASN as a novel prognostic factor and potential therapeutic target for LNM in CC. Title: Polylactic acid microplastics induce higher biotoxicity of decabromodiphenyl ethane on earthworms (Eisenia fetida) compared to polyethylene and polypropylene microplastics Han Y, Fu M, Wu J, Zhou S, Qiao Z, Peng C, Zhang W, Liu F, Ye C, Yang J Ref: Sci Total Environ, :160909, 2022 : PubMed ESTHER: Han_2022_Sci.Total.Environ__160909 PubMedSearch: Han 2022 Sci.Total.Environ 160909 PubMedID: 36526185 Abstract Decabromodiphenyl ethane (DBDPE) and microplastics (MPs), such as fossil-based polymers polyethylene (PE), polypropylene (PP), and bio-based plastics polylactic acid (PLA) are abundant in e-waste dismantling areas. However, the information on the effects of DBDPE combined with MPs (DBDPE-MPs) on earthworms is still limited. In this study, we explored the impacts of DBDPE-MPs on neurotoxic biomarkers, tissue damage, and transcriptomics of Eisenia fetida by simulating different exposure patterns of 10 mg kg(-1) DBDPE and 10 mg kg(-1) DBDPE-MPs (PLA, PP, and PE). Results showed that the activities of acetylcholinesterase, Na(+)/K(+)-ATPase, Ca(2+)/Mg(2+)-ATPase, carboxylate enzyme, and the contents of calcium and glutamate were significantly stimulated. DBDPE-MP co-exposure caused more severe damage to the epidermis, muscles, and tissues. Transcriptomic analysis revealed that differentially expressed genes (DEGs) of DBDPE-MPs were mainly related to inflammation, the immune system, digestive system, endocrine system, and metabolism. DBDPE and PP-MPs had similar influences on immunity and metabolism. However, DBDPE-PLA and DBDPE-PE further affected the endocrine system and signaling pathways. Specific DEGs showed that detoxification systems in the case of MPs were significantly upregulated. The study indicated that MPs exacerbated DBDPE toxicity in the nervous system, epidermis, and gene regulation of E. fetida, helping to assess the ecological risks of e-wastes and microplastics in soil. Title: [Characterization of Humicola insolens cutinase-tachystatin A2 fusion protein and its application in treatment of recycled paper stickies] Li G, Liu Z, Zhang Y, Wu J Ref: Sheng Wu Gong Cheng Xue Bao, 38:207, 2022 : PubMed ESTHER: Li_2022_Sheng.Wu.Gong.Cheng.Xue.Bao_38_207 PubMedSearch: Li 2022 Sheng.Wu.Gong.Cheng.Xue.Bao 38 207 PubMedID: 35142131 Abstract With the decrease of forest timber resources, the recycling of waste paper has received increasing attention. However, the stickies produced in the process of waste paper recycling may negatively affect the production of recycled paper. The biological decomposition of stickies, which has the advantages of high efficiency, high specificity and pollution-free, is achieved mainly through the enzymatic cleavage of the ester bond in the stickies components to prevent flocculation. Cutinase is a serine esterase that can degrade some components of the stickies. Previous research indicated that the anchor peptide tachystatin A2 (TA2) is able to bind polyurethane. In this study, the cutinase HiC derived from Humicola insolens was used to construct a fusion protein HiC-TA2 by megaprimer PCR of the whole plasmid (MEGAWHOP). The enzymatic properties and the degradation efficiency of the fusion protein on poly(ethyl acrylate) (PEA), a model substrate of stickies component, were determined. The results showed that the degradation efficiency, the size decrease of PEA particle, and the amount of ethanol produced by HiC-TA2 were 1.5 times, 6.8 times, and 1.4 times of that by HiC, respectively. These results demonstrated that TA2 improved the degradation efficiency of HiC on PEA. This study provides a useful reference for biological decomposition of stickies produced in the process of recycled paper production. Title: Enhancement of PET biodegradation by anchor peptide-cutinase fusion protein Liu Z, Zhang Y, Wu J Ref: Enzyme Microb Technol, 156:110004, 2022 : PubMed ESTHER: Liu_2022_Enzyme.Microb.Technol_156_110004 PubMedSearch: Liu 2022 Enzyme.Microb.Technol 156 110004 PubMedID: 35217214 Gene_locus related to this paper: thefu-q6a0i3 Abstract With the increasing production of polyethylene terephthalate (PET) plastic products, the problem of PET waste has become a serious threat to ecosystem. PET enzymatic biodegradation, due to its environmental friendliness and sustainability, has gradually attracted attention. As a multifunctional hydrolase, cutinase (EC 3.1.1.74) can not only degrade fatty acid esters, soluble synthetic esters, and emulsified triglycerides, but also exhibit potential for PET degradation. In order to enhance the PET degradation activity of cutinase, we functionally screened several PET binding domains, e.g. carbohydrate binding module, anchor peptide, and hydrophobin, that promote the absorption of enzyme to PET substrate, selected Dermaseptin SI (DSI) and fused it onto the N-terminus of Thermobifida fusca cutinase mutant D204C/E253C (Tfuc2), and finally achieved the PET degradation rate up to 57.9% at 70 degreesC for 96 h, which was 22.7-fold of that of Tfuc2 itself. These results indicate that the fusion of PET binding domain is a promising strategy to enhance PET enzymatic degradation. Title: Enhanced biodegradation activity towards poly(ethyl acrylate) and poly(vinyl acetate) by anchor peptide assistant targeting Liu Z, Li G, Zhang F, Wu J Ref: J Biotechnol, :, 2022 : PubMed ESTHER: Liu_2022_J.Biotechnol__ PubMedSearch: Liu 2022 J.Biotechnol PubMedID: 35292345 Abstract The paper industry is one of the most important basic raw material pillar industries. With the decrease of forest wood resources, the recycling of wastepaper has drawn increasingly attention. However, the stickies generated in the process of wastepaper recycling will flocculate and deposite in the pulp, resulting in production accidents and inferior product quality. The biological enzymatic method, with the advantages of high efficiency, specificity, and pollution-free, can prevent the flocculation of the stickies by enzymatically hydrolyzing the ester bond of the stickies components. Previous studies have demonstrated that cutinase (EC 3.1.1.74) had the ability to degrade polyester components of stickies. Meanwhile, relevant studies have shown that anchor peptides possessed the ability to bind polyester. Herein, the cutinase from Humicola insolens (HiC) was fused with Escherichia coli anchor peptide OMP25, the enzymatic properties of the fusion protein HiC-OMP25 and its degradation efficiency of the stickies model substrate, poly(ethyl acrylate) (PEA) and poly(vinyl acetate) (PVAc), as well as stickies sediment were determined. All of the results demonstrated that OMP25 efficiently enhanced the degradation ability of HiC. Title: Deficiency in endocannabinoid synthase DAGLB contributes to early onset Parkinsonism and murine nigral dopaminergic neuron dysfunction Liu Z, Yang N, Dong J, Tian W, Chang L, Ma J, Guo J, Tan J, Dong A and Tang B <27 more author(s)> Liu Z, Yang N, Dong J, Tian W, Chang L, Ma J, Guo J, Tan J, Dong A, He K, Zhou J, Cinar R, Wu J, Salinas AG, Sun L, Kumar M, Sullivan BT, Oldham BB, Pitz V, Makarious MB, Ding J, Kung J, Xie C, Hawes SL, Wang L, Wang T, Chan P, Zhang Z, Le W, Chen S, Lovinger DM, Blauwendraat C, Singleton AB, Cui G, Li Y, Cai H, Tang B (- 27) Ref: Nat Commun, 13:3490, 2022 : PubMed ESTHER: Liu_2022_Nat.Commun_13_3490 PubMedSearch: Liu 2022 Nat.Commun 13 3490 PubMedID: 35715418 Gene_locus related to this paper: human-DAGLB, mouse-DGLB Abstract Endocannabinoid (eCB), 2-arachidonoyl-glycerol (2-AG), the most abundant eCB in the brain, regulates diverse neural functions. Here we linked multiple homozygous loss-of-function mutations in 2-AG synthase diacylglycerol lipase beta (DAGLB) to an early onset autosomal recessive Parkinsonism. DAGLB is the main 2-AG synthase in human and mouse substantia nigra (SN) dopaminergic neurons (DANs). In mice, the SN 2-AG levels were markedly correlated with motor performance during locomotor skill acquisition. Genetic knockdown of Daglb in nigral DANs substantially reduced SN 2-AG levels and impaired locomotor skill learning, particularly the across-session learning. Conversely, pharmacological inhibition of 2-AG degradation increased nigral 2-AG levels, DAN activity and dopamine release and rescued the locomotor skill learning deficits. Together, we demonstrate that DAGLB-deficiency contributes to the pathogenesis of Parkinsonism, reveal the importance of DAGLB-mediated 2-AG biosynthesis in nigral DANs in regulating neuronal activity and dopamine release, and suggest potential benefits of 2-AG augmentation in alleviating Parkinsonism. Title: Comparative Study of the Molecular Characterization, Evolution, and Structure Modeling of Digestive Lipase Genes Reveals the Different Evolutionary Selection Between Mammals and Fishes Tang SL, Liang XF, He S, Li L, Alam MS, Wu J Ref: Front Genet, 13:909091, 2022 : PubMed ESTHER: Tang_2022_Front.Genet_13_909091 PubMedSearch: Tang 2022 Front.Genet 13 909091 PubMedID: 35991544 Abstract Vertebrates need suitable lipases to digest lipids for the requirement of energy and essential nutrients; however, the main digestive lipase genes of fishes have certain controversies. In this study, two types of digestive lipase genes (pancreatic lipase (pl) and bile salt-activated lipase (bsal)) were identified in mammals and fishes. The neighborhood genes and key active sites of the two lipase genes were conserved in mammals and fishes. Three copies of PL genes were found in mammals, but only one copy of the pl gene was found in most of the fish species, and the pl gene was even completely absent in some fish species (e.g., zebrafish, medaka, and common carp). Additionally, the hydrophobic amino acid residues (Ile and Leu) which are important to pancreatic lipase activity were also absent in most of the fish species. The PL was the main digestive lipase gene in mammals, but the pl gene seemed not to be the main digestive lipase gene in fish due to the absence of the pl gene sequence and the important amino acid residues. In contrast, the bsal gene existed in all fish species, even two to five copies of bsal genes were found in most of the fishes, but only one copy of the BSAL gene was found in mammals. The amino acid residues of bile salt-binding sites and the three-dimensional (3D) structure modeling of Bsal proteins were conserved in most of the fish species, so bsal might be the main digestive lipase gene in fish. The phylogenetic analysis also indicated that pl or bsal showed an independent evolution between mammals and fishes. Therefore, we inferred that the evolutionary selection of the main digestive lipase genes diverged into two types between mammals and fishes. These findings will provide valuable evidence for the study of lipid digestion in fish. Title: Substrate-dependent inhibition of hypericin on human carboxylesterase 2: implications for herb-drug combination Wang D, Zhao T, Zhao S, Chen J, Dou T, Ge G, Wang C, Sun H, Liu K and Wu J <1 more author(s)> Wang D, Zhao T, Zhao S, Chen J, Dou T, Ge G, Wang C, Sun H, Liu K, Meng Q, Wu J (- 1) Ref: Curr Drug Metab, :, 2022 : PubMed ESTHER: Wang_2022_Curr.Drug.Metab__ PubMedSearch: Wang 2022 Curr.Drug.Metab PubMedID: 35114918 Abstract BACKGROUND: Hypericin is the main active ingredient of St. John's wort, a Chinese herb commonly used in treating depression. Previous studies have shown that hypericin can strongly inhibit human cytochrome P450 (CYP) enzyme activities; however, its potential interactions that inhibit human carboxylesterases 2 (hCE2) were unclear. PURPOSE: The study aimed to investigate the inhibition of hypericin on hCE2. METHODS: The inhibition of hypericin on hCE2 was studied by using N-(2-butyl-1,3-dioxo-2,3-dihydro-1H-phenalen-6-yl)-2-chloroacetamide (NCEN). The type of inhibition of hypericin on hCE2 and the corresponding inhibition constant (Ki) value were determined. The inhibition of hypericin on hCE2 in living cells was discussed. The herb-drug interactions (HDI) risk of hypericin and hCE2 in vivo was predicted by estimating the drug concentration-time curve (AUC) ratio of hypericin and hypericin free. To understand the inhibition mechanism of hypericin on the activity of hCE2 in-depth, molecular docking was performed. RESULTS: The half-maximal inhibitory concentration (IC50) values of hypericin against the hydrolysis of NCEN and irinotecan (CPT-11) were calculated to be 26.59 microM and 112.8 microM, respectively. Hypericin inhibited the hydrolysis of NCEN and CPT-11. Their Ki values were 10.53 microM and 81.77 microM, respectively. Moreover, hypericin distinctly suppressed hCE2 activity in living cells. In addition, the AUC of hCE2 metabolic drugs with metabolic sites similar to NCEN was estimated to increase by up to 5%, in the presence of hypericin. More importantly, the exposure of CPT-11 in the intestinal epithelium was predicted to increase by 2%-69% following the oral co-administration of hypericin. Further, molecular simulations indicated that hypericin could strongly interact with ASP98, PHE307, and ARG355 to form four hydrogen bonds within hCE2. CONCLUSION: These findings are of considerable clinical significance to the combination of hypericin-containing herbs and drugs metabolized by hCE2. Title: The Impact of Catalpol on Proliferation, Apoptosis, Migration, and Oxidative Stress of Lung Cancer Cells Based on Nrf2/ARE Signaling Wang H, Wu J, Fan H, Ji Y, Han C, Li C, Jiang S Ref: Biomed Res Int, 2022:5621341, 2022 : PubMed ESTHER: Wang_2022_Biomed.Res.Int_2022_5621341 PubMedSearch: Wang 2022 Biomed.Res.Int 2022 5621341 PubMedID: 35898682 Abstract The effects of catalpol on lung cancer cell proliferation, apoptosis, migration, and oxidative stress via the Nrf2/ARE signaling pathway are investigated in this work. Catalpol-12 g/mL group, catalpol-24 g/mL group, catalpol-48 g/mL group, catalpol - 48 g/mL + vector group, catalpol - 48 g/mL + Nrf2 group, si-NC group, and si-Nrf2 group were used to split lung cancer cells A549 into control groups. Proliferation was detected using the CCK-8 assay; apoptosis was detected using flow cytometry; migration was detected using the transwell chamber; ROS was distinguished using the DCFHDA method; MDA, SOD, and GSH were detected using the microvolume method; and Cleaved Caspase-3, Cleaved Caspase-9, Nrf2, HO-1, MMP-9, and MMP-2 were detected using the Western blot method. Catalpol 12 g/mL and 24 g/mL-48 g/mL treatment decreased the proliferation activity, migration number, and Nrf2, HO-1, MMP-9, and MMP-2 protein levels of lung cancer cells when compared to the control group. SOD and GSH levels of lung cancer cells were decreased, and MDA and ROS levels were increased. Cleaved caspase-3, cleaved caspase-9 protein expression levels, and apoptosis were boosted (P < 0.05). The proliferation activity, migration number, and protein levels of Nrf2, HO-1, MMP-9, and MMP-2 in the catalpol - 48 g/mL + Nrf2 group were raised compared to the catalpol - 48 g/mL + vector group, whereas there was an apparent drop in the Cleaved Caspase-3, Cleaved Caspase-9, and apoptosis rate. Similarly, SOD and GSH contents increased, whereas MDA and ROS decreased (P < 0.05). The proliferation activity, migration number, and Nrf2, HO-1, MMP-9, and MMP-2 protein levels of lung cancer cells in the si-Nrf2 group were all decreased when compared to the si-NC and control groups. Cleaved Caspase-3 and Cleaved Caspase-9 protein expression, on the other hand, increased as MDA and ROS levels were raised while SOD and GSH levels dropped (P < 0.05). It reveals that catalpol inhibits the Nrf2/ARE signaling pathway, which causes antiproliferation, migration, apoptosis, and oxidative stress in cancer cells of lungs. The rate of apoptosis was also lowered. Title: A narrative review: The pharmaceutical evolution of phenolic syringaldehyde Wu J, Fu YS, Lin K, Huang X, Chen YJ, Lai D, Kang N, Huang L, Weng CF Ref: Biomed Pharmacother, 153:113339, 2022 : PubMed ESTHER: Wu_2022_Biomed.Pharmacother_153_113339 PubMedSearch: Wu 2022 Biomed.Pharmacother 153 113339 PubMedID: 35780614 Abstract To better understand the pharmacological characters of syringaldehyde (SA), which is a key-odorant compound of whisky and brandy, this review article is the first to compile the published literature for molecular docking that were subsequently validated by in vitro and in vivo assays to predict and develop insights into the medicinal properties of SA in terms of anti-oxidation, anti-inflammation, and anti-diabetes. The molecular docking displayed significantly binding affinity for SA towards tumor necrosis factor-alpha, interleukin-6, and antioxidant enzymes when inflammation from myocardial infarction and spinal cord ischemia. Moreover, SA nicely docked with dipeptidyl peptidase-IV, glucagon-like peptide 1 receptor, peroxisome proliferator-activated receptor, acetylcholine M2 receptor, and acetylcholinesterase in anti-diabetes investigations. These are associated with (1) an increase glucose utilization and insulin sensitivity to an anti-hyperglycemic effect; and (2) to potentiate intestinal contractility to abolish the alpha-amylase reaction when concurrently reducing retention time and glucose absorption of the intestinal tract to achieve a glucose-lowering effect. In silico screening of multi-targets concomitantly with preclinical tests could provide a potential exploration for new indications for drug discovery and development. Title: Lethal and Sublethal Toxicity Assessment of Cyclosporin C (a Fungal Toxin) against Plutella xylostella (L.) Wu J, Zhang X, Bashir MH, Ali S Ref: Toxins (Basel), 14:, 2022 : PubMed ESTHER: Wu_2022_Toxins.(Basel)_14_ PubMedSearch: Wu 2022 Toxins.(Basel) 14 PubMedID: 36006176 Abstract Secondary metabolites/toxins produced by Purpeocillium lilacinum (Hypocreales; Phiocordycipitaceae), a well-known insect pathogen, can be used for the management of different insect pests. We report the lethal and sublethal effects of cyclosporin C (a toxin produced by P. lilacinum) against a major vegetable pest, Plutella xylostella, at specific organismal (feeding rate, larval growth, adult emergence, fecundity, and adult longevity) and sub-organismal levels (changes in antioxidant and neurophysiological enzyme activities). The toxicity of cyclosporin C against different larval instars of P. xylostella increased with increasing concentrations of the toxin and the maximum percent mortality rates for different P. xylostella larval instars at different times were observed for the 300 microg/mL cyclosporin C treatment, with an average mortality rate of 100% for all larval instars. The median lethal concentrations (LC(50)) of cyclosporin C against the first, second, third, and fourth larval instars of P. xylostella 72 h post-treatment were 78.05, 60.42, 50.83, and 83.05 microg/mL, respectively. Different concentrations of cyclosporin C caused a reduction in the average leaf consumption and average larval weight. Different life history parameters, such as the pupation rate (%), adult emergence (%), female fecundity, and female longevity were also inhibited when different concentrations of cyclosporin C were applied topically. The cyclosporin C concentrations inhibited the activities of different detoxifying (glutathione S-transferase, carboxylesterase, and acetylcholinesterase) and antioxidant enzyme (superoxide dismutase, catalase, and peroxidase) activities of P. xylostella when compared to the control. These findings can serve as baseline information for the development of cyclosporin C as an insect control agent, although further work on mass production, formulation, and field application is still required. Title: A novel approach for on-site screening of organophosphorus nerve agents based on DTNB modified AgNPs using surface-enhanced Raman spectrometry Wu J, Zhu Y, Liu Y, Chen J, Guo L, Xie J Ref: Anal Methods, :, 2022 : PubMed ESTHER: Wu_2022_Anal.Methods__ PubMedSearch: Wu 2022 Anal.Methods PubMedID: 36285727 Abstract Organophosphorus nerve agents (OPNAs), such as Sarin (GB), Tabun (GA), Soman (GD) and VX, would cause tremendous harm in military and terrorist attacks, and thus the development of simple methods for the rapid and efficient detection of these hazardous substances is of great necessity. Herein, we present a novel approach for the facile, rapid and sensitive detection of real OPNAs. The detection substrate is fabricated using functionalized silver nanoparticles (AgNPs) immobilized with acetylcholinesterase (AChE) and 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB). In the absence of OPs, AChE catalyzes the hydrolysis of acetylthiocholine (ATCh) to form thiocholine (TCh), which continues to interact quickly with DTNB to produce a very sensitive Raman probing molecule, TNB. The inhibition of the activity of AChE by OPs could induce an obvious decrease of characteristic Raman peaks of 5-thio-2-nitrobenzoic acid (TNB) at 1335 cm(-1). The introduction of DTNB as an enzyme activity indicator significantly improves the detection sensitivity with distinct characteristic Raman peaks. The LOD of GD, which is one of the most easily aged OPNAs, could reach 0.1 nM due to its strongest inhibition of AChE. Moreover, various OPNAs exhibit different SERS intensities due to their different inhibition capacities of AChE. Hence, the new strategy has great potential in public security early warning and environmental analysis. Title: From tryptamine to the discovery of efficient multi-target directed ligands against cholinesterase-associated neurodegenerative disorders Wu J, Zhang H, Wang Y, Yin G, Li Q, Zhuo L, Chen H, Wang Z Ref: Front Pharmacol, 13:1036030, 2022 : PubMed ESTHER: Wu_2022_Front.Pharmacol_13_1036030 PubMedSearch: Wu 2022 Front.Pharmacol 13 1036030 PubMedID: 36518670 Abstract A novel class of benzyl-free and benzyl-substituted carbamylated tryptamine derivatives (CDTs) was designed and synthesized to serve as effective building blocks for the development of novel multi-target directed ligands (MTDLs) for the treatment of neurological disorders linked to cholinesterase (ChE) activity. The majority of them endowed butyrylcholinesterase (BuChE) with more substantial inhibition potency than acetylcholinesterase (AChE), according to the full study of ChE inhibition. Particularly, hybrids with dibenzyl groups (2b-2f, 2j, 2o, and 2q) showed weak or no neuronal toxicity and hepatotoxicity and single-digit nanomolar inhibitory effects against BuChE. Through molecular docking and kinetic analyses, the potential mechanism of action on BuChE was first investigated. In vitro H(2)O(2)-induced HT-22 cells assay demonstrated the favorable neuroprotective potency of 2g, 2h, 2j, 2m, 2o, and 2p. Besides, 2g, 2h, 2j, 2m, 2o, and 2p endowed good antioxidant activities and COX-2 inhibitory effects. This study suggested that this series of hybrids can be applied to treat various ChE-associated neurodegenerative disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD), as well as promising building blocks for further structure modification to develop efficient MTDLs. Title: Synergism Between Multi-Pseudomonas and Cutinase for Biodegradation of Crude Oil-Based Derivatives Yan ZF, Xu KW, Wu J Ref: Curr Microbiol, 80:30, 2022 : PubMed ESTHER: Yan_2022_Curr.Microbiol_80_30 PubMedSearch: Yan 2022 Curr.Microbiol 80 30 PubMedID: 36474116 Abstract Polyethylene terephthalate (PET) as one of the main crude oil-based derivatives, produces a significant amount of waste that is difficult to degrade. Currently, microbial degradation of PET is an eco-friendly, efficient, and economical method. This study was conducted to propose a novel screening strategy for PET-degrading bacteria, and evaluate their degradation efficiency of PET. Two strains, Pseudomonas nitroreducens S8 and Pseudomonas monteilii S17, were isolated and could utilize PET as a carbon source by co-culture. The combined use of both bacteria gave a synergistic effect on the disruption of the PET surface through colonization behavior, which could enhance the subsequent degradation of PET. Its time of reaching a peak value of PET degradation rate (94.5% at 6 d) was 2sdays earlier than these of single bacteria. A similar synergistic effect was also observed in the metabolization of PET monomers, and the metabolic rate was expressed as 82.4% of bis (2-hydroxyethyl) terephthalate (BHET), 64.0% of mono (2-hydroxyethyl) terephthalate (MHET), and 20.0% of terephthalic acid (TPA), respectively. This study is novel in showing the degradation of PET waste by combinations of bacterial pretreatment and enzymatic treatment, which can be a promising method. Title: A multifunctional anti-AD approach: Design, synthesis, X-ray crystal structure, biological evaluation and molecular docking of chrysin derivatives Yang A, Liu C, Zhang H, Wu J, Shen R, Kou X Ref: Eur Journal of Medicinal Chemistry, 233:114216, 2022 : PubMed ESTHER: Yang_2022_Eur.J.Med.Chem_233_114216 PubMedSearch: Yang 2022 Eur.J.Med.Chem 233 114216 PubMedID: 35227980 Abstract With the aging of the population intensifying, finding a cure or reasonable treatment for Alzheimer' disease (AD) has become an urgent priority. To target the multi-facets of AD, a class of chrysin derivatives (1-4) were rationally designed and synthesized by the multi-target-directed ligands (MTDLs) strategy, which were characterized by (1)H NMR, (13)C NMR, MS and elemental analysis. 1-4 showed inhibitory activities on reactive oxygen species, Abeta(1-42) aggregation (self-, Cu(2+)-induced, AChE-induced). They were also potent inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) with selectivity toward BuChE. Compound 1 as the most promising candidate exhibited the highest selective BuChE inhibition (SI = 15). Furthermore, the kinetic study suggested compound 1 to be a mixed type inhibitor. The results of docking study were consistent with the in vitro results. In addition, compound 1-4 showed favorable blood-brain barrier (BBB) penetration and drug-like property in silico prediction. The corresponding copper complexes of 1-4 have also been synthesized. 1-4 selectively chelated Cu(2+), Fe(2+), Zn(2+) and Al(3+) ions, while had no chelating ability to other biometals. The copper complexes also showed good AChE, BuChE and reactive oxygen species inhibitory activities. Notably, the single crystals of 1-Cu(II) complex [Cu(C(19)H(18)NO(4))(2)] were prepared for the first time and characterized by X-ray single crystal diffraction. X-ray crystallography analysis of 1-Cu(II) complex provided a reliable structure-activity insight at the molecular level about the antioxidative and Abeta(1-42) disaggregation activities. Compound 1 might be a good lead compound to develop promising candidate analogs as AD therapeutics. Title: Hepatitis B virus genotype is an independent prognostic factor of telbivudine and tenofovir treatment in hepatitis B surface antigen-positive pregnant women Zhang B, Yu L, Cheng M, Zhang Q, Wu J, Yang J, Liu Q, Lu S, Zhao X and Zhao P <3 more author(s)> Zhang B, Yu L, Cheng M, Zhang Q, Wu J, Yang J, Liu Q, Lu S, Zhao X, Deng K, Liu Y, Wang J, Zhao P (- 3) Ref: Food Sci Nutr, 10:3, 2022 : PubMed ESTHER: Zhang_2022_Food.Sci.Nutr_10_3 PubMedSearch: Zhang 2022 Food.Sci.Nutr 10 3 PubMedID: 35035905 Abstract To investigate whether HBV genotype influences the effect of tenofovir and telbivudine on HBV DNA and RNA levels in HBsAg-positive pregnant women. This was a retrospective study of 74 HBsAg-positive pregnant women in Guizhou of China. All patients were treated with telbivudine or tenofovir from 12 weeks of pregnancy and HBV infection to the date of delivery. Blood samples were collected at 12-24, 28-32, and 36-40 weeks of pregnancy for the measurement of genotype, HBsAg, hepatitis B e antigen (HBeAg), HBV DNA, HBV RNA, and liver function, including alanine transaminase, aspartate transaminase, total bilirubin, total bile acids, cholinesterase, alkaline phosphatase (ALP), and gamma-glutamyl transferase. All women with HBsAg were followed up. The HBV genotype was B in 64.9% and C in 35.1%. There were 37 patients of telbivudine and tenofovir group respectively. The telbivudine and tenofovir groups showed no differences in demographic and clinical characteristics, including liver function tests, HBsAg, HBeAg, log(10)(HBV DNA), and log(10)(HBV RNA). Compared with baseline (12-24 weeks), telbivudine group showed a significant increase in ALP and significant reductions in HBsAg, HBeAg, log(10)(HBV DNA), and log(10)(HBV RNA) at 36-40 weeks (p < .05). Tenofovir group exhibited a significant increase in ALP and significant reductions in HBeAg, log(10)(HBV DNA), and log(10)(HBV RNA) at 36-40 weeks, compared with baseline (p < .05). HBV genotype (B vs. C) was independently associated with HBV DNA change after therapy (p = .005). In telbivudine group, log(10) (HBV DNA) increased from 3.38 (2.00-7.30) to 7.43 (4.68-8.70). In tenofovir group, log(10) (HBV DNA) decreased from 7.52 (3.32-8.70) to 2.98 (2.00-5.01). HBV genotype was independently associated with HBV DNA change response to telbivudine or tenofovir in pregnant women with hepatitis B. These findings might be helpful for risk assessment regarding vertical transmission of HBV in HBeAg-positive mothers treated with nucleos(t)ide analogues. Title: Chain-locked precursor ion scanning based HPLC-MS/MS for in-depth molecular analysis of lipase-catalyzed transesterification of structured phospholipids containing w-3 fatty acyl chains Zhang M, Wang P, Jin D, Jian S, Wu J, Huang M, Xie H, Zhao Q, Yang H and Shen Q <3 more author(s)> Zhang M, Wang P, Jin D, Jian S, Wu J, Huang M, Xie H, Zhao Q, Yang H, Luo P, Yuan H, Xue J, Shen Q (- 3) Ref: Food Chem, 399:133982, 2022 : PubMed ESTHER: Zhang_2022_Food.Chem_399_133982 PubMedSearch: Zhang 2022 Food.Chem 399 133982 PubMedID: 36027811 Abstract Lipase-catalyzed transesterification of structured phospholipids (sPLs) is a hot topic, but the structural variation of the fatty acyl chains in intact phospholipids at the molecular level remains unclear to date. The present study explored the detailed characteristics of synthesized phospholipids through high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in precursor ion scan mode. The optimal conditions were in-depth inspected and determined for the reaction system, including phospholipase A1 as catalyst, 15% lipase loading, and 1% water content. The sPLs enriched with EPA/DHA were structurally and quantitatively characterized by focusing on the fragments of m/z 301.6 (eicosapentaenoic acid, EPA) and m/z 327.6 (docosahexaenoic acid, DHA), and the results were statistically analyzed using partial least squares discriminant analysis and clustered heatmap hierarchical clustering analysis. PC 38:6 (18:1/20:5), PC 38:7 (18:2/20:5), PC o-40:6 (o-18:0/22:6), and PE 40:8 (18:2/22:6) etc. were revealed as the main variables that were active in the reaction. Title: Toxicological and Biochemical Description of Synergism of Beauveria bassiana and Emamectin Benzoate against Megalurothrips usitatus (Bagrall) Zhang Y, Zhang X, Tian Q, Ali S, Tang L, Wu J Ref: J Fungi (Basel), 8:, 2022 : PubMed ESTHER: Zhang_2022_J.Fungi.(Basel)_8_ PubMedSearch: Zhang 2022 J.Fungi.(Basel) 8 PubMedID: 36135641 Abstract The prophylactic application of synthetic insecticides to manage Megalurothrips usitatus (Bagrall) has resulted in insecticide resistance and negative impacts upon natural ecosystems. This has driven the need for developing alternative pest control strategies. In the present study, we investigated the synergistic interaction between the entomopathogenic fungus Beauveria bassiana and the insecticide emamectin benzoate on M. usitatus. The results of our research exhibited that higher doses of emamectin benzoate inhibited the germination rate and colony growth of B. bassiana. The percentage of M. usitatus mortality following B. bassiana and emamectin benzoate treatment indicated a dose-mortality effect. All concentrations of emamectin benzoate combined with different concentrations of B. bassiana demonstrated a synergistic effect five days post-treatment. When B. bassiana and emamectin benzoate were applied alone or in combination, antioxidant enzyme activities, including acetylcholinesterase, catalase, superoxide dismutase, and peroxidase, were significantly lower in M. usiatus than in the controls at the end of the experimental period. The findings of our study confirm the synergistic effect of B. bassiana and emamectin benzoate on M. usitatus, as well as the biochemical process that might be involved in the regulation of the synergistic effect. Title: Construction of a red emission fluorescent protein chromophore-based probe for detection of carboxylesterase 1 and carbamate pesticide in culture cells Dai J, Hou Y, Wu J, Zhong G, Gao R, Shen B, Huang H Ref: Talanta, 223:121744, 2021 : PubMed ESTHER: Dai_2021_Talanta_223_121744 PubMedSearch: Dai 2021 Talanta 223 121744 PubMedID: 33298268 Abstract Designing fluorescent probe for detecting carboxylesterase 1 is remains challenging. Herein, a red emission human carboxylesterase 1 (CES1) probe (CAE-FP) was synthesized based on fluorescent protein chromophore. Probe CAE-FP can specific detect human CES1 with high selectively. The fluorescence quantum yield was calucated as 0.19. The carboxylic acid ester in CAE-FP could be easily hydrolyzed by CES1 under physiological conditions, and this process could induce the obvious fluorescence signal in red emission region. The detection limit of CES1 was calculated as 84.5 ng/mL. Due to the biological detoxification mechanism of carboxylesterase and the obvious inhibitory effect of pesticides on its activity, CAE-FP was applied to detect carbamate pesticide and have achieved good application results. Since fluorescent protein chromophore has excellent biocompatibility, probe CAE-FP with good cell membrane permeable and was successfully applied to monitor the real activities of CES1 in living cells. In summary, this is one of the few reported fluorescent probes that can specific detect the real-time activity of CES1 in biological samples. Besides, we first applied the fluorescent protein chromophore to construct the specific target enzyme probe. This work would contribute to further investigate CES1-associated physiological and pathological processe. Title: Detection of carboxylesterase 1 and carbamates with a novel fluorescent protein chromophore based probe Dai J, Zhao Y, Hou Y, Zhong G, Gao R, Wu J, Shen B, Zhang X Ref: Dyes and Pigments, 192:109444, 2021 : PubMed ESTHER: Dai_2021_Dyes.Pigments_192_109444 PubMedSearch: Dai 2021 Dyes.Pigments 192 109444 Abstract An aggregation-induced emission (AIE) fluorescent protein chromophore-based probe (CBZ-FP) for detection of human carboxylesterases (CESs) was designed and synthesized. CBZ -FP exhibited good cell permeability with a large stokes shift (116snm) and can be applied to reveal the actual activities of CES1 in living cells associated with pesticide s detoxification process. CBZ-FP can also serve as a fluorescence indicator of pesticide exposure in the way of hydrolyzing the carboxylic acid ester group in CBZ-FP. There fore, CBZ-FP has high selectivity for CESs and can detect real-time activity of CES1 in biological samples. Molecular docking study was used to explore the binding of CESs and CBZ-FP. Finding that only one specific activity site of CESs can bind with probe. In view of the fact that, the biotransformation of drugs and poisons containing ester groups can carry out normally depending on CESs, Carboxylesterase probes are expected to contribute to the characterization of relevant disease. Title: Accelerated biodegradation of polyethylene terephthalate by Thermobifida fusca cutinase mediated by Stenotrophomonas pavanii Huang QS, Yan ZF, Chen XQ, Du YY, Li J, Liu ZZ, Xia W, Chen S, Wu J Ref: Sci Total Environ, :152107, 2021 : PubMed ESTHER: Huang_2021_Sci.Total.Environ__152107 PubMedSearch: Huang 2021 Sci.Total.Environ 152107 PubMedID: 34864034 Abstract Polyethylene terephthalate (PET) is a general plastic that produces a significant amount of waste due to its non-degradable properties. We obtained four bacteria (Stenotrophomonas pavanii JWG-G1, Comamonas thiooxydans CG-1, Comamonas koreensis CG-2 and Fulvimonas soli GM-1) that utilize PET as a sole carbon source through a novel stepwise screening and verification strategy. PET films pretreated with S. pavanii JWG-G1 exhibited weight loss of 91.4% following subsequent degradation by Thermobifida fusca cutinase (TfC). S. pavanii JWG-G1 was able to colonize the PET surface and maintain high cell viability (over 50%) in biofilm, accelerating PET degradation. Compared with PET films with no pretreatment, pretreatment with S. pavanii JWG-G1 caused the PET surface to be significantly rougher with greater hydrophilicity (contact angle of 86.3 +/- 2 degrees vs. 96.6 +/- 2 degrees), providing better opportunities for TfC to contact and act on PET. Our study indicates that S. pavanii JWG-G1 could be used as a novel pretreatment for efficiently accelerating PET biodegradation by TfC. Title: Application of deep eutectic solvent-based extraction coupled with an S-CQD fluorescent sensor for the determination of pirimicarb in cereals Jing X, Wu J, Wang H, Guo L, Zheng X, Wang X, Wang S Ref: Food Chem, 370:131360, 2021 : PubMed ESTHER: Jing_2021_Food.Chem_370_131360 PubMedSearch: Jing 2021 Food.Chem 370 131360 PubMedID: 34662796 Abstract A novel deep eutectic solvent-based extraction and sulfur-doped carbon quantum dots (S-CQDs) serving as fluorescence probes to detect pirimicarb in cereals were established. The deep eutectic solvent was synthesized using choline chloride and butanediol, achieving direct and efficient extraction of pirimicarb residue in the cereals. The fluorescence quenching of S-CQDs was caused by the electrostatic interaction between the negatively charged S-CQDs and positively charged thiocholine, which was the hydrolysate of acetylthiocholine. The fluorescence of S-CQDs was enhanced as the activity of acetylcholinesterase (AChE) was inhibited by pirimicarb, achieving the detection of pirimicarb in the cereal samples. The limit of detection (LOD) was 0.006 microg mL(-1). The recovery ranged from 96.6% to 108.2%. This extraction and detection method of pirimicarb based on an environmentally friendly DES and S-CQD fluorescent sensor maintains good stability and convenience, offering a promising strategy for extracting and testing harmful substances in food samples. Title: pH and Redox Dual-Response Disulfide Bond-Functionalized Red-Emitting Gold Nanoclusters for Monitoring the Contamination of Organophosphorus Pesticides in Foods Li Q, Wu J, Yang Q, Li H, Li F Ref: Analytical Chemistry, 93:7362, 2021 : PubMed ESTHER: Li_2021_Anal.Chem_93_7362 PubMedSearch: Li 2021 Anal.Chem 93 7362 PubMedID: 33961403 Abstract Most of the fluorescence sensors require choline oxidase or quenchers to detect organophosphorus pesticides (OPs) based on a single hydrolysate and suffer from high cost, complex procedures, weak stability, and low sensitivity. Here, we proposed a brand-new fluorescence strategy for highly sensitive detection of OPs based on both hydrolysate-response disulfide bond-functionalized gold nanoclusters (S-S-AuNCs) without additional substances. S-S-AuNCs were synthesized via a facile one-step redox reaction and emitted bright red light with ultrasmall size and high water dispersion. Interestingly, S-S-AuNCs displayed a unique response to thiol compounds and low pH values and were thus pioneered as a high-efficiency sensor for OPs based on acetylcholinesterase (AChE)-catalyzed hydrolysis of acetylthiocholine into thiocholine and CH(3)COOH and OP inhibition of AChE activity. Further, S-S-AuNCs were employed to monitor the residue, distribution, and metabolization of methidathion in pakchoi with acceptable results. We believe that this work supplies a simpler and more highly sensitive approach for OP assay than the known ones and opens a new avenue to development of multistimulus-responsive and high-performance fluorescence substances. Title: Novel chrysin derivatives as hidden multifunctional agents for anti-Alzheimer's disease: design, synthesis and in vitro evaluation Liu C, Kou X, Wang X, Wu J, Yang A, Shen R Ref: Eur J Pharm Sci, :105976, 2021 : PubMed ESTHER: Liu_2021_Eur.J.Pharm.Sci__105976 PubMedSearch: Liu 2021 Eur.J.Pharm.Sci 105976 PubMedID: 34419572 Abstract Alzheimer's disease (AD) is the most common type of dementia, the exact etiology of the disease has not been known yet. The use of single-target drugs limits the efficacy of drugs and has certain side effects. In this study, the 'hidden' multi-target strategy was used in combination with chrysin's metal chelating site and rivastigmine's anti-cholinesterase pharmacophore to form an ester, which improves the hydrophobicity and protects the phenolic hydroxyl group at the same time. Four derivatives (1-4) were synthesized as the hidden multifunctional agents for AD therapy. Most of the compounds displayed good activities of anti-cholinesterase, antioxidant, appropriate blood brain barrier (BBB) penetration and certain inhibitory activity of beta-amyloid (Abeta) aggregation. Compound 3 was demonstrated as the highest selective butyrylcholinesterase (BuChE) inhibitor and targeted both the catalytic active site (CAS) and the peripheral anion site (PAS). And it could be hydrolyzed by BuChE to release chrysin with good ability to chelate Cu(2+) and Fe(2+). At the same time, phenol fragment can exert its good antioxidant effect. Overall, these findings demonstrated that compound 3 might be considered as a potential hidden multifunctional candidate in the therapy of AD. Title: Gender Disparities in Anti-dementia Medication Use among Older Adults: Health Equity Considerations and Management of Alzheimer's Disease and Related Dementias Lu ZK, Xiong X, Wang X, Wu J Ref: Front Pharmacol, 12:706762, 2021 : PubMed ESTHER: Lu_2021_Front.Pharmacol_12_706762 PubMedSearch: Lu 2021 Front.Pharmacol 12 706762 PubMedID: 34512340 Abstract Objective: The prevalence of Alzheimer's disease and related dementias (ADRD) in women is higher than men. However, the knowledge of gender disparity in ADRD treatment is limited. Therefore, this study aimed to determine the gender disparities in the receipt of anti-dementia medications among Medicare beneficiaries with ADRD in the U.S. Methods: We used data from the Medicare Current Beneficiary Survey 2016. Anti-dementia medications included cholinesterase inhibitors (ChEIs; including rivastigmine, donepezil, and galantamine) and N-methyl-D-aspartate (NMDA) receptor antagonists (including memantine). Descriptive analysis and multivariate logistic regression models were implemented to determine the possible gender disparities in the receipt of anti-dementia medications. Subgroup analyses were conducted to identify gender disparities among beneficiaries with Alzheimer's disease (AD) and those with only AD-related dementias. Results: Descriptive analyses showed there were statistically significant differences in age, marital status, and Charlson comorbidities index (CCI) between Medicare beneficiaries who received and who did not receive anti-dementia medications. After controlling for covariates, we found that female Medicare beneficiaries with ADRD were 1.7 times more likely to receive anti-dementia medications compared to their male counterparts (odds ratio [OR]: 1.71; 95% confidence interval [CI]: 1.19-2.45). Specifically, among Medicare beneficiaries with AD, females were 1.2 times more likely to receive anti-dementia medications (Odds Radio: 1.20; 95% confidence interval: 0.58-2.47), and among the Medicare beneficiaries with only AD-related dementias, females were 1.9 times more likely to receive anti-dementia medications (OR: 1.90; 95% CI: 1.23-2.95). Conclusion: Healthcare providers should be aware of gender disparities in receiving anti-dementia medications among patients with ADRD, and the need to plan programs of care to support both women and men. Future approaches to finding barriers of prescribing, receiving and, adhering to anti-dementia medications by gender should include differences in longevity, biology, cognition, social roles, and environment. Title: Osteocyte- and late Osteoblast-derived NOTUM Reduces Cortical Bone Mass in Mice Nilsson KH, Henning P, El Shahawy M, Wu J, Koskela A, Tuukkanen J, Perret C, Lerner UH, Ohlsson C, Moverare-Skrtic S Ref: American Journal of Physiology Endocrinol Metab, :, 2021 : PubMed ESTHER: Nilsson_2021_Am.J.Physiol.Endocrinol.Metab__ PubMedSearch: Nilsson 2021 Am.J.Physiol.Endocrinol.Metab PubMedID: 33749332 Gene_locus related to this paper: human-NOTUM, mouse-notum Abstract Osteoporosis is a common skeletal disease, with increased risk of fractures. Currently available osteoporosis treatments reduce the risk of vertebral fractures, mainly dependent on trabecular bone, whereas the effect on non-vertebral fractures, mainly dependent on cortical bone, is less pronounced. WNT signaling is a crucial regulator of bone homeostasis, and the activity of WNTs is inhibited by NOTUM, a secreted WNT lipase. We previously demonstrated that conditional inactivation of NOTUM in all osteoblast lineage cells increases the cortical but not the trabecular bone mass. The aim of the present study was to determine if NOTUM increasing cortical bone is derived from osteoblast precursors/early osteoblasts or from osteocytes/late osteoblasts. First, we demonstrated Notum mRNA expression in Dmp1-expressing osteocytes and late osteoblasts in cortical bone using in situ hybridization. We then developed a mouse model with inactivation of NOTUM in Dmp1 expressing osteocytes and late osteoblasts (Dmp1-creNotum(flox/flox) mice). We observed that the Dmp1-creNotum(flox/flox) mice displayed a substantial reduction of Notum mRNA in cortical bone, resulting in increased cortical bone mass and decreased cortical porosity in femur, but no change in trabecular bone volume fraction (BV/TV) in femur or in the lumbar vertebrae L5 in Dmp1-creNotum(flox/flox) mice as compared to control mice. In conclusion, osteocytes and late osteoblasts are the principal source of NOTUM in cortical bone, and NOTUM derived from osteocytes/late osteoblasts reduces cortical bone mass. These findings demonstrate that inhibition of osteocyte/late osteoblast-derived NOTUM might be an interesting pharmacological target to increase cortical bone mass and reduce non-vertebral fracture risk. Title: Detoxification II Prescription Suppresses the Th-17/IL-17 Inflammatory Axis to Improve the Liver Function of ACLF-Rats via Inactivating the P38MAPK Pathway Shi Q, Bai W, Mao D, Chen Y, Wang K, Qiu H, Wu J Ref: J Healthc Eng, 2021:7563383, 2021 : PubMed ESTHER: Shi_2021_J.Healthc.Eng_2021_7563383 PubMedSearch: Shi 2021 J.Healthc.Eng 2021 7563383 PubMedID: 34900202 Abstract Hepatitis is a metabolic system disease which is a serious challenge to the medical and healthcare system of the world. This study attempted to investigate the therapeutic effect and illustrate the regulation pharmacological mechanism of Detoxification II Prescription on ACLF. In this study, the rats were injected with D-galactosamine to establish ACLF-rat models, and the levels of cholinesterase (CHE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), and total bilirubin (TBiL) were measured with the related kits to reflect the liver functions of the rats. The levels of IL-17, IL-6, and IFN-gamma in the serums of the rats were detected by qRT-PCR, and the percentages of Th-17 cells in CD4(+) cells of the rats were measured by flow cytometry assay. In the results, the increased ALT, AST, TBiL, IL-6, IL-17, IFN-gamma, and percentage of Th-17 cells in CD4(+) and decreased ALB and CHE were found in the serums of the ACLF-rats, while Detoxification II Prescription could partly reverse those indexes of the ACLF-rats. Moreover, it was also found that Detoxification II Prescription could inhibit the expression of P38MAPK, and P38MAPK downregulation obviously improved the liver function indexes of the ACLF-rats including the levels of ALT, AST, TBiL, IL-6, IL-17, IFN-gamma, and percentage of Th-17 cells in CD4(+) cells. In conclusion, this study suggested that Detoxification II Prescription could suppress the Th-17/IL-17 inflammatory axis to improve the liver function of ACLF-rats via inhibiting the activity of the P38MAPK pathway. Title: Japonisine A, a fawcettimine-type Lycopodium alkaloid with an unusual skeleton from Lycopodium japonicum Thunb Wang X, Wang F, Wu J, Chen SQ, Jiang CS, Yang SP, Wang C, Cai YS Ref: Fitoterapia, 156:105069, 2021 : PubMed ESTHER: Wang_2021_Fitoterapia_156_105069 PubMedSearch: Wang 2021 Fitoterapia 156 105069 PubMedID: 34743932 Abstract Japonisine A, a novel fawcettimine-type Lycopodium alkaloid with an unusual skeleton and two new fawcettimine-type ones, along with 20 known Lycopodium alkaloids, were isolated from the whole plants of Lycopodium japonicum Thunb. Their structures were determined by extensive spectroscopic analysis, including 1D and 2D NMR, and HR-ESIMS, as well as by comparison with the literature data. Notably, japonisine A (1) was the first example of fawcettimine-related Lycopodium alkaloid with a 2-oxopropyl attached at C-6. All the isolates were evaluated for their inhibitory effects on acetylcholinesterase (AChE) and alpha-glucosidase. Unfortunately, the results indicated that all the compounds were inactive against the acetylcholinesterase (AChE) and alpha-glucosidase. Title: Two-Dimensional MnO(2) Nanozyme-Mediated Homogeneous Electrochemical Detection of Organophosphate Pesticides without the Interference of H(2)O(2) and Color Wu J, Yang Q, Li Q, Li H, Li F Ref: Analytical Chemistry, :, 2021 : PubMed ESTHER: Wu_2021_Anal.Chem__ PubMedSearch: Wu 2021 Anal.Chem PubMedID: 33588528 Abstract Traditional peroxidase-like nanozyme-based sensors suffer from self-decomposition and high toxicity of H(2)O(2), as well as the interference of color from nanozymes themselves and testing samples. In this work, we adopt nanozymes (two-dimension (2D) MnO(2) sheets, manganese dioxide nanosheets (MnNS)) with oxidase-like and peroxidase-like properties as advanced catalysts to develop a novel homogeneous electrochemical sensor for organophosphate pesticides (OPs) using dissolved O(2) as a coreactant without the interference of H(2)O(2) and color. Owing to the large surface area and unique catalytic activity of MnNS, a large amount of tetramethylbenzidine (TMB) is catalyzed oxidation, leading to a significantly declined differential pulse voltammetry (DPV) current. Obviously, MnNS display an excellent response to thiocholine, deriving from the catalyzing hydrolysis of acetylthiocholine (ATCh) by acetylcholinesterase (AChE), which switches a homogeneous electrochemical OP detection process based on the depressing AChE activity with a limit of detection (LOD) of 0.025 ng mL(-1). The as-proposed strategy on using nanozymes with oxidase-like and peroxidase-like properties to develop a homogeneous electrochemical sensor will provide a new pathway for improving the performance of nanozyme-based sensors, and the established MnNS-based homogeneous electrochemical sensor will find more applications for OP residue determination in food samples. Title: Inhibition of soluble epoxide hydrolase (sEH) protects hippocampal neurons and reduces cognitive decline in type 2 diabetic mice Wu J, Fan Z, Zhao Y, Chen Q, Xiao Q Ref: European Journal of Neuroscience, :, 2021 : PubMed ESTHER: Wu_2021_Eur.J.Neurosci__ PubMedSearch: Wu 2021 Eur.J.Neurosci PubMedID: 33595911 Abstract Diabetes mellitus is a metabolic disorder that can lead to cognitive dysfunction. The hippocampus plays an important role in the cognitive function. Research has identified correlations between hippocampal impairment and diabetes, yet their intermediate remains unclear. Soluble epoxide hydrolase (sEH) is an enzyme that degrades epoxyeicosatrienoic acids (EETs), which have multiple protective effects by suppressing inflammation, apoptosis and oxidative stress. In this study, under diabetic conditions both hippocampal injury, and cognitive decline are accompanied by upregulation of sEH. Moreover, the sEH inhibitor trans-4-[4-(3-adamantan-1-y1-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB) prevents cognitive dysfunction and decreased ROS accumulation and apoptosis in the diabetic hippocampus. t-AUCB treatment restored neuronal synaptic plasticity by restoring the expression of the postsynaptic proteins Postsynaptic density protein-95 (PSD95) and N-methyl-d-aspartate receptor subunit 2B (NR2B), the levels of which were positively correlated with Proline-rich tyrosine kinase 2 (Pyk2) levels under diabetic conditions. Thus, we suggest that hippocampal protection via sEH inhibition might be a potential therapeutic approach to attenuate the progression of cognitive decline in diabetes. Title: Synergistic Interaction between the Entomopathogenic Fungus Akanthomyces attenuatus (Zare & Gams) and the Botanical Insecticide Matrine against Megalurothrips usitatus (Bagrall) Wu J, Yang B, Zhang X, Cuthbertson AGS, Ali S Ref: J Fungi (Basel), 7:, 2021 : PubMed ESTHER: Wu_2021_J.Fungi.(Basel)_7_ PubMedSearch: Wu 2021 J.Fungi.(Basel) 7 PubMedID: 34356915 Abstract The excessive use of synthetic chemicals for Megalurothrips usitatus (Bagrall) management has resulted in the development of insecticide resistance as well as adverse effects to the natural ecosystem. This has driven the need to develop alternative pest control strategies. This study reports a synergistic interaction between the entomopathogenic fungus Akanthomyces attenuatus (Zare & Gams) and the botanical insecticide matrine against M. usitatus. The results revealed that the germination rate and colony growth of A. attenuatus were inhibited by higher matrine concentrations. Percentage mortalities of M. usitatus following application of A. attenuatus and matrine showed a dose mortality effect. After five days of treatment, all concentrations of matrine combined with different concentrations of A. attenuatus, except one combination (matrine 0.25 mg/mL + 1 x 10(7) conidia/mL), showed synergistic effect. The activities of acetylcholinesterase and antioxidant enzymes (superoxide dismutase, catalase and peroxidase) in M. usitatus, in response to individual or combined application of A. attenuatus and matrine at the end of the experimental period, were significantly lower than controls. The findings confirm the synergistic action of A. attenuatus and matrine against M. usitatus along with the biochemical phenomenon possibly regulating the synergistic effect. Title: Design, synthesis and biological evaluation of naringenin carbamate derivatives as potential multifunctional agents for the treatment of Alzheimer's disease Wu J, Kou X, Ju H, Zhang H, Yang A, Shen R Ref: Bioorganic & Medicinal Chemistry Lett, :128316, 2021 : PubMed ESTHER: Wu_2021_Bioorg.Med.Chem.Lett__128316 PubMedSearch: Wu 2021 Bioorg.Med.Chem.Lett 128316 PubMedID: 34391893 Abstract A series of naringenin derivatives were designed and synthesized as multifunctional anti-Alzheimer's disease (AD) agents. The results showed that these derivatives displayed moderate-to-good acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities at the micromolar range (IC(50), 12.91-62.52 microM for AChE and 0.094-13.72 microM for BuChE). Specifically, compound 1 showed the highest inhibitory activity against BuChE with the IC(50) value of (0.094+/-0.0054) microM. A Lineweaver-Burk plot and molecular docking studies demonstrated that 1 targeted both the catalytically active site (CAS) and the peripheral anion site (PAS) of BuChE. Besides, all derivatives showed excellent hydroxyl free radicals (.OH) scavenging ability than vitamin C and cyclic voltammetry results displayed that 1 could effectively scavenge superoxide anion radical (.O(2)(-)). In addition, compound 1 displayed good metal chelating properties and had anti-Abeta aggregation activities. Therefore, compound 1 might be the potential anti-AD agent for further developments. Title: Synergistic biodegradation of poly(ethylene terephthalate) using Microbacterium oleivorans and Thermobifida fusca cutinase Yan ZF, Wang L, Xia W, Liu ZZ, Gu LT, Wu J Ref: Applied Microbiology & Biotechnology, :, 2021 : PubMed ESTHER: Yan_2021_Appl.Microbiol.Biotechnol__ PubMedSearch: Yan 2021 Appl.Microbiol.Biotechnol PubMedID: 34037842 Abstract Poly(ethylene terephthalate) (PET) is a major source of plastic pollution. Biodegradation technologies are of paramount interest in reducing or recycling PET waste. In particular, a synergistic microbe-enzyme treatment may prove to be a promising approach. In this study, a synergistic system composed of Microbacterium oleivorans JWG-G2 and Thermobifida fusca cutinase (referred to as TfC) was employed to degrade bis(hydroxyethyl) terephthalate (BHET) oligomers and a high crystalline PET film. A novel degradation product that was obtained by M. oleivorans JWG-G2 treatment alone was identified as ethylene glycol terephthalate (EGT). With the addition of TfC as a second biocatalyst, the highest synergy degrees for BHET oligomers and PET film degradation were 2.79 and 2.26, respectively. The largest amounts of terephthalic acid (TPA) and mono(2-hydroxyethyl) terephthalate (MHET) (47 nM and 330 nM, respectively) were detected after combined treatment of PET film with M. oleivorans JWG-G2 at 5 x 10(3) microL/cm(2) and TfC at 120 microg/cm(2), and the degree of PET film surface destruction was more significant than those produced by each treatment alone. The presence of extracellular PET hydrolases in M. oleivorans JWG-G2, including three carboxylesterases, an esterase and a lipase, was predicted by whole genome sequencing analysis, and a predicted PET degradation pathway was proposed for the synergistic microbe-enzyme treatment. The results indicated that synergistic microbe-enzyme treatment may serve as a potentially promising tool for the future development of effective PET degradation. KEY POINTS: An ecofriendly synergistic microbe-enzyme PET degradation system operating at room temperature was first introduced for degrading PET. A novel product (EGT) was first identified during PET degradation. Potential PET hydrolases in M. oleivorans JWG-G2 were predicted by whole genome sequencing analysis. Title: A review on alpha-mangostin as a potential multi-target-directed ligand for Alzheimer's disease Yang A, Liu C, Wu J, Kou X, Shen R Ref: European Journal of Pharmacology, :173950, 2021 : PubMed ESTHER: Yang_2021_Eur.J.Pharmacol__173950 PubMedSearch: Yang 2021 Eur.J.Pharmacol 173950 PubMedID: 33607107 Abstract Alzheimer's disease (AD) is an age-related neurodegenerative disease characterized by progressive memory loss, declining language skills and other cognitive disorders. AD has brought great mental and economic burden to patients, families and society. However due to the complexity of AD's pathology, drugs developed for the treatment of AD often fail in clinical or experimental trials. The main problems of current anti-AD drugs are low efficacy due to mono-target method or side effects, especially high hepatotoxicity. To tackle these two main problems, multi-target-directed ligand (MTDL) based on "one molecule, multiple targets" has been studied. MTDLs can regulate multiple biological targets at the same time, so it has shown higher efficacy, better safety. As a natural active small molecule, alpha-mangostin (alpha-M) has shown potential multi-factor anti-AD activities in a series of studies, furthermore it also has a certain hepatoprotective effect. The good availability of alpha-M also provides support for its application in clinical research. In this work, multiple activities of alpha-M related to AD therapy were reviewed, which included anti-cholinesterase, anti-amyloid-cascade, anti-inflammation, anti-oxidative stress, low toxicity, hepatoprotective effects and drug formulation. It shows that alpha-M is a promising candidate for the treatment of AD. Title: Comparison of the Inhibitory Effects of Clotrimazoleand Ketoconazole against Human Carboxylesterase 2 Zhao T, Wang D, Shan Z, Chen J, Dou T, Ge G, Wang C, Meng Q, Sun H and Wu J <1 more author(s)> Zhao T, Wang D, Shan Z, Chen J, Dou T, Ge G, Wang C, Meng Q, Sun H, Liu K, Wu J (- 1) Ref: Curr Drug Metab, :, 2021 : PubMed ESTHER: Zhao_2021_Curr.Drug.Metab__ PubMedSearch: Zhao 2021 Curr.Drug.Metab PubMedID: 33568030 Abstract BACKGROUND: Both clotrimazole and ketoconazole have been verified that they have an inhibitory effect on CYP3A4. hCE2 is an enzyme closely related to the side effects of several anti-cancer drugs. However, the interactions between hCE2 and clotrimazole and ketoconazole remain unclear. OBJECTIVE: The objective of this study was to investigate and compare the inhibition behaviors of these two antifungal agents, ketoconazole and clotrimazole, on the human liver microsome hCE2 and to explore the underlying mechanism. METHODS: The inhibitory effects were investigated in human liver microsomes (HLMs) using fluorescein diacetate (FD), N-(2-butyl-1,3-dioxo-2,3-dihydro-1H-phenalen-6-yl)-2-chloroacetamide (NCEN) and irinotecan (CPT-11) as substrates of hCE2. RESULTS: Clotrimazole significantly inhibited the hCE2 activity, which was manifested by attenuated fluorescence when the substrates were FD and NCEN. The inhibitory effect of clotrimazole towards hCE2 was much stronger than that of ketoconazole, and the inhibitory behaviors displayed substrate-dependent inhibition. The IC50 value of clotrimazole with CPT-11 as the substate increased by 5 and 37 times than that with FD and NCEN respectively. Furthermore, the inhibitions of clotrimazole towards hCE2-mediated hydrolysis of FD, NCEN and CPT-11 were all in competitive mode with the Ki values of 0.483 microM, 8.63 microM and 29.0 microM, respectively. Molecular docking result of clotrimazole binding to hCE2 illustrated that clotrimazole could efficiently orient itself in the Z site cavity of hCE2. CONCLUSION: Clotrimazole displayed a strong inhibitory effect against hCE2, which might be used as a potential combined agent co-administrated with CPT-11 to alleviate the hCE2-mediated severe side effects. Title: Structure-guided engineering of a Thermobifida fusca cutinase for enhanced hydrolysis on natural polyester substrate Dong Q, Yuan S, Wu L, Su L, Zhao Q, Wu J, Huang W, Zhou J Ref: Bioresour. Bioprocess, 7:37, 2020 : PubMed ESTHER: Dong_2020_Bioresour.Bioprocess_7_37 PubMedSearch: Dong 2020 Bioresour.Bioprocess 7 37 Gene_locus related to this paper: thefu-q6a0i4 Abstract Cutinases could degrade insoluble polyester, including natural cutin and synthetic plastic. However, their turnover efficiency for polyester remains too low for industrial application. Herein, we report the 1.54-A resolution X-ray crystal structure of a cutinase from Thermobifida fusca and modeling structure in complex with a cutin mimic oligo-polyester C24H42O8. These efforts subsequently guided our design of cutinase variants with less bulky residues in the vicinity of the substrate binding site. The L90A and I213A variants exhibit increased hydrolysis activity (5- and 2.4-fold, respectively) toward cutin and also showed enhanced cotton scouring efficiency compared with the wild-type enzyme. Title: In Vitro Hepatic Metabolism of Curcumin Diethyl Disuccinate by Liver S9 from Different Animal Species Jithavech P, Ratnatilaka Na Bhuket P, Supasena W, Qiu G, Ye S, Wu J, Wong TW, Rojsitthisak P Ref: Front Pharmacol, 11:577998, 2020 : PubMed ESTHER: Jithavech_2020_Front.Pharmacol_11_577998 PubMedSearch: Jithavech 2020 Front.Pharmacol 11 577998 PubMedID: 33312126 Abstract Liver S9 (LS9) is a nearly complete collection of all hepatic drug-metabolizing enzymes. It is a low-cost model for predicting drug metabolic activity. This study aimed to identify the suitability of using LS9 of different animal sources in drug metabolism profiling with respect to the possible translation of the in vitro outcomes to clinical studies. The in vitro hepatic metabolism of curcumin diethyl disuccinate (CDD) in LS9 of rats, dogs, monkeys, and humans was evaluated. The identity of CDD metabolites and the metabolism kinetic parameters, including degradation rate constant, in vitro/in vivo intrinsic clearance, and half-life, were determined. CDD was rapidly metabolized into monoethylsuccinyl curcumin and curcumin in LS9 of all tested species mainly by carboxylesterases (CESs), including CES1 and CES2, and butyrylcholinesterase. The in vitro intrinsic clearance of CDD was in the order of human > dog > monkey > rat, whereas that of monoethylsuccinyl curcumin in the order of dog > monkey > human > rat; this parameter was not correlated with their respective in vivo clearance, which followed the order of dog > monkey > rat > human. Therefore, in vitro drug metabolism data inferred from LS9 of nonhuman origin, especially from monkeys and dogs, cannot be used as preclinical data for human trials, as humans have a smaller liver-to-body weight ratio than monkeys, dogs, and rats. The in vivo drug metabolism is dictated by the anatomical factors of the test subject. Title: Relief of Cadmium-Induced Intestinal Motility Disorder in Mice by Lactobacillus plantarum CCFM8610 Liu Y, Wu J, Xiao Y, Liu Q, Yu L, Tian F, Zhao J, Zhang H, Chen W, Zhai Q Ref: Front Immunol, 11:619574, 2020 : PubMed ESTHER: Liu_2020_Front.Immunol_11_619574 PubMedSearch: Liu 2020 Front.Immunol 11 619574 PubMedID: 33362802 Abstract Cadmium (Cd) is a toxic metal inducing a range of adverse effects on organs including liver and kidneys. However, the underlying molecular mechanisms of Cd-induced intestinal toxicity through dietary intake is poorly studied. This study evaluated the toxic effects of Cd on intestinal physiology and confirmed the effectiveness of the protective mechanism of the probiotic Lactobacillus plantarum CCFM8610 against chronic Cd toxicity. After treatment with Cd, the HT-29 cell line was subjected to iTRAQ analysis, which revealed that changes in the proteomic profiles after Cd exposure were related to pathways involved in the stress response and carbohydrate metabolism. The results of an animal trial also indicated that 10 weeks of Cd exposure decreased the fecal water content and contractile response of colonic muscle strips in mice, and delayed the excretion time of the first black feces. L. plantarum CCFM8610 treatment provided protective effects against these Cd-induced intestinal motility dysfunctions by recovering the levels of neurotransmitters, including substance P, acetyl cholinesterase, vasoactive intestinal peptide, 5-hydroxytryptamine, calcitonin gene-related peptide, and nitric oxide, and suppressing the cellular stress response in mice (e.g., the inhibition of mitogen-activated protein kinase pathways). The administration of this probiotic was also observed to reduce Cd levels in the tissues and blood of the mice. Our results suggest a newly identified protective mechanism of probiotics against Cd toxicity that involves the recovery of intestinal motility and increase in fecal cadmium excretion. Title: The toxicity assessment of extract of Peganum harmala L. seeds in Caenorhabditis elegans Miao X, Zhang X, Yuan Y, Zhang Y, Gao J, Kang N, Liu X, Wu J, Liu Y, Tan P Ref: BMC Complement Med Ther, 20:256, 2020 : PubMed ESTHER: Miao_2020_BMC.Complement.Med.Ther_20_256 PubMedSearch: Miao 2020 BMC.Complement.Med.Ther 20 256 PubMedID: 32807143 Abstract BACKGROUND: Peganum harmala L. is a medicinal herb extensively used in traditional Chinese medicine (TCM). So far, relevant reports on the toxicity of Peganum harmala L. seeds (PHS) are hardly available. Especially, we still know little about the in vivo mechanism for PHS toxicity. This study aims to evaluate the toxicity effects of PHS in Caenorhabditis elegans (C. elegans), investigate the possible mechanism of the toxicity effects of PHS, and provide reference for the pharmacological research of PHS. METHODS: In the present study, the C. elegans was exposed to 0.25, 0.50, 1.00 mg/mL of PHS in nematode growth medium (NGM) at 22 degC in the presence of food. Lethality, lifespan, growth, reproduction, and locomotion behavior assays were performed to evaluate the toxicity effects of PHS in C. elegans. We then determined the mechanism of the toxicity effect of PHS by quantitative real-time polymerase chain reaction (qRT-PCR), acetylcholinesterase (AChE) activity assay, and oxidative stress resistance assays. The main components of PHS were detected by high performance liquid chromatography (HPLC). RESULTS: Compared with the control group, the lethality of C. elegans was significantly increased when they were exposed to the ethanol extract of PHS at 0.25, 0.50 and 1.00 mg/mL (P < 0.01), and the mean lifespan was significantly decreased (P < 0.01). We also observed that PHS exposure could induce the toxicity on body length, brood size, and locomotion behavior. CONCLUSION: Our study shows that the ethanol extract of PHS exerts obvious toxic effects on C. elegans, which would provide new ideas and methods for the biological evaluation of the toxicity of Chinese medicinal materials. Title: Prophylactic cognitive enhancers for improvement of cognitive function in patients undergoing electroconvulsive therapy: A systematic review and meta-analysis Niu Y, Ye D, You Y, Wu J Ref: Medicine (Baltimore), 99:e19527, 2020 : PubMed ESTHER: Niu_2020_Medicine.(Baltimore)_99_e19527 PubMedSearch: Niu 2020 Medicine.(Baltimore) 99 e19527 PubMedID: 32176105 Abstract OBJECTIVE: Cognitive enhancers, including cholinesterase inhibitors and memantine, are used to treat dementia, but their effect for reducing post-electroconvulsive therapy (post-ECT) cognitive side effects is unclear. We conducted a systematic review and meta-analysis to assess the effectiveness of cognitive enhancers in the prevention of cognitive side effects due to ECT. METHODS: We identified relevant studies by searching electronic databases (e.g., PubMed, EMBASE, Web of Science, Cochrane Library). Only studies published up to October 2019 comparing cognitive enhancer vs placebo for cognitive function after ECT were included. The primary outcome extracted from the studies was cognitive function score. RESULTS: Five studies with 202 patients were included in this study. The cognitive enhancer group (CEG) had a significantly higher cognitive function score. Moreover, sensitivity analysis showed that no individual study had a significant impact on the overall results. CONCLUSIONS: This meta-analysis revealed that cognitive enhancers might improve cognitive function and reduce ECT-induced cognitive side effects. Nevertheless, more high-quality randomized controlled trials (RCTs) with long-term follow-up are still needed to make the final conclusion. Title: Discovery of Thai mangrove tetranortriterpenoids as agonists of human pregnane-X-receptor and inhibitors against human carboxylesterase 2 Ren YX, Zou XP, Li WS, Wu J, Shen L Ref: Bioorg Chem, 107:104599, 2020 : PubMed ESTHER: Ren_2020_Bioorg.Chem_107_104599 PubMedSearch: Ren 2020 Bioorg.Chem 107 104599 PubMedID: 33421954 Abstract Human pregnane-X-receptor (hPXR) is considered to be the key target for the treatment of cholestasis and liver injury. Agonists of hPXR are potential drug leads. Potent and selective inhibitors of human carboxylesterase 2 (hCES2) could be utilized to alleviate the toxicity induced by ester drugs. In this work, fifteen new tetranortriterpenoids with structure diversity, named thaigranatins F-T (1-15), including four limonoids containing a C(1)-O-C(29) bridge (1-4), four mexicanolides (5-8), three phragmalins (9-11), two limonoids belonging to the small group of trichiliton A (12-13), and two apotirucallanes (14-15), were isolated from seeds of the Thai mangrove, Xylocarpus granatum. The structures of these compounds were established by high resolution-electrospray ionization mass spectroscopy, extensive NMR spectroscopic investigations, single-crystal X-ray diffraction analyses, and the comparison of experimental electronic circular dichroism spectra. Most notably, thaigranatins L (7) and P (11) exhibited agonistic effects on hPXR at the concentration of 10.0 microM and 10.0 nM, respectively, whereas thaigranatins J (5), M (8), and T (15) showed inhibitory activities against hCES2 with IC(50) values of 6.63, 11.35, and 5.05 microM, respectively. The 8alpha,30alpha-epoxy moiety of mexicanolide and the delta(8,14) double bond of phragmalin are pivotal for agonistic effects of these limonoids on hPXR, whereas the 6-OAc group of mexicanolide is crucial for its inhibitory activity against hCES2. Additionally, the flexible C-17-side-chain with appropriate hydroxy groups is considered to be important for the inhibitory activity of apotirucallane against hCES2. Title: Limonoids with diverse structures of rings-A,B from the Thai mangrove, Xylocarpus moluccensis Shen L, Liao Q, Zhang M, Wu J Ref: Fitoterapia, :104737, 2020 : PubMed ESTHER: Shen_2020_Fitoterapia__104737 PubMedSearch: Shen 2020 Fitoterapia 104737 PubMedID: 33022332 Abstract Nine new limonoids, named thaixylomolins S-Z (1-8) and 2-O-acetylthaixylomolin Z (9), were isolated from seeds of the mangrove, Xylocarpus moluccensis, collected in the mangrove swamp of Trang Province, Thailand. Thaixylomolin S (1) is the fourth member of the khayalactone class of limonoids containing a hexahydro-2H-2,5- propanocyclopenta[b]furan motif. Thaixylomolins T-Y (2-7) are structurally diverse mexicanolides; whereas thaixylomolin Z (8) and 2-O-acetylthaixylomolin Z (9) are phragmalin 8,9,30-orthoesters. The structures of these compounds were established by HRESIMS and extensive 1D and 2D NMR investigations. The absolute configurations of thaixylomolins S (1), U (3), and Z (8) were unambiguously established by single-crystal X-ray diffraction analyses, conducted with Cu Kalpha radiation; whereas that of 2-O-acetylthaixylomolin Z (9) was determined to be the same as that of thaixylomolin Z (8) by the accurate fit of their experimental electronic circular dichroism spectra. Thaixylomolin S (1), featuring the presence of a 30-(2'-methyl)butyryloxy group, is the first limonoid of the khayalactone class, whose constitution and absolute configuration are unequivocally determined by X-ray crystallography. The inhibitory activities of all the compounds, except for the epimers 4, were assayed against human carboxylesterase 2. All the tested compounds exhibited inhibition rates in the range of 16-65% at the concentration of 100.0muM. Title: Enhanced activity towards polyacrylates and poly(vinyl acetate) by site-directed mutagenesis of Humicola insolens cutinase Su L, Hong R, Kong D, Wu J Ref: Int J Biol Macromol, 162:1752, 2020 : PubMed ESTHER: Su_2020_Int.J.Biol.Macromol_162_1752 PubMedSearch: Su 2020 Int.J.Biol.Macromol 162 1752 PubMedID: 32771512 Gene_locus related to this paper: humin-cut Abstract Previous studies on the hydrolysis of polyacrylates by cutinase have found that cutinase from Humicola insolens can fulfill the requirement for a thermostable cutinase in the treatment of stickies from papermaking, but it has poor hydrolysis ability. To further improve its ability to hydrolyze the polymers in papermaking, we analyzed the structure of cutinase from H. insolens, and constructed three mutants L66A, I169A, and L66A/I169A to reduce the steric hindrance of the substrate binding region. The hydrolysis results for poly(methyl acrylate), poly(ethyl acrylate), and poly(vinyl acetate) showed the catalytic ability of the mutant L66A/I169A most significantly improved. Using polymer macroporous resin composites as substrate, the released products of L66A/I169A were 1.3-4.4 times higher than that of the wild-type enzyme. When polymer suspensions were no longer being deposited, that is, when the turbidity decrease was less than 1%, the amount of L66A/I169A added was reduced by 19%-51% compared with that of the wild-type enzyme. These results indicated that the removal of the gatekeeper structure above the substrate binding region of H. insolens cutinase enhances its ability to hydrolyze polymers, and provided a basis for the application of cutinase in the practical treatment of stickies. Title: Molecular Mechanism for Antibody-Dependent Enhancement of Coronavirus Entry Wan Y, Shang J, Sun S, Tai W, Chen J, Geng Q, He L, Chen Y, Wu J and Li F <3 more author(s)> Wan Y, Shang J, Sun S, Tai W, Chen J, Geng Q, He L, Chen Y, Wu J, Shi Z, Zhou Y, Du L, Li F (- 3) Ref: J Virol, 94:, 2020 : PubMed ESTHER: Wan_2020_J.Virol_94_ PubMedSearch: Wan 2020 J.Virol 94 PubMedID: 31826992 Abstract Antibody-dependent enhancement (ADE) of viral entry has been a major concern for epidemiology, vaccine development, and antibody-based drug therapy. However, the molecular mechanism behind ADE is still elusive. Coronavirus spike protein mediates viral entry into cells by first binding to a receptor on the host cell surface and then fusing viral and host membranes. In this study, we investigated how a neutralizing monoclonal antibody (MAb), which targets the receptor-binding domain (RBD) of Middle East respiratory syndrome (MERS) coronavirus spike, mediates viral entry using pseudovirus entry and biochemical assays. Our results showed that MAb binds to the virus surface spike, allowing it to undergo conformational changes and become prone to proteolytic activation. Meanwhile, MAb binds to cell surface IgG Fc receptor, guiding viral entry through canonical viral-receptor-dependent pathways. Our data suggest that the antibody/Fc-receptor complex functionally mimics viral receptor in mediating viral entry. Moreover, we characterized MAb dosages in viral-receptor-dependent, Fc-receptor-dependent, and both-receptors-dependent viral entry pathways, delineating guidelines on MAb usages in treating viral infections. Our study reveals a novel molecular mechanism for antibody-enhanced viral entry and can guide future vaccination and antiviral strategies.IMPORTANCE Antibody-dependent enhancement (ADE) of viral entry has been observed for many viruses. It was shown that antibodies target one serotype of viruses but only subneutralize another, leading to ADE of the latter viruses. Here we identify a novel mechanism for ADE: a neutralizing antibody binds to the surface spike protein of coronaviruses like a viral receptor, triggers a conformational change of the spike, and mediates viral entry into IgG Fc receptor-expressing cells through canonical viral-receptor-dependent pathways. We further evaluated how antibody dosages impacted viral entry into cells expressing viral receptor, Fc receptor, or both receptors. This study reveals complex roles of antibodies in viral entry and can guide future vaccine design and antibody-based drug therapy. Title: Cascade Reaction System Integrating Single-Atom Nanozymes with Abundant Cu Sites for Enhanced Biosensing Wu Y, Wu J, Jiao L, Xu W, Wang H, Wei X, Gu W, Ren G, Zhang N and Zhu C <3 more author(s)> Wu Y, Wu J, Jiao L, Xu W, Wang H, Wei X, Gu W, Ren G, Zhang N, Zhang Q, Huang L, Gu L, Zhu C (- 3) Ref: Analytical Chemistry, :, 2020 : PubMed ESTHER: Wu_2020_Anal.Chem__ PubMedSearch: Wu 2020 Anal.Chem PubMedID: 31941278 Abstract Single-atom nanozymes (SAzymes), as novel nanozymes with atomically dispersed active sites, are of great importance in the de-velopment of nanozymes for their high catalytic activities, the maximum utilization efficiency of metal atoms, and the simple mod-el of active sites. Herein, the peroxidase-like SAzymes with high-concentration Cu sites on carbon nanosheets (Cu-N-C) were syn-thesized through a salt-template strategy. With the densely distributed active Cu atoms (~5.1 wt%), the Cu-N-C SAzymes exhibit remarkable activity to mimic natural peroxidase. Integrating Cu-N-C SAzymes with natural acetylcholinesterase and choline oxi-dase, three-enzyme-based cascade reaction system was constructed for the colorimetric detection of acetylcholine and organo-phosphorus pesticides. This work not only provides a strategy to synthesize SAzymes with abundant active sites but also gives some new insights for robust nanozyme biosensing systems. Title: Soluble epoxide hydrolase inhibitor protects against blood-brain barrier dysfunction in a mouse model of type 2 diabetes via the AMPK/HO-1 pathway Wu J, Zhao Y, Fan Z, Chen Q, Chen J, Sun Y, Jiang X, Xiao Q Ref: Biochemical & Biophysical Research Communications, :, 2020 : PubMed ESTHER: Wu_2020_Biochem.Biophys.Res.Commun__ PubMedSearch: Wu 2020 Biochem.Biophys.Res.Commun PubMedID: 32001002 Abstract Diabetes mellitus is a metabolic disorder that can lead to blood-brain barrier (BBB) disruption and cognitive decline. However, the mechanisms of BBB breakdown in diabetes are still unclear. Soluble epoxide hydrolase (sEH) is an enzyme that degrades epoxyeicosatrienoic acids (EETs), which have multiple protective effects on vascular structure and functions. In the current study, we showed increased vascular permeability of the BBB, which was accompanied by upregulation of sEH and downregulation of 14,15-EET. Moreover, the sEH inhibitor t-AUCB restored diabetic BBB integrity in vivo, and 14,15-EET prevented ROS accumulation and MEC injury in vitro. t-AUCB or 14,15-EET treatment provoked AMPK/HO-1 activation under diabetic conditions in vivo and in vitro. Thus, we suggest that decreased EET degradation by sEH inhibition might be a potential therapeutic approach to attenuate the progression of BBB injury in diabetic mice via AMPK/HO-1 pathway activation. Title: Design, synthesis and biological evaluation of novel carbamates as potential inhibitors of acetylcholinesterase and butyrylcholinesterase Wu J, Pistolozzi M, Liu S, Tan W Ref: Bioorganic & Medicinal Chemistry, :115324, 2020 : PubMed ESTHER: Wu_2020_Bioorg.Med.Chem__115324 PubMedSearch: Wu 2020 Bioorg.Med.Chem 115324 PubMedID: 32008882 Abstract Rivastigmine, a dual inhibitor of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), has been approved by U.S. Food and Drug Administration to treat Alzheimer's disease (AD) and Parkinson's disease (PD) dementia. In the current work, a bambuterol derivative lacking one of the carbamoyloxy groups on the benzene ring (BMC-1) and its analogues were synthesized using 1-(3-hydroxyphenyl) ethan-1-one and 1-(4-hydroxyphenyl) ethan-1-one as starting materials. In-vitro cholinesterase assay established that nine compounds were more potent to inhibit both electric eel AChE and equine serum BChE than rivastigmine under the same experimental conditions. Further study confirmed that among the nine carbamates, BMC-3 (IC50(AChE) = 792 nM, IC50(BChE) = 2.2 nM) and BMC-16 (IC50(AChE) = 266 nM, IC50(BChE) = 10.6 nM) were excellent cholinesterase inhibitors with potential of permeating through the blood-brain barrier. These carbamates could be used as potential dual inhibitors of AChE and BChE and to discover novel drugs for the treatment of AD and PD dementia. Title: Biological Impact and Enzyme Activities of Spodoptera litura (Lepidoptera: Noctuidae) in Response to Synergistic Action of Matrine and Beauveria brongniartii Wu J, Li J, Zhang C, Yu X, Cuthbertson AGS, Ali S Ref: Front Physiol, 11:584405, 2020 : PubMed ESTHER: Wu_2020_Front.Physiol_11_584405 PubMedSearch: Wu 2020 Front.Physiol 11 584405 PubMedID: 33224038 Abstract Matrine, a naturally occurring heterocyclic compound, has been shown to enhance the pathogenicity of the entomopathogenic fungus Beauveria brongniartii against Spodoptera litura. In the current study, the biological impacts and synergism activities of these two agents on nutritional efficiency and antioxidant enzymes in S. litura were explored. Our results showed a high antifeedant activity of B. brongniartii and matrine on S. litura. The S. litura larvae were unable to pupate and emerge when treated with combinations of matrine and B. brongniartii. Following on, we measured the activities of five important antioxidant enzymes [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), acetylcholinesterase (AChE), and glutathione-S-transferase (GST)] when treated with B. brongniartii SB010 (1 x 10(9) spores/ml), matrine (0.5 mg/ml), and B. brongniartii SB010 (1 x 10(9) spores/ml) + matrine (0.5 mg/ml). The results indicated the detoxification activity of the five enzymes in the fat body and hemolymph of S. litura when facing a combined B. brongniartii and matrine challenge. The activities of the enzymes were significantly lower than that of the control group 7 days post-treatment, indicating the inhibitory effect of the two xenobiotics. Matrine had better inhibition effects than B. brongniartii in a majority of the trials. The improved detoxification activity of the five enzymes may be the internal mechanism of synergism of matrine on B. brongniartii. Title: Patatin primary structural properties and effects on lipid metabolism Wu J, Wu Q, Yang D, Zhou M, Xu J, Wen Q, Cui Y, Bai Y, Xu S and Wang S <1 more author(s)> Wu J, Wu Q, Yang D, Zhou M, Xu J, Wen Q, Cui Y, Bai Y, Xu S, Wang Z, Wang S (- 1) Ref: Food Chem, :128661, 2020 : PubMed ESTHER: Wu_2020_Food.Chem__128661 PubMedSearch: Wu 2020 Food.Chem 128661 PubMedID: 33272761 Abstract Patatin, the major protein found in potatoes, was purified and shows several isoforms. The essential amino acid content of patatin was ashighas 76%, indicating that it is a valuable protein source. Patatin was an O-linked glycoprotein that contained fucose monosaccharides, as well as mannose, rhamnose, glucose, galactose, xylose, and arabinose. Patatin had a fucosylated glycan structural feature, which strongly bound AAL (Aleuria aurantia Leukoagglutinin), a known fucose binding lectin. Moreover, thelipid metabolism regulatory effects of patatin on the fat catabolism, fat absorption, and inhibition of lipase activity were measured after high-fat feeding of zebrafish larvae. Results revealed that 37.0 g/mL patatin promoted 23% lipid decomposition metabolism. Meanwhile patatin could inhibite lipase activity and fat absorption, whose effects accounted for half that of a positive control drug. Our findings suggest that patatin, a fucosylated glycoprotein, could potentially be used as a naturalactiveconstituent with anti-obesity effects. Title: Tacrine-hydroxamate derivatives as multitarget-directed ligands for the treatment of Alzheimer's disease: Design, synthesis, and biological evaluation Xu A, He F, Zhang X, Li X, Ran Y, Wei C, James Chou C, Zhang R, Wu J Ref: Bioorg Chem, 98:103721, 2020 : PubMed ESTHER: Xu_2020_Bioorg.Chem_98_103721 PubMedSearch: Xu 2020 Bioorg.Chem 98 103721 PubMedID: 32193030 Abstract In order to develop multitarget-directed ligands as potential treatments for Alzheimer's disease, twenty-eight new tacrine-hydroxamate derivatives were designed, synthesized, and biologically evaluated. As expected, most of the compounds exhibited inhibitory activities against cholinesterases (ChEs) and histone deacetylase (HDACs). Among the tested compounds, A10 showed not only potent and selective inhibition on AChE at sub-nanomolar potency (AChEIC50 = 0.12 nM, BChEIC50 = 361.52 nM) but also potent inhibition on HDAC (IC50 = 0.23 nM). Moreover, A10 exhibited inhibitory activity on Abeta1-42 self-aggregation as well as disaggregation activity on pre-formed Abeta fibrils. Furthermore, A10 exhibited antioxidant activity and metal chelating properties. Further mechanistic studies demonstrated that A10 is a pan-inhibitor of HDACs and a mixed-type inhibitor for AChE. It shown that A10 is a BBB penetrant by online prediction. Taken together, the results indicate that A10 can serve as a lead compound to develop promising candidate analogs as AD therapeutics. Title: GDSL esterase/lipases OsGELP34 and OsGELP110/OsGELP115 are essential for rice pollen development Zhang H, Wang M, Li Y, Yan W, Chang Z, Ni H, Chen Z, Wu J, Xu C and Tang X <1 more author(s)> Zhang H, Wang M, Li Y, Yan W, Chang Z, Ni H, Chen Z, Wu J, Xu C, Deng XW, Tang X (- 1) Ref: J Integr Plant Biol, :, 2020 : PubMed ESTHER: Zhang_2020_J.Integr.Plant.Biol__ PubMedSearch: Zhang 2020 J.Integr.Plant.Biol PubMedID: 32068333 Abstract Pollen exine contains complex biopolymers of aliphatic lipids and phenolics. Abnormal development of pollen exine often leads to plant sterility. Molecular mechanisms regulating exine formation have been studied extensively but remain ambiguous. Here we report the analyses of three GDSL esterase/lipase protein genes, OsGELP34, OsGELP110, and OsGELP115, for rice exine formation. OsGELP34 was identified by cloning of a male sterile mutant gene. OsGELP34 encodes an endoplasmic reticulum protein and was mainly expressed in anthers during pollen exine formation. osgelp34 mutant displayed abnormal exine and altered expression of a number of key genes required for pollen development. OsGELP110 was previously identified as a gene differentially expressed in meiotic anthers. OsGELP110 was most homologous to OsGELP115, and the two genes showed similar gene expression patterns. Both OsGELP110 and OsGELP115 proteins were localized in peroxisomes. Individual knockout of OsGELP110 and OsGELP115 did not affect the plant fertility, but double knockout of both genes altered the exine structure and rendered the plant male sterile. OsGELP34 is distant from OsGELP110 and OsGELP115 in sequence, and osgelp34 and osgelp110/osgelp115 mutants were different in anther morphology despite both were male sterile. These results suggested that OsGELP34 and OsGELP110/OsGELP115 catalyze different compounds for pollen exine development. This article is protected by copyright. All rights reserved. Title: Twenty-five limonoids from the Hainan mangrove, Xylocarpus granatum Zhang JC, Liao Q, Shen L, Wu J Ref: Bioorg Chem, 100:103903, 2020 : PubMed ESTHER: Zhang_2020_Bioorg.Chem_100_103903 PubMedSearch: Zhang 2020 Bioorg.Chem 100 103903 PubMedID: 32413629 Abstract Twenty-four new limonoids (1-24), named hainanxylogranins A-X, were isolated from leaves and barks of the Hainan mangrove, Xylocarpus granatum, together with a known compound, tabulvelutin B (25). The structures of these compounds were established by high resolution electrospray ionization mass spectroscopy (HRESIMS), extensive NMR spectroscopic investigations, single-crystal X-ray diffraction analyses, and the comparison of experimental electronic circular dichroism (ECD) spectra. Most notably, the absolute configurations of seven compounds, viz., 1, 2, 6, 16, 17, 22, and 25, were unambiguously determined by single-crystal X-ray diffraction analyses, conducted with Cu Kalpha radiation. Compounds 1-4 belong to a unique group of mexicanolides containing a C3-O-C8 bridge and a C-17 substituted gamma(21)-hydroxybutenolide moiety, whereas 5-9 are mexicanolides comprising a C1-O-C8 bridge. Compounds 10-16 are typical mexicanolides, among which 14 and 15 contain a C-17 substituted gamma(23)-hydroxybutenolide moiety. Compounds 17 and 18 are phragmalin 8,9,30-orthoesters, whereas 19 and 20 are phragmalin 1,8,9-orthoesters. Compound 21 consists of a C1-O-C29 bridge, while 22-24 are derivatives of azadirone. The inhibitory activities of 1, 5-8, 11, 17, 19, 21-23, and 25 against human carboxylesterase 2 (CES2) were assayed. All the tested compounds exhibited inhibition rates of 30-64% at the concentration of 100.0 microM. Title: Assessment of phthalate ester residues and distribution patterns in Baijiu raw materials and Baijiu Dong W, Guo R, Sun X, Li H, Zhao M, Zheng F, Sun J, Huang M, Wu J Ref: Food Chem, 283:508, 2019 : PubMed ESTHER: Dong_2019_Food.Chem_283_508 PubMedSearch: Dong 2019 Food.Chem 283 508 PubMedID: 30722905 Abstract Phthalate esters (PAEs) are harmful to human health and have been repeatedly identified in Baijiu samples. In our study, the distribution and degradation characteristics of 14 PAEs in Baijiu raw materials (BRMs) and Baijiu during distillation were detected using QuEChERS or vortex-assisted surfactant-enhanced-emulsification liquid-liquid micro-extraction (VSLLME) methods coupled with gas chromatography-mass spectrometry. The same five PAEs were detected in all tested samples, values ranged from 0.003 to 0.292 mg/kg; however, higher concentrations existed in BRMs compared to Baijiu samples. Using multivariate statistical analysis, detailed distinctions between different varieties of Baijiu and BRMs and separation-related PAE markers were revealed. PAEs concentration during Baijiu distillation showed a decreasing trend. The highest concentrations detected in distillate heads, were 1.6-, 2.3-, and 8.1-fold higher than those in heart1, heart2, and tail distillates, respectively. These findings revealed that PAEs may migrate from BRMs; moreover, that PAEs content can be regulated by distillation. Title: High-level expression of Humicola insolens cutinase in Pichia pastoris without carbon starvation and its use in cotton fabric bioscouring Hong R, Sun Y, Su L, Gu L, Wang F, Wu J Ref: J Biotechnol, 304:10, 2019 : PubMed ESTHER: Hong_2019_J.Biotechnol_304_10 PubMedSearch: Hong 2019 J.Biotechnol 304 10 PubMedID: 31400343 Abstract Huimcola insolens cutinase (HiC) was heterologously expressed in Pichia pastoris. To avoid a carbon starvation step, fermentation was conducted using combinations of sorbitol with glycerol and methanol in the cell growth and induction phases, respectively. The cutinase productivity (27.71 U mL(-1) h(-1)) was 9.93 U mL(-1) h(-1) greater than that achieved using traditional two-phase methods, and a cutinase activity of 2660 U mL(-1), using p-nitrophenyl butyrate as substrate, was achieved after only 96 h in a 3-L bioreactor. Subsequently, the combination of HiC with Thermobifida fusca cutinase (TfC) in cotton fabric bioscouring was evaluated by monitoring the wettability and dyeability of the fabric. Treatment with 20 U mL(-1) of HiC at 80 degrees C for 5 min followed by 30 U mL(-1) of TfC at 50 degrees C for 1 h gave the best results. The total treatment time was shorter and performance was better than those seen with the alkali method. Title: A Novel Dipeptidyl Peptidase IV Inhibitory Tea Peptide Improves Pancreatic beta-Cell Function and Reduces alpha-Cell Proliferation in Streptozotocin-Induced Diabetic Mice Lu Y, Lu P, Wang Y, Fang X, Wu J, Wang X Ref: Int J Mol Sci, 20:, 2019 : PubMed ESTHER: Lu_2019_Int.J.Mol.Sci_20_ PubMedSearch: Lu 2019 Int.J.Mol.Sci 20 PubMedID: 30646613 Abstract Dipeptidyl peptidase IV (DPP-IV) inhibitors occupy a growing place in the drugs used for the management of type 2 diabetes. Recently, food components, including food-derived bioactive peptides, have been suggested as sources of DPP-IV inhibitors without side effects. Chinese black tea is a traditional health beverage, and it was used for finding DPP-IV inhibitory peptides in this study. The ultra-filtrated fractions isolated from the aqueous extracts of black tea revealed DPP-IV inhibitory activity in vitro. Four peptides under 1 kDa were identified by SDS-PAGE and LC-MS/MS (Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) from the ultra-filtrate. The peptide II (sequence: AGFAGDDAPR), with a molecular mass of 976 Da, showed the greatest DPP-IV inhibitory activity (in vitro) among the four peptides. After administration of peptide II (400 mg/day) for 57 days to streptozotocin (STZ)-induced hyperglycemic mice, the concentration of glucagon-like peptide-1 (GLP-1) in the blood increased from 9.85 +/- 1.96 pmol/L to 19.22 +/- 6.79 pmol/L, and the insulin level was increased 4.3-fold compared to that in STZ control mice. Immunohistochemistry revealed the improved function of pancreatic beta-cells and suppressed proliferation of pancreatic alpha-cells. This study provides new insight into the use of black tea as a potential resource of food-derived DPP-IV inhibitory peptides for the management of type 2 diabetes. Title: Exosomes from activated hepatic stellate cells contain GLUT1 and PKM2: a role for exosomes in metabolic switch of liver nonparenchymal cells Wan L, Xia T, Du Y, Liu J, Xie Y, Zhang Y, Guan F, Wu J, Wang X, Shi C Ref: FASEB Journal, :fj201802675R, 2019 : PubMed ESTHER: Wan_2019_FASEB.J__fj201802675R PubMedSearch: Wan 2019 FASEB.J fj201802675R PubMedID: 30970216 Abstract The mechanism of exosomes derived from activated hepatic stellate cells (HSCs) involved in liver fibrosis is poorly understood. We previously reported that hypoxia-inducible factor 1 (Hif-1) regulated HSC activation, and, therefore, we investigated in current work whether Hif-1 regulates exosome secretion and the metabolic switch of HSCs, thus affecting the metabolism of liver nonparenchymal cells. In this study, the characteristics of exosomes from HSCs were assessed via electron microscopy, Western blot analysis, and acetylcholinesterase activity. Confocal microscopy was used to measure the uptake of exosomes by quiescent HSCs, Kupffer cells (KCs), and liver sinusoidal endothelial cells (LSECs). Hif-1alpha was inhibited via 2-ME or specific small interfering RNAs to investigate its role in exosomes derived from HSCs. It was determined that glucose transporter 1 and pyruvate kinase M2 were increasingly expressed in fibrotic liver samples, cell lysates, and exosomes derived from activated HSCs. Exosomes released from HSCs were associated with activation and glucose uptake of HSCs. Delivery of exosomes from activated HSCs induced glycolysis of quiescent HSCs, KCs, and LSECs. Disruption of Hif-1 expression suppressed the glycolysis effect delivered by exosomes. Conclusively, our results demonstrated that exosomes secreted by activated HSCs affect the metabolic switch of liver nonparenchymal cells via delivery of glycolysis-related proteins. These findings represent a novel mechanism that contributes to liver fibrosis and has significant implications for new diagnosis and treatment of liver diseases.-Wan, L., Xia, T., Du, Y., Liu, J., Xie, Y., Zhang, Y., Guan, F., Wu, J., Wang, X., Shi, C. Exosomes from activated hepatic stellate cells contain GLUT1 and PKM2: a role for exomes in metabolic switch of liver nonparenchymal cells. Title: Electropolymerization-Induced Positively Charged Phenothiazine Polymer Photoelectrode for Highly Sensitive Photoelectrochemical Biosensing Wang J, Lv W, Wu J, Li H, Li F Ref: Analytical Chemistry, 91:13831, 2019 : PubMed ESTHER: Wang_2019_Anal.Chem_91_13831 PubMedSearch: Wang 2019 Anal.Chem 91 13831 PubMedID: 31560517 Abstract Exploring the fabrication of an electrode with high photoelectric conversion efficiency and abundant functional groups for ideal photoelectrochemical (PEC) sensor development is highly urgent but faces a significant challenge. Herein we report an electropolymerization strategy for the preparation of phenothiazine polymeric film on an indium tin oxide (ITO) surface (PPT/ITO), within only a few seconds, and monomers. The fabricated PPT/ITO electrode possessed excellent stability and abundant quaternary ammonium salt groups for developing a highly sensitive PEC sensor through electrostatic binding with negatively charged materials. In this context, a CdS QDs-functionalized PPT/ITO electrode (CdS/PPT/ITO) was proposed and applied to the analysis of chlorpyrifos, used as a model target organophosphorous pesticide (OP). The thiocholine generated from acetylcholinesterase (AChE)-induced catalyzed hydrolysis of acetylthiocholine (ATCh) efficiently directed CdS QDs away from PPT/ITO via electrostatic repulsion, subsequently decreasing PEC current, whereas chlorpyrifos prohibited the generation of thiocholine through inhibiting AChE activity. As compared to the case where chlorpyrifos is absent, significantly enhanced PEC current is determined and is proportional to chlorpyrifos amounts. Thus, the developed CdS/PPT/ITO-based PEC sensor achieved excellent chlorpyrifos biosensing with improved sensitivity down to approximately ng/mL level with good specificity. We envision the proposed strategy will provide a new path to conveniently fabricate photoelectrodes possessing high performance, which will have more useful applications in PEC sensing. Title: HEV-LFS : A novel scoring model for patients with hepatitis E virus-related liver failure Wu J, Guo N, Zhang X, Xiong C, Liu J, Xu Y, Fan J, Yu J, Zhao X and Li L <4 more author(s)> Wu J, Guo N, Zhang X, Xiong C, Liu J, Xu Y, Fan J, Yu J, Zhao X, Liu B, Wang W, Zhang J, Cao H, Li L (- 4) Ref: J Viral Hepat, 26:1334, 2019 : PubMed ESTHER: Wu_2019_J.Viral.Hepat_26_1334 PubMedSearch: Wu 2019 J.Viral.Hepat 26 1334 PubMedID: 31294523 Abstract A noninvasive assessment method for acute or acute-on-chronic liver failure in patients with hepatitis E virus (HEV) infection is urgently needed. We aimed to develop a scoring model for diagnosing HEV patients who developed liver failure (HEV-LF) at different stages. A cross-sectional set of 350 HEV-LF patients were identified and enrolled, and the Guidelines for Diagnosis and Treatment of Liver Failure in China and the Asian Pacific Association for the Study of the Liver were adopted as references. HEV-LFS , a novel scoring model that incorporates data on cholinesterase (CHE), urea nitrogen (UREA), platelets and international normalized ratio was developed using a derived dataset. For diagnosing HEV-LF stages F1 to F3, the HEV-LFS scoring model (F1: 0.87; F2: 0.90; F3: 0.92) had a significantly higher AUROC than did the CLIF-C-ACLFs (F1: 0.65; F2: 0.56; F3: 0.51) and iMELD (F1: 0.70; F2: 0.57; F3: 0.51) scoring models, of which the HEV-LFS scoring model had the best sensitivity and specificity. In addition, the HEV-LFS scoring model was correlated with mortality, length of hospitalization and ICU stay. As the GDTLF score increased, the CHE level decreased and the UREA increased gradually. Encouragingly, a calibration curve showed good agreement between the derivation and validation sets. Notably, we also established a nomogram to facilitate the practical operability of the HEV-LFS scoring model in clinical settings. In conclusion, both CHE and UREA may be indicators for HEV-LF patients. The HEV-LFS scoring model is an efficient and accessible model for classifying HEV-LF at different stages. Title: LIPG SNPs, their haplotypes and gene-environment interactions on serum lipid levels Yang S, Yin RX, Miao L, Zhou YG, Wu J, Zhang QH Ref: Lipids Health Dis, 18:10, 2019 : PubMed ESTHER: Yang_2019_Lipids.Health.Dis_18_10 PubMedSearch: Yang 2019 Lipids.Health.Dis 18 10 PubMedID: 30621702 Gene_locus related to this paper: human-LIPG Abstract BACKGROUND: Maonan nationality is a relatively conservative and isolated minority in the Southwest of China. Little is known about the association of endothelial lipase gene (LIPG) single nucleotide polymorphisms (SNPs) and serum lipid levels in the Chinese populations. METHODS: A total of 1280 subjects of Maonan nationality and 1218 participants of Han nationality were randomly selected from our previous stratified randomized samples. Genotypes of the four LIPG SNPs were determined by polymerase chain reaction-restriction fragment length polymorphism, and then confirmed by direct sequencing. RESULTS: Several SNPs were associated with high-density lipoprotein cholesterol (rs3813082, rs2000813 and rs2097055) in the both ethnic groups; total cholesterol and apolipoprotein (Apo) A1 (rs2000813) in Han nationality; and low-density lipoprotein cholesterol, ApoB, triglyceride (rs2097055) and ApoA1 (rs3819166) in Maonan minority (P < 0.0125 for all after Bonferroni correction). The commonest haplotype was rs3813082T-rs2000813C-rs2097055T-rs3819166A (Han, 44.2% and Maonan, 48.7%). The frequencies of the T-C-T-A, T-C-T-G, T-T-C-G and G-T-C-G haplotypes were different between the Maonan and Han populations (P < 0.05-0.001). The associations between haplotypes and dyslipidemia were also different in the Han and/or Maonan populations (P < 0.05-0.001). CONCLUSIONS: The differences in serum lipid profiles between the two ethnic groups might partly be attributed to these LIPG SNPs, their haplotypes and gene-environmental interactions. TRIAL REGISTRATION: Retrospectively registered. Title: Association between the LIPG polymorphisms and serum lipid levels in the Maonan and Han populations Yang S, Yin RX, Miao L, Zhang QH, Zhou YG, Wu J Ref: J Gene Med, :e3071, 2019 : PubMed ESTHER: Yang_2019_J.Gene.Med__e3071 PubMedSearch: Yang 2019 J.Gene.Med e3071 PubMedID: 30657227 Abstract BACKGROUD: Maonan nationality is a relatively isolated minority in China. Little is known about the endothelial lipase gene (LIPG) single nucleotide polymorphisms (SNPs) and serum lipid levels in the Chinese populations. The present study was undertaken to detect the association of several LIPG SNPs and environmental factors with serum lipid levels in the Chinese Maonan and Han populations. METHODS: A total of 773 subjects of Maonan nationality and 710 participants of Han nationality were randomly selected from our previous stratified randomized samples. Genotypes of the LIPG rs2156552, rs4939883 and rs7241918 SNPs were determined by polymerase chain reaction-restriction fragment length polymorphism, and then confirmed by direct sequencing. RESULTS: The allelic (rs2156552, rs4939883 and rs7241918) and genotypic (rs2156552 and rs4939883) frequencies were different between the two ethnic groups (P < 0.05-0.01). The minor allele carriers had lower ApoA1/ApoB ratio (rs2156552 and rs7241918), high-density lipoprotein cholesterol (HDL-C) and apolipoprotein (Apo) A1 (rs2156552) levels and higher ApoB levels (rs4939883) in Han nationality; and lower HDL-C (rs2156552, rs4939883 and rs7241918) levels in Maonan minority than the minor allele non-carriers (P < 0.0167 after the Bonferroni correction). Subgroup analyses according to sex showed that the minor allele carriers had lower ApoA1/ApoB ratio (rs2156552 and rs7241918) and higher ApoB levels (rs7241918) in Han males; and lower ApoA1 and HDL-C levels in Maonan famales than the minor allele non-carriers (P < 0.0167-0.001). CONCLUSIONS: The present study demonstrates the association between the LIPG polymorphsims and serum lipid levels in the two ethnic groups. These associations might have an ethnic-and or/sex-specificity. Title: Mori Ramulus (Chin.Ph.)-the Dried Twigs of Morus alba L./Part 1: Discovery of Two Novel Coumarin Glycosides from the Anti-Hyperuricemic Ethanol Extract Yao J, He H, Xue J, Wang J, Jin H, Wu J, Hu J, Wang R, Kuchta K Ref: Molecules, 24:, 2019 : PubMed ESTHER: Yao_2019_Molecules_24_ PubMedSearch: Yao 2019 Molecules 24 PubMedID: 30754654 Abstract In Traditional Chinese Medicine (TCM), Mori ramulus (Chin.Ph.)-the dried twigs of Morus alba L.-is extensively used as an antirheumatic agent and also finds additional use in asthma therapy. As a pathological high xanthine oxidase (XO, EC 1.1.3.22) activity is strongly correlated to hyperuricemy and gout, standard anti-hyperuremic therapy typically involves XO inhibitors like allopurinol, which often cause adverse effects by inhibiting other enzymes involved in purine metabolism. Mori ramulus may therefore be a promissing source for the development of new antirheumatic therapeutics with less side effects. Coumarins, one of the dominant groups of bioactive constituents of M. alba, have been demonstrated to possess anti-inflammatory, antiplatelet aggregation, antitumor, and acetylcholinesterase (AChE) inhibitory activities. The combination of HPLC (DAD) and Q-TOF technique could give excellent separating and good structural characterization abilities which make it suitable to analyze complex multi-herbal extracts in TCM. The aim of this study was to develop a HPLC (DAD)/ESI-Q-TOF-MS/MS method for the identification and profiling of pharmacologically active coumarin glycosides in Mori ramulus refined extracts for used in TCM. This HPLC (DAD)/ESI-Q-TOF-MS/MS method provided a rapid and accurate method for identification of coumarin glycosides-including new natural products described here for the first time-in the crude extract of M. alba L. In the course of this project, two novel natural products moriramulosid A (umbelliferone-6-beta-d-apiofuranosyl-(1-->6)-beta-d-glucopyranoside) and moriramulosid B (6-[[6-O-(6-deoxy-alpha-l-mannopyranosyl)-beta-d-glucopyranosyl]oxy]-2H-1-benzopy ran-1-one) were newly discovered and the known natural product Scopolin was identified in M. alba L. for the first time. Title: Surface functionalization of graphene oxide by amino acids for Thermomyces lanuginosus lipase adsorption Zhou W, Zhuang W, Ge L, Wang Z, Wu J, Niu H, Liu D, Zhu C, Chen Y, Ying H Ref: J Colloid Interface Sci, 546:211, 2019 : PubMed ESTHER: Zhou_2019_J.Colloid.Interface.Sci_546_211 PubMedSearch: Zhou 2019 J.Colloid.Interface.Sci 546 211 PubMedID: 30921675 Abstract Graphene oxide (GO) with oxygen containing functional groups can be selectively modified by small biomolecules to achieve heterogeneous surface properties. To achieve a hyper-enzymatic activity, the surface functionality of GO should be tailored to the orientation adsorption of the Thermomyces lanuginosus (TL) lipase, and the active center can be covered by a relatively hydrophobic helical lid for protection. In this work, amino acids were used to interact with GO through reduction reaction, hydrophobic forces, electrostatic forces, or hydrogen bonding to alter the surface hydrophobicity and charge density. Characterization of the structure and surface properties confirmed that the GO samples decorated with phenylalanine (Phe) and glutamic acid (Glu) exhibited superior hydrophobicity than other modifications, whereas tryptophan (Trp) and cysteine (Cys) provided weaker reduction effects on GO. Moreover, the zeta potential of the samples modified by amino acids of lysine (Lys) and arginine (Arg) is higher than other modified samples. The adsorption amount of lipase on Glu-GO reached 172mg/g and the relative enzymatic activity reached up to 200%. The thermodynamic data and the Freundlich isotherm model fitting showed that the lipase adsorption process on modified samples was spontaneous, endothermic and entropy increase. Title: Psychosocial interventions for Alzheimer's disease cognitive symptoms: a Bayesian network meta-analysis Duan Y, Lu L, Chen J, Wu C, Liang J, Zheng Y, Wu J, Rong P, Tang C Ref: BMC Geriatr, 18:175, 2018 : PubMed ESTHER: Duan_2018_BMC.Geriatr_18_175 PubMedSearch: Duan 2018 BMC.Geriatr 18 175 PubMedID: 30086714 Abstract BACKGROUND: Alzheimer disease (AD) is the most common type of dementia with cognitive decline as one of the core symptoms in older adults. Numerous studies have suggested the value of psychosocial interventions to improve cognition in this population, but which one should be preferred are still matters of controversy. Consequently, we aim to compare and rank different psychosocial interventions in the management of mild to moderate AD with cognitive symptoms. METHODS: We did a network meta-analysis to identify both direct and indirect evidence in relevant studies. We searched MEDLINE, EMBASE, PsycINFO through the OVID database, CENTRAL through the Cochrane Library for clinical randomized controlled trials investigating psychosocial interventions of cognitive symptoms in patients with Alzheimer disease, published up to August 31, 2017. We included trials of home-based exercise(HE), group exercise(GE), walking program(WP), reminiscence therapy(RT), art therapy(AT) or the combination of psychosocial interventions and acetylcholinesterase inhibitor (ChEIs). We extracted the relevant information from these trials with a predefined data extraction sheet and assessed the risk of bias with the Cochrane risk of bias tool. The outcomes investigated were Mini-Mental State Examination (MMSE) and compliance. We did a pair-wise meta-analysis using the fixed-effects model and then did a random-effects network meta-analysis within a Bayesian framework. RESULTS: We deemed 10 trials eligible, including 682 patients and 11 treatments. The quality of included study was rated as low in most comparison with Cochrane tools. Treatment effects from the network meta-analysis showed WP was better than control (SMD 4.89, 95% CI -0.07 to 10.00) while cognitive training and acetylcholinesterase inhibitor (CT + ChEIs) was significantly better than the other treatments, when compared with simple ChEIs treatment, assessed by MMSE. In terms of compliance, the pair-wise meta-analysis indicated that WP and HE are better than GE and AT, while CT + ChEIs, CST + ChEIs are better than other combined interventions. CONCLUSION: Our study confirmed the effectiveness of psychosocial interventions for improving cognition or slowing down the progression of cognitive impairment in AD patients and recommended several interventions for clinical practice. Title: Lychee seed extract protects against neuronal injury and improves cognitive function in rats with type II diabetes mellitus with cognitive impairment Tang Y, Yu C, Wu J, Chen H, Zeng Y, Wang X, Yang L, Mei Q, Cao S, Qin D Ref: Int J Mol Med, 41:251, 2018 : PubMed ESTHER: Tang_2018_Int.J.Mol.Med_41_251 PubMedSearch: Tang 2018 Int.J.Mol.Med 41 251 PubMedID: 29138799 Abstract Lychee seed is a traditional Chinese medicine and has many beneficial effects such as modulation of blood sugar and lipids, antioxidation, antivirus and antitumor. Studies have indicated that type II diabetes mellitus (T2DM) and Alzheimer's disease (AD) share common biological mechanisms including insulin resistance, impaired glucose metabolism, betaamyloid (Abeta) formation, oxidative stress and presence of advanced glycation end products (AGEs). The present study investigated the effects of lychee seed extract (LSE) on neuroprotection, cognitive function improvement and possible underlying mechanisms in a rat model of T2DM with cognitive impairment. We analyzed the chemical profile of LSE using a UHPLCSPD chromatogram and evaluated its effect on the improvement of spatial learning and memory of rats by a Morris water maze. The levels of glucose, insulin, Abeta, AGEs, Tau protein and acetylcholinesterase in the blood and/or hippocampus of rats were determined by bloodglucose meter, radioimmunoassay, chemical chromatometry, enzymelinked immunosorbent assay (ELISA) and immunohistochemical analysis, respectively. Results demonstrated that LSE consists of eight major and around 20 minor ingredients, and it remarkably prevents neuronal injury and improves cognitive functions in T2DM rats. The levels of glucose, insulin, Abeta, AGEs and Tau protein were significantly increased in the blood and/or hippocampus of T2DM rats, while LSE remarkably decreased their levels compared to vehicle treatment (P<0.01). The possible mechanisms may be associated with IR improvement and decreased formations of Abeta, AGEs and Tau protein in the hippocampus of T2DM rats. LSE may be developed as the agent for the treatment of T2DM and/or AD clinically. Title: Bioinformatics Analysis and Characterization of Highly Efficient Polyvinyl Alcohol (PVA)-Degrading Enzymes from the Novel PVA Degrader Stenotrophomonas rhizophila QL-P4 Wei Y, Fu J, Wu J, Jia X, Zhou Y, Li C, Dong M, Wang S, Zhang J, Chen F Ref: Applied Environmental Microbiology, 84:, 2018 : PubMed ESTHER: Wei_2018_Appl.Environ.Microbiol_84_ PubMedSearch: Wei 2018 Appl.Environ.Microbiol 84 PubMedID: 29079625 Abstract Polyvinyl alcohol (PVA) is used widely in industry, and associated environmental pollution is a serious problem. Herein, we report a novel, efficient PVA degrader, Stenotrophomonas rhizophila QL-P4, isolated from fallen leaves from a virgin forest in the Qinling Mountains. The complete genome was obtained using single-molecule real-time (SMRT) technology and corrected using Illumina sequencing. Bioinformatics analysis revealed eight PVA/vinyl alcohol oligomer (OVA)-degrading genes. Of these, seven genes were predicted to be involved in the classic intracellular PVA/OVA degradation pathway, and one (BAY15_3292) was identified as a novel PVA oxidase. Five PVA/OVA-degrading enzymes were purified and characterized. One of these, BAY15_1712, a PVA dehydrogenase (PVADH), displayed high catalytic efficiency toward PVA and OVA substrate. All reported PVADHs only have PVA-degrading ability. Most importantly, we discovered a novel PVA oxidase (BAY15_3292) that exhibited higher PVA-degrading efficiency than the reported PVADHs. Further investigation indicated that BAY15_3292 plays a crucial role in PVA degradation in S. rhizophila QL-P4. Knocking out BAY15_3292 resulted in a significant decline in PVA-degrading activity in S. rhizophila QL-P4. Interestingly, we found that BAY15_3292 possesses exocrine activity, which distinguishes it from classic PVADHs. Transparent circle experiments further proved that BAY15_3292 greatly affects extracellular PVA degradation in S. rhizophila QL-P4. The exocrine characteristics of BAY15_3292 facilitate its potential application to PVA bioremediation. In addition, we report three new efficient secondary alcohol dehydrogenases (SADHs) with OVA-degrading ability in S. rhizophila QL-P4; in contrast, only one OVA-degrading SADH was reported previously.IMPORTANCE With the widespread application of PVA in industry, PVA-related environmental pollution is an increasingly serious issue. Because PVA is difficult to degrade, it accumulates in aquatic environments and causes chronic toxicity to aquatic organisms. Biodegradation of PVA, as an economical and environment-friendly method, has attracted much interest. To date, effective and applicable PVA-degrading bacteria/enzymes have not been reported. Herein, we report a new efficient PVA degrader (S. rhizophila QL-P4) that has five PVA/OVA-degrading enzymes with high catalytic efficiency, among which BAY15_1712 is the only reported PVADH with both PVA- and OVA-degrading abilities. Importantly, we discovered a novel PVA oxidase (BAY15_3292) that is not only more efficient than other reported PVA-degrading PVADHs but also has exocrine activity. Overall, our findings provide new insight into PVA-degrading pathways in microorganisms and suggest S. rhizophila QL-P4 and its enzymes have the potential for application to PVA bioremediation to reduce or eliminate PVA-related environmental pollution. Title: Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality Wei C, Yang H, Wang S, Zhao J, Liu C, Gao L, Xia E, Lu Y, Tai Y and Wan X <33 more author(s)> Wei C, Yang H, Wang S, Zhao J, Liu C, Gao L, Xia E, Lu Y, Tai Y, She G, Sun J, Cao H, Tong W, Gao Q, Li Y, Deng W, Jiang X, Wang W, Chen Q, Zhang S, Li H, Wu J, Wang P, Li P, Shi C, Zheng F, Jian J, Huang B, Shan D, Shi M, Fang C, Yue Y, Li F, Li D, Wei S, Han B, Jiang C, Yin Y, Xia T, Zhang Z, Bennetzen JL, Zhao S, Wan X (- 33) Ref: Proc Natl Acad Sci U S A, 115:E4151, 2018 : PubMed ESTHER: Wei_2018_Proc.Natl.Acad.Sci.U.S.A_115_E4151 PubMedSearch: Wei 2018 Proc.Natl.Acad.Sci.U.S.A 115 E4151 PubMedID: 29678829 Gene_locus related to this paper: camsi-a0a4s4dr18, camsi-a0a4s4etg9, camsi-a0a4s4e3j5, camsi-a0a4s4d2s5, camsi-a0a4s4duc4, camsi-a0a4v3wr80, camsi-a0a4v3wpu4 Abstract Tea, one of the world's most important beverage crops, provides numerous secondary metabolites that account for its rich taste and health benefits. Here we present a high-quality sequence of the genome of tea, Camellia sinensis var. sinensis (CSS), using both Illumina and PacBio sequencing technologies. At least 64% of the 3.1-Gb genome assembly consists of repetitive sequences, and the rest yields 33,932 high-confidence predictions of encoded proteins. Divergence between two major lineages, CSS and Camellia sinensis var. assamica (CSA), is calculated to approximately 0.38 to 1.54 million years ago (Mya). Analysis of genic collinearity reveals that the tea genome is the product of two rounds of whole-genome duplications (WGDs) that occurred approximately 30 to 40 and approximately 90 to 100 Mya. We provide evidence that these WGD events, and subsequent paralogous duplications, had major impacts on the copy numbers of secondary metabolite genes, particularly genes critical to producing three key quality compounds: catechins, theanine, and caffeine. Analyses of transcriptome and phytochemistry data show that amplification and transcriptional divergence of genes encoding a large acyltransferase family and leucoanthocyanidin reductases are associated with the characteristic young leaf accumulation of monomeric galloylated catechins in tea, while functional divergence of a single member of the glutamine synthetase gene family yielded theanine synthetase. This genome sequence will facilitate understanding of tea genome evolution and tea metabolite pathways, and will promote germplasm utilization for breeding improved tea varieties. Title: Proteolytic maturation of Drosophila Neuroligin 3 by tumor necrosis factor alpha-converting enzyme in the nervous system Wu J, Tao N, Tian Y, Xing G, Lv H, Han J, Lin C, Xie W Ref: Biochimica & Biophysica Acta, 1862:440, 2018 : PubMed ESTHER: Wu_2018_Biochim.Biophys.Acta_1862_440 PubMedSearch: Wu 2018 Biochim.Biophys.Acta 1862 440 PubMedID: 29107812 Abstract BACKGROUND: The functions of autism-associated Neuroligins (Nlgs) are modulated by their post-translational modifications, such as proteolytic cleavage. A previous study has shown that there are different endogenous forms of DNlg3 in Drosophila, indicating it may undergo proteolytic processing. However, the molecular mechanism underlying DNlg3 proteolytic processing is unknown. Here, we report a novel proteolytic mechanism that is essential for DNlg3 maturation and function in the nervous system. METHODS: Molecular cloning, cell culture, immunohistochemistry, western blotting and genetic studies were employed to map the DNlg3 cleavage region, identify the protease and characterize the cleavage manner. Behavior analysis, immunohistochemistry and genetic manipulations were employed to study the functions of different DNlg3 forms in the nervous system and neuromuscular junction (NMJs). RESULTS: Tumor necrosis factor alpha-converting enzyme (TACE) cleaved DNlg3 exclusively at its extracellular acetylcholinesterase-like domain to generate the N-terminal fragment and the short membrane-anchored fragment (sDNlg3). DNlg3 was constitutively processed in an activity-independent manner. Interestingly, DNlg3 was cleaved intracellularly in the Golgi apparatus before it arrived at the cell surface, a unique cleavage mechanism that is distinct from 'conventional' ectodomain shedding of membrane proteins, including rodent Nlg1. Genetic studies showed that sDNlg3 was essential for maintaining proper locomotor activity in Drosophila. CONCLUSIONS: Our results revealed a unique cleavage mechanism of DNlg3 and a neuron-specific role for DNlg3 maturation which is important in locomotor activity. GENERAL SIGNIFICANCE: Our study provides a new insight into a cleavage mechanism of Nlgs maturation in the nervous system. Title: Effect of food matrices on the in vitro bioavailability and oxidative damage in PC12 cells of lead Xia J, Fang Y, Shi Y, Shen X, Wu J, Xie M, Li P, Pei F, Hu Q Ref: Food Chem, 266:397, 2018 : PubMed ESTHER: Xia_2018_Food.Chem_266_397 PubMedSearch: Xia 2018 Food.Chem 266 397 PubMedID: 30381204 Abstract The bioavailability and oxidative damage toxicity of lead (Pb) in seven food matrices, including rice, milk, tomato, garlic, apple, kelp and pork, were determined using an in vitro digestion/Caco-2 cell model and a rat pheochromocytoma (PC12) oxidative damage model. Results showed that Pb bioaccessibility and bioavailability in the apple and kelp groups were significantly lower than other food matrix groups, with corresponding values of 11.05-28.31% and 1.57-8.81%, respectively. Oxidative damage assays showed that digestion products of apple polyphenol extract, which was selected from seven food matrices, could increase the oxidation resistance and the levels of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and acetyl cholinesterase (AChE) by 32.23%, 39.02%, 27.14% and 30.90%, respectively. Additionally, malondialdehyde (MDA) and reactive oxygen species (ROS) levels could be decreased by 59.66% and 40.21%, respectively. In conclusion, phenolics were an important food matrix that could decrease the bioavailability and oxidative damage of Pb. Title: Novel 8-hydroxyquinoline derivatives targeting beta-amyloid aggregation, metal chelation and oxidative stress against Alzheimer's disease Yang X, Cai P, Liu Q, Wu J, Yin Y, Wang X, Kong L Ref: Bioorganic & Medicinal Chemistry, 26:3191, 2018 : PubMed ESTHER: Yang_2018_Bioorg.Med.Chem_26_3191 PubMedSearch: Yang 2018 Bioorg.Med.Chem 26 3191 PubMedID: 29729985 Abstract A series of multitargeted 8-hydroxyquinoline derivatives were designed and synthesized for the treatment of Alzheimer's disease (AD). In vitro studies indicated that most of the prepared compounds exhibited significant inhibitory effects against self-induced Abeta(1-42) aggregation and potential antioxidant properties especially compound 5b (IC(50) = 5.64 microM for self-induced Abeta aggregation; the oxygen radical absorbance capacity using fluorescein (ORAC-FL) value is 2.63 Trolox equivalents). Notably, 5b can chelate biometals and inhibit Cu(2+)/Zn(2+)-induced Abeta(1-42) aggregation. The cell assays showed that 5b had excellent protective effects against oxidative toxin H(2)O(2) and presented low neurotoxicity in PC12 cells. Furthermore, 5b could penetrate the blood-brain barrier (BBB) in vitro and did not show any acute toxicity in mice at doses up to 2000 mg/kg in vivo. Our findings provide a rationale for the potential application of compound 5b as a lead compound in AD therapy. Title: [Effects of tenofovir and telbivudine on HBV RNA in pregnant women with different genotypes of HBeAg-positive hepatitis B in Guizhou Province] Zhang BF, Cheng ML, Lu S, Wu J, Wu YY, Liu Q, Zhao XK, Li YY, Hu YX, Liu W Ref: Zhonghua Yi Xue Za Zhi, 98:3503, 2018 : PubMed ESTHER: Zhang_2018_Zhonghua.Yi.Xue.Za.Zhi_98_3503 PubMedSearch: Zhang 2018 Zhonghua.Yi.Xue.Za.Zhi 98 3503 PubMedID: 30481899 Abstract Objective: To investigate whether HBV genotype influences HBV DNA and RNA responses to tenofovir(TDF) and telbivudine(LDT) in pregnant women with HBeAg-positive in Guizhou. Methods: This was a retrospective analysis of 75 pregnant women hepatitis B with HBsAg and HBeAg double-positive(19-38 years old, median age 26 years old), who were enrolled in the Department of Infectious Diseases and Obstetrics Clinic of the Affiliated Hospital of Guizhou Medical University from May 2016 to July 2017.Blood samples were collected at 12-24, 28-32 and 36-40 weeks of pregnancy for analyses of genotype, including hepatitis B surface antigen(HBsAg), hepatitis B e antigen (HBeAg), HBV DNA, HBV RNA and liver function, alanine transaminase(ALT), aspartate transaminase(AST), total bilirubin(TBiL), total bile acids(TBA), cholinesterase(CHE), alkaline phosphatase (ALP). Continuous variable was adopted by means of mean+/-standard deviation, and categorical variables were used for statistical analysis. Results: The HBV genotype was B in 64.0%(48/75)and C in 36.0%(27/75). The TDF and LDT groups showed no differences in demographic and clinical characteristics, including liver function tests, HBsAg, HBeAg, log(10)HBV DNA and log(10)HBV RNA.TDF groups, pre-treatment: HBV DNA (4.8+/-2.0), HBV RNA (6.4+/-1.1); at 4 weeks of treatment: HBV DNA (4.0+/-0.8), HBV RNA (6.0+/-0.9); at the end of treatment: HBV DNA (3.1+/-0.7), HBV RNA (5.5+/-0.8). LDT groups, pre-treatment HBV DNA (5.1+/-2.0), HBV RNA(6.5+/-0.9); at 4 weeks of treatment: HBV DNA (4.4+/-1.2), HBV RNA(6.5+/-0.8); at the end of treatment: HBV DNA(3.5+/-1.2), HBV RNA (6.1+/-0.7). Compared with pre-treatment (12-24 weeks), the TDF and LDT group showed significant reductions in log(10)(HBV DNA) and log(10)(HBV RNA) at 36-40 weeks ( P<0.05). Under the influence of excluding other variables, the genotype had a certain influence on the HBV RNA load.That was, HBV RNA in patients with the C genome decreased by 0.54 units(log(10)) at the end of the treatment compared to patients with the B genome, and the P value was less than 0.05. Conclusion: B and C genotypes are predominant in pregnant women with hepatitis B in Guizhou Province. B-type viruses are more easily controlled when different genotypes are treated with nucleotide analogues. Title: Genomic adaptation to polyphagy and insecticides in a major East Asian noctuid pest Cheng T, Wu J, Wu Y, Chilukuri RV, Huang L, Yamamoto K, Feng L, Li W, Chen Z and Mita K <28 more author(s)> Cheng T, Wu J, Wu Y, Chilukuri RV, Huang L, Yamamoto K, Feng L, Li W, Chen Z, Guo H, Liu J, Li S, Wang X, Peng L, Liu D, Guo Y, Fu B, Li Z, Liu C, Chen Y, Tomar A, Hilliou F, Montagne N, Jacquin-Joly E, d'Alencon E, Seth RK, Bhatnagar RK, Jouraku A, Shiotsuki T, Kadono-Okuda K, Promboon A, Smagghe G, Arunkumar KP, Kishino H, Goldsmith MR, Feng Q, Xia Q, Mita K (- 28) Ref: Nat Ecol Evol, 1:1747, 2017 : PubMed ESTHER: Cheng_2017_Nat.Ecol.Evol_1_1747 PubMedSearch: Cheng 2017 Nat.Ecol.Evol 1 1747 PubMedID: 28963452 Abstract The tobacco cutworm, Spodoptera litura, is among the most widespread and destructive agricultural pests, feeding on over 100 crops throughout tropical and subtropical Asia. By genome sequencing, physical mapping and transcriptome analysis, we found that the gene families encoding receptors for bitter or toxic substances and detoxification enzymes, such as cytochrome P450, carboxylesterase and glutathione-S-transferase, were massively expanded in this polyphagous species, enabling its extraordinary ability to detect and detoxify many plant secondary compounds. Larval exposure to insecticidal toxins induced expression of detoxification genes, and knockdown of representative genes using short interfering RNA (siRNA) reduced larval survival, consistent with their contribution to the insect's natural pesticide tolerance. A population genetics study indicated that this species expanded throughout southeast Asia by migrating along a South India-South China-Japan axis, adapting to wide-ranging ecological conditions with diverse host plants and insecticides, surviving and adapting with the aid of its expanded detoxification systems. The findings of this study will enable the development of new pest management strategies for the control of major agricultural pests such as S. litura. Title: A pH-responsive colorimetric strategy for DNA detection by acetylcholinesterase catalyzed hydrolysis and cascade amplification Guo Y, Yang K, Sun J, Wu J, Ju H Ref: Biosensors & Bioelectronics, 94:651, 2017 : PubMed ESTHER: Guo_2017_Biosens.Bioelectron_94_651 PubMedSearch: Guo 2017 Biosens.Bioelectron 94 651 PubMedID: 28388529 Abstract A pH-responsive colorimetric strategy was designed for sensitive and convenient biosensing by introducing acetylcholinesterase (AChE) catalyzed hydrolysis of acetylcholine to change solution pH and phenol red as an indicator. Using DNA as a target model, this technique was successfully employed for sensitive DNA analysis by labeling AChE to DNA. The sensitivity could be greatly improved by coupling a newly designed magnetic probe with target DNA-triggered nonenzymatic cascade amplification. In the presence of a help DNA (H) and the functional probe, the cascade assembly via toehold-mediated strand displacement released the AChE-conjugated sequence from magnetic beads, which could be simply separated from the reaction mixture to catalyze the hydrolysis of ACh in detection solution. The color change of detection solution from pink to orange-red, orange-yellow and ultimately yellow could be used for target DNA detection by naked eye and colorimetry with the absorbance ratio of detection solution at 558nm to 432nm as the signal. The nonenzymatically sensitized colorimetric strategy showed a linear range from 50pM to 50nM with a detection limit of 38pM, indicating a promising application in DNA analysis. Title: The Role of Oxidative Stress in Decreased Acetylcholinesterase Activity at the Neuromuscular Junction of the Diaphragm during Sepsis Liu H, Wu J, Yao JY, Wang H, Li ST Ref: Oxid Med Cell Longev, 2017:9718615, 2017 : PubMed ESTHER: Liu_2017_Oxid.Med.Cell.Longev_2017_9718615 PubMedSearch: Liu 2017 Oxid.Med.Cell.Longev 2017 9718615 PubMedID: 29230271 Abstract Our recent study demonstrated that acetylcholinesterase (AChE) activity at the neuromuscular junction (NMJ) of the diaphragm decreased during sepsis. However, the mechanisms were not clearly identified. In this study, we aimed to investigate whether the decreased AChE activity was related to oxidative stress by observing AChE activity in different grades of sepsis induced by caecal ligation and puncture (CLP). At 24 h after surgery, an assay of thiobarbituric acid reactive species (TBARS) and protein carbonyls, as well as the myeloperoxidase (MPO), superoxide dismutase (SOD), and catalase (CAT) activity, was conducted. AChE activity was measured by biochemical and histological detection. AChE and CAT activity in the diaphragm decreased, while the contents of TBARS and protein carbonyls, the activity of MPO and SOD, and the SOD/CAT ratios increased. The above changes were much more significant in the mid-grade septic group than in the low-grade septic group. The colour of the AChE activity staining at the NMJ gradually lightened from the sham surgery group to the mid-grade septic group. AChE activity was significantly negatively correlated with the levels of TBARS and protein carbonyls. We consider that oxidative stress might be responsible for decreased AChE activity in the diaphragms of rats induced with sepsis. Title: Sepsis decreases the activity of acetylcholinesterase by reducing its expression at the neuromuscular junction Wu J, Jin T, Wang H, Li ST Ref: Mol Med Rep, 16:5263, 2017 : PubMed ESTHER: Wu_2017_Mol.Med.Rep_16_5263 PubMedSearch: Wu 2017 Mol.Med.Rep 16 5263 PubMedID: 28849127 Abstract Our previous study demonstrated that sepsis may decrease the activity of acetylcholinesterase (AChE) at the neuromuscular junction (NMJ) of the diaphragm at 24 h, and thus improve the antagonistic action of neostigmine on rocuronium. The present study aimed to determine the effects of sepsis on AChE activity over 2 weeks, which is a more clinically relevant time period. Furthermore, the present study aimed to elucidate the association between AChE activity and its expression at the NMJ during sepsis. Male adult SpragueDawley rats were randomly divided into the sham or sepsis groups. Sepsis was induced by cecal ligation and puncture. On days 1, 3, 7 and 14 after surgery, AChE activity at the NMJ of the diaphragm was detected using a modified Karnovsky and Roots method. Furthermore, AChE expression levels at the NMJ, and in the whole muscle fibers of the diaphragm, were detected by immunohistofluorescence staining and western blot analysis, respectively. AChE activity was significantly decreased in the sepsis group, with its lowest level detected on day 7; however, its activity had partially recovered on day 14 (P<0.01). AChE activity was positively correlated (r=0.975, P=0.025) with its expression at the NMJ, which showed a similar trend over 2 weeks of sepsis. The protein expression levels of AChE in the whole muscle fibers of the diaphragm were significantly decreased on days 1, 3 and 7 in the sepsis group (P<0.01), with the lowest level observed on day 3. In conclusion, sepsis decreased AChE activity by reducing its expression at the NMJ over 14 days; the reduced expression of AChE at the NMJ might be as a result of its reduced muscular production. Title: Sequence Assembly of Yarrowia lipolytica Strain W29/CLIB89 Shows Transposable Element Diversity Magnan C, Yu J, Chang I, Jahn E, Kanomata Y, Wu J, Zeller M, Oakes M, Baldi P, Sandmeyer S Ref: PLoS ONE, 11:e0162363, 2016 : PubMed ESTHER: Magnan_2016_PLoS.ONE_11_E0162363 PubMedSearch: Magnan 2016 PLoS.ONE 11 E0162363 PubMedID: 27603307 Gene_locus related to this paper: yarli-q6c4p0 Abstract Yarrowia lipolytica, an oleaginous yeast, is capable of accumulating significant cellular mass in lipid making it an important source of biosustainable hydrocarbon-based chemicals. In spite of a similar number of protein-coding genes to that in other Hemiascomycetes, the Y. lipolytica genome is almost double that of model yeasts. Despite its economic importance and several distinct strains in common use, an independent genome assembly exists for only one strain. We report here a de novo annotated assembly of the chromosomal genome of an industrially-relevant strain, W29/CLIB89, determined by hybrid next-generation sequencing. For the first time, each Y. lipolytica chromosome is represented by a single contig. The telomeric rDNA repeats were localized by Irys long-range genome mapping and one complete copy of the rDNA sequence is reported. Two large structural variants and retroelement differences with reference strain CLIB122 including a full-length, novel Ty3/Gypsy long terminal repeat (LTR) retrotransposon and multiple LTR-like sequences are described. Strikingly, several of these are adjacent to RNA polymerase III-transcribed genes, which are almost double in number in Y. lipolytica compared to other Hemiascomycetes. In addition to previously-reported dimeric RNA polymerase III-transcribed genes, tRNA pseudogenes were identified. Multiple full-length and truncated LINE elements are also present. Therefore, although identified transposons do not constitute a significant fraction of the Y. lipolytica genome, they could have played an active role in its evolution. Differences between the sequence of this strain and of the existing reference strain underscore the utility of an additional independent genome assembly for this economically important organism. Title: Alpha-asarone improves striatal cholinergic function and locomotor hyperactivity in Fmr1 knockout mice Qiu G, Chen S, Guo J, Wu J, Yi YH Ref: Behavioural Brain Research, 312:212, 2016 : PubMed ESTHER: Qiu_2016_Behav.Brain.Res_312_212 PubMedSearch: Qiu 2016 Behav.Brain.Res 312 212 PubMedID: 27316341 Abstract Hyperactivity is a symptom found in several neurological and psychiatric disorders, including Fragile X syndrome (FXS). The animal model of FXS, fragile X mental retardation gene (Fmr1) knockout (KO) mouse, exhibits robust locomotor hyperactivity. Alpha (alpha)-asarone, a major bioactive component isolated from Acorus gramineus, has been shown in previous studies to improve various disease conditions including central nervous system disorders. In this study, we show that treatment with alpha-asarone alleviates locomotor hyperactivity in Fmr1 KO mice. To elucidate the mechanism underlying this improvement, we evaluated the expressions of various cholinergic markers, as well as acetylcholinesterase (AChE) activity and acetylcholine (ACh) levels, in the striatum of Fmr1 KO mice. We also analyzed the AChE-inhibitory activity of alpha-asarone. Striatal samples from Fmr1 KO mice showed decreased m1 muscarinic acetylcholine receptor (m1 mAChR) expression, increased AChE activity, and reduced ACh levels. Treatment with alpha-asarone improved m1 mAChR expression and ACh levels, and attenuated the increased AChE activity. In addition, alpha-asarone dose-dependently inhibited AChE activity in vitro. These results indicate that direct inhibition of AChE activity and up-regulation of m1 mAChR expression in the striatum might contribute to the beneficial effects of alpha-asarone on locomotor hyperactivity in Fmr1 KO mice. These findings might improve understanding of the neurobiological mechanisms responsible for locomotor hyperactivity. Title: Short-chain aliphatic ester synthesis using Thermobifida fusca cutinase Su L, Hong R, Guo X, Wu J, Xia Y Ref: Food Chem, 206:131, 2016 : PubMed ESTHER: Su_2016_Food.Chem_206_131 PubMedSearch: Su 2016 Food.Chem 206 131 PubMedID: 27041308 Abstract Short-chain aliphatic esters are commonly used as fruit flavorings in the food industry. In this study, Thermobifida fusca (T. fusca) cutinase was used for the synthesis of aliphatic esters, and the maximum yield of ethyl caproate reached 99.2% at a cutinase concentration of 50U/ml, 40 degrees C, and water content of 0.5%, representing the highest ester yield to date. The cutinase-catalyzed esterification displayed strong tolerance for water content (up to 8%) and acid concentration (up to 0.8M). At substrate concentrations 0.8M, the ester yield remained above 80%. Moreover, ester yields of more than 98% and 95% were achieved for acids of C3-C8 and alcohols of C1-C6, respectively, indicating extensive chain length selectivity of the cutinase. These results demonstrate the superior ability of T. fusca cutinase to catalyze the synthesis of short-chain esters. This study provides the basis for industrial production of short-chain esters using T. fusca cutinase. Title: Comparative Genomics Analysis of Streptomyces Species Reveals Their Adaptation to the Marine Environment and Their Diversity at the Genomic Level Tian X, Zhang Z, Yang T, Chen M, Li J, Chen F, Yang J, Li W, Zhang B and Xiao J <3 more author(s)> Tian X, Zhang Z, Yang T, Chen M, Li J, Chen F, Yang J, Li W, Zhang B, Wu J, Zhang C, Long L, Xiao J (- 3) Ref: Front Microbiol, 7:998, 2016 : PubMed ESTHER: Tian_2016_Front.Microbiol_7_998 PubMedSearch: Tian 2016 Front.Microbiol 7 998 PubMedID: 27446038 Gene_locus related to this paper: 9actn-a0a1e7k9d4, 9actn-a0a1e7l883, 9actn-a0a1e7kw84, 9actn-a0a1e7kkk9, 9actn-a0a1e7jip1, 9actn-a0a1e7jjj1, 9actn-a0a1e7k159, 9actn-a0a1e7kfw2, 9actn-a0a1e7l1n0, 9actn-a0a1e7jf99 Abstract Over 200 genomes of streptomycete strains that were isolated from various environments are available from the NCBI. However, little is known about the characteristics that are linked to marine adaptation in marine-derived streptomycetes. The particularity and complexity of the marine environment suggest that marine streptomycetes are genetically diverse. Here, we sequenced nine strains from the Streptomyces genus that were isolated from different longitudes, latitudes, and depths of the South China Sea. Then we compared these strains to 22 NCBI downloaded streptomycete strains. Thirty-one streptomycete strains are clearly grouped into a marine-derived subgroup and multiple source subgroup-based phylogenetic tree. The phylogenetic analyses have revealed the dynamic process underlying streptomycete genome evolution, and lateral gene transfer is an important driving force during the process. Pan-genomics analyses have revealed that streptomycetes have an open pan-genome, which reflects the diversity of these streptomycetes and guarantees the species a quick and economical response to diverse environments. Functional and comparative genomics analyses indicate that the marine-derived streptomycetes subgroup possesses some common characteristics of marine adaptation. Our findings have expanded our knowledge of how ocean isolates of streptomycete strains adapt to marine environments. The availability of streptomycete genomes from the South China Sea will be beneficial for further analysis on marine streptomycetes and will enrich the South China Sea's genetic data sources. Title: SAR11 bacteria linked to ocean anoxia and nitrogen loss Tsementzi D, Wu J, Deutsch S, Nath S, Rodriguez RL, Burns AS, Ranjan P, Sarode N, Malmstrom RR and Stewart FJ <8 more author(s)> Tsementzi D, Wu J, Deutsch S, Nath S, Rodriguez RL, Burns AS, Ranjan P, Sarode N, Malmstrom RR, Padilla CC, Stone BK, Bristow LA, Larsen M, Glass JB, Thamdrup B, Woyke T, Konstantinidis KT, Stewart FJ (- 8) Ref: Nature, 536:179, 2016 : PubMed ESTHER: Tsementzi_2016_Nature_536_179 PubMedSearch: Tsementzi 2016 Nature 536 179 PubMedID: 27487207 Gene_locus related to this paper: 9prot-a0a1c2agj3, 9prot-a0a1c2azw4 Abstract Bacteria of the SAR11 clade constitute up to one half of all microbial cells in the oxygen-rich surface ocean. SAR11 bacteria are also abundant in oxygen minimum zones (OMZs), where oxygen falls below detection and anaerobic microbes have vital roles in converting bioavailable nitrogen to N2 gas. Anaerobic metabolism has not yet been observed in SAR11, and it remains unknown how these bacteria contribute to OMZ biogeochemical cycling. Here, genomic analysis of single cells from the world's largest OMZ revealed previously uncharacterized SAR11 lineages with adaptations for life without oxygen, including genes for respiratory nitrate reductases (Nar). SAR11 nar genes were experimentally verified to encode proteins catalysing the nitrite-producing first step of denitrification and constituted ~40% of OMZ nar transcripts, with transcription peaking in the anoxic zone of maximum nitrate reduction activity. These results link SAR11 to pathways of ocean nitrogen loss, redefining the ecological niche of Earth's most abundant organismal group. Title: Sepsis Strengthens Antagonistic Actions of Neostigmine on Rocuronium in a Rat Model of Cecal Ligation and Puncture Wu J, Jin T, Wang H, Li ST Ref: Chinese Medical Journal (Engl), 129:1477, 2016 : PubMed ESTHER: Wu_2016_Chin.Med.J.(Engl)_129_1477 PubMedSearch: Wu 2016 Chin.Med.J.(Engl) 129 1477 PubMedID: 27270546 Inhibitor(s) related to this paper: Rocuronium Abstract BACKGROUND: The antagonistic actions of anticholinesterase drugs on non-depolarizing muscle relaxants are theoretically related to the activity of acetylcholinesterase (AChE) in the neuromuscular junction (NMJ). However, till date the changes of AChE activity in the NMJ during sepsis have not been directly investigated. We aimed to investigate the effects of sepsis on the antagonistic actions of neostigmine on rocuronium (Roc) and the underlying changes of AChE activity in the NMJ in a rat model of cecal ligation and puncture (CLP). Title: Chemical Constituents of Plants from the Genus Phlegmariurus Yang Y, Wang Z, Wu J, Chen Y Ref: Chem Biodivers, 13:269, 2016 : PubMed ESTHER: Yang_2016_Chem.Biodivers_13_269 PubMedSearch: Yang 2016 Chem.Biodivers 13 269 PubMedID: 26916276 Abstract Phlegmariurus is a genus of ca. 200 species in the family Huperziaceae. Up to now, six species of the Phlegmariurus genus have been chemically investigated, and 89 compounds, including Lycopodium alkaloids possessing diverse structures and serratane-type triterpenes, have been isolated. These compounds show potent bioactivities, such as acetylcholinesterase inhibitory and cytotoxic activities. Title: A nano-silver enzyme electrode for organophosphorus pesticide detection Zheng Q, Yu Y, Fan K, Ji F, Wu J, Ying Y Ref: Anal Bioanal Chem, 408:5819, 2016 : PubMed ESTHER: Zheng_2016_Anal.Bioanal.Chem_408_5819 PubMedSearch: Zheng 2016 Anal.Bioanal.Chem 408 5819 PubMedID: 27342792 Abstract A nano-silver electrode immobilizing acetylcholinesterase (AChE) for the detection of organophosphorus (OPPs) pesticides is reported. Scanning electron microscopy (SEM) was used to characterize the surface structure of two kinds of electrodes fabricated with different sizes of silver powders and the interface between chitosan layer and nano-silver powder layer. Cyclic voltammetry was carried out to characterize the response of silver/chitosan electrode in the absence and in the presence of thiocholine (TCh). It was also used to evaluate the insulativity of the chitosan layer. An amperometric method was performed to measure the response of the electrode to TCh, which is the product of the enzymatic reaction for detecting organophosphorus pesticides indirectly. Although there are many kinds of nanoparticles, silver was chosen for its internal advantage in detecting TCh at low potential without further modification. The result shows nano-silver powder has better performance than usual silver powder, and the limit of detection of paraoxon is 4 ppb under optimized conditions. One percent (w/v) chitosan solution was used as binder for the immobilization of nano-silver powder and AChE, which made it possible for independent electrode fabrication at room temperature, whereas 3% (w/v) chitosan solution was used as insulating compound for controlling the electrode area. Unlike traditional organic insulating ink, chitosan is safe and environmentally friendly, and it is used as insulating material for the first time. The flexible nano-silver/AChE/chitosan electrode was evaluated in Chinese chives and cabbage, and the recoveries of standard addition were 105.11 and 96.41%, respectively. Owing to the antibacterial property of nano-silver and the biocompatibility, safety, and biodegradability of chitosan, the proposed method is safe, facile, environmentally friendly, and has great potential in organophosphorus pesticide detection for food safety. Graphical Abstract Current response of nano-silver electrode (a) and silver electrode (b) to thiocholine in 0.02 M PBS + KCl at 0.15 V; addition of thiocholine (0.09 mM) every 50 s ( downward arrow); inset: calibration curve of nano-silver () and silver () electrode. Title: Genome sequence of cultivated Upland cotton (Gossypium hirsutum TM-1) provides insights into genome evolution Li F, Fan G, Lu C, Xiao G, Zou C, Kohel RJ, Ma Z, Shang H, Ma X and Yu S <30 more author(s)> Li F, Fan G, Lu C, Xiao G, Zou C, Kohel RJ, Ma Z, Shang H, Ma X, Wu J, Liang X, Huang G, Percy RG, Liu K, Yang W, Chen W, Du X, Shi C, Yuan Y, Ye W, Liu X, Zhang X, Liu W, Wei H, Wei S, Zhu S, Zhang H, Sun F, Wang X, Liang J, Wang J, He Q, Huang L, Cui J, Song G, Wang K, Xu X, Yu JZ, Zhu Y, Yu S (- 30) Ref: Nat Biotechnol, 33:524, 2015 : PubMed ESTHER: Li_2015_Nat.Biotechnol_33_524 PubMedSearch: Li 2015 Nat.Biotechnol 33 524 PubMedID: 25893780 Gene_locus related to this paper: gosra-a0a0d2rxs2, gosra-a0a0d2tng2, gosra-a0a0d2twz7, goshi-a0a1u8hr03, gosra-a0a0d2vdc5, goshi-a0a1u8ljh5, gosra-a0a0d2vj24, goshi-a0a1u8pxd3, gosra-a0a0d2sr31, goshi-a0a1u8knd1, goshi-a0a1u8nhw9, goshi-a0a1u8mt09, goshi-a0a1u8kis4, goshi-a0a1u8ibk3, goshi-a0a1u8ieg2, goshi-a0a1u8iki6, goshi-a0a1u8jvp4, goshi-a0a1u8jw35, gosra-a0a0d2pzd7, goshi-a0a1u8ied7 Abstract Gossypium hirsutum has proven difficult to sequence owing to its complex allotetraploid (AtDt) genome. Here we produce a draft genome using 181-fold paired-end sequences assisted by fivefold BAC-to-BAC sequences and a high-resolution genetic map. In our assembly 88.5% of the 2,173-Mb scaffolds, which cover 89.6% approximately 96.7% of the AtDt genome, are anchored and oriented to 26 pseudochromosomes. Comparison of this G. hirsutum AtDt genome with the already sequenced diploid Gossypium arboreum (AA) and Gossypium raimondii (DD) genomes revealed conserved gene order. Repeated sequences account for 67.2% of the AtDt genome, and transposable elements (TEs) originating from Dt seem more active than from At. Reduction in the AtDt genome size occurred after allopolyploidization. The A or At genome may have undergone positive selection for fiber traits. Concerted evolution of different regulatory mechanisms for Cellulose synthase (CesA) and 1-Aminocyclopropane-1-carboxylic acid oxidase1 and 3 (ACO1,3) may be important for enhanced fiber production in G. hirsutum. Title: Extracellular expression of Thermobifida fusca cutinase with pelB signal peptide depends on more than type II secretion pathway in Escherichia coli Su L, Yu L, Xu C, Wu J Ref: J Biotechnol, 204:47, 2015 : PubMed ESTHER: Su_2015_J.Biotechnol_204_47 PubMedSearch: Su 2015 J.Biotechnol 204 47 PubMedID: 25863154 Abstract Our previous studies demonstrated that Thermobifida fusca cutinase is released into culture medium when expressed without a signal peptide in Escherichia coli, and this extracellular expression results from an enhanced membrane permeability caused by cutinase's phospholipid hydrolase activity. The present study investigated whether this phenomenon would also occur during the expression of cutinase fused to pelB signal peptide (pelB-cutinase). Secretion of fusion proteins of this type is generally believed to occur via type II secretion pathway. The results showed that when pelB-cutinase was expressed in a secB knockout strain, which has a defective type II secretion pathway, there was still a large amount of cutinase in the culture medium. Additional experiments confirmed that the periplasmic and cytoplasmic fractions of the expressing cells had hydrolytic activity toward phosphatidyl ethanolamine, and the recombinant cells showed correspondingly improved membrane permeability. All these phenomena were also observed in the parent E. coli strain. Moreover, the secretion efficiency of the inactive cutinase mutant was found to be significantly lower than that of pelB-cutinase in the parent E. coli. Based on these results, the phospholipid hydrolase activity of pelB-cutinase must play a larger role in its extracellular production than does type II secretion pathway. Title: Extracellular expression of natural cytosolic arginine deiminase from Pseudomonas putida and its application in the production of l-citrulline Su L, Ma Y, Wu J Ref: Bioresour Technol, 196:176, 2015 : PubMed ESTHER: Su_2015_Bioresour.Technol_196_176 PubMedSearch: Su 2015 Bioresour.Technol 196 176 PubMedID: 26233330 Abstract The Pseudomonas putida arginine deiminase (ADI), a natural cytosolic enzyme, and Thermobifida fusca cutinase were co-expressed in Escherichia coli, and the optimized cutinase gene was used for increasing its expression level. 90.9% of the total ADI protein was released into culture medium probably through a nonspecific leaking mechanism caused by the co-expressed cutinase. The enzymatic properties of the extracellular ADI were found to be similar to those of ADI prepared by conventional cytosolic expression. Extracellular production of ADI was further scaled up in a 3-L fermentor. When the protein expression was induced by IPTG (25.0muM) and lactose (0.1gL-1h-1) at 30 degrees C, the extracellular ADI activity reached 101.2UmL-1, which represented the highest ADI production ever reported. In addition, the enzymatic synthesis of l-citrulline was performed using the extracellularly expressed ADI, and the conversion rate reached 100% with high substrate concentration at 650gL-1. Title: alpha7 and beta2 Nicotinic Acetylcholine Receptor Subunits Form Heteromeric Receptor Complexes that Are Expressed in the Human Cortex and Display Distinct Pharmacological Properties Thomsen MS, Zwart R, Ursu D, Jensen MM, Pinborg LH, Gilmour G, Wu J, Sher E, Mikkelsen JD Ref: PLoS ONE, 10:e0130572, 2015 : PubMed ESTHER: Thomsen_2015_PLoS.One_10_e0130572 PubMedSearch: Thomsen 2015 PLoS.One 10 e0130572 PubMedID: 26086615 Abstract The existence of alpha7beta2 nicotinic acetylcholine receptors (nAChRs) has recently been demonstrated in both the rodent and human brain. Since alpha7-containing nAChRs are promising drug targets for schizophrenia and Alzheimer's disease, it is critical to determine whether alpha7beta2 nAChRs are present in the human brain, in which brain areas, and whether they differ functionally from alpha7 nAChR homomers. We used alpha-bungarotoxin to affinity purify alpha7-containing nAChRs from surgically excised human temporal cortex, and found that alpha7 subunits co-purify with beta2 subunits, indicating the presence of alpha7beta2 nAChRs in the human brain. We validated these results by demonstrating co-purification of beta2 from wild-type, but not alpha7 or beta2 knock-out mice. The pharmacology and kinetics of human alpha7beta2 nAChRs differed significantly from that of alpha7 homomers in response to nAChR agonists when expressed in Xenopus oocytes and HEK293 cells. Notably, alpha7beta2 heteromers expressed in HEK293 cells display markedly slower rise and decay phases. These results demonstrate that alpha7 subunits in the human brain form heteromeric complexes with beta2 subunits, and that human alpha7beta2 nAChR heteromers respond to nAChR agonists with a unique pharmacology and kinetic profile. alpha7beta2 nAChRs thus represent an alternative mechanism for the reported clinical efficacy of alpha7 nAChR ligands. Title: Perilipin 5 improves hepatic lipotoxicity by inhibiting lipolysis Wang C, Zhao Y, Gao X, Li L, Yuan Y, Liu F, Zhang L, Wu J, Hu P and Ye J <6 more author(s)> Wang C, Zhao Y, Gao X, Li L, Yuan Y, Liu F, Zhang L, Wu J, Hu P, Zhang X, Gu Y, Xu Y, Wang Z, Li Z, Zhang H, Ye J (- 6) Ref: Hepatology, 61:870, 2015 : PubMed ESTHER: Wang_2015_Hepatology_61_870 PubMedSearch: Wang 2015 Hepatology 61 870 PubMedID: 25179419 Abstract Abnormal metabolism of nonesterified fatty acids (NEFAs) and their derivatives has been reported to be the main cause of intracellular lipotoxic injury. Normally, NEFAs are stored in lipid droplets (LDs) in the form of triglyceride (TG), which could reduce the lipotoxicity of cytosolic NEFAs. Previous studies have implicated that Perilipin 5 (Plin5), an LD-binding protein, regulates the storage and hydrolysis of TG in LD. However, its roles and underlying mechanisms in the liver remain unknown. Here we found that Plin5 expression was increased in steatotic livers. Using Plin5 knockout mice, we found that Plin5 deficiency resulted in reduced hepatic lipid content and smaller-sized LDs, which was due to the elevated lipolysis rate and fatty acid utilization. Plin5-deficient hepatocytes showed increased mitochondria proliferation, which could be explained by the increased expression and activity of PPARalpha stimulated by the increased NEFA levels. Meanwhile, Plin5-deficient livers also exhibited enhanced mitochondrial oxidative capacity. We also found that Plin5 deficiency induces lipotoxic injury in hepatocytes, attributed to lipid peroxidation. Mechanistically, we found that Plin5 blocks adipose triglyceride lipase (ATGL)-mediated lipolysis by competitively binding to comparative gene identification-58 (CGI-58) and disrupting the interaction between CGI-58 and ATGL. CONCLUSION: Plin5 is an important protective factor against hepatic lipotoxicity induced by NEFAs generated from lipolysis. This provides an important new insight into the regulation of hepatic lipid storage and relation between lipid storage and lipotoxicity. Title: The halo-substituent effect on Pseudomonas cepacia lipase-mediated regioselective acylation of nucleosides: A comparative investigation Wang ZY, Bi YH, Yang RL, Duan ZQ, Nie LH, Li XQ, Zong MH, Wu J Ref: J Biotechnol, 212:153, 2015 : PubMed ESTHER: Wang_2015_J.Biotechnol_212_153 PubMedSearch: Wang 2015 J.Biotechnol 212 153 PubMedID: 26325198 Abstract In this work, comparative experiments were explored to investigate the substrate specificity of Pseudomonas cepacia lipase in regioselective acylation of nucleosides carrying various substituents (such as the H, F, Cl, Br, I) at 2'- and 5-positions. Experimental data indicated that the catalytic performance of the enzyme depended very much on the halo-substituents in nucleosides. The increased bulk of 2'-substituents in ribose moiety of the nucleoside might contribute to the improved 3'-regioselectivity (90-98%, nucleosides a-d) in enzymatic decanoylation, while the enhancement of regioselectivity (93-99%) in 3'-O-acylated nucleosides e-h could be attributable to the increasing hydrophobicity of the halogen atoms at 5-positions. With regard to the chain-length selectivity, P. cepacia lipase displayed the highest 3'-regioselectivity toward the longer chain (C14) as compared to shorter (C6 and C10) ones. The position, orientation and property of the substituent, specific structure of the lipase's active site, and acyl structure could account for the diverse results. Title: FgRIC8 is involved in regulating vegetative growth, conidiation, deoxynivalenol production and virulence in Fusarium graminearum Wu J, Liu Y, Lv W, Yue X, Que Y, Yang N, Zhang Z, Ma Z, Talbot NJ, Wang Z Ref: Fungal Genet Biol, 83:92, 2015 : PubMed ESTHER: Wu_2015_Fungal.Genet.Biol_83_92 PubMedSearch: Wu 2015 Fungal.Genet.Biol 83 92 PubMedID: 26341536 Abstract Proteins of the resistance to inhibitors of cholinesterase 8 (Ric8) group act as guanine nucleotide exchange factors (GEFs) and play important roles in regulating G-protein signaling in animals. In filamentous fungi, putative Ric8 orthologs have so far been identified in Magnaporthe oryzae, Neurospora crassa, Aspergillus nidulans and Aspergillus fumigatus. Here, we report the functional investigation of a potential RIC8 ortholog (FgRIC8) in the wheat head blight pathogen Fusarium graminearum. Targeted gene deletion mutants of FgRIC8 exhibited a significant reduction in vegetative growth, conidiation, pigment production as well as deoxynivalenol (DON) biosynthesis. Pathogenicity assays using a point-inoculated spikelet approach showed that the mutants were severely impaired in virulence on flowering wheat heads. Quantitative RT-PCR analysis revealed that genes encoding F. graminearum Galpha (FgGpa1 and FgGpa3), Gbeta (FgGpb1) and Ggamma (FgGpg1) subunits were significantly down-regulated in Fgric8 mutants. Moreover, we showed that FgRic8 physically interacts with both FgGpa1 and FgGpa3, but not FgGpa2, in yeast two-hybrid assays. The intracellular cAMP levels in Fgric8 mutants were significantly decreased compared to the isogenic wild-type strain. Taken together, our results indicate that FgRic8 plays critical roles in fungal development, secondary metabolism and virulence in F. graminearum and may act as a regulator of G protein alpha subunits. Title: Anticancer drugs induce hypomethylation of the acetylcholinesterase promoter via a phosphorylated-p38-DNMT1-AChE pathway in apoptotic hepatocellular carcinoma cells Xi Q, Gao N, Yang Y, Ye W, Zhang B, Wu J, Jiang G, Zhang X Ref: International Journal of Biochemistryistry & Cell Biology, 68:21, 2015 : PubMed ESTHER: Xi_2015_Int.J.Biochem.Cell.Biol_68_21 PubMedSearch: Xi 2015 Int.J.Biochem.Cell.Biol 68 21 PubMedID: 26299326 Abstract Apoptosis, also known as programmed cell death, plays an essential role in eliminating excessive, damaged or harmful cells. Previous work has demonstrated that anticancer drugs induce cell apoptosis by inducing cytotoxicity. In recent years, several reports demonstrated modulated expression of DNA methyltransferases 1 (DNMT1) and acetylcholinesterase (AChE) in a variety of tumors. In this study, we showed that the expression of DNMT1 was decreased and the methylation of CpGs in the promoter of AChE was reduced in anticancer drugs-induced apoptotic hepatocellular carcinoma cells. Silencing of DNMT1 expression by AZA or RNA interference (RNAi) restored AChE production and inhibition of AChE expression by RNAi protected HCC cells from anticancer drugs-induced apoptosis. Furthermore, we demonstrated that the regulation of AChE by DNMT1 was involved in the phosphorylated p38 pathway in anticancer drugs-induced apoptosis. In addition, immunohistochemical staining showed that P-p38, DNMT1 and AChE were aberrantly expressed in a subset of HCC tumors. Taken together, we demonstrated the regulation of AChE by DNMT1 and further, we found that this regulation was involved in the phosphorylated p38 pathway in anticancer drugs-induced apoptosis. Title: Discovery and expression of a Pseudomonas sp. esterase as a novel biocatalyst for the efficient biosynthesis of a chiral intermediate of pregabalin Xu F, Chen S, Xu G, Wu J, Yang L Ref: Biotechnol Bioprocess Eng, 20:473, 2015 : PubMed ESTHER: Xu_2015_Biotechnol.Bioprocess.Eng_20_473 PubMedSearch: Xu 2015 Biotechnol.Bioprocess.Eng 20 473 Title: Epigenetic suppression of neuroligin 1 underlies amyloid-induced memory deficiency Bie B, Wu J, Yang H, Xu JJ, Brown DL, Naguib M Ref: Nat Neurosci, 17:223, 2014 : PubMed ESTHER: Bie_2014_Nat.Neurosci_17_223 PubMedSearch: Bie 2014 Nat.Neurosci 17 223 PubMedID: 24441681 Abstract Amyloid-induced microglial activation and neuroinflammation impair central synapses and memory function, although the mechanism remains unclear. Neuroligin 1 (NLGN1), a postsynaptic protein found in central excitatory synapses, governs excitatory synaptic efficacy and plasticity in the brain. Here we found, in rodents, that amyloid fibril-induced neuroinflammation enhanced the interaction between histone deacetylase 2 and methyl-CpG-binding protein 2, leading to suppressed histone H3 acetylation and enhanced cytosine methylation in the Nlgn1 promoter region and decreased NLGN1 expression, underlying amyloid-induced memory deficiency. Manipulation of microglia-associated neuroinflammation modulated the epigenetic modification of the Nlgn1 promoter, hippocampal glutamatergic transmission and memory function. These findings link neuroinflammation, synaptic efficacy and memory, thus providing insight into the pathogenesis of amyloid-associated diseases. Title: Omarigliptin (MK-3102): a novel long-acting DPP-4 inhibitor for once-weekly treatment of type 2 diabetes Biftu T, Sinha-Roy R, Chen P, Qian X, Feng D, Kuethe JT, Scapin G, Gao YD, Yan Y and Weber AE <19 more author(s)> Biftu T, Sinha-Roy R, Chen P, Qian X, Feng D, Kuethe JT, Scapin G, Gao YD, Yan Y, Krueger D, Bak A, Eiermann G, He J, Cox J, Hicks J, Lyons K, He H, Salituro G, Tong S, Patel S, Doss G, Petrov A, Wu J, Xu SS, Sewall C, Zhang X, Zhang B, Thornberry NA, Weber AE (- 19) Ref: Journal of Medicinal Chemistry, 57:3205, 2014 : PubMed ESTHER: Biftu_2014_J.Med.Chem_57_3205 PubMedSearch: Biftu 2014 J.Med.Chem 57 3205 PubMedID: 24660890 Gene_locus related to this paper: human-DPP4 Inhibitor(s) related to this paper: Omarigliptin Abstract In our effort to discover DPP-4 inhibitors with added benefits over currently commercially available DPP-4 inhibitors, MK-3102 (omarigliptin), was identified as a potent and selective dipeptidyl peptidase 4 (DPP-4) inhibitor with an excellent pharmacokinetic profile amenable for once-weekly human dosing and selected as a clinical development candidate. This manuscript summarizes the mechanism of action, scientific rationale, medicinal chemistry, pharmacokinetic properties, and human efficacy data for omarigliptin, which is currently in phase 3 clinical development. Title: Identification and characterization of a new erythromycin biosynthetic gene cluster in Actinopolyspora erythraea YIM90600, a novel erythronolide-producing halophilic actinomycete isolated from salt field Chen D, Feng J, Huang L, Zhang Q, Wu J, Zhu X, Duan Y, Xu Z Ref: PLoS ONE, 9:e108129, 2014 : PubMed ESTHER: Chen_2014_PLoS.ONE_9_E108129 PubMedSearch: Chen 2014 PLoS.ONE 9 E108129 PubMedID: 25250723 Gene_locus related to this paper: 9acto-a0a099d7w4, 9actn-a0a099d934 Abstract Erythromycins (Ers) are clinically potent macrolide antibiotics in treating pathogenic bacterial infections. Microorganisms capable of producing Ers, represented by Saccharopolyspora erythraea, are mainly soil-dwelling actinomycetes. So far, Actinopolyspora erythraea YIM90600, a halophilic actinomycete isolated from Baicheng salt field, is the only known Er-producing extremophile. In this study, we have reported the draft genome sequence of Ac. erythraea YIM90600, genome mining of which has revealed a new Er biosynthetic gene cluster encoding several novel Er metabolites. This Er gene cluster shares high identity and similarity with the one of Sa. erythraea NRRL2338, except for two absent genes, eryBI and eryG. By correlating genotype and chemotype, the biosynthetic pathways of 3'-demethyl-erythromycin C, erythronolide H (EH) and erythronolide I have been proposed. The formation of EH is supposed to be sequentially biosynthesized via C-6/C-18 epoxidation and C-14 hydroxylation from 6-deoxyerythronolide B. Although an in vitro enzymatic activity assay has provided limited evidence for the involvement of the cytochrome P450 oxidase EryFAc (derived from Ac. erythraea YIM90600) in the catalysis of a two-step oxidation, resulting in an epoxy moiety, the attempt to construct an EH-producing Sa. erythraea mutant via gene complementation was not successful. Characterization of EryKAc (derived from Ac. erythraea YIM90600) in vitro has confirmed its unique role as a C-12 hydroxylase, rather than a C-14 hydroxylase of the erythronolide. Genomic characterization of the halophile Ac. erythraea YIM90600 will assist us to explore the great potential of extremophiles, and promote the understanding of EH formation, which will shed new insights into the biosynthesis of Er metabolites. Title: Characterization and structure basis of Pseudomonas alcaligenes lipase's enantiopreference towards d,l-menthyl propionate Chen H, Wu J, Yang L, Xu G Ref: J Mol Catal B Enzym, 102:81, 2014 : PubMed ESTHER: Chen_2014_J.Mol.Catal.B.Enzym_102_81 PubMedSearch: Chen 2014 J.Mol.Catal.B.Enzym 102 81 Gene_locus related to this paper: stema-s4tny8 Abstract In this work, a lipase from Pseudomonas alcaligenes CGMCC4405 (PaL) was cloned and expressed. It was very attractive that the recombinant PaL exhibited excellent enantioselectivity (E > 200) in the resolution of racemic d,l-menthyl propionate to produce l-menthol. The structure basis of enantiopreference is a fundamental scientific problem which needs to be resolved. In our research, molecular dynamic simulation (MD) research was employed to research the different binding modes of d and l-menthyl propionate. The results showed that when bound with slow-reacting enantiomer (d-menthyl propionate), the steric requirements of the large substituent (isopropyl) of the d-menthyl propionate force a rotation of the imidazole ring of catalytic residue His271 and further pushed the active site His271 away from its proper orientation. Moreover, the average distance between alcohol oxygen (Oalc) and HNe of catalytic His271 increased to 3.7 A, which was too far to form an essential hydrogen bond and further prevented efficient catalysis of slow enantiomer. This correlation of the distance between alcohol oxygen (Oalc) and HNe of catalytic His271 and the enantioselectivity was also confirmed by the result of site-directed mutagenesis. Title: The role of synaptic activity in the regulation of amyloid beta levels in Alzheimer's disease Cheng X, Wu J, Geng M, Xiong J Ref: Neurobiology of Aging, 35:1217, 2014 : PubMed ESTHER: Cheng_2014_Neurobiol.Aging_35_1217 PubMedSearch: Cheng 2014 Neurobiol.Aging 35 1217 PubMedID: 24368087 Abstract Alzheimer's disease (AD) is the most common form of dementia. Accumulation of amyloid-beta (Abeta) peptides is regarded as the critical component associated with AD pathogenesis, which is derived from the amyloid precursor protein (APP) cleavage. Recent studies suggest that synaptic activity is one of the most important factors that regulate Abeta levels. It has been found that synaptic activity facilitates APP internalization and influences APP cleavage. Glutamatergic, cholinergic, serotonergic, leptin, adrenergic, orexin, and gamma-amino butyric acid receptors, as well as the activity-regulated cytoskeleton-associated protein (Arc) are all involved in these processes. The present review summarizes the evidence for synaptic activity-modulated Abeta levels and the mechanisms underlying this regulation. Interestingly, the immediate early gene product Arc may also be the downstream signaling molecule of several receptors in the synaptic activity-modulated Abeta levels. Elucidating how Abeta levels are regulated by synaptic activity may provide new insights in both the understanding of the pathogenesis of AD and in the development of therapies to slow down the progression of AD. Title: GW1929 inhibits alpha7 nAChR expression through PPARgamma-independent activation of p38 MAPK and inactivation of PI3-K/mTOR: The role of Egr-1 Hahn SS, Tang Q, Zheng F, Zhao S, Wu J Ref: Cell Signal, 26:730, 2014 : PubMed ESTHER: Hahn_2014_Cell.Signal_26_730 PubMedSearch: Hahn 2014 Cell.Signal 26 730 PubMedID: 24412748 Abstract Studies demonstrated that peroxisome proliferator-activated receptor gamma (PPARgamma) ligands reduce nicotine-induced non small cell lung carcinoma (NSCLC) cell growth through inhibition of nicotinic acetylcholine receptor (nAChR) mediated signaling pathways. However, the mechanisms by which PPARgamma ligands inhibited nAChR expression remain elucidated. Here, we show that GW1929, a synthetic PPARgamma ligand, not only inhibited but also antagonized the stimulatory effect of acetylcholine on NSCLC cell proliferation. Interestingly, GW1929 inhibited alpha7 nAChR expression, which was not blocked by GW9662, an antagonist of PPARgamma, or by PPARgamma siRNA, but was abrogated by the p38 MPAK inhibitor SB239063. GW1929 reduced the promoter activity of alpha7 nAChR and induced early growth response-1 (Egr-1) protein expression, which was overcame by SB239063, but enhanced by inhibitors of PI3-K and mTOR. Silencing of Egr-1 blocked, while overexpression of Egr-1 enhanced, the effect of GW1929 on alpha7 nAChR expression and promoter activity. Finally, GW1929 induced Egr-1 bound to specific DNA areas in the alpha7 nAChR gene promoter. Collectively, these results demonstrate that GW1929 not only inhibits but also antagonizes Ach-induced NSCLC cell growth by inhibition of alpha7 nAChR expression through PPARgamma-independent signals that are associated with activation of p38 MPAK and inactivation of PI3-K/mTOR, followed by inducing Egr-1 protein and Egr-1 binding activity in the alpha7 nAChR gene promoter. By downregulation of the alpha7 nAchR, GW1929 blocks cholinergic signaling and inhibits NSCLC cell growth. Title: A unique mono- and diacylglycerol lipase from Penicillium cyclopium: heterologous expression, biochemical characterization and molecular basis for its substrate selectivity Tan ZB, Li JF, Li XT, Gu Y, Wu MC, Wu J, Wang JQ Ref: PLoS ONE, 9:e102040, 2014 : PubMed ESTHER: Tan_2014_PLoS.One_9_e102040 PubMedSearch: Tan 2014 PLoS.One 9 e102040 PubMedID: 25051359 Abstract A cDNA gene encoding a mature peptide of the mono- and diacylglycerol lipase (abbreviated to PcMdl) from Penicillium cyclopium PG37 was cloned and expressed in Pichia pastoris GS115. The recombinant PcMdl (rePcMdl) with an apparent molecular weight of 39 kDa showed the highest activity (40.5 U/mL of culture supernatant) on 1,2-dibutyrin substrate at temperature 35 degrees C and pH 7.5. The rePcMdl was stable at a pH range of 6.5-9.5 and temperatures below 35 degrees C. The activity of rePcMdl was inhibited by Hg2+ and Fe3+, but not significantly affected by EDTA or the other metal ions such as Na+, K+, Li+, Mg2+, Zn2+, Ca2+, Mn2+, Cu2+, and Fe2+. PcMdl was identified to be strictly specific to mono- and diacylglycerol, but not triacylglycerol. Stereographic view of PcMdl docked with substrate (tri- or diacylglycerol) analogue indicated that the residue Phe256 plays an important role in conferring the substrate selectivity. Phe256 projects its side chain towards the substrate binding groove and makes the sn-1 moiety difficult to insert in. Furthermore, sn-1 moiety prevents the phosphorus atom (substitution of carboxyl carbon) from getting to the Ogamma of Ser145, which results in the failure of triacylglycerol hydrolysis. These results should provide a basis for molecular engineering of PcMdl and expand its applications in industries. Title: Complete Genome Sequence of Acinetobacter baumannii ZW85-1 Wang X, Zhang Z, Hao Q, Wu J, Xiao J, Jing H Ref: Genome Announc, 2:e01083, 2014 : PubMed ESTHER: Wang_2014_Genome.Announc_2_e01083 PubMedSearch: Wang 2014 Genome.Announc 2 e01083 PubMedID: 24459253 Gene_locus related to this paper: aciba-a0a009wzt4 Abstract Acinetobacter baumannii is an aerobic, nonmotile Gram-negative bacterium that causes nosocomial infections worldwide. Here, we report the complete genome sequence of Acinetobacter baumannii strain ZW85-1 and its two plasmids. One of the plasmids carries genes for NDM-1, which can hydrolyze a wide range of antibiotics. Title: [(1)(2)(5)I]AT-1012, a new high affinity radioligand for the alpha3beta4 nicotinic acetylcholine receptors Wu J, Perry DC, Bupp JE, Jiang F, Polgar WE, Toll L, Zaveri NT Ref: Neuropharmacology, 77:193, 2014 : PubMed ESTHER: Wu_2014_Neuropharmacol_77_193 PubMedSearch: Wu 2014 Neuropharmacol 77 193 PubMedID: 24095990 Abstract Recent genetic and pharmacological studies have implicated the alpha3, beta4 and alpha5 subunits of the nicotinic acetylcholine receptor (nAChR) in dependence to nicotine and other abused drugs and nicotine withdrawal. The alpha3beta4* nAChR subtype has been shown to co-assemble with the alpha5 or beta3 nAChR subunits, and is found mainly in the autonomic ganglia and select brain regions. It has been difficult to study the alpha3beta4 nAChR because there have been no selective nonpeptidic ligands available to independently examine its pharmacology. We recently reported the synthesis of a [(125)I]-radiolabeled analog of a high affinity, selective small-molecule alpha3beta4 nAChR ligand, AT-1012. We report here the vitro characterization of this radioligand in receptor binding and in vitro autoradiographic studies targeting the alpha3beta4* nAChR. Binding of [(125)I]AT-1012 was characterized at the rat alpha3beta4 and alpha4beta2 nAChR transfected into HEK cells, as well as at the human alpha3beta4alpha5 nAChR in HEK cells. Binding affinity of [(125)I]AT-1012 at the rat alpha3beta4 nAChR was 1.4 nM, with a B(max) of 10.3 pmol/mg protein, similar to what was determined for unlabeled AT-1012 using [(3)H]epibatidine. Saturation isotherms suggested that [(125)I]AT-1012 binds to a single site on the alpha3beta4 nAChR. Similar high binding affinity was also observed for [(125)I]AT-1012 at the human alpha3beta4alpha5 nAChR transfected into HEK cells. [(125)I]AT-1012 did not bind with high affinity to membranes from alpha4beta2 nAChR-transfected HEK cells. Binding studies with [(3)H]epibatidine further confirmed that AT-1012 had over 100-fold binding selectivity for alpha3beta4 over alpha4beta2 nAChR. K(i) values determined for known nAChR compounds using [(125)I]AT-1012 as radioligand were comparable to those obtained with [(3)H]epibatidine. [(125)I]AT-1012 was also used to label alpha3beta4 nAChR in rat brain slices in vitro using autoradiography, which showed highly localized binding of the radioligand in brain regions consistent with the discreet localization of the alpha3beta4 nAChR. We demonstrate that [(125)I]AT-1012 is an excellent tool for labeling the alpha3beta4 nAChR in the presence of other nAChR subtypes. Title: Neuronal nicotinic acetylcholine receptors are important targets for alcohol reward and dependence Wu J, Gao M, Taylor DH Ref: Acta Pharmacol Sin, 35:311, 2014 : PubMed ESTHER: Wu_2014_Acta.Pharmacol.Sin_35_311 PubMedSearch: Wu 2014 Acta.Pharmacol.Sin 35 311 PubMedID: 24464050 Abstract Neuronal nicotinic acetylcholine receptors are important targets for alcohol reward and dependence. Alcoholism is a serious public health problem and has been identified as the third major cause of preventable mortality in the world. Worldwide, about 2 billion people consume alcohol, with 76.3 million having diagnosable alcohol use disorders. Alcohol is currently responsible for the death of 4% of adults worldwide (about 2.5 million deaths each year), and this number will be significantly increased by 2020 unless effective action is taken. Alcohol is the most commonly abused substance by humans. Ethanol (EtOH) is the intoxicating agent in alcoholic drinks that can lead to abuse and dependence. Although it has been extensively studied, the mechanisms of alcohol reward and dependence are still poorly understood. The major reason is that, unlike other addictive drugs (eg, morphine, cocaine or nicotine) that have specific molecular targets, EtOH affects much wider neuronal functions. These functions include phospholipid membranes, various ion channels and receptors, synaptic and network functions, and intracellular signaling molecules. The major targets in the brain that mediate EtOH's effects remain unclear. This knowledge gap results in a therapeutic barrier in the treatment of alcoholism. Interestingly, alcohol and nicotine are often co-abused, which suggests that neuronal nicotinic acetylcholine receptors (nAChRs), the molecular targets for nicotine, may also contribute to alcohol's abusive properties. Here, we briefly summarize recent lines of evidence showing how EtOH modulates nAChRs in the mesolimbic pathway, which provides a perspective that nAChRs are important targets mediating alcohol abuse. Title: In Vitro Metabolism and Drug-Drug Interaction Potential of UTL-5g, a Novel Chemo- and Radioprotective Agent Wu J, Shaw J, Dubaisi S, Valeriote F, Li J Ref: Drug Metabolism & Disposition: The Biological Fate of Chemicals, 42:2058, 2014 : PubMed ESTHER: Wu_2014_Drug.Metab.Dispos_42_2058 PubMedSearch: Wu 2014 Drug.Metab.Dispos 42 2058 PubMedID: 25249693 Abstract N-(2,4-dichlorophenyl)-5-methyl-1,2-oxazole-3-carboxamide (UTL-5g), a potential chemo- and radioprotective agent, acts as a prodrug requiring bioactivation to the active metabolite 5-methylisoxazole-3-carboxylic acid (ISOX). UTL-5g hydrolysis to ISOX and 2,4-dichloroaniline (DCA) has been identified in porcine and rabbit liver esterases. The purpose of this study was to provide insights on the metabolism and drug interaction potential of UTL-5g in humans. The kinetics of UTL-5g hydrolysis was determined in human liver microsomes (HLM) and recombinant human carboxylesterases (hCE1b and hCE2). The potential of UTL-5g and its metabolites for competitive inhibition and time-dependent inhibition of microsomal cytochrome P450 (P450) was examined in HLM. UTL-5g hydrolysis to ISOX and DCA in HLM were NADPH-independent, with a maximum rate of reaction (Vmax) of 11.1 nmol/min per mg and substrate affinity (Km) of 41.6 microM. Both hCE1b and hCE2 effectively catalyzed UTL-5g hydrolysis, but hCE2 exhibited approximately 30-fold higher catalytic efficiency (Vmax/Km) than hCE1b. UTL-5g and DCA competitively inhibited microsomal CYP1A2, CYP2B6, and CYP2C19 (IC50 values <50 microM), and exhibited time-dependent inhibition of microsomal CYP1A2 with the inactivation efficiency (kinact/KI) of 0.68 and 0.51 minute(-1).mM(-1), respectively. ISOX did not inhibit or inactivate any tested microsomal P450. In conclusion, hCE1b and hCE2 play a key role in the bioactivation of UTL-5g. Factors influencing carboxylesterase activities may have a significant impact on the pharmacological and therapeutic effects of UTL-5g. UTL-5g has the potential to inhibit P450-mediated metabolism through competitive inhibition or time-dependent inhibition. Caution is particularly needed for potential drug interactions involving competitive inhibition or time-dependent inhibition of CYP1A2 in the future clinical development of UTL-5g. Title: A natural antisense transcript regulates acetylcholinesterase gene expression via epigenetic modification in Hepatocellular Carcinoma Xi Q, Gao N, Zhang X, Zhang B, Ye W, Wu J Ref: International Journal of Biochemistry & Cell Biology, 55C:242, 2014 : PubMed ESTHER: Xi_2014_Int.J.Biochem.Cell.Biol_55C_242 PubMedSearch: Xi 2014 Int.J.Biochem.Cell.Biol 55C 242 PubMedID: 25240585 Abstract In recent years, widespread antisense transcripts have been identified systematically in mammalian cells and are known to regulate gene expression, although their functional significance remains largely unknown. Previous work has identified that acetylcholinesterase (AChE) is expressed aberrantly in various malignant tumors and function as a tumor growth suppressor. However, the mechanism of AChE gene regulation in tumors remains unclear. In this study, we show that the AChE antisense RNA (AChE-AS) play an important role in AChE expression regulation. An inverse relationship was identified between AChE-AS and AChE expression in hepatocellular carcinoma and hepatoma cells. The silenced AChE-AS corresponds to elevated expression of AChE. Furthermore, we demonstrated that reduced AChE-AS increased H3K4 methylation and decreased H3K9 methylation in the AChE promoter region. As expected, elevated AChE levels induced by inhibition of AChE-AS enhanced anticarcinogen-induced apoptosis. These observations demonstrated that AChE-AS modulates AChE expression and exerts an anti-apoptotic effect through direct repression of AChE expression in HCC cells. Thus, natural antisense RNA may play an important role in AChE regulation via affecting the epigenetic modification in the AChE promoter region. Title: Hsa-miR-132 Regulates Apoptosis in Non-Small Cell Lung Cancer Independent of Acetylcholinesterase Zhang B, Lu L, Zhang X, Ye W, Wu J, Xi Q Ref: Journal of Molecular Neuroscience, 53:335, 2014 : PubMed ESTHER: Zhang_2014_J.Mol.Neurosci_53_335 PubMedSearch: Zhang 2014 J.Mol.Neurosci 53 335 PubMedID: 24158730 Abstract MiR-132 is enriched in the central nerve system and is thought to be involved in neuronal development, maturation and function, and to be associated with several neurological disorders including Alzheimer's disease. In addition to its documented neuronal functions, an emerging role for miR-132 in tumorigenesis has been suggested. Recently, hsa-miR-132 was shown to be modulated in different tumor types. However, its role in non-small cell lung cancer (NSCLC) remains unclear. Here, we show that hsa-miR-132 can initiate apoptosis in NSCLC cells to dramatically attenuate tumor formation in nude mice independent of its effect on the proliferation/apoptosis-associated gene, acetylcholinesterase (AChE). Interestingly, hsa-miR-132 has no pro-apoptotic effect in normal pulmonary trachea epithelium. Taken together, these results suggest that hsa-miR-132 represses NSCLC growth by inducing apoptosis independent of AChE. Title: Cutinase: Characteristics, preparation, and application Chen S, Su L, Chen J, Wu J Ref: Biotechnol Adv, 31:1754, 2013 : PubMed ESTHER: Chen_2013_Biotechnol.Adv_31_1754 PubMedSearch: Chen 2013 Biotechnol.Adv 31 1754 PubMedID: 24055682 Abstract Cutinases (E.C. 3.1.1.74) belong to the alpha/beta-hydrolase superfamily. They were initially discovered because they are secreted by fungi to hydrolyze the ester bonds of the plant polymer cutin. Since then, they have been shown to catalyze the hydrolysis of a variety of polymers, insoluble triacylglycerols, and low-molecular-weight soluble esters. Cutinases are also capable of catalyzing esterification and transesterification reactions. These relatively small, versatile, secreted catalysts have shown promise in a number of industrial applications. This review begins by describing the characteristics of cutinases, pointing out key differences among cutinases, esterases and lipases, and reviewing recent progress in engineering improved cutinases. It continues with a review of the methods used to produce cutinases, with the goal of obtaining sufficient quantities of material for use in industrial processes. Finally, the uses of cutinases in the textile industry are described. The studies presented here demonstrate that the cutinases are poised to become important industrial catalysts, replacing older technologies with more environmentally friendly processes. Title: Poster: Nicotine perturbs PFC-VTA coupling: A new mechanism for nicotine addiction Gao M, Lukas RJ, Wu J Ref: Biochemical Pharmacology, 86:1234, 2013 : PubMed Title: DPP8 and DPP9 expression in cynomolgus monkey and Sprague Dawley rat tissues Harstad EB, Rosenblum JS, Gorrell MD, Achanzar WE, Minimo L, Wu J, Rosini-Marthaler L, Gullo R, Ordway ND and Moyer CF <3 more author(s)> Harstad EB, Rosenblum JS, Gorrell MD, Achanzar WE, Minimo L, Wu J, Rosini-Marthaler L, Gullo R, Ordway ND, Kirby MS, Chadwick KD, Cosma GN, Moyer CF (- 3) Ref: Regul Pept, 186C:26, 2013 : PubMed ESTHER: Harstad_2013_Regul.Pept_186C_26 PubMedSearch: Harstad 2013 Regul.Pept 186C 26 PubMedID: 23850796 Gene_locus related to this paper: rat-dpp9 Abstract Dipeptidyl peptidases (DPPs) are proteolytic enzymes that regulate many physiological systems by degrading signaling peptides. DPP8 and DPP9 are distinct from DPP4 in sequence, cellular localization and expression levels, thus implying distinct functions. However, DPP8 and DPP9 expression needs further delineation. We evaluated DPP4, DPP8 and DPP9 expression using three independent methods at the mRNA, protein, and functional levels to better understand the local physiological contribution of each enzyme. Sprague Dawley rats and cynomolgus monkeys were selected for DPP4, DPP8 and DPP9 expression profiling to represent animal species commonly utilized for drug preclinical safety evaluation. A novel Xhibit assay of DPP protease activity was applied in addition to newly available antibodies for immunohistochemical localization. This combined approach can facilitate a functional evaluation of protease expression, which is important for understanding physiological relevance. Few inter-species differences were observed. Tissue mRNA and protein levels generally correlated to functional DPP4 and DPP8/9 enzymatic activity. All three proteins were seen in epithelial cells, lymphoid cells and some endothelial and vascular smooth muscle cells. Combined DPP8/DPP9 enzymatic activity was uniformly intracellular across tissues at approximately 10-fold lower levels than non-renal DPP4. Consistent levels of each DPP were detected among most non-renal tissues in rats and monkeys. DPP4 was ubiquitous, principally detected on cell membranes of epithelial and endothelial cells and was greatest in the kidney. The expression patterns suggest that DPP8 and DPP9 may act similarly across tissues, and that their actions might in part overlap with DPP4. Title: Identification and characterization of a cold-active phthalate esters hydrolase by screening a metagenomic library derived from biofilms of a wastewater treatment plant Jiao Y, Chen X, Wang X, Liao X, Xiao L, Miao A, Wu J, Yang L Ref: PLoS ONE, 8:e75977, 2013 : PubMed ESTHER: Jiao_2013_PLoS.One_8_e75977 PubMedSearch: Jiao 2013 PLoS.One 8 e75977 PubMedID: 24116085 Gene_locus related to this paper: acilw-ESTA, 9bact-u5qec2 Abstract A cold-active phthalate esters hydrolase gene (designated dphB) was identified through functional screening of a metagenomic library derived from biofilms of a wastewater treatment plant. The enzyme specifically catalyzed the hydrolysis of dipropyl phthalate, dibutyl phthalate, and dipentyl phthalate to the corresponding monoalkyl phthalate esters at low temperatures. The catalytic triad residues of DphB were proposed to be Ser159, Asp251, and His281. Title: Lithium treatment induces proteasomal degradation of over-expressed acetylcholinesterase (AChE-S) and inhibit GSK3beta Jing P, Zhang JY, Ouyang Q, Wu J, Zhang XJ Ref: Chemico-Biological Interactions, 203:309, 2013 : PubMed ESTHER: Jing_2013_Chem.Biol.Interact_203_309 PubMedSearch: Jing 2013 Chem.Biol.Interact 203 309 PubMedID: 22944069 Abstract Lithium is one of the most widely used mood-stabilizing agents for the treatment of bipolar disorder. Lithium is also a potent inhibitor of glycogen synthase kinase-3beta (GSK3beta) activity, which is linked to Alzheimer's disease (AD). In experiments with cultured HEK293T cells, we show here that GSK3beta stabilizes synaptic acetylcholinesterase (AChE-S), a critical component of AD development. Cells treated with lithium exhibited rapid proteasomal degradation of AChE-S. Furthermore treatment of the cells with MG132, an inhibitor of the 26S proteasome, prevented the destabilizing effect of lithium on AChE-S. Taken together, these findings suggest that regulation of AChE-S protein stability may be an important biological target of lithium therapy. Title: Improvement of the Oryza sativa Nipponbare reference genome using next generation sequence and optical map data Kawahara Y, de la Bastide M, Hamilton JP, Kanamori H, McCombie WR, Ouyang S, Schwartz DC, Tanaka T, Wu J and Matsumoto T <12 more author(s)> Kawahara Y, de la Bastide M, Hamilton JP, Kanamori H, McCombie WR, Ouyang S, Schwartz DC, Tanaka T, Wu J, Zhou S, Childs KL, Davidson RM, Lin H, Quesada-Ocampo L, Vaillancourt B, Sakai H, Lee SS, Kim J, Numa H, Itoh T, Buell CR, Matsumoto T (- 12) Ref: Rice (N Y), 6:4, 2013 : PubMed ESTHER: Kawahara_2013_Rice.(N.Y)_6_4 PubMedSearch: Kawahara 2013 Rice.(N.Y) 6 4 PubMedID: 24280374 Gene_locus related to this paper: orysa-Q0JK71, orysj-q6yse8, orysa-q6yzk1, orysa-q7xej4, orysa-q7xem8, orysa-q7xr64, orysj-a0a0p0y6l9, orysj-pla4, orysj-pla1, orysj-q2r2z8 Abstract BACKGROUND: Rice research has been enabled by access to the high quality reference genome sequence generated in 2005 by the International Rice Genome Sequencing Project (IRGSP). To further facilitate genomic-enabled research, we have updated and validated the genome assembly and sequence for the Nipponbare cultivar of Oryza sativa (japonica group). RESULTS: The Nipponbare genome assembly was updated by revising and validating the minimal tiling path of clones with the optical map for rice. Sequencing errors in the revised genome assembly were identified by re-sequencing the genome of two different Nipponbare individuals using the Illumina Genome Analyzer II/IIx platform. A total of 4,886 sequencing errors were identified in 321 Mb of the assembled genome indicating an error rate in the original IRGSP assembly of only 0.15 per 10,000 nucleotides. A small number (five) of insertions/deletions were identified using longer reads generated using the Roche 454 pyrosequencing platform. As the re-sequencing data were generated from two different individuals, we were able to identify a number of allelic differences between the original individual used in the IRGSP effort and the two individuals used in the re-sequencing effort. The revised assembly, termed Os-Nipponbare-Reference-IRGSP-1.0, is now being used in updated releases of the Rice Annotation Project and the Michigan State University Rice Genome Annotation Project, thereby providing a unified set of pseudomolecules for the rice community. CONCLUSIONS: A revised, error-corrected, and validated assembly of the Nipponbare cultivar of rice was generated using optical map data, re-sequencing data, and manual curation that will facilitate on-going and future research in rice. Detection of polymorphisms between three different Nipponbare individuals highlights that allelic differences between individuals should be considered in diversity studies. Title: Human alpha4beta2 nicotinic acetylcholine receptor as a novel target of oligomeric alpha-synuclein Liu Q, Emadi S, Shen JX, Sierks MR, Wu J Ref: PLoS ONE, 8:e55886, 2013 : PubMed ESTHER: Liu_2013_PLoS.One_8_e55886 PubMedSearch: Liu 2013 PLoS.One 8 e55886 PubMedID: 23437071 Abstract Cigarette smoking is associated with a decreased incidence of Parkinson disease (PD) through unknown mechanisms. Interestingly, a decrease in the numbers of alpha4beta2 nicotinic acetylcholine receptors (alpha4beta2-nAChRs) in PD patients suggests an alpha4beta2-nAChR-mediated cholinergic deficit in PD. Although oligomeric forms of alpha-synuclein have been recognized to be toxic and involved in the pathogenesis of PD, their direct effects on nAChR-mediated cholinergic signaling remains undefined. Here, we report for the first time that oligomeric alpha-synuclein selectively inhibits human alpha4beta2-nAChR-mediated currents in a dose-dependent, non-competitive and use-independent manner. We show that pre-loading cells with guanyl-5'-yl thiophosphate fails to prevent this inhibition, suggesting that the alpha-synuclein-induced inhibition of alpha4beta2-nAChR function is not mediated by nAChR internalization. By using a pharmacological approach and cultures expressing transfected human nAChRs, we have shown a clear effect of oligomeric alpha-synuclein on alpha4beta2-nAChRs, but not on alpha4beta4- or alpha7-nAChRs, suggesting nAChR subunit selectivity of oligomeric alpha-synuclein-induced inhibition. In addition, by combining the size exclusion chromatography and atomic force microscopy (AFM) analyses, we find that only large (>4 nm) oligomeric alpha-synuclein aggregates (but not monomeric, small oligomeric or fibrillar alpha-synuclein aggregates) exhibit the inhibitory effect on human alpha4beta2-nAChRs. Collectively, we have provided direct evidence that alpha4beta2-nAChR is a sensitive target to mediate oligomeric alpha-synuclein-induced modulation of cholinergic signaling, and our data imply that therapeutic strategies targeted toward alpha4beta2-nAChRs may have potential for developing new treatments for PD. Title: Synaptic acetylcholinesterase targeted by microRNA-212 functions as a tumor suppressor in non-small cell lung cancer Lu L, Zhang X, Zhang B, Wu J Ref: International Journal of Biochemistry & Cell Biology, 45:2530, 2013 : PubMed ESTHER: Lu_2013_Int.J.Biochem.Cell.Biol_45_2530 PubMedSearch: Lu 2013 Int.J.Biochem.Cell.Biol 45 2530 PubMedID: 23974008 Abstract Acetylcholinesterase expression is modulated in various types of tumor, which suggests it is associated with tumor development; however, the mechanism of acetylcholinesterase gene regulation in tumors remains unclear. Here, we report that acetylcholinesterase is aberrantly expressed in non-small cell lung cancer and is an evolutionarily conserved functional target of miR-212. Acetylcholinesterase expression was negatively regulated by miR-212 in vitro and was inversely correlated with miR-212 expression in vivo. In addition, acetylcholinesterase levels were increased, and miR-212 levels decreased, in non-small cell lung cancer cells during cisplatin-induced apoptosis. We further determined that acetylcholinesterase acted as a pro-apoptotic gene in non-small cell lung cells; and attenuated the growth of xenografts in nude mice when upregulated. In contrast, elevated miR-212 levels preserved the protective effect of acetylcholinesterase silencing by RNA interference against cisplatin-induced apoptosis, whereas restoration of miR-212-resistant synaptic acetylcholinesterase expression inhibited the miR-212 anti-apoptotic function. The results demonstrated that miR-212 exerted an anti-apoptotic effect through direct repression of synaptic acetylcholinesterase expression in non-small cell lung cancer cells. Taken together, our study revealed that synaptic acetylcholinesterase may be a tumor suppressor and is modulated by miR-212 in non-small cell lung cancer. Title: Genome of the long-living sacred lotus (Nelumbo nucifera Gaertn.) Ming R, VanBuren R, Liu Y, Yang M, Han Y, Li LT, Zhang Q, Kim MJ, Schatz MC and Shen-Miller J <60 more author(s)> Ming R, VanBuren R, Liu Y, Yang M, Han Y, Li LT, Zhang Q, Kim MJ, Schatz MC, Campbell M, Li J, Bowers JE, Tang H, Lyons E, Ferguson AA, Narzisi G, Nelson DR, Blaby-Haas CE, Gschwend AR, Jiao Y, Der JP, Zeng F, Han J, Min XJ, Hudson KA, Singh R, Grennan AK, Karpowicz SJ, Watling JR, Ito K, Robinson SA, Hudson ME, Yu Q, Mockler TC, Carroll A, Zheng Y, Sunkar R, Jia R, Chen N, Arro J, Wai CM, Wafula E, Spence A, Xu L, Zhang J, Peery R, Haus MJ, Xiong W, Walsh JA, Wu J, Wang ML, Zhu YJ, Paull RE, Britt AB, Du C, Downie SR, Schuler MA, Michael TP, Long SP, Ort DR, Schopf JW, Gang DR, Jiang N, Yandell M, dePamphilis CW, Merchant SS, Paterson AH, Buchanan BB, Li S, Shen-Miller J (- 60) Ref: Genome Biol, 14:R41, 2013 : PubMed ESTHER: Ming_2013_Genome.Biol_14_R41 PubMedSearch: Ming 2013 Genome.Biol 14 R41 PubMedID: 23663246 Gene_locus related to this paper: nelnu-a0a1u8aj84, nelnu-a0a1u8bpe4, nelnu-a0a1u7z9m9, nelnu-a0a1u7ywy5, nelnu-a0a1u8aik2, nelnu-a0a1u7zmb5, nelnu-a0a1u8a7m7, nelnu-a0a1u8b0n9, nelnu-a0a1u8b461, nelnu-a0a1u7zzj3, nelnu-a0a1u8ave7, nelnu-a0a1u7yn26 Abstract BACKGROUND: Sacred lotus is a basal eudicot with agricultural, medicinal, cultural and religious importance. It was domesticated in Asia about 7,000 years ago, and cultivated for its rhizomes and seeds as a food crop. It is particularly noted for its 1,300-year seed longevity and exceptional water repellency, known as the lotus effect. The latter property is due to the nanoscopic closely packed protuberances of its self-cleaning leaf surface, which have been adapted for the manufacture of a self-cleaning industrial paint, Lotusan. RESULTS: The genome of the China Antique variety of the sacred lotus was sequenced with Illumina and 454 technologies, at respective depths of 101x and 5.2x. The final assembly has a contig N50 of 38.8 kbp and a scaffold N50 of 3.4 Mbp, and covers 86.5% of the estimated 929 Mbp total genome size. The genome notably lacks the paleo-triplication observed in other eudicots, but reveals a lineage-specific duplication. The genome has evidence of slow evolution, with a 30% slower nucleotide mutation rate than observed in grape. Comparisons of the available sequenced genomes suggest a minimum gene set for vascular plants of 4,223 genes. Strikingly, the sacred lotus has 16 COG2132 multi-copper oxidase family proteins with root-specific expression; these are involved in root meristem phosphate starvation, reflecting adaptation to limited nutrient availability in an aquatic environment. CONCLUSIONS: The slow nucleotide substitution rate makes the sacred lotus a better resource than the current standard, grape, for reconstructing the pan-eudicot genome, and should therefore accelerate comparative analysis between eudicots and monocots. Title: Fusion of binding domains to Thermobifida cellulosilytica cutinase to tune sorption characteristics and enhancing PET hydrolysis Ribitsch D, Yebra AO, Zitzenbacher S, Wu J, Nowitsch S, Steinkellner G, Greimel K, Doliska A, Oberdorfer G and Guebitz GM <5 more author(s)> Ribitsch D, Yebra AO, Zitzenbacher S, Wu J, Nowitsch S, Steinkellner G, Greimel K, Doliska A, Oberdorfer G, Gruber CC, Gruber K, Schwab H, Stana-Kleinschek K, Herrero Acero E, Guebitz GM (- 5) Ref: Biomacromolecules, 14:1769, 2013 : PubMed ESTHER: Ribitsch_2013_Biomacromolecules_14_1769 PubMedSearch: Ribitsch 2013 Biomacromolecules 14 1769 PubMedID: 23718548 Gene_locus related to this paper: thefu-q6a0i4 Abstract A cutinase from Thermomyces cellullosylitica (Thc_Cut1), hydrolyzing the synthetic polymer polyethylene terephthalate (PET), was fused with two different binding modules to improve sorption and thereby hydrolysis. The binding modules were from cellobiohydrolase I from Hypocrea jecorina (CBM) and from a polyhydroxyalkanoate depolymerase from Alcaligenes faecalis (PBM). Although both binding modules have a hydrophobic nature, it was possible to express the proteins in E. coli . Both fusion enzymes and the native one had comparable kcat values in the range of 311 to 342 s(-1) on pNP-butyrate, while the catalytic efficiencies kcat/Km decreased from 0.41 s(-1)/ muM (native enzyme) to 0.21 and 0.33 s(-1)/muM for Thc_Cut1+PBM and Thc_Cut1+CBM, respectively. The fusion enzymes were active both on the insoluble PET model substrate bis(benzoyloxyethyl) terephthalate (3PET) and on PET although the hydrolysis pattern was differed when compared to Thc_Cut1. Enhanced adsorption of the fusion enzymes was visible by chemiluminescence after incubation with a 6xHisTag specific horseradish peroxidase (HRP) labeled probe. Increased adsorption to PET by the fusion enzymes was confirmed with Quarz Crystal Microbalance (QCM-D) analysis and indeed resulted in enhanced hydrolysis activity (3.8x for Thc_Cut1+CBM) on PET, as quantified, based on released mono/oligomers. Title: Extracellular location of Thermobifida fusca cutinase expressed in Escherichia coli BL21(DE3) without mediation of a signal peptide Su L, Woodard RW, Chen J, Wu J Ref: Applied Environmental Microbiology, 79:4192, 2013 : PubMed ESTHER: Su_2013_Appl.Environ.Microbiol_79_4192 PubMedSearch: Su 2013 Appl.Environ.Microbiol 79 4192 PubMedID: 23603671 Abstract Cutinase is a multifunctional esterase with potential industrial applications. In the present study, a truncated version of the extracellular Thermobifida fusca cutinase without a signal peptide (referred to as cutinase(NS)) was heterologously expressed in Escherichia coli BL21(DE3). The results showed that the majority of the cutinase activity was located in the culture medium. In a 3-liter fermentor, the cutinase activity in the culture medium reached 1,063.5 U/ml (2,380.8 mg/liter), and the productivity was 40.9 U/ml/h. Biochemical characterization of the purified cutinase(NS) showed that it has enzymatic properties similar to those of the wild-type enzyme. In addition, E. coli cells producing inactive cutinase(NS)S130A were constructed, and it was found that the majority of the inactive enzyme was located in the cytoplasm. Furthermore, T. fusca cutinase was confirmed to have hydrolytic activity toward phospholipids, an important component of the cell membrane. Compared to the cells expressing the inactive cutinase(NS)S130A, the cells expressing cutinase(NS) showed increased membrane permeability and irregular morphology. Based on these results, a hypothesis of "cell leakage induced by the limited phospholipid hydrolysis of cutinase(NS)" was proposed to explain the underlying mechanism for the extracellular release of cutinase(NS). Title: A novel strategy for enhancing extracellular secretion of recombinant proteins in Escherichia coli Su L, Xu C, Woodard RW, Chen J, Wu J Ref: Applied Microbiology & Biotechnology, 97:6705, 2013 : PubMed ESTHER: Su_2013_Appl.Microbiol.Biotechnol_97_6705 PubMedSearch: Su 2013 Appl.Microbiol.Biotechnol 97 6705 PubMedID: 23722267 Abstract Secretion of cytoplasmic expressed proteins into culture medium has significant commercial advantages in large-scale production of proteins. Our previous study demonstrated that the membrane permeability of Escherichia coli could be significantly improved when Thermobifida fusca cutinase, without a signal peptide, was expressed in cytoplasm. This study investigated the extracellular production of other recombinant proteins, including both secretory and cytosolic proteins, with co-expression of cutinase. When the secretory enzymes, xylanase and alpha-amylase, were co-expressed with cutinase, the culture period was shortened by half, and the productivity was 7.9 and 2.0-fold to that of their individual control without co-expression, respectively. When the normally cytosolic proteins, xylose isomerase and trehalose synthase, were co-expressed with cutinase, more than half of the target proteins were "secreted" into the culture medium. Moreover, by using beta-galactosidase to detect membrane leakage, the improved secretion of the above model proteins was confirmed not to be due to cell lysis. The study provides a novel strategy for enhancing extracellular secretion of recombinant proteins in E. coli. Title: Cortical control of VTA function and influence on nicotine reward Wu J, Gao M, Shen JX, Shi WX, Oster AM, Gutkin BS Ref: Biochemical Pharmacology, 86:1173, 2013 : PubMed ESTHER: Wu_2013_Biochem.Pharmacol_86(8)_1173 PubMedSearch: Wu 2013 Biochem.Pharmacol 86(8) 1173 PubMedID: 23933294 Abstract Tobacco use is a major public health problem. Nicotine acts on widely distributed nicotinic acetylcholine receptors (nAChRs) in the brain and excites dopamine (DA) neurons in the ventral tegmental area (VTA). The elicited increase of DA neuronal activity is thought to be an important mechanism for nicotine reward and subsequently the transition to addiction. However, the current understanding of nicotine reward is based predominantly on the data accumulated from in vitro studies, often from VTA slices. Isolated VTA slices artificially terminate communications between neurons in the VTA and other brain regions that may significantly alter nicotinic effects. Consequently, the mechanisms of nicotinic excitation of VTA DA neurons under in vivo conditions have received only limited attention. Building upon the existing knowledge acquired in vitro, it is now time to elucidate the integrated mechanisms of nicotinic reward on intact systems that are more relevant to understanding the action of nicotine or other addictive drugs. In this review, we summarize recent studies that demonstrate the impact of prefrontal cortex (PFC) on the modulation of VTA DA neuronal function and nicotine reward. Based on existing evidence, we propose a new hypothesis that PFC-VTA functional coupling serves as an integration mechanism for nicotine reward. Moreover, addiction may develop due to nicotine perturbing the PFC-VTA coupling and thereby eliminating the PFC-dependent cognitive control over behavior. Title: Cloning of a dibutyl phthalate hydrolase gene from Acinetobacter sp. strain M673 and functional analysis of its expression product in Escherichia coli Wu J, Liao X, Yu F, Wei Z, Yang L Ref: Applied Microbiology & Biotechnology, 97:2483, 2013 : PubMed ESTHER: Wu_2013_Appl.Microbiol.Biotechnol_97_2483 PubMedSearch: Wu 2013 Appl.Microbiol.Biotechnol 97 2483 PubMedID: 22729233 Gene_locus related to this paper: acilw-ESTA Abstract A dibutyl phthalate (DBP) transforming bacterium, strain M673, was isolated and identified as Acinetobacter sp. This strain could not grow on dialkyl phthalates, including dimethyl, diethyl, dipropyl, dibutyl, dipentyl, dihexyl, di(2-ethylhexyl), di-n-octyl, and dinonyl phthalate, but suspensions of cells could transform these compounds to phthalate via corresponding monoalkyl phthalates. During growth in Luria-Bertani medium, M673 produced the high amounts of non-DBP-induced intracellular hydrolase in the stationary phase. One DBP hydrolase gene containing an open reading frame of 1,095 bp was screened from a genomic library, and its expression product hydrolyzed various dialkyl phthalates to the corresponding monoalkyl phthalates. Title: Acetylcholinesterase deficiency decreases apoptosis in dopaminergic neurons in the neurotoxin model of Parkinson's disease Zhang X, Lu L, Liu S, Ye W, Wu J Ref: International Journal of Biochemistry & Cell Biology, 45:265, 2013 : PubMed ESTHER: Zhang_2013_Int.J.Biochem.Cell.Biol_45_265 PubMedSearch: Zhang 2013 Int.J.Biochem.Cell.Biol 45 265 PubMedID: 23201480 Abstract The apoptosis pathway has been proposed to be involved in causing neuronal cell death in the pathogenesis of Parkinson's disease. However, the details of this pathway are poorly understood. Previous research has shown increased acetylcholinesterase expression during apoptosis in various cell types, which suggests that acetylcholinesterase has a potential role in neuronal cell death. In this study, we found that acetylcholinesterase protein expression increased and caspase-3 was activated in PC12 cells treated with 1-methyl-4-phenylpyridinium. Furthermore, the genetic or pharmacological inhibition of acetylcholinesterase was shown to protect PC12 cells from MPP+ induced apoptotic cell death. To study the function of acetylcholinesterase as a mechanism of neuronal cell death in vivo, we subsequently established a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine Parkinson's disease mouse model utilizing acetylcholinesterase-deficient mice. Studies in these mice revealed reduced dopaminergic neuron loss and lower expression levels of apoptotic proteins in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated heterozygous mice compared to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated wild-type mice. We conclude that it is highly probable that acetylcholinesterase is involved in the pathogenesis of the neurotoxin model of Parkinson's disease via apoptosis. Specifically, a deficiency or inhibition of acetylcholinesterase can decrease apoptosis and protect dopaminergic neurons in the neurotoxin model of Parkinson's disease. Title: Enhanced activity toward PET by site-directed mutagenesis of Thermobifida fusca cutinase-CBM fusion protein Zhang Y, Wang L, Chen J, Wu J Ref: Carbohydr Polym, 97:124, 2013 : PubMed ESTHER: Zhang_2013_Carbohydr.Polym_97_124 PubMedSearch: Zhang 2013 Carbohydr.Polym 97 124 PubMedID: 23769527 Abstract In the present study, cutinase-CBMCenA fusion protein was genetically modified in the carbohydrate-binding module (CBM) binding sites, by site-directed mutagenesis, to enhance its activity toward polyethylene terephthalate (PET) fiber. The effects of tryptophan at particular positions of CBMCenA on the binding and hydrolysis of polyester substrate were investigated by replacing each of Trp14, Trp50 and Trp68 with leucine or tyrosine, respectively. All the mutants were expressed in Escherichia coli and purified to homogeneity. Enzyme characterization showed that the mutants displayed similar thermostability and pH stabilities in response to the native enzyme. Furthermore, W68L and W68Y, among all the mutants, exhibited significant improvement in binding and catalytic efficiency (1.4-1.5 fold) toward PET fiber when compared to that of the native enzyme. The enhanced binding and hydrolytic activity might be a result of creating new hydrogen bond or hydrophobic interaction between the enzyme and PET fiber. Title: Functional alpha7beta2 nicotinic acetylcholine receptors expressed in hippocampal interneurons exhibit high sensitivity to pathological level of amyloid beta peptides Liu Q, Huang Y, Shen J, Steffensen S, Wu J Ref: BMC Neurosci, 13:155, 2012 : PubMed ESTHER: Liu_2012_BMC.Neurosci_13_155 PubMedSearch: Liu 2012 BMC.Neurosci 13 155 PubMedID: 23272676 Abstract BACKGROUND: beta-amyloid (Abeta) accumulation is described as a hallmark of Alzheimer's disease (AD). Abeta perturbs a number of synaptic components including nicotinic acetylcholine receptors containing alpha7 subunits (alpha7-nAChRs), which are abundantly expressed in the hippocampus and found on GABAergic interneurons. We have previously demonstrated the existence of a novel, heteromeric alpha7beta2-nAChR in basal forebrain cholinergic neurons that exhibits high sensitivity to acute Abeta exposure. To extend our previous work, we evaluated the expression and pharmacology of alpha7beta2-nAChRs in hippocampal interneurons and their sensitivity to Abeta. Title: alpha7 nicotinic acetylcholine receptor-mediated neuroprotection against dopaminergic neuron loss in an MPTP mouse model via inhibition of astrocyte activation Liu Y, Hu J, Wu J, Zhu C, Hui Y, Han Y, Huang Z, Ellsworth K, Fan W Ref: J Neuroinflammation, 9:98, 2012 : PubMed ESTHER: Liu_2012_J.Neuroinflammation_9_98 PubMedSearch: Liu 2012 J.Neuroinflammation 9 98 PubMedID: 22624500 Abstract BACKGROUND: Although evidence suggests that the prevalence of Parkinson's disease (PD) is lower in smokers than in non-smokers, the mechanisms of nicotine-induced neuroprotection remain unclear. Stimulation of the alpha7 nicotinic acetylcholine receptor (alpha7-nAChR) seems to be a crucial mechanism underlying the anti-inflammatory potential of cholinergic agonists in immune cells, including astrocytes, and inhibition of astrocyte activation has been proposed as a novel strategy for the treatment of neurodegenerative disorders such as PD. The objective of the present study was to determine whether nicotine-induced neuroprotection in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model occurs via alpha7-nAChR-mediated inhibition of astrocytes. Title: alpha7beta2 nicotinic acetylcholine receptors assemble, function, and are activated primarily via their alpha7-alpha7 interfaces Murray TA, Bertrand D, Papke RL, George AA, Pantoja R, Srinivasan R, Liu Q, Wu J, Whiteaker P and Lukas RJ <1 more author(s)> Murray TA, Bertrand D, Papke RL, George AA, Pantoja R, Srinivasan R, Liu Q, Wu J, Whiteaker P, Lester HA, Lukas RJ (- 1) Ref: Molecular Pharmacology, 81:175, 2012 : PubMed ESTHER: Murray_2012_Mol.Pharmacol_81_175 PubMedSearch: Murray 2012 Mol.Pharmacol 81 175 PubMedID: 22039094 Abstract We investigated assembly and function of nicotinic acetylcholine receptors (nAChRs) composed of alpha7 and beta2 subunits. We measured optical and electrophysiological properties of wild-type and mutant subunits expressed in cell lines and Xenopus laevis oocytes. Laser scanning confocal microscopy indicated that fluorescently tagged alpha7 and beta2 subunits colocalize. Forster resonance energy transfer between fluorescently tagged subunits strongly suggested that alpha7 and beta2 subunits coassemble. Total internal reflection fluorescence microscopy revealed that assemblies localized to filopodia-like processes of SH-EP1 cells. Gain-of-function alpha7 and beta2 subunits confirmed that these subunits coassemble within functional receptors. Moreover, alpha7beta2 nAChRs composed of wild-type subunits or fluorescently tagged subunits had pharmacological properties similar to those of alpha7 nAChRs, although amplitudes of alpha7beta2 nAChR-mediated, agonist-evoked currents were generally ~2-fold lower than those for alpha7 nAChRs. It is noteworthy that alpha7beta2 nAChRs displayed sensitivity to low concentrations of the antagonist dihydro-beta-erythroidine that was not observed for alpha7 nAChRs at comparable concentrations. In addition, cysteine mutants revealed that the alpha7-beta2 subunit interface does not bind ligand in a functionally productive manner, partly explaining lower alpha7beta2 nAChR current amplitudes and challenges in identifying the function of native alpha7beta2 nAChRs. On the basis of our findings, we have constructed a model predicting receptor function that is based on stoichiometry and position of beta2 subunits within the alpha7beta2 nAChRs. Title: Synthesis and structure-activity relationship of (1-halo-2-naphthyl) carbamate-based inhibitors of KIAA1363 (NCEH1/AADACL1) Shreder KR, Lin EC, Wu J, Cajica J, Amantea CM, Hu Y, Okerberg E, Brown HE, Pham LM and Kozarich JW <4 more author(s)> Shreder KR, Lin EC, Wu J, Cajica J, Amantea CM, Hu Y, Okerberg E, Brown HE, Pham LM, Chung de M, Fraser AS, McGee E, Rosenblum JS, Kozarich JW (- 4) Ref: Bioorganic & Medicinal Chemistry Lett, 22:5748, 2012 : PubMed ESTHER: Shreder_2012_Bioorg.Med.Chem.Lett_22_5748 PubMedSearch: Shreder 2012 Bioorg.Med.Chem.Lett 22 5748 PubMedID: 22877630 Gene_locus related to this paper: human-NCEH1 Abstract KIAA1363 is a serine hydrolase whose activity has been shown to be positively associated with tumor cell invasiveness. Thus, inhibitors of KIAA1363 represent a novel targeted therapy approach towards cancer. AX11890 ((1-bromo-2-naphthyl) N,N-dimethylcarbamate) was identified as a KIAA1363 inhibitor with an IC(50) value of 1.2 muM and was shown using ESI-MS to carbamylate the catalytic residue Ser(191). SAR studies explored both substitution of the 1-bromo group and derivatization of the 6-position. Activity-based protein profiling demonstrated AX13057 inhibited tumor-localized KIAA1363 in SK-OV-3 xenograft-bearing mice. Title: Molecular recognition of esterase plays a major role on the removal of fatty soils during detergency Silva C, Azoia NG, Martins M, Matama T, Wu J, Cavaco-Paulo A Ref: J Biotechnol, 161:228, 2012 : PubMed ESTHER: Silva_2012_J.Biotechnol_161_228 PubMedSearch: Silva 2012 J.Biotechnol 161 228 PubMedID: 22750090 Abstract In this work it is describe for the first time the use of an esterase with null activity Tfu_0883 bacterial cutinase from Thermobifida fusca on the removal of fat from the surface of a cotton substrate Similar levels of fat removal were found for both null and wild-type proteins despite that only wild type protein yielded fatty acids Our results show that molecular recognition of esterase plays a major role on the removal of fatty soils allowing important guidelines for the design of detergent enzymes Furthermore the advantage of using null esterase enzymes lies in the avoidance of the rancid smell of short chained fatty acids typical after esterase treatment. Title: Extracellular overexpression of recombinant Thermobifida fusca cutinase by alpha-hemolysin secretion system in E. coli BL21(DE3) Su L, Chen S, Yi L, Woodard RW, Chen J, Wu J Ref: Microb Cell Fact, 11:8, 2012 : PubMed ESTHER: Su_2012_Microb.Cell.Fact_11_8 PubMedSearch: Su 2012 Microb.Cell.Fact 11 8 PubMedID: 22239833 Abstract BACKGROUND: Extracellular expression of proteins has an absolute advantage in a large-scale industrial production. In our previous study, Thermobifida fusca cutinase, an enzyme mainly utilized in textile industry, was expressed via type II secretory system in Escherichia coli BL21(DE3), and it was found that parts of the expressed protein was accumulated in the periplasmic space. Due to the fact that alpha-hemolysin secretion system can export target proteins directly from cytoplasm across both cell membrane of E. coli to the culture medium, thus in the present study we investigated the expression of cutinase using this alpha-hemolysin secretion system. RESULTS: T. fusca cutinase was fused with the specific signal peptide of alpha-hemolysin scretion system and expressed in E. coli BL21(DE3). In addition, HlyB and HlyD, strain-specific translocation components of alpha-hemolysin secretion system, were coexpressed to facilitate the enzyme expression. The cultivation of this engineered cell showed that cutinase activity in the culture medium reached 334 U/ml, which is 2.5 times that from type II secretion pathway under the same culture condition. The recombinant cutinase was further purified. Biochemical characterization of purified enzyme, which had an alpha-hemolysin secretion pathway signal peptide attached, had substrate specificity, pH and temperature profile, as well as application capability in bioscouring similar to that of wild-type cutinase. CONCLUSIONS: In the present study, T. fusca cutinase was successfully secreted to the culture media by alpha-hemolysin secretion system. This is the first report of cutinase being efficiently secreted by this pathway. Due to the limited cases of successful expression of industrial enzyme by E. coli alpha-hemolysin secretion system, our study further explored the utilization of this pathway in industrial enzymes. Title: Inactivation of chloramphenicol and florfenicol by a novel chloramphenicol hydrolase Tao W, Lee MH, Wu J, Kim NH, Kim JC, Chung E, Hwang EC, Lee SW Ref: Applied Environmental Microbiology, 78:6295, 2012 : PubMed ESTHER: Tao_2012_Appl.Environ.Microbiol_78_6295 PubMedSearch: Tao 2012 Appl.Environ.Microbiol 78 6295 PubMedID: 22752166 Gene_locus related to this paper: 9bact-g3cr00, 9bact-g3cr02 Abstract Chloramphenicol and florfenicol are broad-spectrum antibiotics. Although the bacterial resistance mechanisms to these antibiotics have been well documented, hydrolysis of these antibiotics has not been reported in detail. This study reports the hydrolysis of these two antibiotics by a specific hydrolase that is encoded by a gene identified from a soil metagenome. Hydrolysis of chloramphenicol has been recognized in cell extracts of Escherichia coli expressing a chloramphenicol acetate esterase gene, estDL136. A hydrolysate of chloramphenicol was identified as p-nitrophenylserinol by liquid chromatography-mass spectroscopy and proton nuclear magnetic resonance spectroscopy. The hydrolysis of these antibiotics suggested a promiscuous amidase activity of EstDL136. When estDL136 was expressed in E. coli, EstDL136 conferred resistance to both chloramphenicol and florfenicol on E. coli, due to their inactivation. In addition, E. coli carrying estDL136 deactivated florfenicol faster than it deactivated chloramphenicol, suggesting that EstDL136 hydrolyzes florfenicol more efficiently than it hydrolyzes chloramphenicol. The nucleotide sequences flanking estDL136 encode proteins such as amidohydrolase, dehydrogenase/reductase, major facilitator transporter, esterase, and oxidase. The most closely related genes are found in the bacterial family Sphingomonadaceae, which contains many bioremediation-related strains. Whether the gene cluster with estDL136 in E. coli is involved in further chloramphenicol degradation was not clear in this study. While acetyltransferases for chloramphenicol resistance and drug exporters for chloramphenicol or florfenicol resistance are often detected in numerous microbes, this is the first report of enzymatic hydrolysis of florfenicol resulting in inactivation of the antibiotic. Title: Residues and dissipation dynamics of fosthiazate in tomato and soil Wu J, Wang K, Zhang H Ref: Bulletin of Environmental Contamination & Toxicology, 89:664, 2012 : PubMed ESTHER: Wu_2012_Bull.Environ.Contam.Toxicol_89_664 PubMedSearch: Wu 2012 Bull.Environ.Contam.Toxicol 89 664 PubMedID: 22801926 Abstract Residue dynamics of fosthiazate in tomato and soil was studied in this paper utilizing liquid chromatography with tandem mass spectrometry (LC-MS/MS). The field trial was conducted in three sites: Beijing, Liaoning, Hubei in China. Fosthiazate dissipated with the half-life 0.75-2.6 days in tomato or tomato plants and 2.5-11.6 days in soil. In the terminal residue experiment, no higher residue than 0.023 mg kg(-1) in tomato and 0.27 mg kg(-1) in soil was detected. Residues of fosthiazte in tomato were far below Japan maximum residue levels (0.2 mg kg(-1)). Title: Impact of prefrontal cortex in nicotine-induced excitation of ventral tegmental area dopamine neurons in anesthetized rats Zhang D, Gao M, Xu D, Shi WX, Gutkin BS, Steffensen SC, Lukas RJ, Wu J Ref: Journal of Neuroscience, 32:12366, 2012 : PubMed ESTHER: Zhang_2012_J.Neurosci_32_12366 PubMedSearch: Zhang 2012 J.Neurosci 32 12366 PubMedID: 22956827 Abstract Systemic administration of nicotine increases dopaminergic (DA) neuron firing in the ventral tegmental area (VTA), which is thought to underlie nicotine reward. Here, we report that the medial prefrontal cortex (mPFC) plays a critical role in nicotine-induced excitation of VTA DA neurons. In chloral hydrate-anesthetized rats, extracellular single-unit recordings showed that VTA DA neurons exhibited two types of firing responses to systemic nicotine. After nicotine injection, the neurons with type-I response showed a biphasic early inhibition and later excitation, whereas the neurons with type-II response showed a monophasic excitation. The neurons with type-I, but not type-II, response exhibited pronounced slow oscillations (SOs) in firing. Pharmacological or structural mPFC inactivation abolished SOs and prevented systemic nicotine-induced excitation in the neurons with type-I, but not type-II, response, suggesting that these VTA DA neurons are functionally coupled to the mPFC and nicotine increases firing rate in these neurons in part through the mPFC. Systemic nicotine also increased the firing rate and SOs in mPFC pyramidal neurons. mPFC infusion of a non-alpha7 nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine blocked the excitatory effect of systemic nicotine on the VTA DA neurons with type-I response, but mPFC infusion of nicotine failed to excite these neurons. These results suggest that nAChR activation in the mPFC is necessary, but not sufficient, for systemic nicotine-induced excitation of VTA neurons. Finally, systemic injection of bicuculline prevented nicotine-induced firing alterations in the neurons with type-I response. We propose that the mPFC plays a critical role in systemic nicotine-induced excitation of VTA DA neurons. Title: Acetylcholinesterase is associated with apoptosis in beta cells and contributes to insulin-dependent diabetes mellitus pathogenesis Zhang B, Yang L, Yu L, Lin B, Hou Y, Wu J, Huang Q, Han Y, Guo L and Zhang X <2 more author(s)> Zhang B, Yang L, Yu L, Lin B, Hou Y, Wu J, Huang Q, Han Y, Guo L, Ouyang Q, Lu L, Zhang X (- 2) Ref: Acta Biochim Biophys Sin (Shanghai), 44:207, 2012 : PubMed ESTHER: Zhang_2012_Acta.Biochim.Biophys.Sin.(Shanghai)_44_207 PubMedSearch: Zhang 2012 Acta.Biochim.Biophys.Sin.(Shanghai) 44 207 PubMedID: 22236578 Abstract Acetylcholinesterase (AChE) expression is pivotal during apoptosis. Indeed, AChE inhibitors partially protect cells from apoptosis. Insulin-dependent diabetes mellitus (IDDM) is characterized in part by pancreatic beta-cell apoptosis. Here, we investigated the role of AChE in the development of IDDM and analyzed protective effects of AChE inhibitors. Multiple low-dose streptozotocin (MLD-STZ) administration resulted in IDDM in a mouse model. Western blot analysis, cytochemical staining, and immunofluorescence staining were used to detect AChE expression in MIN6 cells, primary beta cells, and apoptotic pancreatic beta cells of MLD-STZ-treated mice. AChE inhibitors were administered intraperitoneally to the MLD-STZ mice for 30 days. Blood glucose, plasma insulin, and creatine levels were measured, and glucose tolerance tests were performed. The effects of AChE inhibitors on MIN6 cells were also evaluated. AChE expression was induced in the apoptotic MIN6 cells and primary beta cells in vitro and pancreatic islets in vivo when treated with STZ. Induction and progressive accumulation of AChE in the pancreatic islets were associated with apoptotic beta cells during IDDM development. The administration of AChE inhibitors effectively decreased hyperglycemia and incidence of diabetes, and restored plasma insulin levels and plasma creatine clearance in the MLD-STZ mice. AChE inhibitors partially protected MIN6 cells from the damage caused by STZ treatment. Induction and accumulation of AChE in pancreatic islets and the protective effects of AChE inhibitors on the onset and development of IDDM indicate a close relationship between AChE and IDDM. Title: [Construction of Aspergillus niger lipase mutants with oil-water interface independence] Chen D, Shu Z, Xue L, Lin R, Wu J, Jiang Y, Li X, Lin Y, Huang J Ref: Sheng Wu Gong Cheng Xue Bao, 27:860, 2011 : PubMed ESTHER: Chen_2011_Sheng.Wu.Gong.Cheng.Xue.Bao_27_860 PubMedSearch: Chen 2011 Sheng.Wu.Gong.Cheng.Xue.Bao 27 860 PubMedID: 22034814 Abstract Based on previous bioinformational analysis results, two Aspergillus niger lipase (ANL) mutants, ANL-Ser84Gly and ANL-Asp99Pro were constructed to screen ANL mutants with oil-water interface independence. ANL-Ser84Gly still displayed a pronounced interfacial activation, while ANL-Asp99Pro displayed no interfacial activation. The specific activity of ANL-Ser84Gly towards p-nitrophenyl palmitate (-myristate, -laurate and -decanoate) decreased by 29.8% (53.1, 60.1 and 77.1, respectively) than that of ANL, while the specific activity of ANL-Asp99Pro towards p-nitrophenyl palmitate increased by 2.2-fold. The mutation in the hinge region at both sides of the lid domain also destabilized various secondary structure factors of ANL-S84G and ANL-D99P, which resulted in a substantial decrease in thermostability. The achievement to construct oil-water interface-independent ANL mutants would help to further understand lipase interfacial activation mechanism. Title: Study on improvement of extracellular production of recombinant Thermobifida fusca cutinase by Escherichia coli Chen S, Liu Z, Chen J, Wu J Ref: Appl Biochem Biotechnol, 165:666, 2011 : PubMed ESTHER: Chen_2011_Appl.Biochem.Biotechnol_165_666 PubMedSearch: Chen 2011 Appl.Biochem.Biotechnol 165 666 PubMedID: 21594592 Abstract Escherichia coli is one of the most commonly used host strains for recombinant protein production. More and more research works on the production of recombinant protein indicate that extracellular production throughout a culture medium is more convenient and attractive compared to intracellular production. In present work, inducing temperature and isopropyl beta-D: -1-thiogalactopyranoside (IPTG) concentration were investigated to decrease the formation of inclusion body and increase the amount of soluble recombinant cutinase initially. Enzyme activity in the culture medium reached to 118.9 U/ml at 64 h of culture, and no inclusion body was detected in cytoplasm under the inducement condition of 0.2 mM IPTG and 30 degrees C. In addition, it was found that a large amount of cutinase had been accumulated in periplasm since 16-h cultivation under the same inducement condition. Therefore, glycine and surfactant sodium taurodeoxycholate (TDOC) were further used to promote the leakage of recombinant cutinase from periplasm. Supplied with 100 mM glycine and 1 mM TDOC, the amount of cutinase in periplasm decreased remarkably, and the activity in the culture medium reached to 146.2 and 149.2 U/ml after 54 h of culturing, respectively. Title: Attenuation of CNS inflammatory responses by nicotine involves alpha7 and non-alpha7 nicotinic receptors Hao J, Simard AR, Turner GH, Wu J, Whiteaker P, Lukas RJ, Shi FD Ref: Experimental Neurology, 227:110, 2011 : PubMed ESTHER: Hao_2011_Exp.Neurol_227_110 PubMedSearch: Hao 2011 Exp.Neurol 227 110 PubMedID: 20932827 Abstract A considerable number of in vivo studies have demonstrated that the cholinergic system can dampen the peripheral immune response, and it is thought that the alpha7-nicotinic acetylcholine receptor (nAChR) subtype is a key mediator of this process. The goal of the present study was to determine if nicotine modulates immunological mechanisms known to be involved in the development of experimental autoimmune encephalomyelitis (EAE), a mouse model for CNS autoimmune disease, via alpha7-nAChRs. Here we show that nicotine exposure attenuates EAE severity and that this effect is largely abolished in nAChR alpha7 subunit knock-out mice. However, nicotine exposure partially retains the ability to reduce lymphocyte infiltration into the CNS, inhibit auto-reactive T cell proliferation and helper T cell cytokine production, down-regulate co-stimulatory protein expression on myeloid cells, and increase the differentiation and recruitment of regulatory T cells, even in the absence of alpha7-nAChRs. Diverse effects of nicotine on effector and regulatory T cells, as well as antigen-presenting cells, may be linked to differential expression patterns of nAChR subunits across these cell types. Taken together, our data show that although alpha7-nAChRs indeed seem to play an important role in nicotine-conferred reduction of the CNS inflammatory response and protection against EAE, other nAChR subtypes also are involved in the anti-inflammatory properties of the cholinergic system. Title: Cardiac abnormalities in severe acute dichlorvos poisoning He X, Li C, Wei D, Wu J, Shen L, Wang T Ref: Critical Care Medicine, 39:1906, 2011 : PubMed ESTHER: He_2011_Crit.Care.Med_39_1906 PubMedSearch: He 2011 Crit.Care.Med 39 1906 PubMedID: 21516037 Abstract OBJECTIVE: Patients with organophosphorus poisoning sometimes die suddenly during rigorous treatment, possibly from myocardial injury. This study sought to elucidate the mechanisms underlying organophosphorus poisoning-induced cardiotoxicity. DESIGN: Prospective observational study. SETTING: Urban, tertiary teaching hospital emergency intensive care unit with 10 beds. PATIENTS: Forty-one patients with severe acute dichlorvos poisoning were consecutively enrolled (n = 92) at emergency intensive care unit and followed for 3 months. MEASUREMENTS AND MAIN RESULTS: Levels of serum creatine kinase isoenzyme myocardium, cardiac troponin I, acetylcholinesterase, acetylcholine, epinephrine, and norepinephrine were tested on hospital days 1, 3, and 5 and on discharge day. Electrocardiography was recorded on admission and then every other day. Transthoracic echocardiography was performed at admission, in the acute phase, before discharge, and during follow-up. Technetium 99m-sestamibi myocardial single photon emission computed tomography was conducted in four patients. Thirty-seven (90.2%) patients survived and four (9.8%) patients died during treatment. We observed sinus tachycardia in 37 (90.2%) patients and ST-T changes in 33 (80.4%) patients. Creatine kinase isoenzyme myocardium and cardiac troponin I levels peaked at day 3 postadmission and then decreased to normal levels. Serum acetylcholine, epinephrine, and norepinephrine peaked at day 1 after admission and then decreased. Echocardiography revealed marked decreases in wall motion of the interventricular septum and left ventricle in the acute phase but returned to normal in the recovery phase. The left ventricular ejection fraction improved significantly from 42 +/- 5% to 59 +/- 4% (p = .001). Single photon emission computed tomography showed abnormal left ventricle perfusion. CONCLUSION: Severe acute dichlorvos poisoning is associated with reversible myocardial dysfunction, possibly through an increase in catecholamine levels. Title: Dynamic alterations of gene expression of nicotinic acetylcholine receptor alpha7, alpha4 and beta2 subunits in an acute MPTP-lesioned mouse model Hu J, Zhu C, Liu Y, Wang F, Huang Z, Fan W, Wu J Ref: Neuroscience Letters, 494:232, 2011 : PubMed ESTHER: Hu_2011_Neurosci.Lett_494_232 PubMedSearch: Hu 2011 Neurosci.Lett 494 232 PubMedID: 21406211 Abstract Epidemiologic studies show that the prevalence of Parkinson's disease (PD) is lower in smokers than in nonsmokers. Nicotine, a potent agonist of nicotinic acetylcholine receptors (nAChRs), excites midbrain dopaminergic neurons and this may contribute to the anti-parkinsonian effects. However, the alterations in gene expression of nAChR subunits using an acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse PD model remain unclear. In the present study, we profile the time course of nAChR alpha7, alpha4 and beta2 subunit expression levels using a comparative RT-PCR approach after acute MPTP injection. The results fall into four categories. (1) MPTP treatment transiently increased nAChR alpha7 (after last injection of MPTP 3 and 24 h), alpha4 and beta2 (24 h) mRNA expression in the substantia nigra (SN) and striatum. (2) Compared to cortical and hippocampal tissues, this transient increase of nAChR subunit expression specifically occurred in the SN and striatum. (3) In the acute MPTP model, time-courses of altered expression for nAChR alpha7, alpha4 and beta2 subunits closely mirrored the deficits observed in animal motor activity. (4) Stereological data showed that after administration of MPTP for 24h, there was a robust astrogliosis in the SN associated with significant dopaminergic neurodegeneration. These changes followed or paralleled MPTP-induced elevation in the levels of alpha7, alpha4 and beta2 mRNAs. Collectively, our results demonstrate that nAChRs are important targets in the MPTP neurotoxic process. These data suggest that therapeutic strategies targeted toward nAChR alpha7, alpha4 and beta2 subunits may have potential for developing new treatments for PD. Title: Complete genome sequence of Streptococcus suis serotype 14 strain JS14 Hu P, Yang M, Zhang A, Wu J, Chen B, Hua Y, Yu J, Xiao J, Jin M Ref: Journal of Bacteriology, 193:2375, 2011 : PubMed ESTHER: Hu_2011_J.Bacteriol_193_2375 PubMedSearch: Hu 2011 J.Bacteriol 193 2375 PubMedID: 21398551 Gene_locus related to this paper: strej-e8uk64, strsu-a4vws4, strsu-q673u2, strsy-a4vus4 Abstract Streptococcus suis is an important zoonotic agent leading to a variety of diseases in swine and can be transmitted to human beings upon close contact. Here, we report the complete genome sequence of S. suis serotype 14 strain JS14 which was isolated from a diseased pig in Jiangsu Province, China. Title: Complete genome sequence of Streptococcus suis serotype 3 strain ST3 Hu P, Yang M, Zhang A, Wu J, Chen B, Hua Y, Yu J, Chen H, Xiao J, Jin M Ref: Journal of Bacteriology, 193:3428, 2011 : PubMed ESTHER: Hu_2011_J.Bacteriol_193_3428 PubMedSearch: Hu 2011 J.Bacteriol 193 3428 PubMedID: 21572001 Gene_locus related to this paper: strsu-b9wvz1 Abstract Streptococcus suis is a zoonotic pathogen causing economic loss in the swine industry and is also a threat to human health. To date, the mechanism of pathogenesis is not fully understood. Here, we report the complete genome sequence of S. suis strain ST3 of serotype 3, which provides opportunities to reveal genetic basis of infection of S. suis non-serotype 2 strains. Title: Exposure of nicotine to ventral tegmental area slices induces glutamatergic synaptic plasticity on dopamine neurons Jin Y, Yang K, Wang H, Wu J Ref: Synapse, 65:332, 2011 : PubMed ESTHER: Jin_2011_Synapse_65_332 PubMedSearch: Jin 2011 Synapse 65 332 PubMedID: 20730803 Abstract Nicotine promotes glutamatergic synaptic plasticity in dopaminergic (DA) neurons in the ventral tegmental area (VTA), which is thought to be an important mechanism underlying nicotine reward. However, it is unclear whether exposure of nicotine alone to VTA slice is sufficient to increase glutamatergic synaptic strength on DA neurons and which nicotinic acetylcholine receptor (nAChR) subtype mediates this effect. Here, we report that the incubation of rat VTA slices with 500 nM nicotine induces glutamatergic synaptic plasticity in DA neurons. We measure the ratio of AMPA and NMDA receptor-mediated currents (AMPA/NMDA) and compare these ratios between nicotine-treated and -untreated slices. Our results demonstrate that the incubation of VTA slices with 500 nM nicotine for 1 h (but not for 10 min) significantly increases the AMPA/NMDA ratio when compared with controls. Preincubation with 10 nM of the alpha7-nAChR antagonist, methyllycaconitine (MLA) but not 1 muM alpha4-containing nAChR antagonist, dihydro-beta-erythroidine (DHbetaE) prevents nicotinic effect, suggesting that alpha7-nAChRs are mainly mediated this nicotinic effect. This finding is further supported by the disappearance of this nicotinic effect in nAChR alpha7 knockout (KO) mice. Furthermore, nicotine reduced paired-pulse ratio (PPR) of evoked excitatory postsynaptic potential (eEPSP) in the VTA slices prepared from wild-type (WT) mice but not alpha7 KO mice. Collectively, these findings suggest that exposure of smoking-relevant concentrations of nicotine to VTA slices is sufficient to increase glutamatergic synaptic strength on DA neurons and that alpha7-nAChRs likely mediate this nicotinic effect through increasing presynaptic release of glutamate. Title: Effect of Silent Mutations in Translational Initial Region on the Production of Recombinant Cutinase in Escherichia coli Liu ZG, Zhu L, Zhu KL, Chen S, Chen J, Wu J Ref: Curr Microbiol, 62:1302, 2011 : PubMed ESTHER: Liu_2011_Curr.Microbiol_62_1302 PubMedSearch: Liu 2011 Curr.Microbiol 62 1302 PubMedID: 21212953 Abstract Translational Initial Region (TIR) is the threshold of an intracellular translation process, and tiny alterations in this region are reported to intensely influence the downstream expression. Such property provides a potential utilization in extracellular production of recombinant enzyme. As an esterase, cutinase is an essential catalyst in the process of textile scouring, and has a potential application in food and chemical industry. In the present study, a bacterial cutinase (Tfu_0883) from Thermobifida fusca was expressed in Escherichia coli with pelB as its signal peptide using trc as its promoter. A subsequent TIR degeneracy mutagenesis was then carried out in the initial sequence of pelB. A fast screening method for these mutants was developed and a series of strains with different expression strengths were accordingly obtained. Among these mutants, a high cutinase production level of 38.0 U/ml was achieved, which is three times that of the control group. This study explored the potential utilization of TIR degeneracy mutagenesis in the production of industrial enzymes. Title: Poster: Tethered pentamersLow sensitivity alpha4/beta2-nicotinic acetylcholine receptors Lucero L, Bhakta M, Liu G, Wu J, Hauser TA, Bencherif M, Bermudez I, Whiteaker P, Lukas RJ Ref: Biochemical Pharmacology, 82:1025, 2011 : PubMed ESTHER: Lucero_2011_Biochem.Pharmacol_82(8)_1025 PubMedSearch: Lucero 2011 Biochem.Pharmacol 82(8) 1025 Title: Construction of Aspergillus niger lipase mutants with oil-water interface independence Shu Z, Wu J, Xue L, Lin R, Jiang Y, Tang L, Li X, Huang J Ref: Enzyme Microb Technol, 48:129, 2011 : PubMed ESTHER: Shu_2011_Enzyme.Microb.Technol_48_129 PubMedSearch: Shu 2011 Enzyme.Microb.Technol 48 129 PubMedID: 22112821 Abstract Based on previous bioinformational analytical results [Shu ZY, et al. Biotechnol Prog 2009;25:409-16], four A. niger lipase (ANL) mutants, ANL-Ser84Gly, ANL-Asp99Pro, ANL-Lys108Glu and ANL-EalphaH (obtained by replacing the lid domain of ANL with the corresponding domain from A. niger feruloyl esterase), were constructed to screen out ANL mutants with oil-water interface independence. ANL-S84G displayed a pronounced interfacial activation, while ANL-D99P and ANL-K108E displayed no interfacial activation. The specific activity of ANL-S84G towards p-nitrophenyl esters decreased from 29.8% to 76.5% compared with that of ANL, while the specific activity of ANL-D99P towards p-nitrophenyl palmitate increased 2.2-fold. The thermostability of ANL-K108E was almost unchanged, while the thermostability of ANL-S84G and ANL-D99P significantly decreased compared with that of ANL. The construction of oil-water interface-independent ANL mutants would help to further understand the mechanism of lipase interfacial activation. Title: Engineered Thermobifida fusca cutinase with increased activity on polyester substrates Silva C, Da S, Silva N, Matama T, Araujo R, Martins M, Chen S, Chen J, Wu J and Cavaco-Paulo A <1 more author(s)> Silva C, Da S, Silva N, Matama T, Araujo R, Martins M, Chen S, Chen J, Wu J, Casal M, Cavaco-Paulo A (- 1) Ref: Biotechnol J, 6:1230, 2011 : PubMed ESTHER: Silva_2011_Biotechnol.J_6_1230 PubMedSearch: Silva 2011 Biotechnol.J 6 1230 PubMedID: 21751386 Gene_locus related to this paper: thefu-q6a0i4 Abstract A bacterial cutinase from Thermobifida fusca, named Tfu_0883, was genetically modified by site-directed mutagenesis to enhance its activity on poly(ethylene terephthalate) (PET). The new mutations tailored the catalytic site for PET, increasing the affinity of cutinase to this hydrophobic substrate and the ability to hydrolyze it. The mutation I218A was designed to create space and the double mutation Q132A/T101A was designed both to create space and to increase hydrophobicity. The activity of the double mutant on the soluble substrate p-nitrophenyl butyrate increased two-fold compared to wild-type cutinase, while on PET both single and double mutants exhibited considerably higher hydrolysis efficiency. The replacement of specific amino acids at the active site was an effective approach for the improvement of the Tfu_0883 cutinase capacity to hydrolyze polyester surfaces. Thus, this study provides valuable insight on how the function and stability of enzymes can be improved by molecular engineering for their application in synthetic fiber biotransformation. Title: Poster: Acetylcholine binding protein-nicotinic receptor chimeras for delineating structure and determinants of ligand selectivity Talley TT, Nemecz A, Yamauchi JG, Wu J, Ho KY, Sankaran B, Taylor P Ref: Biochemical Pharmacology, 82:1028, 2011 : PubMed ESTHER: Talley_2011_Biochem.Pharmacol_82(8)_1028 PubMedSearch: Talley 2011 Biochem.Pharmacol 82(8) 1028 Title: Characterization of two metagenome-derived esterases that reactivate chloramphenicol by counteracting chloramphenicol acetyltransferase Tao W, Lee MH, Yoon MY, Kim JC, Malhotra S, Wu J, Hwang EC, Lee SW Ref: J Microbiol Biotechnol, 21:1203, 2011 : PubMed ESTHER: Tao_2011_J.Microbiol.Biotechnol_21_1203 PubMedSearch: Tao 2011 J.Microbiol.Biotechnol 21 1203 PubMedID: 22210605 Gene_locus related to this paper: 9bact-g3cr00, 9bact-g3cr02 Abstract Function-driven metagenomic analysis is a powerful approach to screening for novel biocatalysts. In this study, we investigated lipolytic enzymes selected from an alluvial soil metagenomic library, and identified two novel esterases, EstDL26 and EstDL136. EstDL26 and EstDL136 reactivated chloramphenicol from its acetyl derivates by counteracting the chloramphenicol acetyltransferase (CAT) activity in Escherichia coli. These two enzymes showed only 27% identity in amino acid sequence to each other; however both preferentially hydrolyzed short-chain p-nitrophenyl esters (< or =C5) and showed mesophilic properties. In vitro, EstDL136 catalyzed the deacetylation of 1- and 3- acetyl and 1,3-diacetyl derivates; in contrast, EstDL26 was not capable of the deacetylation at C1, indicating a potential regioselectivity. EstDL26 and EstDL136 were similar to microbial hormone-sensitive lipase (HSL), and since chloramphenicol acetate esterase (CAE) activity was detected from two other soil esterases in the HSL family, this suggests a distribution of CAE among the soil microorganisms. The isolation and characterization of EstDL26 and EstDL136 in this study may be helpful in understanding the diversity of CAE enzymes and their potential role in releasing active chloramphenicol in the producing bacteria. Title: Isolation and characterization of a family VII esterase derived from alluvial soil metagenomic library Tao W, Lee MH, Wu J, Kim NH, Lee SW Ref: J Microbiol, 49:178, 2011 : PubMed ESTHER: Tao_2011_J.Microbiol_49_178 PubMedSearch: Tao 2011 J.Microbiol 49 178 PubMedID: 21538236 Gene_locus related to this paper: 9bact-g1ard1 Abstract A novel esterase gene, estDL30, was isolated from an alluvial metagenomic library using function-driven screening. estDL30 consisted of 1,524 nucleotides and encoded a 507-amino acid protein. Sequence analysis revealed that EstDL30 is similar to many type B carboxylesterases, containing a G-E-S-A-G pentapeptide with a catalytic Ser residue. Phylogenetic analysis suggested that EstDL30 belongs to the family VII lipases, together with esterases from Bacillus subtilis (P37967), Streptomyces coelicolor A3(2) (CAA22794), and Arthrobacter oxydans (Q01470). Purified EstDL30 showed its highest catalytic efficiency toward p-nitrophenyl butyrate, with a k (cat) of 2293 s(-1) and k (cat)/K (m) of 176.4 s(-1)mM(-1); however, little activity was detected when the acyl chain length exceeded C(8). Biochemical characterization of EstDL30 revealed that it is an alkaline esterase that possesses maximal activity at pH 8 and 40 degrees C. The effects of denaturants and divalent cations were also investigated. EstDL30 tolerated well the presence of methanol and Tween 20. Its activity was strongly inhibited by 1 mM Cu(2+) and Zn(2+), but stimulated by Fe(2+). The unique properties of EstDL30, its high activity under alkaline conditions and stability in the presence of organic solvents, may render it applicable to organic synthesis. Title: The genome of the mesopolyploid crop species Brassica rapa Wang X, Wang H, Wang J, Sun R, Wu J, Liu S, Bai Y, Mun JH, Bancroft I and Zhang Z <88 more author(s)> Wang X, Wang H, Wang J, Sun R, Wu J, Liu S, Bai Y, Mun JH, Bancroft I, Cheng F, Huang S, Li X, Hua W, Freeling M, Pires JC, Paterson AH, Chalhoub B, Wang B, Hayward A, Sharpe AG, Park BS, Weisshaar B, Liu B, Li B, Tong C, Song C, Duran C, Peng C, Geng C, Koh C, Lin C, Edwards D, Mu D, Shen D, Soumpourou E, Li F, Fraser F, Conant G, Lassalle G, King GJ, Bonnema G, Tang H, Belcram H, Zhou H, Hirakawa H, Abe H, Guo H, Jin H, Parkin IA, Batley J, Kim JS, Just J, Li J, Xu J, Deng J, Kim JA, Yu J, Meng J, Min J, Poulain J, Hatakeyama K, Wu K, Wang L, Fang L, Trick M, Links MG, Zhao M, Jin M, Ramchiary N, Drou N, Berkman PJ, Cai Q, Huang Q, Li R, Tabata S, Cheng S, Zhang S, Sato S, Sun S, Kwon SJ, Choi SR, Lee TH, Fan W, Zhao X, Tan X, Xu X, Wang Y, Qiu Y, Yin Y, Li Y, Du Y, Liao Y, Lim Y, Narusaka Y, Wang Z, Li Z, Xiong Z, Zhang Z (- 88) Ref: Nat Genet, 43:1035, 2011 : PubMed ESTHER: Wang_2011_Nat.Genet_43_1035 PubMedSearch: Wang 2011 Nat.Genet 43 1035 PubMedID: 21873998 Gene_locus related to this paper: braol-Q8GTM3, braol-Q8GTM4, brarp-m4ei94, brarp-m4c988, brana-a0a078j4a9, brana-a0a078e1m0, brana-a0a078cd75, brarp-m4dwa6, brana-a0a078j4f0, brana-a0a078cus4, brana-a0a078f8c2, brana-a0a078jql1, brana-a0a078dgj3, brana-a0a078hw50, brana-a0a078cuu0, brana-a0a078dfa9, brana-a0a078ic91, brarp-m4ctw3, brana-a0a078ca65, brana-a0a078ctc8, brana-a0a078h021, brana-a0a078jx23, brarp-m4da84, brarp-m4dwr7, brana-a0a078dh94, brana-a0a078h612, brana-a0a078j2t3, braol-a0a0d3dpb2, braol-a0a0d3dx76, brana-a0a078jxa8, brana-a0a078i2k3, brarp-m4cwq4, brarp-m4dcj8, brarp-m4eh17, brarp-m4eey4, brarp-m4dnj8, brarp-m4ey83, brarp-m4ey84 Abstract We report the annotation and analysis of the draft genome sequence of Brassica rapa accession Chiifu-401-42, a Chinese cabbage. We modeled 41,174 protein coding genes in the B. rapa genome, which has undergone genome triplication. We used Arabidopsis thaliana as an outgroup for investigating the consequences of genome triplication, such as structural and functional evolution. The extent of gene loss (fractionation) among triplicated genome segments varies, with one of the three copies consistently retaining a disproportionately large fraction of the genes expected to have been present in its ancestor. Variation in the number of members of gene families present in the genome may contribute to the remarkable morphological plasticity of Brassica species. The B. rapa genome sequence provides an important resource for studying the evolution of polyploid genomes and underpins the genetic improvement of Brassica oil and vegetable crops. Title: TiO2-decorated graphene nanohybrids for fabricating an amperometric acetylcholinesterase biosensor Wang K, Li HN, Wu J, Ju C, Yan JJ, Liu Q, Qiu B Ref: Analyst, 136:3349, 2011 : PubMed ESTHER: Wang_2011_Analyst_136_3349 PubMedSearch: Wang 2011 Analyst 136 3349 PubMedID: 21738917 Abstract This work describes a highly sensitive and rapid amperometric biosensor for organophosphate compounds (OPs) based on immobilization of acetylcholinesterase (AChE) on a novel TiO(2)-decorated graphene (TiO(2)-G) nanohybrid, which was constructed by in situ growth of TiO(2) nanoparticles (NPs) on the graphene sheet. The well-dispersed TiO(2) NPs eliminated the restacking of TiO(2)-G nanohybrids. Due to the integrating of TiO(2)-G nanohybrids, the as-prepared biosensor showed high affinity to acetylthiocholine (ATCl) with a Michaelis-Menten constant (K(m)) value of 0.22 mM, and rapid inhibition time (3 min). Further, based on the inhibition of OPs on the enzymatic activity of the immobilized AChE, and using carbaryl as a model compound, the inhibition of carbaryl was proportional to its concentration ranging from 0.001 to 0.015 and 0.015 to 2 mug mL(-1) with a detection limit of 0.3 ng mL(-1) (S/N = 3). The developed biosensor exhibited a good performance for organophosphate pesticide detection, including good reproducibility and acceptable stability, which provided a new and promising tool for the analysis of enzyme inhibitors. Title: Naturally-expressed nicotinic acetylcholine receptor subtypes Wu J, Lukas RJ Ref: Biochemical Pharmacology, 82:800, 2011 : PubMed ESTHER: Wu_2011_Biochem.Pharmacol_82(8)_800 PubMedSearch: Wu 2011 Biochem.Pharmacol 82(8) 800 PubMedID: 21787755 Abstract Nicotinic acetylcholine receptors (nAChRs) warrant attention, as they play many critical roles in brain and body function and have been implicated in a number of neurological and psychiatric disorders, including nicotine dependence. nAChRs are composed as diverse subtypes containing specific combinations of genetically-distinct subunits and that have different functional properties, distributions, and pharmacological profiles. There had been confidence that the rules that define ranges of assembly partners for specific subunits were well-established, especially for the more prominent nAChR subtypes. However, we review here some newer findings indicating that nAChRs having largely the same, major subunits exist as isoforms with unexpectedly different properties. Moreover, we also summarize our own studies indicating that novel nAChR subtypes exist and/or have distributions not heretofore described. Importantly, the nAChRs that exist as new isoforms or subtypes or have interesting distributions require alteration in thinking about their roles in health and disease. Title: Induction of a 55 kDa acetylcholinesterase protein during apoptosis and its negative regulation by the Akt pathway Xie J, Jiang H, Wan YH, Du AY, Guo KJ, Liu T, Ye WY, Niu X, Wu J and Zhang XJ <1 more author(s)> Xie J, Jiang H, Wan YH, Du AY, Guo KJ, Liu T, Ye WY, Niu X, Wu J, Dong XQ, Zhang XJ (- 1) Ref: J Molecular & Cellular Biology, 3:250, 2011 : PubMed ESTHER: Xie_2011_J.Mol.Cell.Biol_3_250 PubMedSearch: Xie 2011 J.Mol.Cell.Biol 3 250 PubMedID: 21377978 Abstract Acetylcholinesterase (AChE) is emerging as an important contributor to apoptosis in various cell types. However, overexpression of AChE does not initiate apoptosis, and cells which express AChE at basal levels grow normally, suggesting that AChE may function differently between normal and apoptotic conditions. In this study, we determined that an AChE-derived protein ( approximately 55 kDa) positively correlated with cellular apoptotic levels. The 55 kDa AChE protein was not a result of a novel splice variant of the AChE primary transcript. Instead, it was determined to be a cleaved fragment of the full-length 68 kDa AChE protein that could not be inhibited by cycloheximide (CHX) but could be suppressed by caspase inhibitors in apoptotic PC-12 cells. Furthermore, activation of the Akt cascade abolished the 55 kDa protein, and both AChE protein forms (68 and 55 kDa) accumulated in the nucleus during apoptosis. In a mouse model for ischemia/reperfusion (I/R)-induced acute renal failure, the 55 kDa AChE protein was detected in the impaired organs but not in the normal ones, and its levels correlated with the genotype of the mice. In summary, a 55 kDa AChE protein resulting from the cleavage of 68 kDa AChE is induced during apoptosis, and it is negatively regulated by the Akt pathway. This study suggests that an alternative form of AChE may play a role in apoptosis. Title: Functional nicotinic acetylcholine receptors containing alpha6 subunits are on GABAergic neuronal boutons adherent to ventral tegmental area dopamine neurons Yang K, Buhlman L, Khan GM, Nichols RA, Jin G, McIntosh JM, Whiteaker P, Lukas RJ, Wu J Ref: Journal of Neuroscience, 31:2537, 2011 : PubMed ESTHER: Yang_2011_J.Neurosci_31_2537 PubMedSearch: Yang 2011 J.Neurosci 31 2537 PubMedID: 21325521 Abstract Diverse nicotinic acetylcholine receptor (nAChR) subtypes containing different subunit combinations can be placed on nerve terminals or soma/dendrites in the ventral tegmental area (VTA). nAChR alpha6 subunit message is abundant in the VTA, but alpha6*-nAChR cellular localization, function, pharmacology, and roles in cholinergic modulation of dopaminergic (DA) neurons within the VTA are not well understood. Here, we report evidence for alpha6beta2*-nAChR expression on GABA neuronal boutons terminating on VTA DA neurons. alpha-Conotoxin (alpha-Ctx) MII labeling coupled with immunocytochemical staining localizes putative alpha6*-nAChRs to presynaptic GABAergic boutons on acutely dissociated, rat VTA DA neurons. Functionally, acetylcholine (ACh) induces increases in the frequency of bicuculline-, picrotoxin-, and 4-aminopyridine-sensitive miniature IPSCs (mIPSCs) mediated by GABA(A) receptors. These increases are abolished by alpha6*-nAChR-selective alpha-Ctx MII or alpha-Ctx PIA (1 nm) but not by alpha7 (10 nm methyllycaconitine) or alpha4* (1 mum dihydro-beta-erythroidine)-nAChR-selective antagonists. ACh also fails to increase mIPSC frequency in VTA DA neurons prepared from nAChR beta2 knock-out mice. Moreover, ACh induces an alpha-Ctx PIA-sensitive elevation in intraterminal Ca(2+) in synaptosomes prepared from the rat VTA. Subchronic exposure to 500 nm nicotine reduces ACh-induced GABA release onto the VTA DA neurons, as does 10 d of systemic nicotine exposure. Collectively, these results indicate that alpha6beta2*-nAChRs are located on presynaptic GABAergic boutons within the VTA and modulate GABA release onto DA neurons. These presynaptic alpha6beta2*-nAChRs likely play important roles in nicotinic modulation of DA neuronal activity. Title: Genomic and proteomic analyses of the fungus Arthrobotrys oligospora provide insights into nematode-trap formation Yang J, Wang L, Ji X, Feng Y, Li X, Zou C, Xu J, Ren Y, Mi Q and Zhang KQ <16 more author(s)> Yang J, Wang L, Ji X, Feng Y, Li X, Zou C, Xu J, Ren Y, Mi Q, Wu J, Liu S, Liu Y, Huang X, Wang H, Niu X, Li J, Liang L, Luo Y, Ji K, Zhou W, Yu Z, Li G, Li L, Qiao M, Feng L, Zhang KQ (- 16) Ref: PLoS Pathog, 7:e1002179, 2011 : PubMed ESTHER: Yang_2011_PLoS.Pathog_7_E1002179 PubMedSearch: Yang 2011 PLoS.Pathog 7 E1002179 PubMedID: 21909256 Gene_locus related to this paper: artoa-g1wyr4, artoa-g1x1a7, artoa-g1x3f4, artoa-g1x3h6, artoa-g1x9s5, artoa-g1x9z4, artoa-g1xcb5, artoa-g1xhl6, artoa-g1xjb3, artoa-g1xjy0, artoa-g1xkw3, artoa-g1xnf2, artoa-g1xnf8, artoa-g1xqd4, artoa-g1xqt1, artoa-g1xte8, artoa-g1xu91, artoa-g1xv59, artoa-g1x382, artoa-g1x3q3, artoa-g1wxl5, artoa-g1xj75, artoa-g1xd25, artoa-g1wzu7, artoa-g1xt42, artoa-g1xhm8, artoa-g1wy43 Abstract Nematode-trapping fungi are "carnivorous" and attack their hosts using specialized trapping devices. The morphological development of these traps is the key indicator of their switch from saprophytic to predacious lifestyles. Here, the genome of the nematode-trapping fungus Arthrobotrys oligospora Fres. (ATCC24927) was reported. The genome contains 40.07 Mb assembled sequence with 11,479 predicted genes. Comparative analysis showed that A. oligospora shared many more genes with pathogenic fungi than with non-pathogenic fungi. Specifically, compared to several sequenced ascomycete fungi, the A. oligospora genome has a larger number of pathogenicity-related genes in the subtilisin, cellulase, cellobiohydrolase, and pectinesterase gene families. Searching against the pathogen-host interaction gene database identified 398 homologous genes involved in pathogenicity in other fungi. The analysis of repetitive sequences provided evidence for repeat-induced point mutations in A. oligospora. Proteomic and quantitative PCR (qPCR) analyses revealed that 90 genes were significantly up-regulated at the early stage of trap-formation by nematode extracts and most of these genes were involved in translation, amino acid metabolism, carbohydrate metabolism, cell wall and membrane biogenesis. Based on the combined genomic, proteomic and qPCR data, a model for the formation of nematode trapping device in this fungus was proposed. In this model, multiple fungal signal transduction pathways are activated by its nematode prey to further regulate downstream genes associated with diverse cellular processes such as energy metabolism, biosynthesis of the cell wall and adhesive proteins, cell division, glycerol accumulation and peroxisome biogenesis. This study will facilitate the identification of pathogenicity-related genes and provide a broad foundation for understanding the molecular and evolutionary mechanisms underlying fungi-nematodes interactions. Title: Comparative genomic analysis of Streptococcus suis reveals significant genomic diversity among different serotypes Zhang A, Yang M, Hu P, Wu J, Chen B, Hua Y, Yu J, Chen H, Xiao J, Jin M Ref: BMC Genomics, 12:523, 2011 : PubMed ESTHER: Zhang_2011_BMC.Genomics_12_523 PubMedSearch: Zhang 2011 BMC.Genomics 12 523 PubMedID: 22026465 Gene_locus related to this paper: strsu-b9wvz1 Abstract BACKGROUND: Streptococcus suis (S. suis) is a major swine pathogen and an emerging zoonotic agent. Serotypes 1, 2, 3, 7, 9, 14 and 1/2 are the most prevalent serotypes of this pathogen. However, almost all studies were carried out on serotype 2 strains. Therefore, characterization of genomic features of other serotypes will be required to better understand their virulence potential and phylogenetic relationships among different serotypes. Title: Biochemical characterization of the cutinases from Thermobifida fusca Chen S, Su L, Billig S, Zimmermann W, Chen J, Wu J Ref: J Mol Catal B Enzym, 63:121, 2010 : PubMed ESTHER: Chen_2010_J.Mol.Catal.B.Enzym_63_121 PubMedSearch: Chen 2010 J.Mol.Catal.B.Enzym 63 121 Gene_locus related to this paper: thefu-q6a0i4, thefu-q6a0i3 Abstract Thermobifida fusca produces two cutinases which share 93% identity in amino acid sequence. In the present study, we investigated the detailed biochemical properties of T. fusca cutinases for the first time. For a better comparison between bacterial and fungal cutinases, recombinant Fusarium solani pisi cutinase was subjected to the similar analysis. The results showed that both bacterial and fungal cutinases are monomeric proteins in solution. The bacterial cutinases exhibited a broad substrate specificity against plant cutin, synthetic polyesters, insoluble triglycerides, and soluble esters. In addition, the two isoenzymes of T. fusca and the F.solani pisi cutinase are similar in substrate kinetics, the lack of interfacial activation, and metal ion requirements. However, the T.fusca cutinases showed higher stability in the presence of surfactants and organic solvents. Considering the versatile hydrolytic activity, good tolerance to surfactants, superior stability in organic solvents, and thermostability demonstrated by T. fusca cutinases, they may have promising applications in related industries. Title: Decoding state transitions in hippocampal oscillatory activity in mice Dragomir A, Akay YM, Wang K, Wu J, Akay M Ref: Conf Proc IEEE Eng Med Biol Soc, 2010:2822, 2010 : PubMed ESTHER: Dragomir_2010_Conf.Proc.IEEE.Eng.Med.Biol.Soc_2010_2822 PubMedSearch: Dragomir 2010 Conf.Proc.IEEE.Eng.Med.Biol.Soc 2010 2822 PubMedID: 21096221 Abstract Understanding the intricate dynamics of the hippocampal neural network, from which several types of neural oscillation rhythms arise, is an important step in uncovering the role of the hippocampus in the formation of memory. The different oscillation types commonly recorded in the hippocampus are thought to correspond to several states of neural network synchronization. Therefore, accurate segmentation and decoding of these underlying states provide useful insight on the rhythms' generation. In this study we use a framework based on Hidden Markov Models, coupled with a nonlinear dynamics method based on the Lempel-Ziv estimator. The method allows us to decode and model the neural state transitions. Network synchronization was induced by acute exposure to cholinergic agonist carbachol and oscillations were recorded from the Cornu Ammonis (CA1) region of the mouse hippocampus. Our results prove that deficits in cholinergic neuro-transmission found in triple transgenic mice (3xTG, as Alzheimer's disease animal model) lead to increased instability in the hippocampal neural network synchronization. Title: Mechanisms involved in systemic nicotine-induced glutamatergic synaptic plasticity on dopamine neurons in the ventral tegmental area Gao M, Jin Y, Yang K, Zhang D, Lukas RJ, Wu J Ref: Journal of Neuroscience, 30:13814, 2010 : PubMed ESTHER: Gao_2010_J.Neurosci_30_13814 PubMedSearch: Gao 2010 J.Neurosci 30 13814 PubMedID: 20943922 Abstract Systemic exposure to nicotine induces glutamatergic synaptic plasticity on dopamine (DA) neurons in the ventral tegmental area (VTA), but mechanisms are largely unknown. Here, we report that single, systemic exposure in rats to nicotine (0.17 mg/kg free base) increases the ratio of DA neuronal currents mediated by AMPA relative to NMDA receptors (AMPA/NMDA ratio) assessed 24 h later, based on slice-patch recording. The AMPA/NMDA ratio increase is evident within 1 h and lasts for at least 72 h after nicotine exposure (and up to 8 d after repeated nicotine administration). This effect cannot be prevented by systemic injection of either alpha7-nAChR (nicotinic ACh receptor)-selective [methyllycaconitine (MLA)] or beta2*-nAChR-selective [mecamylamine (MEC)] antagonists but is prevented by coinjection of MLA and MEC. In either nAChR alpha7 or beta2 subunit knock-out mice, systemic exposure to nicotine still increases the AMPA/NMDA ratio. Preinjection in rats of a NMDA receptor antagonist MK-801((+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate), but neither DA receptor antagonists [SCH-23390 (R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine) plus haloperidol] nor a calcineurin inhibitor (cyclosporine), prevents the nicotine-induced increase in AMPA/NMDA ratio. After systemic exposure to nicotine, glutamatergic (but not GABAergic) transmission onto rat VTA DA neuronal inputs is enhanced. Correspondingly, DA neuronal firing measured 24 h after nicotine exposure using extracellular single-unit recording in vivo is significantly faster, and there is conversion of silent to active DA neurons. Collectively, these findings demonstrate that systemic nicotine acting via either alpha7- or beta2*-nAChRs increases presynaptic and postsynaptic glutamatergic function, and consequently initiates glutamatergic synaptic plasticity, which may be an important, early neuronal adaptation in nicotine reward and reinforcement. Title: Characterization of [(3)H]CHIBA-1001 binding to alpha7 nicotinic acetylcholine receptors in the brain from rat, monkey, and human Tanibuchi Y, Wu J, Toyohara J, Fujita Y, Iyo M, Hashimoto K Ref: Brain Research, 1348:200, 2010 : PubMed ESTHER: Tanibuchi_2010_Brain.Res_1348_200 PubMedSearch: Tanibuchi 2010 Brain.Res 1348 200 PubMedID: 20537987 Abstract Accumulating evidence suggests that the alpha7 subtype of nicotinic acetylcholine receptors (nAChRs) plays a role in the pathophysiology of neuropsychiatric diseases, including schizophrenia and Alzheimer's disease. Currently, there are no suitable small molecule radioligands for alpha7 nAChRs in the brain, although [(125)I]alpha-bungarotoxin has been widely used as a radioligand for alpha7 nAChRs. In the present study, we characterized a new radioligand, 4-[(3)H]methylphenyl 2,5-diazabicyclo[3.2.2]nonane-2-carboxylate ([(3)H]CHIBA-1001), a derivative of the selective alpha7 nAChR agonist SSR180711, in brain membranes from rat, monkey, and human. Scatchard analysis revealed an apparent equilibrium dissociation constant (Kd) of 193.4nM in rat brain membranes at 4 degrees C, and the maximal number of binding sites (Bmax) was 346.2fmol/mg protein. The order of drugs for the inhibition of [(3)H]CHIBA-1001 binding to rat brain membranes is SSR180711>A-844606>MG624>epibatidine>DMAB>A-582941, suggesting a similarity of alpha7 nAChR pharmacological profiles. In contrast, alpha-bungarotoxin, MLA, and nicotine were found to be very weak. The distribution of [(3)H]CHIBA-1001 binding to crude membranes from dissected regions of rat, monkey, and human brain was different from that of [(125)I]alpha-bungarotoxin binding, suggesting that [(3)H]CHIBA-1001 binding sites may not be identical to [(125)I]alpha-bungarotoxin binding in the brain. In summary, [(3)H]CHIBA-1001 would be a useful radioligand for alpha7 nAChRs in the brains of rodents, non-human primates, and humans. Title: Genome sequencing and analysis of the model grass Brachypodium distachyon. Vogel JP, Garvin DF, Mockler TC, Schmutz J, Rokhsar D, Bevan MW, Barry K, Lucas S, Harmon-Smith M and Baxter I <127 more author(s)> Vogel JP, Garvin DF, Mockler TC, Schmutz J, Rokhsar D, Bevan MW, Barry K, Lucas S, Harmon-Smith M, Lail K, Tice H, Grimwood J, McKenzie N, Huo N, Gu YQ, Lazo GR, Anderson OD, You FM, Luo MC, Dvorak J, Wright J, Febrer M, Idziak D, Hasterok R, Lindquist E, Wang M, Fox SE, Priest HD, Filichkin SA, Givan SA, Bryant DW, Chang JH, Wu H, Wu W, Hsia AP, Schnable PS, Kalyanaraman A, Barbazuk B, Michael TP, Hazen SP, Bragg JN, Laudencia-Chingcuanco D, Weng Y, Haberer G, Spannagl M, Mayer K, Rattei T, Mitros T, Lee SJ, Rose JK, Mueller LA, York TL, Wicker T, Buchmann JP, Tanskanen J, Schulman AH, Gundlach H, Bevan M, de Oliveira AC, Maia Lda C, Belknap W, Jiang N, Lai J, Zhu L, Ma J, Sun C, Pritham E, Salse J, Murat F, Abrouk M, Bruggmann R, Messing J, Fahlgren N, Sullivan CM, Carrington JC, Chapman EJ, May GD, Zhai J, Ganssmann M, Gurazada SG, German M, Meyers BC, Green PJ, Tyler L, Wu J, Thomson J, Chen S, Scheller HV, Harholt J, Ulvskov P, Kimbrel JA, Bartley LE, Cao P, Jung KH, Sharma MK, Vega-Sanchez M, Ronald P, Dardick CD, De Bodt S, Verelst W, Inz D, Heese M, Schnittger A, Yang X, Kalluri UC, Tuskan GA, Hua Z, Vierstra RD, Cui Y, Ouyang S, Sun Q, Liu Z, Yilmaz A, Grotewold E, Sibout R, Hematy K, Mouille G, Hofte H, Michael T, Pelloux J, O'Connor D, Schnable J, Rowe S, Harmon F, Cass CL, Sedbrook JC, Byrne ME, Walsh S, Higgins J, Li P, Brutnell T, Unver T, Budak H, Belcram H, Charles M, Chalhoub B, Baxter I (- 127) Ref: Nature, 463:763, 2010 : PubMed ESTHER: Vogel_2010_Nature_463_763 PubMedSearch: Vogel 2010 Nature 463 763 PubMedID: 20148030 Gene_locus related to this paper: bradi-i1grm0, bradi-i1gx82, bradi-i1hb80, bradi-i1hkv6, bradi-i1hpu6, bradi-i1i3e4, bradi-i1i9i0, bradi-i1i435, bradi-i1ix93, bradi-i1gsk6, bradi-i1hk44, bradi-i1hk45, bradi-i1hnk7, bradi-i1hsd5, bradi-i1huy4, bradi-i1huy9, bradi-i1huz0, bradi-i1gxx9, bradi-i1hl25, bradi-i1hcw7, bradi-i1hyv6, bradi-i1hyb5, bradi-i1hvr8, bradi-i1hmu2, bradi-i1hf05, bradi-i1gry7, bradi-i1hf06, bradi-i1i5z8, bradi-i1icy3, bradi-i1j1h3, bradi-i1h1e3, bradi-i1hvr9, bradi-a0a0q3r7i7, bradi-i1i377, bradi-i1hjg5, bradi-i1h3i9, bradi-i1gsg5, bradi-a0a0q3mph9, bradi-i1h682, bradi-a0a0q3lc91, bradi-i1gx49, bradi-i1i839, bradi-a0a2k2dsp5, bradi-i1gsb5 Title: Double target concept for smoking cessation Wu J Ref: Acta Pharmacol Sin, 31:1015, 2010 : PubMed ESTHER: Wu_2010_Acta.Pharmacol.Sin_31_1015 PubMedSearch: Wu 2010 Acta.Pharmacol.Sin 31 1015 PubMedID: 20711220 Abstract Tobacco use is estimated to be the largest single cause of premature death in the world. Nicotine is the major addictive substance in tobacco products. After cigarette smoking, nicotine quickly acts on its target, nicotinic acetylcholine receptors (nAChRs), which are widely distributed throughout the mammalian central nervous system and are expressed as diverse subtypes on cell bodies, dendrites and/or nerve terminals. Through the nAChRs in brain reward circuits, nicotine alters dopaminergic (DA) neuronal function in the ventral tegmental area (VTA) and increases dopamine release from VTA to nuclear accumbens (NA), which leads to nicotine reward, tolerance and dependence. After quitting smoking, smokers experience withdrawal symptoms, including depression, irritability, difficulty concentrating or sleeping, headache, and tiredness. Recently, evidence has been accumulated to reveal the molecular and cellular mechanisms of nicotine reward, tolerance and dependence. The outcomes of these investigations provide pharmacological basis for smoking cessation. Here, I briefly summarize recent advancements of our understanding of nicotine reward, tolerance and dependence. Based on these understandings, I propose a double target hypothesis, in which nAChRs and dopamine release process are two important targets for smoking cessation. Dysfunction of nAChRs (antagonism or desensitization) is crucial to abolish nicotine dependence and the maintenance of an appropriate level of extracellular dopamine eliminates nicotine withdrawal syndromes. Therefore, the medications simultaneously act on these two targets should have the desired effect for smoking cessation. I discuss how to use this double target concept to interpret recent therapies and to develop new candidate compounds for smoking cessation. Title: Pharmacological characterization of [(125)I]CHIBA-1006 binding, a new radioligand for alpha7 nicotinic acetylcholine receptors, to rat brain membranes Wu J, Toyohara J, Tanibuchi Y, Fujita Y, Zhang J, Chen H, Matsuo M, Wang RF, Hashimoto K Ref: Brain Research, 1360:130, 2010 : PubMed ESTHER: Wu_2010_Brain.Res_1360_130 PubMedSearch: Wu 2010 Brain.Res 1360 130 PubMedID: 20816767 Abstract The alpha7 nicotinic acetylcholine receptors (nAChRs) play an important role in the pathophysiology of neuropsychiatric diseases such as schizophrenia and Alzheimer's disease. However, there are currently no suitable small molecule radioligands for imaging alpha7 nAChRs in the brain. In this study, we synthesized the novel radioligand [(125)I]4-iodophenyl 1,4-diazaicyclo[3.2.2]nonane-4-carboxylate ([(125)I]CHIBA-1006), a iodine-derivative of the selective alpha7 nAChR agonist SSR180711, and studied the characterization of [(125)I]CHIBA-1006 binding to rat brain membranes. The assays of [(125)I]CHIBA-1006 binding to rat brain membranes were performed at 4 degrees C. The presence of a single saturable high-affinity binding component for [(125)I]CHIBA-1006 in the rat brain was shown. Scatchard analysis revealed an apparent equilibrium dissociation constant (K(d)) of 88.2+/-21.4nM and a maximal number of binding sites (B(max)) of 65.4+/-6.8fmol/mg protein (mean+/-SEM, n=4). The specific binding of [(125)I]CHIBA-1006 was inhibited by a number of alpha7 nAChR-selective ligands (e.g., unlabeled CHIBA-1006, SSR180711, CHIBA-1001, MG624 and A844606), suggesting a similarity among alpha7 nAChR pharmacological profiles. In contrast, alpha-bungarotoxin, MLA, and nicotine showed very weak affinity for [(125)I]CHIBA-1006 binding. The regional distribution of [(125)I]CHIBA-1006 binding to crude membranes from dissected regions of the rat brain was different from that of [(125)I]alpha-bungarotoxin binding, suggesting that [(125)I]CHIBA-1006 binding sites may not be identical to [(125)I]alpha-bungarotoxin binding sites in the rat brain. The present findings suggest that [(125)I]CHIBA-1006 would be a useful new small molecule radioligand for alpha7 nAChRs in the brain. Title: Comparative study of properties of immobilized lipase onto glutaraldehyde-activated amino-silica gel via different methods Yang G, Wu J, Xu G, Yang L Ref: Colloids Surf B Biointerfaces, 78:351, 2010 : PubMed ESTHER: Yang_2010_Colloids.Surf.B.Biointerfaces_78_351 PubMedSearch: Yang 2010 Colloids.Surf.B.Biointerfaces 78 351 PubMedID: 20399626 Abstract The enzyme-aggregate coating method was performed to immobilize Arthrobacter sp. lipase in order to achieve better catalytic properties comparable to the conventional covalent attachment and covalent attachment plus cross-linking. The glutaraldehyde-activated amino-silica gel which was synthesized by sol-gel technique was used as the support, and the catalytic characteristics of the lipase preparations were tested in the asymmetric acylation of 4-hydroxy-3-methyl-2-(2-propenyl)-2-cyclopenten-1-one (HMPC) in organic solvents. The results showed that the immobilized lipase by enzyme-aggregate coating possessed both higher activity and stability than those by other methods, e.g. it obtained an activity of 82.6 U/g and remained 42% and 93% of the original activity after incubation in vinyl acetate at 60 degrees C for 16 h and 9 times recycles, respectively, while the covalently attached lipase got an activity of 67.4 U/g and left 33% and 73% of the original under the same conditions, and the enzyme prepared by covalent attachment plus cross-linking exhibited the lowest activity yield. Moreover, excellent enantioselectivity (E > or =400) was achieved by all the three prepared lipases in our paper (E=85 for the free enzyme). Title: AChE deficiency or inhibition decreases apoptosis and p53 expression and protects renal function after ischemia/reperfusion Ye W, Gong X, Xie J, Wu J, Zhang X, Ouyang Q, Zhao X, Shi Y Ref: Apoptosis, 15:474, 2010 : PubMed ESTHER: Ye_2010_Apoptosis_15_474 PubMedSearch: Ye 2010 Apoptosis 15 474 PubMedID: 20054652 Abstract We recently reported that the expression of the synaptic form of acetylcholinesterase (AChE) is induced during apoptosis in various cell types in vitro. Here, we provide evidence to confirm that AChE is expressed during ischemia-reperfusion (I/R)-induced apoptosis in vivo. Renal I/R is a major cause of acute renal failure (ARF), resulting in injury and the eventual death of renal cells due to a combination of apoptosis and necrosis. Using AChE-deficient mice and AChE inhibitors, we investigated whether AChE deficiency or inhibition can protect against apoptosis caused by I/R in a murine kidney model. Unilateral clamping of renal pedicles for 90 min followed by reperfusion for 24 h caused significant renal dysfunction and injury. Both genetic AChE deficiency and chemical inhibition of AChE (provided by huperzine A, tacrine and donepezil) significantly reduced the biochemical and histological evidence of renal dysfunction following I/R. Activation of caspases-8, -9, -12, and -3 in vivo were prevented and associated with reduced levels of cell apoptosis and cell death. A further investigation also confirmed that AChE deficiency down-regulated p53 induction and phosphorylation at serine-15, and decreased the Bax/Bcl-2 ratio during I/R. In conclusion, our study demonstrates that AChE may be a pro-apoptotic factor and the inhibition of AChE reduces renal I/R injury. These findings suggest that AChE inhibitors may represent a therapeutic strategy for protection against ischemic acute renal failure. Title: Sequence analysis of pKF3-70 in Klebsiella pneumoniae: probable origin from R100-like plasmid of Escherichia coli Yi H, Xi Y, Liu J, Wang J, Wu J, Xu T, Chen W, Chen B, Lin M and Bao Q <5 more author(s)> Yi H, Xi Y, Liu J, Wang J, Wu J, Xu T, Chen W, Chen B, Lin M, Wang H, Zhou M, Li J, Xu Z, Jin S, Bao Q (- 5) Ref: PLoS ONE, 5:e8601, 2010 : PubMed ESTHER: Yi_2010_PLoS.One_5_e8601 PubMedSearch: Yi 2010 PLoS.One 5 e8601 PubMedID: 20066042 Abstract BACKGROUND: Klebsiella pneumoniae is a clinically significant species of bacterium which causes a variety of diseases. Clinical treatment of this bacterial infection is greatly hindered by the emergence of multidrug-resistant strains. The resistance is largely due to the acquisition of plasmids carrying drug-resistant as well as pathogenic genes, and its conjugal transfer facilitates the spread of resistant phenotypes. METHODOLOGY/PRINCIPAL FINDINGS: The 70,057 bp plasmid pKF3-70, commonly found in Klebsiella pneumoniae, is composed of five main functional modules, including regions involved in replication, partition, conjugation, transfer leading, and variable regions. This plasmid is more similar to several Escherichia coli plasmids than any previously reported K. pneumoniae plasmids and pKF3-70 like plasmids share a common and conserved backbone sequence. The replication system of the pKF3-70 is 100% identical to that of RepFII plasmid R100 from E. coli. A beta-lactamase gene ctx-m-14 with its surrounding insertion elements (ISEcp1, truncated IS903 and a 20 bp inverted repeat sequence) may compose an active transposon which is directly bordered by two putative target repeats "ATTAC." CONCLUSIONS/SIGNIFICANCE: The K. pneumoniae plasmid pKF3-70 carries an extended-spectrum beta-lactamase gene, ctx-m-14. The conjugative characteristic makes it a widespread plasmid among genetically relevant genera which poses significant threat to public health. Title: Characterization of Thermobifida fusca cutinase-carbohydrate-binding module fusion proteins and their potential application in bioscouring Zhang Y, Chen S, Xu M, Cavaco-Paulo A, Wu J, Chen J Ref: Applied Environmental Microbiology, 76:6870, 2010 : PubMed ESTHER: Zhang_2010_Appl.Environ.Microbiol_76_6870 PubMedSearch: Zhang 2010 Appl.Environ.Microbiol 76 6870 PubMedID: 20729325 Abstract Cutinase from Thermobifida fusca is thermally stable and has potential application in the bioscouring of cotton in the textile industry. In the present study, the carbohydrate-binding modules (CBMs) from T. fusca cellulase Cel6A (CBM(Cel6A)) and Cellulomonas fimi cellulase CenA (CBM(CenA)) were fused, separately, to the carboxyl terminus of T. fusca cutinase. Both fusion enzymes, cutinase-CBM(Cel6A) and cutinase-CBM(CenA), were expressed in Escherichia coli and purified to homogeneity. Enzyme characterization showed that both displayed similar catalytic properties and pH stabilities in response to T. fusca cutinase. In addition, both fusion proteins displayed an activity half-life of 53 h at their optimal temperature of 50 degrees C. Compared to T. fusca cutinase, in the absence of pectinase, the binding activity on cotton fiber was enhanced by 2% for cutinase-CBM(Cel6A) and by 28% for cutinase-CBM(CenA), whereas in the presence of pectinase, the binding activity was enhanced by 40% for the former and 45% for the latter. Notably, a dramatic increase of up to 3-fold was observed in the amount of released fatty acids from cotton fiber by both cutinase-CBM fusion proteins when acting in concert with pectinase. This is the first report of improving the scouring efficiency of cutinase by fusing it with CBM. The improvement in activity and the strong synergistic effect between the fusion proteins and pectinase suggest that they may have better applications in textile bioscouring than the native cutinase. Title: The anticonvulsive drug lamotrigine blocks neuronal {alpha}4{beta}2 nicotinic acetylcholine receptors Zheng C, Yang K, Liu Q, Wang MY, Shen J, Valles AS, Lukas RJ, Barrantes FJ, Wu J Ref: Journal of Pharmacology & Experimental Therapeutics, 335:401, 2010 : PubMed ESTHER: Zheng_2010_J.Pharmacol.Exp.Ther_335_401 PubMedSearch: Zheng 2010 J.Pharmacol.Exp.Ther 335 401 PubMedID: 20688974 Abstract Lamotrigine (LTG), an anticonvulsive drug, is often used for the treatment of a variety of epilepsies. In addition to block of sodium channels, LTG may act on other targets to exert its antiepileptic effect. In the present study, we evaluated the effects of LTG on neuronal nicotinic acetylcholine receptors (nAChRs) using the patch-clamp technique on human alpha4beta2-nAChRs heterologously expressed in the SH-EP1 cell line and on native alpha4beta2-nAChRs in dopaminergic (DA) neurons in rat ventral tegmental area (VTA). In SH-EP1 cells, LTG diminished the peak and steady-state components of the inward alpha4beta2-nAChR-mediated currents. This effect exhibited concentration-, voltage- and use-dependent behavior. Nicotine dose-response curves showed that in the presence of LTG, the nicotine-induced maximal current was reduced, suggesting a noncompetitive inhibition. These findings suggest that LTG inhibits human neuronal alpha4beta2-nAChR function through an open-channel blocking mechanism. LTG-induced inhibition in alpha4beta2-nAChRs was more profound when preceded by a 2-min pretreatment, after which the nicotine-induced current was reduced even without coapplication of LTG, suggesting that LTG is also able to inhibit alpha4beta2-nAChRs without channel activation. In freshly dissociated VTA DA neurons, LTG inhibited alpha4beta2-nAChR-mediated currents but did not affect glutamate- or GABA-induced currents, indicating that LTG selectively inhibits nAChR function. Collectively, our data suggest that the neuronal alpha4beta2-nAChR is likely an important target for mediating the anticonvulsive effect of LTG and the blockade of alpha4beta2-nAChR possibly underlying the mechanism through which LTG effectively controls some types of epilepsy, such as autosomal dominant nocturnal frontal lobe epilepsy or juvenile myoclonic epilepsy. Title: Nonlinear dynamical analysis of carbachol induced hippocampal oscillations in mice Akay M, Wang K, Akay YM, Dragomir A, Wu J Ref: Acta Pharmacol Sin, 30:859, 2009 : PubMed ESTHER: Akay_2009_Acta.Pharmacol.Sin_30_859 PubMedSearch: Akay 2009 Acta.Pharmacol.Sin 30 859 PubMedID: 19498425 Abstract AIM: Hippocampal neuronal network and synaptic impairment underlie learning and memory deficit in Alzheimer's disease (AD) patients and animal models. In this paper, we analyzed the dynamics and complexity of hippocampal neuronal network synchronization induced by acute exposure to carbachol, a nicotinic and muscarinic receptor co-agonist, using the nonlinear dynamical model based on the Lempel-Ziv estimator. We compared the dynamics of hippocampal oscillations between wild-type (WT) and triple-transgenic (3xTg) mice, as an AD animal model. We also compared these dynamic alterations between different age groups (5 and 10 months). We hypothesize that there is an impairment of complexity of CCh-induced hippocampal oscillations in 3xTg AD mice compared to WT mice, and that this impairment is age-dependent. Title: RanBPM is an acetylcholinesterase-interacting protein that translocates into the nucleus during apoptosis Gong X, Ye W, Zhou H, Ren X, Li Z, Zhou W, Wu J, Gong Y, Ouyang Q and Zhang X <1 more author(s)> Gong X, Ye W, Zhou H, Ren X, Li Z, Zhou W, Wu J, Gong Y, Ouyang Q, Zhao X, Zhang X (- 1) Ref: Acta Biochim Biophys Sin (Shanghai), 41:883, 2009 : PubMed ESTHER: Gong_2009_Acta.Biochim.Biophys.Sin.(Shanghai)_41_883 PubMedSearch: Gong 2009 Acta.Biochim.Biophys.Sin.(Shanghai) 41 883 PubMedID: 19902122 Abstract Acetylcholinesterase (AChE) expression may be induced during apoptosis in various cell types. Here, we used the C-terminal of AChE to screen the human fetal brain library and found that it interacted with Ran-binding protein in the microtubule-organizing center (RanBPM). This interaction was further confirmed by coimmunoprecipitation analysis. In HEK293T cells, RanBPM and AChE were heterogeneously expressed in the cisplatin-untreated cytoplasmic extracts and in the cisplatin-treated cytoplasmic or nuclear extracts. Our previous studies performed using morphologic methods have shown that AChE translocates from the cytoplasm to the nucleus during apoptosis. Taken together, these results suggest that RanBPM is an AChE-interacting protein that is translocated from the cytoplasm into the nucleus during apoptosis, similar to the translocation observed in case of AChE. Title: The genome of the cucumber, Cucumis sativus L Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B and Du Y <77 more author(s)> Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EA, Wu Y, Guo J, He J, Jia Z, Tian G, Lu Y, Ruan J, Qian W, Wang M, Huang Q, Li B, Xuan Z, Cao J, Asan, Wu Z, Zhang J, Cai Q, Bai Y, Zhao B, Han Y, Li Y, Li X, Wang S, Shi Q, Liu S, Cho WK, Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H, Li M, Liang H, Ren X, Shi Z, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Ma L, Liu H, Zhou Y, Zhao J, Fang X, Fang L, Liu D, Zheng H, Zhang Y, Qin N, Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Li S, Zhang X, Wang J, Sun R, Zhang B, Jiang S, Du Y (- 77) Ref: Nat Genet, 41:1275, 2009 : PubMed ESTHER: Huang_2009_Nat.Genet_41_1275 PubMedSearch: Huang 2009 Nat.Genet 41 1275 PubMedID: 19881527 Gene_locus related to this paper: cucsa-a0a0a0ktw5, cucsa-a0a0a0lnt6, cucsa-a0a0a0kpn7, cucsa-a0a0a0lvt9, cucsa-a0a0a0kdx8, cucsa-a0a0a0m228, cucsa-a0a0a0kz31, cucsa-a0a0a0k5t5, cucsa-a0a0a0kfs7, cucsa-a0a0a0kjj7, cucsa-a0a0a0kzs7, cucsa-a0a0a0l0a6, cucsa-a0a0a0l4w4, cucsa-a0a0a0lpz0, cucsa-a0a0a0ls66 Abstract Cucumber is an economically important crop as well as a model system for sex determination studies and plant vascular biology. Here we report the draft genome sequence of Cucumis sativus var. sativus L., assembled using a novel combination of traditional Sanger and next-generation Illumina GA sequencing technologies to obtain 72.2-fold genome coverage. The absence of recent whole-genome duplication, along with the presence of few tandem duplications, explains the small number of genes in the cucumber. Our study establishes that five of the cucumber's seven chromosomes arose from fusions of ten ancestral chromosomes after divergence from Cucumis melo. The sequenced cucumber genome affords insight into traits such as its sex expression, disease resistance, biosynthesis of cucurbitacin and 'fresh green' odor. We also identify 686 gene clusters related to phloem function. The cucumber genome provides a valuable resource for developing elite cultivars and for studying the evolution and function of the plant vascular system. Title: An organophosphorus hapten used in the preparation of monoclonal antibody and as an active immunization vaccine in the detoxication of soman poisoning Jia P, Wang Y, Yu M, Wu J, Yang R, Zhao Y, Zhou L Ref: Toxicol Lett, 187:45, 2009 : PubMed ESTHER: Jia_2009_Toxicol.Lett_187_45 PubMedSearch: Jia 2009 Toxicol.Lett 187 45 PubMedID: 19429243 Abstract Soman is an organophosphorus neurotoxin which inhibits the activity of acetylcholinesterase (AChE). The goal of this work was to find out whether antibodies against an organophosphorus hapten could protect mice from soman toxicity. An organophosphorus hapten P6 was synthesized. Its chemical conjugates with limulus polyphemus hemocyanin and bovine serum albumin were used as immune antigen (P6-LPH) and detection antigen (P6-BSA), respectively. Eight hybridoma cell lines secreting monoclonal antibodies (Mabs) were established. The binding reactivities of Mabs with P6 and soman were determined by competitive inhibition enzyme immunoassay (CIEIA). All antibodies recognized P6 and four of them (2C10, 3G1, 3B9 and 3C11) combined with soman. The IC(50) was 10(-6.5) to 10(-5.3)mol/l for P6 and 10(-5) to 10(-3.5)mol/l for soman. Furthermore, Mab 3G1 reduced the inhibition of AChE activity by soman in vitro. When soman was pre-incubated with Mabs before being injected into mice, soman potency was reduced, indicating that Mabs could protect mice from soman toxicity. In an active immunization regimen, mice immunized with P6-LPH and challenged with 0.15mg/kg soman injected subcutaneously, had fewer signs of intoxication and a higher survival rate compared with control mice. These results demonstrate that the anti-soman antibodies have proper characteristics as scavengers in the detoxication of soman poisoning. Title: A novel nicotinic acetylcholine receptor subtype in basal forebrain cholinergic neurons with high sensitivity to amyloid peptides Liu Q, Huang Y, Xue F, Simard AR, DeChon J, Li G, Zhang J, Lucero L, Wang M and Wu J <4 more author(s)> Liu Q, Huang Y, Xue F, Simard AR, DeChon J, Li G, Zhang J, Lucero L, Wang M, Sierks M, Hu G, Chang Y, Lukas RJ, Wu J (- 4) Ref: Journal of Neuroscience, 29:918, 2009 : PubMed ESTHER: Liu_2009_J.Neurosci_29_918 PubMedSearch: Liu 2009 J.Neurosci 29 918 PubMedID: 19176801 Abstract Nicotinic acetylcholine receptors (nAChRs) containing alpha7 subunits are thought to assemble as homomers. alpha7-nAChR function has been implicated in learning and memory, and alterations of alpha7-nAChR have been found in patients with Alzheimer's disease (AD). Here we report findings consistent with a novel, naturally occurring nAChR subtype in rodent, basal forebrain cholinergic neurons. In these cells, alpha7 subunits are coexpressed, colocalize, and coassemble with beta2 subunit(s). Compared with homomeric alpha7-nAChRs from ventral tegmental area neurons, functional, presumably heteromeric alpha7beta2-nAChRs on cholinergic neurons freshly dissociated from medial septum/diagonal band (MS/DB) exhibit relatively slow kinetics of whole-cell current responses to nicotinic agonists and are more sensitive to the beta2 subunit-containing nAChR-selective antagonist, dihydro-beta-erythroidine (DHbetaE). Interestingly, presumed, heteromeric alpha7beta2-nAChRs are highly sensitive to functional inhibition by pathologically relevant concentrations of oligomeric, but not monomeric or fibrillar, forms of amyloid beta(1-42) (Abeta(1-42)). Slow whole-cell current kinetics, sensitivity to DHbetaE, and specific antagonism by oligomeric Abeta(1-42) also are characteristics of heteromeric alpha7beta2-nAChRs, but not of homomeric alpha7-nAChRs, heterologously expressed in Xenopus oocytes. Moreover, choline-induced currents have faster kinetics and less sensitivity to Abeta when elicited from MS/DB neurons derived from nAChR beta2 subunit knock-out mice rather than from wild-type mice. The presence of novel, functional, heteromeric alpha7beta2-nAChRs on basal forebrain cholinergic neurons and their high sensitivity to blockade by low concentrations of oligomeric Abeta(1-42) suggests possible mechanisms for deficits in cholinergic signaling that could occur early in the etiopathogenesis of AD and might be targeted by disease therapies. Title: Defining pre-synaptic nicotinic receptors regulated by beta amyloid in mouse cortex and hippocampus with receptor null mutants Mehta TK, Dougherty JJ, Wu J, Choi CH, Khan GM, Nichols RA Ref: Journal of Neurochemistry, 109:1452, 2009 : PubMed ESTHER: Mehta_2009_J.Neurochem_109_1452 PubMedSearch: Mehta 2009 J.Neurochem 109 1452 PubMedID: 19457164 Abstract Disruption of neuronal signaling by soluble beta-amyloid has been implicated in deficits in short-term recall in the early stages of Alzheimer's disease. One potential target for beta-amyloid is the synapse, with evidence for differential interaction with both pre- and post-synaptic elements. Our previous work revealed an agonist-like action of soluble beta-amyloid (pM to nM) on isolated pre-synaptic terminals to increase [Ca(2+)]i, with apparent involvement of pre-synaptic nicotinic receptors. To directly establish the role of nicotinic receptors in pre-synaptic Ca(2+) regulation, we investigated the pre-synaptic action of beta-amyloid on terminals isolated from mice harboring either beta2 or alpha7 nicotinic receptor null mutants (knockouts). Average pre-synaptic responses to beta-amyloid in hippocampal terminals of alpha7 knockout mice were unchanged, whereas responses in hippocampal terminals from beta2 knockout mice were strongly attenuated. In contrast, pre-synaptic responses to soluble beta-amyloid were strongly attenuated in cortical terminals from alpha7 knockout mice but were moderately attenuated in cortical terminals from beta2 knockout mice. The latter responses, having distinct kinetics, were completely blocked by alpha-bungarotoxin. The use of receptor null mutants thus permitted direct demonstration of the involvement of specific nicotinic receptors in pre-synaptic Ca(2+) regulation by soluble beta-amyloid, and also indicated differential neuromodulation by beta-amyloid of synapses in hippocampus and cortex. Title: Nicotine and inflammatory neurological disorders Piao WH, Campagnolo D, Dayao C, Lukas RJ, Wu J, Shi FD Ref: Acta Pharmacol Sin, 30:715, 2009 : PubMed ESTHER: Piao_2009_Acta.Pharmacol.Sin_30_715 PubMedSearch: Piao 2009 Acta.Pharmacol.Sin 30 715 PubMedID: 19448649 Abstract Cigarette smoke is a major health risk factor which significantly increases the incidence of diseases including lung cancer and respiratory infections. However, there is increasing evidence that smokers have a lower incidence of some inflammatory and neurodegenerative diseases. Nicotine is the main immunosuppressive constituent of cigarette smoke, which inhibits both the innate and adaptive immune responses. Unlike cigarette smoke, nicotine is not yet considered to be a carcinogen and may, in fact, have therapeutic potential as a neuroprotective and anti-inflammatory agent. This review provides a synopsis summarizing the effects of nicotine on the immune system and its (nicotine) influences on various neurological diseases. Title: Cutinase-like proteins of Mycobacterium tuberculosis: characterization of their variable enzymatic functions and active site identification West NP, Chow FM, Randall EJ, Wu J, Chen J, Ribeiro JM, Britton WJ Ref: FASEB Journal, 23:1694, 2009 : PubMed ESTHER: West_2009_Faseb.J_23_1694 PubMedSearch: West 2009 Faseb.J 23 1694 PubMedID: 19225166 Gene_locus related to this paper: myctu-cut3, myctu-cutas1, myctu-cutas2, myctu-RV1758, myctu-RV3452, myctu-RV3724, myctu-Rv3802c Abstract Discovery and characterization of novel secreted enzymes of Mycobacterium tuberculosis are important for understanding the pathogenesis of one of the most important human bacterial pathogens. The proteome of M. tuberculosis contains over 400 potentially secreted proteins, the majority of which are uncharacterized. A family of seven cutinase-like proteins (CULPs) was identified by bioinformatic analysis, expressed and purified from Escherichia coli, and characterized in terms of their enzymatic activities. These studies revealed a functional diversity of enzyme classes based on differential preferences for substrate chain length. One member, Culp1, exhibited strong esterase activity, 40-fold higher than that of Culp6, which had strong activity as a lipase. Another, Culp4, performed moderately as an esterase and weakly as a lipase. Culp6 lipase activity was optimal above pH 7.0, and fully maintained to pH 8.5. None of the CULP members exhibited cutinase activity. Site-directed mutagenesis of each residue of the putative catalytic triad in Culp6 confirmed that each was essential for activity toward all fatty acid chain lengths of nitrophenyl esters and lipolytic function. Culp1 and Culp2 were present only in culture supernatants of M. tuberculosis, while Culp6, which is putatively essential for mycobacterial growth, was retained in the cell wall, suggesting the proteins play distinct roles in mycobacterial biology. Title: Understanding of nicotinic acetylcholine receptors Wu J Ref: Acta Pharmacol Sin, 30:653, 2009 : PubMed ESTHER: Wu_2009_Acta.Pharmacol.Sin_30_653 PubMedSearch: Wu 2009 Acta.Pharmacol.Sin 30 653 PubMedID: 19498413 Title: Nicotine modulates GABAergic transmission to dopaminergic neurons in substantia nigra pars compacta Xiao C, Yang KC, Zhou CY, Jin GZ, Wu J, Ye JH Ref: Acta Pharmacol Sin, 30:851, 2009 : PubMed ESTHER: Xiao_2009_Acta.Pharmacol.Sin_30_851 PubMedSearch: Xiao 2009 Acta.Pharmacol.Sin 30 851 PubMedID: 19498424 Abstract AIM: Dopaminergic neurons in the substantia nigra pars compacta (SNc) play important roles in motor control and drug addiction. As the major afferent, GABAergic innervation controls the activity of SNc dopaminergic neurons. Although it is clear that nicotine modulates SNc dopaminergic neurons by activating subtypes of somatodendritic nicotinic acetylcholine receptors (nAChRs), the detailed mechanisms of this activation remain to be addressed. Title: Impact of cigarette smoking in type 2 diabetes development Xie XT, Liu Q, Wu J, Wakui M Ref: Acta Pharmacol Sin, 30:784, 2009 : PubMed ESTHER: Xie_2009_Acta.Pharmacol.Sin_30_784 PubMedSearch: Xie 2009 Acta.Pharmacol.Sin 30 784 PubMedID: 19434055 Abstract Many patients with type 2 diabetes mellitus (DM2) are at risk for micro and macro vascular complications, which could be observed in heavy smokers. Cigarette smoking increases the risk for type 2 diabetes incidence. Nicotine, acknowledged as the major pharmacologically active chemical in tobacco, is responsible for the association between cigarette smoking and development of diabetes. This minireview summarized recent studies on nicotine effects on insulin action and insulin secretion, indicating the impact of nicotine on type 2 diabetes development. Title: Isolation and Characterization of a Methomyl-Degrading Paracoccus sp. mdw-1 Xu JL, Wu J, Wang ZC, Wang K, Li MY, Jiang JD, He J, Li SP Ref: Pedosphere, 19:238, 2009 : PubMed ESTHER: Xu_2009_Pedosphere_19_238 PubMedSearch: Xu 2009 Pedosphere 19 238 Abstract Methomyl, an extremely toxic pesticide, is widely used in agriculture. A strain named mdw-1 capable of degrading methomyl rapidly was successfully isolated from activated sludge in this study. It could utilize methomyl as the sole carbon or nitrogen source. The optimal temperature and medium pH for its growth and methomyl biodegradation were 30 C and 7.0, respectively. It was identified as a Paracoccus sp. according to its morphological features, physiological and biochemical characteristics, and phylogenetic analysis based on the sequence of 16S rDNA. Gas chromatography-mass spectrometry (GC-MS) analysis showed that methomyl could be completely transformed to S-methyl-N-hydroxythioacetamidate in 10 h of incubation with the isolate mdw-1. Title: Distinctive nicotinic acetylcholine receptor functional phenotypes of rat ventral tegmental area dopaminergic neurons Yang K, Hu J, Lucero L, Liu Q, Zheng C, Zhen X, Jin G, Lukas RJ, Wu J Ref: Journal of Physiology, 587:345, 2009 : PubMed ESTHER: Yang_2009_J.Physiol_587_345 PubMedSearch: Yang 2009 J.Physiol 587 345 PubMedID: 19047205 Abstract Dopaminergic (DAergic) neuronal activity in the ventral tegmental area (VTA) is thought to contribute generally to pleasure, reward, and drug reinforcement and has been implicated in nicotine dependence. nAChRs expressed in the VTA exhibit diverse subunit compositions, but the functional and pharmacological properties are largely unknown. Here, using patch-clamp recordings in single DAergic neurons freshly dissociated from rat VTA, we clarified three functional subtypes of nAChRs (termed ID, IID and IIID receptors) based on whole-cell current kinetics and pharmacology. Kinetic analysis demonstrated that comparing to ID, IID receptor-mediated current had faster activation and decay constant and IIID receptor-mediated current had larger current density. Pharmacologically, ID receptor-mediated current was sensitive to the alpha4beta2-nAChR agonist RJR-2403 and antagonist dihydro-beta-erythroidine (DHbetaE); IID receptor-mediated current was sensitive to the selective alpha7-nAChR agonist choline and antagonist methyllycaconitine (MLA); while IIID receptor-mediated current was sensitive to the beta4-containing nAChR agonist cytisine and antagonist mecamylamine (MEC). The agonist concentration-response relationships demonstrated that IID receptor-mediated current exhibited the highest EC(50) value compared to ID and IIID receptors, suggesting a relatively low agonist affinity of type IID receptors. These results suggest that the type ID, IID and IIID nAChR-mediated currents are predominately mediated by activation of alpha4beta2-nAChR, alpha7-nAChR and a novel nAChR subtype(s), respectively. Collectively, these findings indicate that the VTA DAergic neurons express diversity and multiplicity of functional nAChR subtypes. Interestingly, each DAergic neuron predominantly expresses only one particularly functional nAChR subtype, which may have distinct but important roles in regulation of VTA DA neuronal function, DA transmission and nicotine dependence. Title: Mysterious alpha6-containing nAChRs: function, pharmacology, and pathophysiology Yang KC, Jin GZ, Wu J Ref: Acta Pharmacol Sin, 30:740, 2009 : PubMed ESTHER: Yang_2009_Acta.Pharmacol.Sin_30_740 PubMedSearch: Yang 2009 Acta.Pharmacol.Sin 30 740 PubMedID: 19498417 Abstract Neuronal nicotinic acetylcholine receptors (nAChRs) are the superfamily of ligand-gated ion channels and widely expressed throughout the central and peripheral nervous systems. nAChRs play crucial roles in modulating a wide range of higher cognitive functions by mediating presynaptic, postsynaptic, and extrasynaptic signaling. Thus far, nine alpha (alpha2-alpha10) and three beta (beta2, beta3, and beta4) subunits have been identified in the CNS, and these subunits assemble to form a diversity of functional nAChRs. Although alpha4beta2- and alpha7-nAChRs are the two major functional nAChR types in the CNS, alpha6*-nAChRs are abundantly expressed in the midbrain dopaminergic (DAergic) system, including mesocorticolimbic and nigrostriatal pathways, and particularly present in presynaptic nerve terminals. Recently, functional and pharmacological profiles of alpha6*-nAChRs have been assessed with the use of alpha6 subunit blockers such as alpha-conotoxin MII and PIA, and also by using alpha6 subunit knockout mice. By modulating DA release in the nucleus accumbens (NAc) and modulating GABA release onto DAergic neurons in the ventral tegmental area (VTA), alpha6*-nAChRs may play important roles in the mediation of nicotine reward and addiction. Furthermore, alpha6*-nAChRs in the nigrostriatal DAergic system may be promising targets for selective preventative treatment of Parkinson's disease (PD). Thus, alpha6*-nAChRs may hold promise for future clinical treatment of human disorders, such as nicotine addiction and PD. In this review, we mainly focus on the recent advances in the understanding of alpha6*-nAChR function, pharmacology and pathophysiology. Title: Kinetics of desensitization and recovery from desensitization for human alpha4beta2-nicotinic acetylcholine receptors stably expressed in SH-EP1 cells Yu KD, Liu Q, Wu J, Lukas RJ Ref: Acta Pharmacol Sin, 30:805, 2009 : PubMed ESTHER: Yu_2009_Acta.Pharmacol.Sin_30_805 PubMedSearch: Yu 2009 Acta.Pharmacol.Sin 30 805 PubMedID: 19498421 Abstract AIM: Studies were conducted to define the kinetics of the onset of and recovery from desensitization for human alpha4beta2-nicotinic acetylcholine receptors (nAChR) heterologously expressed in the SH-EP1 human epithelial cell line. Title: U18666A, a cholesterol-inhibition agent, modulates human neuronal nicotinic acetylcholine receptors heterologously expressed in SH-EP1 cell line Zheng C, Wang MY, Liu Q, Wakui M, Whiteaker P, Lukas RJ, Wu J Ref: Journal of Neurochemistry, 108:1526, 2009 : PubMed ESTHER: Zheng_2009_J.Neurochem_108_1526 PubMedSearch: Zheng 2009 J.Neurochem 108 1526 PubMedID: 19183258 Abstract In this study, we evaluate the effects of (3beta)-3-[2-(diethylamino)ethoxy]androst-5-en-17-one dihydrochloride (U18666A), a cholesterol synthesis/transporter inhibitor, on selected human neuronal nicotinic acetylcholine receptors (nAChRs) heterologously expressed in the SH-EP1 cell line using whole-cell patch-clamp recordings. The results indicate that with 2-min pretreatment, U18666A inhibited different nAChR subtypes with a rank-order of potency (IC(50) of whole-cell peak current): alpha4beta2 (8.0 +/- 3.0 nM) > alpha3beta2 (1.7 +/- 0.4 microM) > alpha4beta4 (26 +/- 7.2 microM) > alpha7 (> 100 microM), suggesting this compound is more selective to alpha4beta2-nAChRs. Thus, the pharmacological profiles and mechanisms of U18666A acting on alpha4beta2-nAChRs were investigated in detail. U18666A suppresses both peak and steady state components of whole-cell currents mediated by human alpha4beta2-nAChRs in response to nicotine. In nicotine-induced concentration-response curves, U18666A reduces nicotine-induced current at maximally effective agonist concentrations without influencing nicotine's EC(50) value, suggesting a non-competitive inhibition. U18666A-induced inhibition of nAChR function is concentration-, voltage-, and use-dependent, suggesting an open channel block. Taken into consideration of approximately 10 000-fold enhancement of the potency of U18666A after 2-min pre-treatment, this compound also likely inhibits alpha4beta2-nAChRs through a close channel block. In addition, the U18666A-induced inhibition in alpha4beta2-nAChRs is not mediated by either increased receptor endocytosis or altered cell cholesterol. These data indicate that U18666A is a potent antagonist of alpha4beta2-nAChRs and may be useful as a tool in the functional characterization and pharmacological profiling of nAChRs, as well as a potential candidate for smoking cessation. Title: Identification and characterization of bacterial cutinase Chen S, Tong X, Woodard RW, Du G, Wu J, Chen J Ref: Journal of Biological Chemistry, 283:25854, 2008 : PubMed ESTHER: Chen_2008_J.Biol.Chem_283_25854 PubMedSearch: Chen 2008 J.Biol.Chem 283 25854 PubMedID: 18658138 Gene_locus related to this paper: thefu-q6a0i4, thefu-q6a0i3 Abstract Cutinase, which exists in both fungi and bacteria, catalyzes the cleavage of the ester bonds of cutin. Fungal cutinases have been extensively studied, however, reports on bacterial cutinases have been limited due to the lack of knowledge concerning the identity of their open reading frames. In the present study, the cutinase from Thermobifida fusca was induced by cutin and purified to homogeneity by following p-nitrophenyl butyrate hydrolyzing activity. Peptide mass fingerprinting analysis of the wild-type enzyme matched two proteins, Tfu_0883 and Tfu_0882, which are 93% identical in sequence. Both proteins were cloned and overexpressed in their mature form. Recombinant Tfu_0883 and Tfu_0882 display very similar enzymatic properties and were confirmed to be cutinases by their capability to hydrolyze the ester bonds of cutin. Comparative characterization of Fusarium solani pisi and T. fusca cutinases indicated that they have similar substrate specificity and catalytic properties except that the T. fusca enzymes are thermally more stable. Homology modeling revealed that T. fusca cutinases adopt an alpha/beta-hydrolase fold that exhibits both similarities and variations from the fungal cutinase structure. A serine hydrolase catalytic mechanism involving a Ser(170)-His(248)-Asp(216) (Tfu_0883 numbering) catalytic triad was supported by active site-directed inhibition studies and mutational analyses. This is the first report of cutinase encoding genes from bacterial sources. Title: Chronic nicotine alters nicotinic receptor-induced presynaptic Ca2+ responses in isolated nerve terminals Dougherty JJ, Wu J, Mehta TK, Brown B, Nichols RA Ref: Neurochem Res, 33:1106, 2008 : PubMed ESTHER: Dougherty_2008_Neurochem.Res_33_1106 PubMedSearch: Dougherty 2008 Neurochem.Res 33 1106 PubMedID: 18095155 Abstract Brain nicotinic receptors display pronounced permeability for Ca2+ and localize to presynaptic nerve terminals, in addition to postsynaptic sites. Chronic exposure to nicotine has been shown to alter brain nicotinic receptor expression, but the functional consequences for presynaptic Ca2+ have not been directly examined. Here, we used confocal imaging to assess Ca2+ responses in individual nerve terminals from cortices of mice treated up to 14 days with nicotine as compared to vehicle-treated controls. Chronic nicotine treatment led to substantially enhanced amplitudes of presynaptic Ca2+ responses to acute application of nicotine at concentrations of 50 nM (2-fold) and 500 nM (1.7-fold), but not 50 microM. In addition, increased expression of high-affinity nicotinic receptors on isolated terminals was observed following chronic treatment, as determined immunocytochemically and pharmacologically. These findings suggest that chronic exposure to nicotine may lead to enhanced sensitivity to nicotine at select presynaptic sites in brain via up-regulation of high-affinity nicotinic receptors. Title: GSK3beta mediates the induced expression of synaptic acetylcholinesterase during apoptosis Jing P, Jin Q, Wu J, Zhang XJ Ref: Journal of Neurochemistry, 104:409, 2008 : PubMed ESTHER: Jing_2008_J.Neurochem_104_409 PubMedSearch: Jing 2008 J.Neurochem 104 409 PubMedID: 17949411 Abstract Besides its role in terminating acetylcholine-mediated neurotransmission, acetylcholinesterase (AChE) is found to be expressed and participate in the process of apoptosis in various cell types. However, the mechanisms underlying AChE up-regulation in neuronal cells remain elusive. Herein we demonstrated that glycogen synthase kinase-3beta (GSK3beta) mediates induced AChE-S expression during apoptosis. In this study, A23187 and thapsigargin (TG) were employed to induce apoptosis in neuroendocrine PC12 cells. The results showed that exposure of PC12 cells to A23187 and TG up-regulated AChE activity significantly. The same treatment also led to activation of GSK3beta. Two different inhibitors of GSK3beta (lithium and GSK3beta-specific inhibitor VIII) could block A23187- or TG-induced up-regulation of AChE activity, AChE-S mRNA level and protein expression. However, lithium could not inhibit the induction of AChE-R mRNA and protein under similar conditions. Taken together, our results show that GSK3beta is specifically involved in the induction of AChE-S expression in PC12 cells during apoptosis. Title: Discovery of new binding elements in DPP-4 inhibition and their applications in novel DPP-4 inhibitor design Liang GB, Qian X, Biftu T, Singh S, Gao YD, Scapin G, Patel S, Leiting B, Patel R and Weber AE <3 more author(s)> Liang GB, Qian X, Biftu T, Singh S, Gao YD, Scapin G, Patel S, Leiting B, Patel R, Wu J, Zhang X, Thornberry NA, Weber AE (- 3) Ref: Bioorganic & Medicinal Chemistry Lett, 18:3706, 2008 : PubMed ESTHER: Liang_2008_Bioorg.Med.Chem.Lett_18_3706 PubMedSearch: Liang 2008 Bioorg.Med.Chem.Lett 18 3706 PubMedID: 18524582 Gene_locus related to this paper: human-DPP4 Inhibitor(s) related to this paper: DB07181 Abstract Probing with tool molecules, and by modeling and X-ray crystallography the binding modes of two structurally distinct series of DPP-4 inhibitors led to the discovery of a rare aromatic fluorine H-bond and the spatial requirement for better biaryl binding in the DPP-4 enzyme active site. These newly found binding elements were successfully incorporated into novel DPP-4 inhibitors. Title: Agonist-induced hump current production in heterologously-expressed human alpha4beta2-nicotinic acetylcholine receptors Liu Q, Yu KW, Chang YC, Lukas RJ, Wu J Ref: Acta Pharmacol Sin, 29:305, 2008 : PubMed ESTHER: Liu_2008_Acta.Pharmacol.Sin_29_305 PubMedSearch: Liu 2008 Acta.Pharmacol.Sin 29 305 PubMedID: 18298895 Abstract AIM: To characterize the functional and pharmacological features of agonist-induced hump currents in human alpha4beta2-nicotinic acetylcholine receptors (nAChR). Title: Encapsulated in silica: genome, proteome and physiology of the thermophilic bacterium Anoxybacillus flavithermus WK1 Saw JH, Mountain BW, Feng L, Omelchenko MV, Hou S, Saito JA, Stott MB, Li D, Zhao G and Alam M <8 more author(s)> Saw JH, Mountain BW, Feng L, Omelchenko MV, Hou S, Saito JA, Stott MB, Li D, Zhao G, Wu J, Galperin MY, Koonin EV, Makarova KS, Wolf YI, Rigden DJ, Dunfield PF, Wang L, Alam M (- 8) Ref: Genome Biol, 9:R161, 2008 : PubMed ESTHER: Saw_2008_Genome.Biol_9_R161 PubMedSearch: Saw 2008 Genome.Biol 9 R161 PubMedID: 19014707 Gene_locus related to this paper: anofw-b7ghw8, anofw-b7gk10, anofw-b7gk45, anofw-b7gln5, anofw-b7glx5, anofw-b7gm70 Abstract BACKGROUND: Gram-positive bacteria of the genus Anoxybacillus have been found in diverse thermophilic habitats, such as geothermal hot springs and manure, and in processed foods such as gelatin and milk powder. Anoxybacillus flavithermus is a facultatively anaerobic bacterium found in super-saturated silica solutions and in opaline silica sinter. The ability of A. flavithermus to grow in super-saturated silica solutions makes it an ideal subject to study the processes of sinter formation, which might be similar to the biomineralization processes that occurred at the dawn of life. RESULTS: We report here the complete genome sequence of A. flavithermus strain WK1, isolated from the waste water drain at the Wairakei geothermal power station in New Zealand. It consists of a single chromosome of 2,846,746 base pairs and is predicted to encode 2,863 proteins. In silico genome analysis identified several enzymes that could be involved in silica adaptation and biofilm formation, and their predicted functions were experimentally validated in vitro. Proteomic analysis confirmed the regulation of biofilm-related proteins and crucial enzymes for the synthesis of long-chain polyamines as constituents of silica nanospheres. CONCLUSIONS: Microbial fossils preserved in silica and silica sinters are excellent objects for studying ancient life, a new paleobiological frontier. An integrated analysis of the A. flavithermus genome and proteome provides the first glimpse of metabolic adaptation during silicification and sinter formation. Comparative genome analysis suggests an extensive gene loss in the Anoxybacillus/Geobacillus branch after its divergence from other bacilli. Title: A monoclonal antibody against synaptic AChE: a useful tool for detecting apoptotic cells Su W, Wu J, Ye WY, Zhang XJ Ref: Chemico-Biological Interactions, 175:101, 2008 : PubMed ESTHER: Su_2008_Chem.Biol.Interact_175_101 PubMedSearch: Su 2008 Chem.Biol.Interact 175 101 PubMedID: 18538755 Abstract The classical function of acetylcholinesterase (AChE) is to terminate synaptic transmission at cholinergic synapses by rapidly hydrolyzing the neurotransmitter acetylcholine (ACh). Non-classical functions of AChE involve accelerating the assembly of Abeta peptide into amyloid fibrils and participating in haematopoiesis and neurite growth. Although numerous antibodies have been raised against AChE, many researchers have questioned their reliability to identify the AChE in situ, especially with the regard to its non-classical roles. Researchers attended the Ninth International Meeting on Cholinesterase raised this question by showing different Western blot patterns of AChE detected by different Abs. Producing more effective and reliable Abs for measuring AChE in vivo or in situ has become an important issue in many scientific fields. In this paper, we introduce a monoclonal antibody raised against synaptic AChE that we identified by Western blot assays, immunofluorescent staining and immunoprecipitation of AChE, and mass spectrometry. Our results strongly demonstrate the specificity of our monoclonal antibody to recognize synaptic AChE; hence our antibody can be used as an effective tool to study the various functions of AChE. Since the apoptosis-related AChE was its synaptic form, our antibody can be used as a tool to detect apoptotic cells. Title: The Rice Annotation Project Database (RAP-DB): 2008 update Tanaka T, Antonio BA, Kikuchi S, Matsumoto T, Nagamura Y, Numa H, Sakai H, Wu J, Itoh T and Echeverria M <80 more author(s)> Tanaka T, Antonio BA, Kikuchi S, Matsumoto T, Nagamura Y, Numa H, Sakai H, Wu J, Itoh T, Sasaki T, Aono R, Fujii Y, Habara T, Harada E, Kanno M, Kawahara Y, Kawashima H, Kubooka H, Matsuya A, Nakaoka H, Saichi N, Sanbonmatsu R, Sato Y, Shinso Y, Suzuki M, Takeda J, Tanino M, Todokoro F, Yamaguchi K, Yamamoto N, Yamasaki C, Imanishi T, Okido T, Tada M, Ikeo K, Tateno Y, Gojobori T, Lin YC, Wei FJ, Hsing YI, Zhao Q, Han B, Kramer MR, McCombie RW, Lonsdale D, O'Donovan CC, Whitfield EJ, Apweiler R, Koyanagi KO, Khurana JP, Raghuvanshi S, Singh NK, Tyagi AK, Haberer G, Fujisawa M, Hosokawa S, Ito Y, Ikawa H, Shibata M, Yamamoto M, Bruskiewich RM, Hoen DR, Bureau TE, Namiki N, Ohyanagi H, Sakai Y, Nobushima S, Sakata K, Barrero RA, Souvorov A, Smith-White B, Tatusova T, An S, An G, S OO, Fuks G, Messing J, Christie KR, Lieberherr D, Kim H, Zuccolo A, Wing RA, Nobuta K, Green PJ, Lu C, Meyers BC, Chaparro C, Piegu B, Panaud O, Echeverria M (- 80) Ref: Nucleic Acids Research, 36:D1028, 2008 : PubMed ESTHER: Tanaka_2008_Nucleic.Acids.Res_36_D1028 PubMedSearch: Tanaka 2008 Nucleic.Acids.Res 36 D1028 PubMedID: 18089549 Gene_locus related to this paper: orysa-Q9FW17, orysa-Q0JK71, orysa-B9EWJ8, orysa-Q5N7L1, orysa-pir7a, orysa-q2qyj1, orysj-q6yse8, orysa-q6yzk1, orysa-Q8S0U8, orysa-q33aq0, orysa-Q0J0A4, orysi-a2z179, orysi-a2zef2, orysi-b8a7e6, orysi-b8a7e7, orysi-b8bfe5, orysi-b8bhp9, orysj-b9fi05, orysj-b9fkb0, orysj-cgep, orysj-q0djj0, orysj-q0dud7, orysj-q0jaf0, orysj-q0jga1, orysj-q5jl22, orysj-q5jlw7, orysj-q6h7q9, orysj-q6yvk6, orysj-q7f8x1, orysj-q7xcx3, orysj-q9fwm6, orysj-q10j20, orysj-q10ss2, orysj-q69uw6, orysj-q94d71, orysj-q0iq98, orysj-b9gbs4, orysj-b9gbs1, orysj-pla4, orysj-pla1 Abstract The Rice Annotation Project Database (RAP-DB) was created to provide the genome sequence assembly of the International Rice Genome Sequencing Project (IRGSP), manually curated annotation of the sequence, and other genomics information that could be useful for comprehensive understanding of the rice biology. Since the last publication of the RAP-DB, the IRGSP genome has been revised and reassembled. In addition, a large number of rice-expressed sequence tags have been released, and functional genomics resources have been produced worldwide. Thus, we have thoroughly updated our genome annotation by manual curation of all the functional descriptions of rice genes. The latest version of the RAP-DB contains a variety of annotation data as follows: clone positions, structures and functions of 31 439 genes validated by cDNAs, RNA genes detected by massively parallel signature sequencing (MPSS) technology and sequence similarity, flanking sequences of mutant lines, transposable elements, etc. Other annotation data such as Gnomon can be displayed along with those of RAP for comparison. We have also developed a new keyword search system to allow the user to access useful information. The RAP-DB is available at: http://rapdb.dna.affrc.go.jp/ and http://rapdb.lab.nig.ac.jp/. Title: Methyl jasmonate-elicited herbivore resistance: does MeJA function as a signal without being hydrolyzed to JA? Wu J, Wang L, Baldwin IT Ref: Planta, 227:1161, 2008 : PubMed ESTHER: Wu_2008_Planta_227_1161 PubMedSearch: Wu 2008 Planta 227 1161 PubMedID: 18214527 Abstract Treatment with methyl jasmonate (MeJA) elicits herbivore resistance in many plant species and over-expression of JA carboxyl methyltransferase (JMT) constitutively increases JA-induced responses in Arabidopsis. When wild-type (WT) Nicotiana attenuata plants are treated with MeJA, a rapid transient endogenous JA burst is elicited, which in turn increases levels of nicotine and trypsin proteinase inhibitors (TPIs) and resistance to larvae of the specialist herbivore, Manduca sexta. All of these responses are impaired in plants silenced in lipoxygenase 3 expression (asLOX3) but are restored to WT levels by MeJA treatment. Whether these MeJA-induced responses are directly elicited by MeJA or by its cleavage product, JA, is unknown. Using virus-induced gene silencing (VIGS), we silenced MeJA-esterase (NaMJE) expression and found this gene responsible for most of the MeJA-cleaving activity in N. attenuata protein extracts. Silencing NaMJE in asLOX3, but not in WT plants, significantly reduced MeJA-induced nicotine levels and resistance to M. sexta, but not TPI levels. MeJA-induced transcript levels of threonine deaminase (NaTD) and phenylalanine ammonia lyase (NaPAL1) were also decreased in VIGS MJE (asLOX3) plants. Finally the performance of M. sexta larvae that fed on plants treated with JA or MeJA demonstrated that silencing NaMJE inhibited MeJA-induced but not JA-induced resistance in asLOX3 plants. From these results, we conclude that the resistance elicited by MeJA treatment is directly elicited not by MeJA but by its de-methylated product, JA. Title: Hepatic lipase gene -514C/T polymorphism in the Guangxi Hei Yi Zhuang and Han populations Wu J, Yin R, Lin W, Pan S, Yang D Ref: Lipids, 43:733, 2008 : PubMed ESTHER: Wu_2008_Lipids_43_733 PubMedSearch: Wu 2008 Lipids 43 733 PubMedID: 18592285 Gene_locus related to this paper: human-LIPC Abstract Hei Yi Zhuang is an isolated subgroup of the Zhuang minority in China. This study was designed to compare the difference in the hepatic lipase gene (LIPC) -514C/T polymorphism and its association with lipid profiles between the Guangxi Hei Yi Zhuang and Han populations. Genotyping of the LIPC -514C/T was performed in 873 subjects of Hei Yi Zhuang and 867 participants of Han Chinese. The frequency of -514T allele was 43.47% in Hei Yi Zhuang, and 36.10% in Han (P < 0.001). The frequencies of CC, CT and TT genotypes were 30.01, 53.04 and 16.95% in Hei Yi Zhuang, and 40.95, 45.91 and 13.14% in Han (P < 0.001); respectively. Serum high-density lipoprotein cholesterol (HDL-C) and apolipoprotein B levels in both ethnic groups were higher in LIPC -514T carriers than in C carriers. In addition, serum triglyceride levels in Han were higher in TT genotype individuals than in CC genotype subjects (P < 0.05). Serum HDL-C levels were positively correlated with age, alcohol consumption and LIPC -514C/T genotypes, and negatively associated with hypertension and cigarette smoking in Hei Yi Zhuang (P < 0.05-0.01), whereas HDL-C levels were positively correlated with age, alcohol consumption and LIPC -514C/T genotypes, and negatively associated with body mass index and cigarette smoking in Han (P < 0.05-0.001). The differences in serum HDL-C levels between the two ethnic groups might partially attribute to the differences in the LIPC -514C/T polymorphism. Title: The JNK/AP1/ATF2 pathway is involved in H2O2-induced acetylcholinesterase expression during apoptosis Zhang JY, Jiang H, Gao W, Wu J, Peng K, Shi YF, Zhang XJ Ref: Cell Mol Life Sciences, 65:1435, 2008 : PubMed ESTHER: Zhang_2008_Cell.Mol.Life.Sci_65_1435 PubMedSearch: Zhang 2008 Cell.Mol.Life.Sci 65 1435 PubMedID: 18385943 Abstract We show that H2O2 increases acetylcholinesterase (AChE) expression via transcriptional activation through c-Jun N-terminal kinase (JNK), since the JNK inhibitor SP600125, but not the extracellular signal-regulated kinase (ERK) pathway inhibitor PD98059 or p38 kinase inhibitor SB203580, attenuated H2O2-induced AChE expression and its promoter activity. Overexpression of hemagglutinin (HA)-JNK increases H2O2-induced AChE expression and its promoter activity, whereas the dominant negative mutant form of JNK suppressed H2O2-induced AChE expression and promoter activity. Mutation analysis indicates that the major response elements for JNK in the AChE promoter are the AP1-like element (TGAGTCT) site, located within the -1565/-1569 region of the AChE promoter, and the ATF2 element (CCACGTCA), within the -2185/-2177 region. The AP1-like element binds to the transcription factors, c-jun and ATF2, while the ATF2 element binds mainly ATF2. Taken together, our results strongly suggest that H2O2 induces AChE expression via the JNK/AP1/ ATF2 signaling pathway. Title: (3R)-4-[(3R)-3-Amino-4-(2,4,5-trifluorophenyl)butanoyl]-3-(2,2,2-trifluoroethyl)- 1,4-diazepan-2-one, a selective dipeptidyl peptidase IV inhibitor for the treatment of type 2 diabetes Biftu T, Feng D, Qian X, Liang GB, Kieczykowski G, Eiermann G, He H, Leiting B, Lyons K and Weber AE <10 more author(s)> Biftu T, Feng D, Qian X, Liang GB, Kieczykowski G, Eiermann G, He H, Leiting B, Lyons K, Petrov A, Sinha-Roy R, Zhang B, Scapin G, Patel S, Gao YD, Singh S, Wu J, Zhang X, Thornberry NA, Weber AE (- 10) Ref: Bioorganic & Medicinal Chemistry Lett, 17:49, 2007 : PubMed ESTHER: Biftu_2007_Bioorg.Med.Chem.Lett_17_49 PubMedSearch: Biftu 2007 Bioorg.Med.Chem.Lett 17 49 PubMedID: 17055272 Gene_locus related to this paper: human-DPP4 Inhibitor(s) related to this paper: Diazepanone-Analog-18, Diazepanone-Analog-1 Abstract Replacement of the triazolopiperazine ring of sitagliptin (DPP-4 IC(50)=18nM) with 3-(2,2,2-trifluoroethyl)-1,4-diazepan-2-one gave dipeptidyl peptidase IV (DPP-4) inhibitor 1 which is potent (DPP-4 IC(50)=2.6nM), selective, and efficacious in an oral glucose tolerance test in mice. It was selected for extensive preclinical development as a potential back-up candidate to sitagliptin. Title: Rational design of a novel, potent, and orally bioavailable cyclohexylamine DPP-4 inhibitor by application of molecular modeling and X-ray crystallography of sitagliptin Biftu T, Scapin G, Singh S, Feng D, Becker JW, Eiermann G, He H, Lyons K, Patel S and Weber AE <7 more author(s)> Biftu T, Scapin G, Singh S, Feng D, Becker JW, Eiermann G, He H, Lyons K, Patel S, Petrov A, Sinha-Roy R, Zhang B, Wu J, Zhang X, Doss GA, Thornberry NA, Weber AE (- 7) Ref: Bioorganic & Medicinal Chemistry Lett, 17:3384, 2007 : PubMed ESTHER: Biftu_2007_Bioorg.Med.Chem.Lett_17_3384 PubMedSearch: Biftu 2007 Bioorg.Med.Chem.Lett 17 3384 PubMedID: 17433672 Gene_locus related to this paper: human-DPP4 Inhibitor(s) related to this paper: CHEMBL233360, Sitagliptin Abstract Molecular modeling was used to design a rigid analog of sitagliptin 1. The X-ray crystal structure of sitagliptin bound to DPP-4 suggested that the central beta-amino butyl amide moiety could be replaced with a cyclohexylamine group. This was confirmed by structural analysis and the resulting analog 2a was synthesized and found to be a potent DPP-4 inhibitor (IC(50)=21 nM) with excellent in vivo activity and pharmacokinetic profile. Title: Optimization of 1,4-diazepan-2-one containing dipeptidyl peptidase IV inhibitors for the treatment of type 2 diabetes Liang GB, Qian X, Feng D, Biftu T, Eiermann G, He H, Leiting B, Lyons K, Petrov A and Weber AE <5 more author(s)> Liang GB, Qian X, Feng D, Biftu T, Eiermann G, He H, Leiting B, Lyons K, Petrov A, Sinha-Roy R, Zhang B, Wu J, Zhang X, Thornberry NA, Weber AE (- 5) Ref: Bioorganic & Medicinal Chemistry Lett, 17:1903, 2007 : PubMed ESTHER: Liang_2007_Bioorg.Med.Chem.Lett_17_1903 PubMedSearch: Liang 2007 Bioorg.Med.Chem.Lett 17 1903 PubMedID: 17291750 Inhibitor(s) related to this paper: Diazepanone-Analog-18, Diazepanone-Analog-1 Abstract Following the discovery of N-acyl-1,4-diazepan-2-one as a novel pharmacophore for potent and selective DPP-4 inhibitors, optimization of this new lead with different substitution on the seven-membered ring resulted in several highly potent and selective, orally bioavailable, and efficacious DPP-4 inhibitors, such as 3R-methyl-1-cyclopropyl-1,4-diazepan-2-one derivative 9i (DPP-4 IC(50)=8.0 nM) and 3R,6R-dimethyl-1,4-diazepan-2-one derivative 14a (DPP-4 IC(50)=9.7 nM). Title: A constitutive, transient receptor potential-like Ca2+ influx pathway in presynaptic nerve endings independent of voltage-gated Ca2+ channels and Na+/Ca2+ exchange Nichols RA, Dengler AF, Nakagawa EM, Bashkin M, Paul BT, Wu J, Khan GM Ref: Journal of Biological Chemistry, 282:36102, 2007 : PubMed ESTHER: Nichols_2007_J.Biol.Chem_282_36102 PubMedSearch: Nichols 2007 J.Biol.Chem 282 36102 PubMedID: 17928293 Abstract Calcium levels in the presynaptic nerve terminal are altered by several pathways, including voltage-gated Ca(2+) channels, the Na(+)/Ca(2+) exchanger, Ca(2+)-ATPase, and the mitochondria. The influx pathway for homeostatic control of [Ca(2+)](i) in the nerve terminal has been unclear. One approach to detecting the pathway that maintains internal Ca(2+) is to test for activation of Ca(2+) influx following Ca(2+) depletion. Here, we demonstrate that a constitutive influx pathway for Ca(2+) exists in presynaptic terminals to maintain internal Ca(2+) independent of voltage-gated Ca(2+) channels and Na(+)/Ca(2+) exchange, as measured in intact isolated nerve endings from mouse cortex and in intact varicosities in a neuronal cell line using fluorescence spectroscopy and confocal imaging. The Mg(2+) and lanthanide sensitivity of the influx pathway, in addition to its pharmacological and short hairpin RNA sensitivity, and the results of immunostaining for transient receptor potential (TRP) channels indicate the involvement of TRPC channels, possibly TRPC5 and TRPC1. This constitutive Ca(2+) influx pathway likely serves to maintain synaptic function under widely varying levels of synaptic activity. Title: Improvement in extraction and catalytic activity of Mucor javanicus lipase by modification of AOT reverse micelle Talukder MR, Susanto D, Feng G, Wu J, Choi WJ, Chow Y Ref: Biotechnol J, 2:1369, 2007 : PubMed ESTHER: Talukder_2007_Biotechnol.J_2_1369 PubMedSearch: Talukder 2007 Biotechnol.J 2 1369 PubMedID: 17639532 Abstract Reverse micelles are formed in apolar solvents by spontaneous aggregation of surfactants. Surfactant sodium bis (2-ethylhexyl) sulfosuccinate (AOT) is most often used for the reverse micellar extraction of enzymes. However, the inactivation of enzyme due to strong interaction with AOT molecules is a severe problem. To overcome this problem, the AOT/water/isooctane reverse micellar system was modified by adding short chain polyethylene glycol 400 (PEG 400). The modified AOT reverse micellar system was used to extract Mucor javanicus lipase from the aqueous phase to the reverse micellar phase. The extraction efficiency (E) increased with the increase in PEG 400 addition and the maximum E in PEG 400 modified system was twofold higher than that in the PEG 400-free system. Upon addition of PEG 400, the water activity (a(w)) of aqueous phase decreased, whereas a(w) of reverse micellar phase increased. The circular dichroism spectroscopy analysis revealed that PEG 400 changes the secondary and tertiary structure of lipase. The maximum specific activity of lipase extracted in PEG 400-modified reverse micellar system was threefold higher than that in the PEG-free system. Title: Dopamine release in prefrontal cortex in response to beta-amyloid activation of alpha7 * nicotinic receptors Wu J, Khan GM, Nichols RA Ref: Brain Research, 1182:82, 2007 : PubMed ESTHER: Wu_2007_Brain.Res_1182_82 PubMedSearch: Wu 2007 Brain.Res 1182 82 PubMedID: 17935702 Abstract The levels of soluble beta-amyloid (Abeta) are correlated with symptom severity in Alzheimer's disease. Soluble Abeta has been shown to disrupt synaptic function and it has been proposed that accumulation of soluble Abeta triggers synapse loss over the course of the disease. Numerous studies indicate that soluble Abeta has multiple targets, one of which appears to be the nicotinic acetylcholine receptor, particularly for Abeta concentrations of pM to nM. Moreover, pM to nM soluble Abeta was found to increase presynaptic Ca(2+) levels, suggesting that it may have an impact on neurotransmitter release. In the present study, soluble Abeta was perfused into mouse prefrontal cortex and the effect on the release of dopamine outflow via microdialysis was assessed. In the presence of tetrodotoxin, Abeta(1-42) at 100 nM evoked the release of dopamine to approximately 170% of basal levels. The Abeta(1-42)-evoked dopamine release was sensitive to antagonists of alpha7 nicotinic receptors and was absent in mice harboring a null mutation for the alpha7 nicotinic subunit, but was intact in mice harboring a null mutation for the beta2 nicotinic subunit. The control peptide Abeta(40-1) was without effect. In contrast, Abeta(1-42) at 1-10 pM caused a profound but slowly developing decrease in dopamine outflow. These results suggest that Abeta alters dopamine release in mouse prefrontal cortex, perhaps involving distinct targets as it accumulates during Alzheimer's disease and leading to disruption of synaptic signaling. Title: A gene linB2 responsible for the conversion of beta-HCH and 2,3,4,5,6-pentachlorocyclohexanol in Sphingomonas sp. BHC-A Wu J, Hong Q, Han P, He J, Li S Ref: Applied Microbiology & Biotechnology, 73:1097, 2007 : PubMed ESTHER: Wu_2007_Appl.Microbiol.Biotechnol_73_1097 PubMedSearch: Wu 2007 Appl.Microbiol.Biotechnol 73 1097 PubMedID: 16977465 Gene_locus related to this paper: sphpi-linb Abstract Commercial formulations of hexachlorocyclohexane (HCH) consist of a mixture of four isomers: alpha, beta, gamma, and delta. All four isomers are toxic and recalcitrant pollutants. beta-HCH is more problematic due to its longer persistence in the environment. Sphingomonas sp. BHC-A was able to degrade not only alpha-, gamma-, and delta-HCH but also beta-HCH. To clone a gene responsible for the degradation of beta-HCH, a Tn5 mutation was introduced into BHC-A, and one mutant BHC-A45 defective in beta-HCH degradation was selected. Sequencing analysis showed this mutant had a Tn5 insertion at the site of one haloalkane dehalogenase gene, designated linB2. linB2 was overexpressed in Escherichia coli and the 32-kDa product LinB2 showed the conversion activity of not only beta-HCH to beta-2,3,4,5,6-pentachlorocyclohexanol (beta-PCHL) but also beta-PCHL to beta-2,3,5,6-tetrachloro-1,4-cyclohexanediol. Title: Analysis of the role of LinA and LinB in biodegradation of delta-hexachlorocyclohexane Wu J, Hong Q, Sun Y, Hong Y, Yan Q, Li S Ref: Environ Microbiol, 9:2331, 2007 : PubMed ESTHER: Wu_2007_Environ.Microbiol_9_2331 PubMedSearch: Wu 2007 Environ.Microbiol 9 2331 PubMedID: 17686029 Abstract Commercial formulations of hexachlorocyclohexane (HCH) consist of a mixture of four isomers, alpha, beta, gamma and delta. All these four isomers are toxic and recalcitrant pollutants. Sphingobium (formerly Sphingomonas) sp. strain BHC-A is able to degrade all four HCH isomers. Eight lin genes responsible for the degradation of gamma-HCH in BHC-A were cloned and analysed for their role in the degradation of delta-HCH, and the initial conversion steps in delta-HCH catabolism by LinA and LinB in BHC-A were found. LinA dehydrochlorinated delta-HCH to produce 1,3,4,6-tetrachloro-1,4-cyclohexadiene (1,4-TCDN) via delta-pentachlorocyclohexene (delta-PCCH). Subsequently, both 1,4-TCDN and delta-PCCH are catalysed by LinB via two successive rounds of hydrolytic dechlorinations to form 2,5-dichloro-2,5-cyclohexadiene-1,4-diol (2,5-DDOL) and 2,3,5-trichloro-5-cyclohexene-1,4-diol (2,3,5-TCDL) respectively. LinB could also catalyse the hydrolytic dechlorination of delta-HCH to 2,3,5,6-tetrachloro-1,4-cyclohexanediol (TDOL) via 2,3,4,5,6-pentachlorocyclohexanol (PCHL). Title: Calcineurin mediates acetylcholinesterase expression during calcium ionophore A23187-induced HeLa cell apoptosis Zhu H, Gao W, Jiang H, Wu J, Shi YF, Zhang XJ Ref: Biochimica & Biophysica Acta, 1773:593, 2007 : PubMed ESTHER: Zhu_2007_Biochim.Biophys.Acta_1773_593 PubMedSearch: Zhu 2007 Biochim.Biophys.Acta 1773 593 PubMedID: 17320203 Abstract We previously reported that acetylcholinesterase plays a critical role in apoptosis and its expression is regulated by Ca(2+) mobilization. In the present study, we show that activated calpain, a cytosolic calcium-activated cysteine protease, and calcineurin, a calcium-dependent protein phosphatase, regulate acetylcholinesterase expression during A23187-induced apoptosis. The calpain inhibitor, calpeptin, and the calcineurin inhibitors, FK506 and cyclosporine A, inhibited acetylcholinesterase expression at both mRNA and protein levels and suppressed the activity of the human acetylcholinesterase promoter. In contrast, overexpression of constitutively active calcineurin significantly activated the acetylcholinesterase promoter. Furthermore, we identify a role for the transcription factor NFAT (nuclear factor of activated T cells), a calcineurin target, in regulating the acetylcholinesterase promoter during ionophore-induced apoptosis. Overexpression of human NFATc3 and NFATc4 greatly increased the acetylcholinesterase promoter activity in HeLa cells treated with A23187. Overexpression of constitutive nuclear NFATc4 activated the acetylcholinesterase promoter independent of A23187, whereas overexpression of dominant-negative NFAT blocked A23187-induced acetylcholinesterase promoter activation. These results indicate that calcineurin mediates acetylcholinesterase expression during apoptosis. Title: Solution structure of GOPC PDZ domain and its interaction with the C-terminal motif of neuroligin Li X, Zhang J, Cao Z, Wu J, Shi Y Ref: Protein Science, 15:2149, 2006 : PubMed ESTHER: Li_2006_Protein.Sci_15_2149 PubMedSearch: Li 2006 Protein.Sci 15 2149 PubMedID: 16882988 Gene_locus related to this paper: human-NLGN1 Abstract GOPC (Golgi-associated PDZ and coiled-coil motif-containing protein) represents a PDZ domain-containing protein associated with the Golgi apparatus, which plays important roles in vesicular trafficking in secretory and endocytic pathways. GOPC interacts with many other proteins, such as the Wnt receptors Frizzled 8 and neuroligin via its PDZ domain. Neuroligin is a neural cell-adhesion molecule of the post-synapse, which binds to the presynapse molecule neurexin to form a heterotypic intercellular junction. Here we report the solution structure of the GOPC PDZ domain by NMR. Our results show that it is a canonical class I PDZ domain, which contains two alpha-helices and six beta-strands. Using chemical shift perturbation experiments, we further studied the binding properties of the GOPC PDZ domain with the C-terminal motif of neuroligin. The observations showed that the ensemble of the interaction belongs to fast exchange with low affinity. The 3D model of the GOPC PDZ domain/neuroligin C-terminal peptide complex was constructed with the aid of the molecular dynamics simulation method. Our discoveries provide insight into the specific interaction of the GOPC PDZ domain with the C-terminal peptide of Nlg and also provide a general insight about the possible binding mode of the interaction of Nlg with other PDZ domain-containing proteins. Title: Iptakalim inhibits nicotine-induced enhancement of extracellular dopamine and glutamate levels in the nucleus accumbens of rats Liu Q, Li Z, Ding JH, Liu SY, Wu J, Hu G Ref: Brain Research, 1085:138, 2006 : PubMed ESTHER: Liu_2006_Brain.Res_1085_138 PubMedSearch: Liu 2006 Brain.Res 1085 138 PubMedID: 16647046 Abstract Iptakalim (Ipt) is a novel ATP-sensitive potassium channel opener. It has been reported that Ipt inhibited cocaine-induced dopamine and glutamate release, suggesting that Ipt may regulate drug addiction. Recently, we found that Ipt blocked nicotinic acetylcholine receptor (nAChR)-mediated currents in a heterologously expressed SH-EP1 cell line and in native midbrain dopamine neurons. In the present study, we examined whether Ipt prevents nicotine-induced neurotransmitter release in the nucleus accumbens (NAc) using in vivo microdialysis methods in awake, freely moving rats. Ipt was administered through a microdialysis probe, following systemic administration of nicotine (0.5 mg/kg, s.c.). The results show that acute nicotine treatment induced an increase of both dopamine and glutamate levels in the rat NAc, and that Ipt significantly attenuated nicotine's effects in a concentration-dependent manner. Therefore, Ipt may serve as a novel compound to block nicotine-induced dopamine and glutamate release in the brain reward center, in turn decreasing nicotine reinforcement and dependence. Title: Neuronal nicotinic acetylcholine receptors serve as sensitive targets that mediate beta-amyloid neurotoxicity Liu Q, Wu J Ref: Acta Pharmacol Sin, 27:1277, 2006 : PubMed ESTHER: Liu_2006_Acta.Pharmacol.Sin_27_1277 PubMedSearch: Liu 2006 Acta.Pharmacol.Sin 27 1277 PubMedID: 17007734 Abstract Alzheimer's disease (AD) is the most common form of brain dementia characterized by the accumulation of beta-amyloid peptides (Abeta) and loss of forebrain cholinergic neurons. Abeta accumulation and aggregation are thought to contribute to cholinergic neuronal degeneration, in turn causing learning and memory deficits, but the specific targets that mediate Abeta neurotoxicity remain elusive. Recently, accumulating lines of evidence have demonstrated that Abeta directly modulates the function of neuronal nicotinic acetylcholine receptors (nAChRs), which leads to the new hypothesis that neuronal nAChRs may serve as important targets that mediate Abeta neurotoxicity. In this review, we summarize current studies performed in our laboratory and in others to address the question of how Abeta modulates neuronal nAChRs, especially nAChR subunit function. Title: Roles of nicotinic acetylcholine receptor beta subunits in function of human alpha4-containing nicotinic receptors Wu J, Liu Q, Yu K, Hu J, Kuo YP, Segerberg M, St John PA, Lukas RJ Ref: Journal of Physiology, 576:103, 2006 : PubMed ESTHER: Wu_2006_J.Physiol_576_103 PubMedSearch: Wu 2006 J.Physiol 576 103 PubMedID: 16825297 Abstract Naturally expressed nicotinic acetylcholine receptors (nAChR) containing alpha4 subunits (alpha4*-nAChR) in combination with beta2 subunits (alpha4beta2-nAChR) are among the most abundant, high-affinity nicotine binding sites in the mammalian brain. beta4 subunits are also richly expressed and colocalize with alpha4 subunits in several brain regions implicated in behavioural responses to nicotine and nicotine dependence. Thus, alpha4beta4-nAChR also may exist and play important functional roles. In this study, properties were determined of human alpha4beta2- and alpha4beta4-nAChR heterologously expressed de novo in human SH-EP1 epithelial cells. Whole-cell currents mediated via human alpha4beta4-nAChR have approximately 4-fold higher amplitude than those mediated via human alpha4beta2-nAChR and exhibit much slower acute desensitization and functional rundown. Nicotinic agonists induce peak whole-cell current responses typically with higher functional potency at alpha4beta4-nAChR than at alpha4beta2-nAChR. Cytisine and lobeline serve as full agonists at alpha4beta4-nAChR but are only partial agonists at alpha4beta2-nAChR. However, nicotinic antagonists, except hexamethonium, have comparable affinities for functional alpha4beta2- and alpha4beta4-nAChR. Whole-cell current responses show stronger inward rectification for alpha4beta2-nAChR than for alpha4beta4-nAChR at a positive holding potential. Collectively, these findings demonstrate that human nAChR beta2 or beta4 subunits can combine with alpha4 subunits to generate two forms of alpha4*-nAChR with distinctive physiological and pharmacological features. Diversity in alpha4*-nAChR is of potential relevance to nervous system function, disease, and nicotine dependence. Title: Nanofibrous poly(acrylonitrile-co-maleic acid) membranes functionalized with gelatin and chitosan for lipase immobilization Ye P, Xu ZK, Wu J, Innocent C, Seta P Ref: Biomaterials, 27:4169, 2006 : PubMed ESTHER: Ye_2006_Biomaterials_27_4169 PubMedSearch: Ye 2006 Biomaterials 27 4169 PubMedID: 16584770 Abstract Nanofibrous membranes with an average diameter of 100 and 180 nm were fabricated from poly(acrylonitrile-co-maleic acid) (PANCMA) by the electrospinning process. These nanofibrous membranes contain reactive groups which can be used to covalently immobilize biomacromolecules. Two natural macromolecules, chitosan and gelatin, were tethered on these nanofibrous membranes to fabricate dual-layer biomimetic supports for enzyme immobilization in the presence of 1-ethyl-3-(dimethyl-aminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxyl succinimide (NHS). Lipase from Candida rugosa was then immobilized on these dual-layer biomimetic supports using glutaraldehyde (GA), and on the nascent PANCMA fibrous membrane using EDC/NHS as coupling agent, respectively. The properties of the immobilized lipases were assayed. It was found that there is an increase of the activity retention of the immobilized lipase on the chitosan-modified nanofibrous membrane (45.6+/-1.8%) and on the gelatin-modified one (49.7+/-1.8%), compared to that on the nascent one (37.6+/-1.8%). The kinetic parameters of the free and immobilized lipases, K(m) and V(max), were also assayed. In comparison with the immobilized lipase on the nascent nanofibrous membrane, there is an increase of the V(max) value for the immobilized lipases on the chitosan- and gelatin-modified nanofibrous membranes. Results also indicate that the pH and thermal stabilities of lipases increase upon immobilization. The residual activities of the immobilized lipases are 55% on the chitosan-modified nanofibrous membrane and 60% on the gelatin-modified one, after 10 uses. Title: A fine physical map of the rice chromosome 5 Cheng CH, Chung MC, Liu SM, Chen SK, Kao FY, Lin SJ, Hsiao SH, Tseng IC, Hsing YI and Chow TY <7 more author(s)> Cheng CH, Chung MC, Liu SM, Chen SK, Kao FY, Lin SJ, Hsiao SH, Tseng IC, Hsing YI, Wu HP, Chen CS, Shaw JF, Wu J, Matsumoto T, Sasaki T, Chen HH, Chow TY (- 7) Ref: Mol Genet Genomics, 274:337, 2005 : PubMed ESTHER: Cheng_2005_Mol.Genet.Genomics_274_337 PubMedSearch: Cheng 2005 Mol.Genet.Genomics 274 337 PubMedID: 16261349 Gene_locus related to this paper: orysa-gid1, orysa-q6f358, orysj-PLA7 Abstract A fine physical map of the rice (Oryza sativa spp. Japonica var. Nipponbare) chromosome 5 with bacterial artificial chromosome (BAC) and PI-derived artificial chromosome (PAC) clones was constructed through integration of 280 sequenced BAC/PAC clones and 232 sequence tagged site/expressed sequence tag markers with the use of fingerprinted contig data of the Nipponbare genome. This map consists of five contigs covering 99% of the estimated chromosome size (30.08 Mb). The four physical gaps were estimated at 30 and 20 kb for gaps 1-3 and gap 4, respectively. We have submitted 42.2-Mb sequences with 29.8 Mb of nonoverlapping sequences to public databases. BAC clones corresponding to telomere and centromere regions were confirmed by BAC-fluorescence in situ hybridization (FISH) on a pachytene chromosome. The genetically centromeric region at 54.6 cM was covered by a minimum tiling path spanning 2.1 Mb with no physical gaps. The precise position of the centromere was revealed by using three overlapping BAC/PACs for approximately 150 kb. In addition, FISH results revealed uneven chromatin condensation around the centromeric region at the pachytene stage. This map is of use for positional cloning and further characterization of the rice functional genomics. Title: Roles for nicotinic acetylcholine receptor subunit large cytoplasmic loop sequences in receptor expression and function Kuo YP, Xu L, Eaton JB, Zhao L, Wu J, Lukas RJ Ref: Journal of Pharmacology & Experimental Therapeutics, 314:455, 2005 : PubMed ESTHER: Kuo_2005_J.Pharmacol.Exp.Ther_314_455 PubMedSearch: Kuo 2005 J.Pharmacol.Exp.Ther 314 455 PubMedID: 15833891 Abstract To evaluate possible physiological roles of the large cytoplasmic loops (C2) and neighboring transmembrane domains of nicotinic acetylcholine receptor (nAChR) subunits, we generated novel fusion constructs in which human nAChR alpha4, beta2, or beta4 subunit C2 or C2 and neighboring sequences were replaced by corresponding sequences from the mouse serotonin type 3A (5-HT3A) receptor subunit. Following stable expression in human SH-EP1 cells, we found that extensive sequence substitutions involving third and fourth transmembrane domains and neighboring "proximal" C2 sequences (e.g., beta2 H322-V335 and V449-R460) did not allow functional expression of nAChR containing chimeric subunits. However, expression of functional nAChR was achieved containing wild-type alpha4 subunits and chimeric beta2 (beta2chi) subunits whose "nested" C2 domain sequences K336-S448 were replaced with the corresponding 5-HT3A subunit sequences. Whereas these findings suggested indispensable roles for M3/M4 transmembrane and/or proximal C2 sequences in alpha4beta2-nAChR function, nested C2 sequences in the beta2 subunit are not essential for functional receptor expression. Ligand-binding analyses also revealed only subtle differences in pharmacological profiles of alpha4beta2-nAChR compared with alpha4beta2chi-nAChR. Nevertheless, there was heightened emergence of agonist-mediated self-inhibition of alpha4beta2chi function, greater sensitivity to functional blockade by a number of antagonists, and faster and more complete acute desensitization of alpha4beta2chi-nAChR than for alpha4beta2-nAChR. These studies are consistent with unexpected roles of nested C2 sequences in nAChR function. Title: High-resolution functional proteomics by active-site peptide profiling Okerberg ES, Wu J, Zhang B, Samii B, Blackford K, Winn DT, Shreder KR, Burbaum JJ, Patricelli MP Ref: Proc Natl Acad Sci U S A, 102:4996, 2005 : PubMed ESTHER: Okerberg_2005_Proc.Natl.Acad.Sci.U.S.A_102_4996 PubMedSearch: Okerberg 2005 Proc.Natl.Acad.Sci.U.S.A 102 4996 PubMedID: 15795380 Abstract Characterization and functional annotation of the large number of proteins predicted from genome sequencing projects poses a major scientific challenge. Whereas several proteomics techniques have been developed to quantify the abundance of proteins, these methods provide little information regarding protein function. Here, we present a gel-free platform that permits ultrasensitive, quantitative, and high-resolution analyses of protein activities in proteomes, including highly problematic samples such as undiluted plasma. We demonstrate the value of this platform for the discovery of both disease-related enzyme activities and specific inhibitors that target these proteins. Title: Role of alpha7-nicotinic acetylcholine receptors in tetanic stimulation-induced gamma oscillations in rat hippocampal slices Song C, Murray TA, Kimura R, Wakui M, Ellsworth K, Javedan SP, Marxer-Miller S, Lukas RJ, Wu J Ref: Neuropharmacology, 48:869, 2005 : PubMed ESTHER: Song_2005_Neuropharmacol_48_869 PubMedSearch: Song 2005 Neuropharmacol 48 869 PubMedID: 15829257 Abstract Hippocampal gamma oscillations, as a form of neuronal network synchronization, are speculated to be associated with learning, memory and attention. Nicotinic acetylcholine receptor alpha7 subtypes (alpha7-nAChRs) are highly expressed in hippocampal neurons and play important roles in modulating neuronal function, synaptic plasticity, learning and memory. However, little is known about the role of alpha7-nAChRs in hippocampal gamma oscillations. Here, we examined the effects of selective alpha7- and non-alpha7-nAChR antagonists on tetanic gamma oscillations in rat hippocampal slices. We found that brief tetanic stimulation-induced gamma oscillations (30-80 Hz) and pharmacological blockade of alpha7-nAChRs using the relatively selective alpha7-nAChR antagonists, methyllycaconitine (10 or 100 nM) or alpha-bungarotoxin (10 nM), significantly reduced the frequency spectrum power, the number of spikes, and burst duration of evoked gamma oscillations. Neither mecamylamine nor dihydro-beta-erythroidine, which are selective antagonists of non-alpha7-nAChRs, demonstrated significant effects on tetanic gamma oscillations. Nicotine exposure promotes hippocampal gamma oscillations in a methyllycaconitine-sensitive manner. It is concluded that alpha7-nAChRs in hippocampal slices play important roles in regulation of gamma oscillations, thus potentially helping to explain roles of nAChRs in cognitive functions such as learning, memory and attention. Title: The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC) Gerhard DS, Wagner L, Feingold EA, Shenmen CM, Grouse LH, Schuler G, Klein SL, Old S, Rasooly R and Malek J <104 more author(s)> Gerhard DS, Wagner L, Feingold EA, Shenmen CM, Grouse LH, Schuler G, Klein SL, Old S, Rasooly R, Good P, Guyer M, Peck AM, Derge JG, Lipman D, Collins FS, Jang W, Sherry S, Feolo M, Misquitta L, Lee E, Rotmistrovsky K, Greenhut SF, Schaefer CF, Buetow K, Bonner TI, Haussler D, Kent J, Kiekhaus M, Furey T, Brent M, Prange C, Schreiber K, Shapiro N, Bhat NK, Hopkins RF, Hsie F, Driscoll T, Soares MB, Casavant TL, Scheetz TE, Brown-stein MJ, Usdin TB, Toshiyuki S, Carninci P, Piao Y, Dudekula DB, Ko MS, Kawakami K, Suzuki Y, Sugano S, Gruber CE, Smith MR, Simmons B, Moore T, Waterman R, Johnson SL, Ruan Y, Wei CL, Mathavan S, Gunaratne PH, Wu J, Garcia AM, Hulyk SW, Fuh E, Yuan Y, Sneed A, Kowis C, Hodgson A, Muzny DM, McPherson J, Gibbs RA, Fahey J, Helton E, Ketteman M, Madan A, Rodrigues S, Sanchez A, Whiting M, Madari A, Young AC, Wetherby KD, Granite SJ, Kwong PN, Brinkley CP, Pearson RL, Bouffard GG, Blakesly RW, Green ED, Dickson MC, Rodriguez AC, Grimwood J, Schmutz J, Myers RM, Butterfield YS, Griffith M, Griffith OL, Krzywinski MI, Liao N, Morin R, Palmquist D, Petrescu AS, Skalska U, Smailus DE, Stott JM, Schnerch A, Schein JE, Jones SJ, Holt RA, Baross A, Marra MA, Clifton S, Makowski KA, Bosak S, Malek J (- 104) Ref: Genome Res, 14:2121, 2004 : PubMed ESTHER: Gerhard_2004_Genome.Res_14_2121 PubMedSearch: Gerhard 2004 Genome.Res 14 2121 PubMedID: 15489334 Gene_locus related to this paper: human-AFMID, human-CES4A, human-CES5A, human-NOTUM, human-SERAC1, human-SERHL2, human-TMEM53, mouse-acot1, mouse-adcl4, mouse-Ces2f, mouse-Ces4a, mouse-notum, mouse-q6wqj1, mouse-Q9DAI6, mouse-rbbp9, mouse-SERHL, mouse-srac1, mouse-tmm53, rat-abhd6, rat-abhda, rat-abhea, rat-abheb, rat-Ldah, rat-cd029, rat-estd, rat-Kansl3, rat-nceh1, ratno-acph, ratno-CMBL, mouse-b2rwd2, rat-b5den3, rat-ab17c Abstract The National Institutes of Health's Mammalian Gene Collection (MGC) project was designed to generate and sequence a publicly accessible cDNA resource containing a complete open reading frame (ORF) for every human and mouse gene. The project initially used a random strategy to select clones from a large number of cDNA libraries from diverse tissues. Candidate clones were chosen based on 5'-EST sequences, and then fully sequenced to high accuracy and analyzed by algorithms developed for this project. Currently, more than 11,000 human and 10,000 mouse genes are represented in MGC by at least one clone with a full ORF. The random selection approach is now reaching a saturation point, and a transition to protocols targeted at the missing transcripts is now required to complete the mouse and human collections. Comparison of the sequence of the MGC clones to reference genome sequences reveals that most cDNA clones are of very high sequence quality, although it is likely that some cDNAs may carry missense variants as a consequence of experimental artifact, such as PCR, cloning, or reverse transcriptase errors. Recently, a rat cDNA component was added to the project, and ongoing frog (Xenopus) and zebrafish (Danio) cDNA projects were expanded to take advantage of the high-throughput MGC pipeline. Title: Electrophysiological, pharmacological, and molecular evidence for alpha7-nicotinic acetylcholine receptors in rat midbrain dopamine neurons Wu J, George AA, Schroeder KM, Xu L, Marxer-Miller S, Lucero L, Lukas RJ Ref: Journal of Pharmacology & Experimental Therapeutics, 311:80, 2004 : PubMed ESTHER: Wu_2004_J.Pharmacol.Exp.Ther_311_80 PubMedSearch: Wu 2004 J.Pharmacol.Exp.Ther 311 80 PubMedID: 15178698 Abstract Dopamine (DA) neurons located in the mammalian midbrain have been generally implicated in reward and drug reinforcement and more specifically in nicotine dependence. However, roles played by nicotinic acetylcholine receptors, including those composed of alpha7-subunits [alpha7-nicotinic acetylcholine receptors (nAChRs)], in modulation of DA signaling and in nicotine dependence are not clearly understood. Although midbrain slice recording has been used previously to identify functional alpha7-nAChRs, these preparations are not optimally designed for extremely rapid and reproducible drug application, and rapidly desensitized, alpha7-nAChR-mediated currents may have been underestimated or not detected. Here, we use patch-clamp, whole-cell current recordings from single neurons acutely dissociated from midbrain nuclei and having features of DA neurons to characterize acetylcholine-induced, inward currents that rapidly activate and desensitize, are mimicked by the alpha7-nAChR-selective agonist, choline, blocked by the alpha7-nAChR-selective antagonists, methyllycaconitine and alpha-bungarotoxin, and are similar to those of heterologously expressed, human alpha7-nAChRs. We also use reverse transcriptase-polymerase chain reaction, in situ hybridization, and immunocytochemical staining to demonstrate nAChR alpha7 subunit gene expression as message and protein in the rat substantia nigra pars compacta and ventral tegmental area. Expression of alpha7 subunit message and of alpha7-nAChR-mediated responses is developmentally regulated, with both being absent in samples taken from rats at postnatal day 7, but later becoming present and increasing over the next 2 weeks. Collectively, this electrophysiological, pharmacological, and molecular evidence indicates that nAChR alpha7 subunits and functional alpha7-nAChRs are expressed somatodendritically by midbrain DA neurons, where they may play important physiological roles and contribute to nicotine reinforcement and dependence. Title: beta-Amyloid directly inhibits human alpha4beta2-nicotinic acetylcholine receptors heterologously expressed in human SH-EP1 cells Wu J, Kuo YP, George AA, Xu L, Hu J, Lukas RJ Ref: Journal of Biological Chemistry, 279:37842, 2004 : PubMed ESTHER: Wu_2004_J.Biol.Chem_279_37842 PubMedSearch: Wu 2004 J.Biol.Chem 279 37842 PubMedID: 15234980 Abstract Amyloid-beta (Abeta) accumulation and aggregation are thought to contribute to the pathogenesis of Alzheimer's disease (AD). In AD, there is a selective decrease in the numbers of radioligand binding sites corresponding to the most abundant nicotinic acetylcholine receptor (nAChR) subtype, which contains human alpha4 and beta2 subunits (halpha4beta2-nAChR). However, the relationships between these phenomena are uncertain, and effects of Abeta on halpha4beta2-nAChR function have not been investigated in detail. We first confirmed expression of halpha4 and hbeta2 subunits as messenger RNA in transfected, human SHEP1 cells by reverse transcription-polymerase chain reaction and mRNA fluorescence in situ hybridization analyses. Immunoprecipitation Western analyses confirmed alpha4 and beta2 subunit protein expression and co-assembly. Whole cell current recording demonstrated heterologous expression in SH-EP1-halpha4beta2 cells of functional halpha4beta2-nAChRs with characteristic responses to nicotinic agonists or antagonists. Nicotine-induced whole cell currents were suppressed by Abeta(1-42) in a dose-dependent manner. Functional inhibition was selective for Abeta(1-42) compared with the functionally inactive, control peptide Abeta(40-1).Abeta(1-42)-mediated inhibition of halpha4beta2-nAChR function was non-competitive, voltage-independent, and use-independent. Pre-loading of cells with guanyl-5'-yl thiophosphate failed to prevent Abeta(1-42)-induced inhibition, suggesting that down-regulation of halpha4beta2-nAChR function by Abeta(1-42) is not mediated by nAChR internalization. Sensitivity to Abeta(1-42) antagonism at 1 nm was evident for halpha4beta2-nAChRs, but not for heterologously expressed human alpha7-nAChRs, although both nAChR subtypes were functionally inhibited by 100 nm Abeta(1-42), with the magnitude of functional block being higher for 100 nm Abeta(1-42) acting on halpha7-nAChRs. These findings suggest that halpha4beta2-nAChRs are sensitive and perhaps pathophysiologically relevant targets for Abeta neurotoxicity in AD. Title: Enhanced activity and enantioselectivity of Candida rugosa lipase immobilized on macroporous adsorptive resins for ibuprofen resolution Yu H, Wu J, Ching CB Ref: Biotechnol Lett, 26:629, 2004 : PubMed ESTHER: Yu_2004_Biotechnol.Lett_26_629 PubMedSearch: Yu 2004 Biotechnol.Lett 26 629 PubMedID: 15200171 Abstract The lipase from Candida rugosa was immobilized on three commercially available macroporous adsorptive resins for kinetic resolution of ibuprofen. One resin, CRB02, increased the enzyme activity by 50% to 0.027 g g(-1) min(-1). The deactivation constant (0.19 h(-1)) of the immobilized enzyme was half of that of the native enzyme and the enantioselectivity (E = 29.2) of the immobilized lipase was 2.2 times as much as that of the native lipase for the kinetic resolution of ibuprofen with 1-propanol in isooctane at 30 degrees C. Title: Beta-amyloid regulation of presynaptic nicotinic receptors in rat hippocampus and neocortex Dougherty JJ, Wu J, Nichols RA Ref: Journal of Neuroscience, 23:6740, 2003 : PubMed ESTHER: Dougherty_2003_J.Neurosci_23_6740 PubMedSearch: Dougherty 2003 J.Neurosci 23 6740 PubMedID: 12890766 Abstract Alteration by beta-amyloid (Abeta) of signaling via nicotinic acetylcholine receptors (nAChRs) has been implicated in the early stages of Alzheimer's disease. nAChRs function both post- and presynaptically in the nervous system; however, little is known about the functional consequence of the interaction of Abeta with these receptors, particularly those on presynaptic nerve terminals. In view of the strong correlation between loss of synaptic terminals and dementia, together with the reduction in nAChRs in Alzheimer's disease, the possibility exists that presynaptic nAChRs may be targets for Abeta. To explore this possibility, we assessed the effect of Abeta peptides on nicotine-evoked changes in presynaptic Ca2+ level via confocal imaging of isolated presynaptic nerve endings from rat hippocampus and neocortex. Abeta1-42 appeared to inhibit presynaptic nAChR activation by nicotine. Surprisingly, picomolar Abeta1-42 was found to directly evoke sustained increases in presynaptic Ca2+ via nAChRs, revealing that the apparent inhibitory action of Abeta1-42 was the result of an occlusion of nicotine to further stimulate the receptors. The direct effect of Abeta was found to be sensitive to alpha-bungarotoxin, mecamylamine, and dihydro-beta-erythroidine, indicating involvement of alpha7-containing nAChRs and non-alpha7-containing nAChRs. Prior depolarization strongly attenuated subsequent Abeta-evoked responses in a manner dependent on the amplitude of the initial presynaptic Ca2+ increase, suggesting that nerve activity or Ca2+ channel density may control the impact of Abeta on presynaptic nerve terminal function. Together, these results suggest that the sustained increases in presynaptic Ca2+ evoked by Abeta may underlie disruptions in neuronal signaling via nAChRs in the early stages of Alzheimer's disease. Title: Evolutionary and biomedical implications of a Schistosoma japonicum complementary DNA resource Hu W, Yan Q, Shen DK, Liu F, Zhu ZD, Song HD, Xu XR, Wang ZJ, Rong YP and Han ZG <14 more author(s)> Hu W, Yan Q, Shen DK, Liu F, Zhu ZD, Song HD, Xu XR, Wang ZJ, Rong YP, Zeng LC, Wu J, Zhang X, Wang JJ, Xu XN, Wang SY, Fu G, Zhang XL, Wang ZQ, Brindley PJ, McManus DP, Xue CL, Feng Z, Chen Z, Han ZG (- 14) Ref: Nat Genet, 35:139, 2003 : PubMed ESTHER: Hu_2003_Nat.Genet_35_139 PubMedSearch: Hu 2003 Nat.Genet 35 139 PubMedID: 12973349 Gene_locus related to this paper: schja-Q86EA4, schja-Q86EV0, schja-Q86F58, schja-Q86F66, schja-Q5DDP9 Abstract Schistosoma japonicum causes schistosomiasis in humans and livestock in the Asia-Pacific region. Knowledge of the genome of this parasite should improve understanding of schistosome-host interactions, biomedical aspects of schistosomiasis and invertebrate evolution. We assigned 43,707 expressed sequence tags (ESTs) derived from adult S. japonicum and their eggs to 13,131 gene clusters. Of these, 35% shared no similarity with known genes and 75% had not been reported previously in schistosomes. Notably, S. japonicum encoded mammalian-like receptors for insulin, progesterone, cytokines and neuropeptides, suggesting that host hormones, or endogenous parasite homologs, could orchestrate schistosome development and maturation and that schistosomes modulate anti-parasite immune responses through inhibitors, molecular mimicry and other evasion strategies. Title: Toxicity of chlorpyrifos adsorbed on humic colloids to larval walleye (Stizostedion vitreum) Phillips TA, Summerfelt RC, Wu J, Laird DA Ref: Archives of Environmental Contamination & Toxicology, 45:258, 2003 : PubMed ESTHER: Phillips_2003_Arch.Environ.Contam.Toxicol_45_258 PubMedSearch: Phillips 2003 Arch.Environ.Contam.Toxicol 45 258 PubMedID: 14565584 Abstract After application, organophosphorus insecticides (OPs) are often strongly adsorbed to soil constituents. Because of their relatively low water solubility, OPs may be transferred from field to stream adsorbed on suspended solids. However, we are not aware of research done to evaluate the bioavailability (i.e., toxicity) of OPs transported on suspended solids to fish. We conducted 48-h static toxicity tests to determine the toxicity of chlorpyrifos in aqueous solution and adsorbed on calcium-saturated humic acid (HA) to three larval stages of walleye (Stizostedion vitreum). Three concentrations of chlorpyrifos adsorbed on HA, a HA control, and a chlorpyrifos-only treatment were tested. Fish that survived the 48-h static toxicity tests were analyzed to determine total cholinesterase (ChE) activity. In general, survival of all larval stages of walleye exposed to chlorpyrifos-HA complexes was less than that of walleye exposed to HA controls and the chlorpyrifos-only treatment, which were not toxic to walleye. Cholinesterase inhibition of larval walleye exposed to chlorpyrifos-HA complexes was similar to the ChE inhibition observed in larval walleye exposed to chlorpyrifos in the aqueous phase. These laboratory experiments indicate potential toxicity of chlorpyrifos-soil complexes to larval fish. Title: Functional properties of homomeric, human alpha 7-nicotinic acetylcholine receptors heterologously expressed in the SH-EP1 human epithelial cell line Zhao L, Kuo YP, George AA, Peng JH, Purandare MS, Schroeder KM, Lukas RJ, Wu J Ref: Journal of Pharmacology & Experimental Therapeutics, 305:1132, 2003 : PubMed ESTHER: Zhao_2003_J.Pharmacol.Exp.Ther_305_1132 PubMedSearch: Zhao 2003 J.Pharmacol.Exp.Ther 305 1132 PubMedID: 12626641 Abstract alpha 7-Nicotinic acetylcholine receptors (alpha 7-nAChRs) are broadly distributed in the central nervous system, where they play important roles in chemical and electrical signaling, and perhaps in neurite outgrowth, synaptic plasticity, and neuronal death/survival. To help elucidate their normal and pathophysiological roles, we have heterologously expressed human alpha 7-nAChR in transfected SH-EP1 human epithelial cells. Reverse transcription-polymerase chain reaction and mRNA fluorescence in situ hybridization analyses demonstrate expression of human alpha 7 subunits as messenger RNA. Patch-clamp recordings exploiting a novel strategy to prevent functional rundown of whole-cell peak current responses to repeated acute challenges with nicotinic agonists show successful expression of functional alpha 7-nAChR that mediate inward currents characterized by rapid phases of activation and inactivation. Concentration-response curves show that nicotine, acetylcholine, and choline are efficacious agonists at human alpha 7-nAChRs. Current-voltage relationships show inward rectification for agonist-induced currents. Human alpha 7-nAChRs exhibit some sensitivity to alpha 7-nAChR antagonists alpha-bungarotoxin (Bgt) or methyllycaconitine (MLA) when applied coincidentally with agonist, but much higher affinity block occurs when cells and alpha 7-nAChRs are pre-exposed to antagonists for 2 min before challenge with agonist. Both Bgt and MLA are competitive inhibitors of alpha 7-nAChR function. Whole-cell current peak amplitudes and half-times for inactivation of alpha 7-nAChR functional responses to nicotine are dramatically reduced in the absence of extracellular Ca2+, suggestive of high Ca2+ permeability of the alpha 7-nAChR channel. Thus, heterologously expressed human alpha 7-nAChR in mammalian cells have properties of native alpha 7-nAChR or of alpha 7-nAChR heterologously expressed in other systems and serve as excellent models for studies of molecular bases of alpha 7-nAChR function. Title: Acute toxicity and cholinesterase inhibition in larval and early juvenile walleye exposed to chlorpyrifos Phillips TA, Wu J, Summerfelt RC, Atchison GJ Ref: Environ Toxicol Chem, 21:1469, 2002 : PubMed ESTHER: Phillips_2002_Environ.Toxicol.Chem_21_1469 PubMedSearch: Phillips 2002 Environ.Toxicol.Chem 21 1469 PubMedID: 12109748 Abstract Application of organophosphorus (OP) insecticides to agricultural fields during the spring often overlaps the period of spawning, egg incubation, and larval stages of fish species such as walleye (Stizostedion vitreum). Because life stage can affect uptake, distribution, and effects of contaminants, our objective was to identify the most sensitive life stage of walleye tochlorpyrifos, a broad-spectrum OP insecticide. Prolarvae (yolk sac, endogenous feeding stage) were least sensitive to chlorpyrifos (median lethal concentration [LC50] = 225-316 microg/L), and postlarvae I (oil globule, exogenous feeding stage) were less sensitive (LC50 = 24-29 microg/L) than postlarvae II (oil globule absent; LC50 = 12-13 microg/L). The differences in sensitivity of the larval stages coincide with stages of gill development. Gill filaments were absent until the end of the prolarval stage, and development of secondary lamellae did not occur until the end of the postlarval I stage. Juvenile fish were less sensitive than postlarval stages, but did not differ significantly among the juvenile ages tested. The LC50s ranged from 37 to 45 microg/L for 30- and 90-d-old juvenile walleye, respectively. Larval walleye survived when cholinesterase (ChE) activity was inhibited by as much as 90%; however, 60- and 90-d-old juvenile walleye did not survive when ChE activity was inhibited more than 71%. Title: The genome sequence and structure of rice chromosome 1 Sasaki T, Matsumoto T, Yamamoto K, Sakata K, Baba T, Katayose Y, Wu J, Niimura Y, Cheng Z and Gojobori T <69 more author(s)> Sasaki T, Matsumoto T, Yamamoto K, Sakata K, Baba T, Katayose Y, Wu J, Niimura Y, Cheng Z, Nagamura Y, Antonio BA, Kanamori H, Hosokawa S, Masukawa M, Arikawa K, Chiden Y, Hayashi M, Okamoto M, Ando T, Aoki H, Arita K, Hamada M, Harada C, Hijishita S, Honda M, Ichikawa Y, Idonuma A, Iijima M, Ikeda M, Ikeno M, Ito S, Ito T, Ito Y, Iwabuchi A, Kamiya K, Karasawa W, Katagiri S, Kikuta A, Kobayashi N, Kono I, Machita K, Maehara T, Mizuno H, Mizubayashi T, Mukai Y, Nagasaki H, Nakashima M, Nakama Y, Nakamichi Y, Nakamura M, Namiki N, Negishi M, Ohta I, Ono N, Saji S, Sakai K, Shibata M, Shimokawa T, Shomura A, Song J, Takazaki Y, Terasawa K, Tsuji K, Waki K, Yamagata H, Yamane H, Yoshiki S, Yoshihara R, Yukawa K, Zhong H, Iwama H, Endo T, Ito H, Hahn JH, Kim HI, Eun MY, Yano M, Jiang J, Gojobori T (- 69) Ref: Nature, 420:312, 2002 : PubMed ESTHER: Sasaki_2002_Nature_420_312 PubMedSearch: Sasaki 2002 Nature 420 312 PubMedID: 12447438 Gene_locus related to this paper: orysa-Q9S7P1, orysa-Q9FYP7, orysa-Q5ZBH3, orysa-Q5NA74, orysa-Q5ZA26, orysa-Q5JLP6, orysa-Q94D81, orysa-cbp, orysa-Q5VQE5, orysa-Q8RZ95, orysa-Q9AWW1, orysa-Q9AS70, orysa-Q0JK71, orysa-Q8S1D9, orysa-Q5N8V4, orysa-Q943F9, orysa-B9EWJ8, orysa-Q5N8H1, orysa-Q5NAI4, orysa-Q94DP8, orysa-Q658B2, orysa-Q5JMQ8, orysa-Q5QMD9, orysa-Q5N7L1, orysa-Q5N7J6, orysa-Q8RYV9, orysa-Q5SNH3, orysa-Q94DD0, orysa-Q8W0F0, orysa-pir7a, orysa-pir7b, orysa-Q4VWY7, orysa-q5jlm9, orysa-q5na00, orysa-q5nbu1, orysa-Q5QLC0, orysa-q5vnp5, orysa-Q5VP27, orysa-Q5ZAM8, orysa-Q5ZBI5, orysa-q5zc23, orysa-Q5ZCR3, orysa-Q8L562, orysa-Q8L570, orysa-Q8LQS5, orysa-Q8RZ40, orysa-Q8RZ79, orysa-Q8S0U8, orysa-Q8S0V0, orysa-Q8S125, orysa-Q9LHX5, orysa-Q94E46, orysa-Q656F2, orysi-a2wn01, orysi-b8a7e6, orysi-b8a7e7, orysj-b9eya5, orysj-q5jl22, orysj-q5jlw7, orysj-q94d71 Abstract The rice species Oryza sativa is considered to be a model plant because of its small genome size, extensive genetic map, relative ease of transformation and synteny with other cereal crops. Here we report the essentially complete sequence of chromosome 1, the longest chromosome in the rice genome. We summarize characteristics of the chromosome structure and the biological insight gained from the sequence. The analysis of 43.3 megabases (Mb) of non-overlapping sequence reveals 6,756 protein coding genes, of which 3,161 show homology to proteins of Arabidopsis thaliana, another model plant. About 30% (2,073) of the genes have been functionally categorized. Rice chromosome 1 is (G + C)-rich, especially in its coding regions, and is characterized by several gene families that are dispersed or arranged in tandem repeats. Comparison with a draft sequence indicates the importance of a high-quality finished sequence. Title: Kinetic Analysis of Acetylcholinesterase in a Propoxur-Resistant Strain of Housefly (Musca domestica) from Shanghai, China Shi MA, Yuan JZ, Wu J, Zhuang PJ, Wu XF, Tang ZH Ref: Pesticide Biochemistry and Physiology, 72:72, 2002 : PubMed ESTHER: Shi_2002_Pestic.Biochem.Physiol_72_72 PubMedSearch: Shi 2002 Pestic.Biochem.Physiol 72 72 Abstract A propoxur-selected resistant (R) strain of housefly (Musca domestica), from Shanghai, China, exhibited >185-, 47.2-, 41.3-, and 50.5-fold greater resistance to propoxur, methomyl, paraoxon, and dichlorvos, respectively, than a susceptible (S) strain. The turnover number (kcat) and catalytic efficiency (kcat/Kmm) of acetylcholinesterase (AChE), determined by 7-[(diethoxyphosphoryl)-oxy]-1-methylquinolinium iodide (DEPQ) as a titrant and acetylthiocholine as a substrate, were 180,805 min-1 and 846.9 x 10 6 M-1 min-1 for the R strain and 119,875 min-1 and 2378.5 x 10 6 M-1 min-1 for the S strain, respectively. The greater maximal velocity of AChE from the R strain appeared to be due to increased turnover number and higher concentration of the AChE active site. AChE from the R strain was 3.4-, 21.0-, and 21.4-fold less sensitive to three carbamates, eserine, methomyl, and propoxur, and 2.6-, 33.1-, and 19.3-fold less sensitive to three organophosphates, DEPQ, dichlorvos, and paraoxon, respectively, based on the bimolecular rate constant (ki). The differences in ki were attributed mainly to the decreased affinity of these compounds for AChE from the R strain as determined by the affinity constant (Ka). All these data suggested that decreased sensitivity of AChE was associated with decreased catalytic efficiency of the enzyme in the R strain. Title: Association between low-density lipoprotein receptor-related protein gene, butyrylcholinesterase gene and Alzheimer' s disease in Chinese Bi S, Zhang Y, Wu J, Wang D, Zhao Q Ref: Chin Med Sci J, 16:71, 2001 : PubMed ESTHER: Bi_2001_Chin.Med.Sci.J_16_71 PubMedSearch: Bi 2001 Chin.Med.Sci.J 16 71 PubMedID: 12901493 Abstract OBJECTIVE: To research the relations between low-density lipoprotein receptor-related protein gene (LRP) polymorphism, butyrylcholinesterase gene (BchE) polymorphism and Alzheimer's disease (AD) in Chinese. Title: [Cloning, expression and preliminary application of a alpha-hydroxynitrile lyase from cassave] Cheng SH, Yan GH, Wu J, Sun WR Ref: Sheng Wu Gong Cheng Xue Bao, 17:78, 2001 : PubMed ESTHER: Cheng_2001_Sheng.Wu.Gong.Cheng.Xue.Bao_17_78 PubMedSearch: Cheng 2001 Sheng.Wu.Gong.Cheng.Xue.Bao 17 78 PubMedID: 11330194 Gene_locus related to this paper: manes-hnl Abstract alpha-Hydroxynitrile lyase (ME-HNLs, E.C. 4.1.2.3.37) from the cyanogenic crop cassava(Manihot esculentz, Crantz) catalyze the condensation of hydrocyanic acid and aldehydes or ketone into (s)-cyanohydrins, which are valuable starting material for various optically active compounds, such as pharmaceuticals and agrochemicals. The cDNA of a ME-HNL were obtained by RT-PCR and cloned. The sequencing result for the cDNA showed that the sequence encoded for the ME-HNL was inconsistent with all those which are published, such as hnl10, hnl24, hnl4. The full sequence analysis demonstrated that the cloned cDNA was about 75.2%, 79.8%, 99.2% homologous to other three reported HNL genes from cassava, respectively, among which the last was the same to the cloned gene except the five base substitution at the site 142, 337, 476, 634 and 636, respectively. The two base substitutions lead to change the amino acid sequence, i.e., Ser113-->Gly113, Phe158-->Tyr158. To construct the recombinant plasmid pET30a-hnl, the cDNA was inserted into an expression vector pET30a. After transformation of pET30a-hnl and induction with IPTG, the ME-HNL was efficiently expressed in E. coli. BL21 (DE3) and reached over 2100 units/L of culture with the specific activity 8.5 u/mg protein. By one simple treatment, incubating 10 minutes at 70 degrees C, the recombinant ME-HNL may be used as an catalyst for production of (S)-mandelonitrile with enantiomeric excess of 95.2% and 98.2% yield. Title: Molecular mechanisms underlying the activity-linked alterations in acetylcholinesterase mRNAs in developing versus adult rat skeletal muscles Boudreau-Lariviere C, Chan RY, Wu J, Jasmin BJ Ref: Journal of Neurochemistry, 74:2250, 2000 : PubMed ESTHER: Boudreau-Lariviere_2000_J.Neurochem_74_2250 PubMedSearch: Boudreau-Lariviere 2000 J.Neurochem 74 2250 PubMedID: 10820184 Abstract The molecular mechanisms underlying the activity-linked plasticity of acetylcholinesterase (AChE) mRNA levels in mammalian skeletal muscle have yet to be established. Here, we demonstrate that denervation of adult muscle induces a dramatic (up to 90%) and rapid (within 24 h) decrease in the abundance of AChE mRNAs. By contrast, denervation of 14-day-old rats leads to a significantly less pronounced reduction (50% of control) in the expression of AChE mRNAs. Assessment of the transcriptional activity of the AChE gene reveals that it remains essentially unchanged in adult denervated muscles, whereas it displays an approximately two- to three-fold increase (p < 0.05) in denervated muscles from 2- to 14-day-old rats. In addition, we observed a higher rate of degradation of in vitro transcribed AChE mRNAs upon incubation with protein extracts from denervated muscles. Finally, UV-crosslinking experiments reveal that denervation increases the abundance of RNA-protein interactions in the 3' untranslated region of AChE transcripts. Taken together, these data suggest that the abundance of AChE transcripts in mature muscles is controlled primarily via posttranscriptional regulatory mechanisms, whereas in neo- and postnatal muscles, both transcriptional and posttranscriptional regulation appears critical in dictating AChE mRNA levels. Accordingly, the activity-linked transcriptional regulation of the AChE gene appears to demonstrate a high level of plasticity during muscle development when maturation of the neuromuscular junctions is still occurring. Title: Carboxyl methylation of the phosphoprotein phosphatase 2A catalytic subunit promotes its functional association with regulatory subunits in vivo Wu J, Tolstykh T, Lee J, Boyd K, Stock JB, Broach JR Ref: EMBO Journal, 19:5672, 2000 : PubMed ESTHER: Wu_2000_EMBO.J_19_5672 PubMedSearch: Wu 2000 EMBO.J 19 5672 PubMedID: 11060018 Gene_locus related to this paper: yeast-ppme1 Abstract The phosphoprotein phosphatase 2A (PP2A) catalytic subunit contains a methyl ester on its C-terminus, which in mammalian cells is added by a specific carboxyl methyltransferase and removed by a specific carboxyl methylesterase. We have identified genes in yeast that show significant homology to human carboxyl methyltransferase and methylesterase. Extracts of wild-type yeast cells contain carboxyl methyltransferase activity, while extracts of strains deleted for one of the methyltransferase genes, PPM1, lack all activity. Mutation of PPM1 partially disrupts the PP2A holoenzyme in vivo and ppm1 mutations exhibit synthetic lethality with mutations in genes encoding the B or B' regulatory subunit. Inactivation of PPM1 or overexpression of PPE1, the yeast gene homologous to bovine methylesterase, yields phenotypes similar to those observed after inactivation of either regulatory subunit. These phenotypes can be reversed by overexpression of the B regulatory subunit. These results demonstrate that Ppm1 is the sole PP2A methyltransferase in yeast and that its activity is required for the integrity of the PP2A holoenzyme. Title: Regulation of utrophin expression during development of skeletal muscle cells Gramolini AO, Lunde JA, Wu J, Jasmin BJ Ref: Journal de Physiologie (Paris), 92:434, 1998 : PubMed Title: Degradation of dimethoate in chrysanthemums and soil Wu J, Fan D Ref: Bulletin of Environmental Contamination & Toxicology, 59:564, 1997 : PubMed ESTHER: Wu_1997_Bull.Environ.Contam.Toxicol_59_564 PubMedSearch: Wu 1997 Bull.Environ.Contam.Toxicol 59 564 PubMedID: 9307420 | |||||||
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