Huang R

References (32)

Title : Identification of Potent and Selective Acetylcholinesterase\/Butyrylcholinesterase Inhibitors by Virtual Screening - Xu_2023_J.Chem.Inf.Model__
Author(s) : Xu T , Li S , Li AJ , Zhao J , Sakamuru S , Huang W , Xia M , Huang R
Ref : J Chem Inf Model , : , 2023
Abstract : Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) play important roles in human neurodegenerative disorders such as Alzheimer's disease. In this study, machine learning methods were applied to develop quantitative structure-activity relationship models for the prediction of novel AChE and BChE inhibitors based on data from quantitative high-throughput screening assays. The models were used to virtually screen an in-house collection of -360K compounds. The optimal models achieved good performance with area under the receiver operating characteristic curve values ranging from 0.83 +/- 0.03 to 0.87 +/- 0.01 for the prediction of AChE/BChE inhibition activity and selectivity. Experimental validation showed that the best-performing models increased the assay hit rate by several folds. We identified 88 novel AChE and 126 novel BChE inhibitors, 25% (AChE) and 53% (BChE) of which showed potent inhibitory effects (IC(50) < 5 microM). In addition, structure-activity relationship analysis of the BChE inhibitors revealed scaffolds for chemistry design and optimization. In conclusion, machine learning models were shown to efficiently identify potent and selective inhibitors against AChE and BChE and novel structural series for further design and development of potential therapeutics against neurodegenerative disorders.
ESTHER : Xu_2023_J.Chem.Inf.Model__
PubMedSearch : Xu_2023_J.Chem.Inf.Model__
PubMedID: 37011147

Title : Ultrasensitive Detection of Organophosphorus Pesticides Using Single-Molecule Conductance Measurement - Dong_2023_Anal.Chem_95_9831
Author(s) : Dong X , Tang Z , Zhang H , Hu Y , Yao Z , Huang R , Bai J , Yang Y , Hong W
Ref : Analytical Chemistry , 95 :9831 , 2023
Abstract : Detection of organophosphorus pesticides (OPs) with high sensitivity in environmental samples is of vital importance for environmental safety and human health. However, it remains a challenge to achieve fM (10(-15) mol/L) sensitivity for detecting OPs. Herein, we developed an acetylcholinesterase sensor based on 3,3',5,5'-tetramethylbenzidine (TMB) combining an enzyme-mediated strategy and scanning tunneling microscopy break junction (STM-BJ). Benefiting from the enzyme inhibition kinetics of OPs and the customized spectral clustering analysis method, our new strategy achieved the detection of methamidophos (MTMP) with a limit of 10 aM (10(-17) mol/L) and 3 times higher selectivity in mixed OPs. As applied to natural lake waters, it also exhibited high reproducibility, high stability, and good recovery. This work paves a new avenue toward the application of single-molecule conductance characterizations for biochemical analysis and environmental monitoring.
ESTHER : Dong_2023_Anal.Chem_95_9831
PubMedSearch : Dong_2023_Anal.Chem_95_9831
PubMedID: 37347983

Title : Enantioconvergent hydrolysis of racemic 1,2-epoxypentane and 1,2-epoxyhexane by an engineered Escherichia coli strain overexpressing a novel Streptomyces fradiae epoxide hydrolase - Huang_2023_Enzyme.Microb.Technol_166_110228
Author(s) : Huang R , Li C , Zhao SG , Liu QT , Liu Y , Xue ZL
Ref : Enzyme Microb Technol , 166 :110228 , 2023
Abstract : In order to excavate microbial epoxide hydrolases (EHs) with desired catalytic properties, a novel EH, SfEH1, was identified based on the genome annotation of Streptomyces fradiae and sequence alignment analysis with local protein library. The SfEH1-encoding gene, sfeh1, was then cloned and over-expressed in soluble form in Escherichia coli/BL21(DE3). The optimal temperature and pH of recombinant SfEH1 (reSfEH1) and reSfEH1-expressing E. coli (E. coli/sfeh1) were both determined as 30 degC and 7.0, also indicating that the influences of temperature and pH on reSfEH1's activities were more obvious than those of E. coli/sfeh1 whole cells. Subsequently, using E. coli/sfeh1 as catalyst, its catalytic properties towards thirteen common mono-substituted epoxides were tested, in which E. coli/sfeh1 had the highest activity of 28.5 U/g dry cells for rac-1,2-epoxyoctane (rac-6a), and (R)-1,2-pentanediol ((R)-3b) (or (R)-1,2-hexanediol ((R)-4b)) with up to 92.5% (or 94.1%) ee(p) was obtained at almost 100% conversion ratio. Regioselectivity coefficients (alpha(S) and beta(R)) displayed in the enantioconvergent hydrolysis of rac-3a (or rac-4a) were calculated to be 98.7% and 93.8% (or 95.2% and 98.9%). Finally, the reason of the high and complementary regioselectivity was confirmed by both kinetic parameter analysis and molecular docking simulations.
ESTHER : Huang_2023_Enzyme.Microb.Technol_166_110228
PubMedSearch : Huang_2023_Enzyme.Microb.Technol_166_110228
PubMedID: 36940599
Gene_locus related to this paper: strfr-SfEH1

Title : Selective inhibition of neurogenic, but not agonist-induced contractions by phospholipase A(2) inhibitors points to presynaptic phospholipase A(2) functions in contractile neurotransmission to human prostate smooth muscle - Hu_2023_Neurourol.Urodyn_42_1522
Author(s) : Hu S , Huang R , Keller P , Gotz M , Tamalunas A , Weinhold P , Waidelich R , Stief CG , Hennenberg M
Ref : Neurourol Urodyn , 42 :1522 , 2023
Abstract : BACKGROUND: Phospholipases A(2) (PLA(2) ) may be involved in alpha(1) -adrenergic contraction by formation of thromboxane A(2) in different smooth muscle types. However, whether this mechanism occurs with alpha(1) -adrenergic contractions of the prostate, is still unknown. While alpha(1) -adrenoceptor antagonists are the first line option for medical treatment of voiding symptoms in benign prostatic hyperplasia (BPH), improvements are limited, probably by nonadrenergic contractions including thromboxane A(2) . Here, we examined effects of PLA(2) inhibitors on contractions of human prostate tissues. METHODS: Prostate tissues were obtained from radical prostatectomy. Contractions were induced by electric field stimulation (EFS) and by alpha(1) -adrenergic agonists in an organ bath, after application of the cytosolic PLA(2) inhibitors ASB14780 and AACOCF3, the secretory PLA(2) inhibitor YM26734, the leukotriene receptor antagonist montelukast, or of solvent to controls. RESULTS: Frequency-dependent contractions of human prostate tissues induced by EFS were inhibited by 25% at 8 Hz, 38% at 16 Hz and 37% at 32 Hz by ASB14780 (1 microM), and by 32% at 16 Hz and 22% at 32 Hz by AACOCF3 (10 microM). None of both inhibitors affected contractions induced by noradrenaline, phenylephrine or methoxamine. YM26734 (3 microM) and montelukast (0.3 and 1 microM) neither affected EFS-induced contractions, nor contractions by alpha(1) -adrenergic agonists, while all contractions were substantially inhibited by silodosin (100 nM). CONCLUSIONS: Our findings suggest presynaptic PLA(2) functions in prostate smooth muscle contraction, while contractions induced by alpha(1) -adrenergic agonists occur PLA(2) -independent. Lacking sensitivity to montelukast excludes an involvement of PLA(2) -derived leukotrienes in promotion of contractile neurotransmission.
ESTHER : Hu_2023_Neurourol.Urodyn_42_1522
PubMedSearch : Hu_2023_Neurourol.Urodyn_42_1522
PubMedID: 37583250

Title : Biodegradation of Free Gossypol by Helicoverpa armigera Carboxylesterase Expressed in Pichia pastoris - Zhang_2022_Toxins.(Basel)_14_816
Author(s) : Zhang L , Yang X , Huang R , Nie C , Niu J , Chen C , Zhang W
Ref : Toxins (Basel) , 14 :816 , 2022
Abstract : Gossypol is a polyphenolic toxic secondary metabolite derived from cotton. Free gossypol in cotton meal is remarkably harmful to animals. Furthermore, microbial degradation of gossypol produces metabolites that reduce feed quality. We adopted an enzymatic method to degrade free gossypol safely and effectively. We cloned the gene cce001a encoding carboxylesterase (CarE) into pPICZalphaA and transformed it into Pichia pastoris GS115. The target protein was successfully obtained, and CarE CCE001a could effectively degrade free gossypol with a degradation rate of 89%. When esterase was added, the exposed toxic groups of gossypol reacted with different amino acids and amines to form bound gossypol, generating substances with (M + H) m/z ratios of 560.15, 600.25, and 713.46. The molecular formula was C(27)H(28)O(13), C(34)H(36)N(2)O(6), and C(47)H(59)N(3)O(3). The observed instability of the hydroxyl groups caused the substitution and shedding of the group, forming a substance with m/z of 488.26 and molecular formula C(31)H(36)O(5). These properties render the CarE CCE001a a valid candidate for the detoxification of cotton meal. Furthermore, the findings help elucidate the degradation process of gossypol in vitro.
ESTHER : Zhang_2022_Toxins.(Basel)_14_816
PubMedSearch : Zhang_2022_Toxins.(Basel)_14_816
PubMedID: 36548713

Title : Knockdown of hepatocyte Perilipin-3 mitigates hepatic steatosis and steatohepatitis caused by hepatocyte CGI-58 deletion in mice - Bao_2022_J.Mol.Cell.Biol__
Author(s) : Bao X , Ma X , Huang R , Chen J , Xin H , Zhou M , Li L , Tong S , Zhang Q , Shui G , Deng F , Yu L , Li MD , Zhang Z
Ref : J Molecular & Cellular Biology , : , 2022
Abstract : Comparative gene identification-58 (CGI-58), also known as alpha/beta hydrolase domain containing 5 (ABHD5), is the co-activator of adipose triglyceride lipase that hydrolyzes triglycerides stored in the cytosolic lipid droplets. Mutations in CGI-58 gene cause Chanarin-Dorfman syndrome (CDS), an autosomal recessive neutral lipid storage disease with ichthyosis. The liver pathology of CDS manifests as steatosis and steatohepatitis, which currently has no effective treatments. Perilipin-3 (Plin3) is a member of the Perilipin-ADRP-TIP47 (PAT) protein family that is essential for lipid droplet biogenesis. The objective of this study was to test a hypothesis that deletion of a major lipid droplet protein alleviates fatty liver pathogenesis caused by CGI-58 deficiency in hepatocytes. Adult CGI-58-floxed mice were injected with adeno-associated vectors simultaneously expressing the Cre recombinase and microRNA against Plin3 under the control of a hepatocyte-specific promoter, followed by high-fat diet (HFD) feeding for 6 weeks. Liver and blood samples were then collected from these animals for histological and biochemical analysis. Plin3 knockdown in hepatocytes prevented steatosis, steatohepatitis, and necroptosis caused by hepatocyte CGI-58 deficiency. Our work is the first to show that inhibiting Plin3 in hepatocytes is sufficient to mitigate hepatocyte CGI-58 deficiency-induced hepatic steatosis and steatohepatitis in mice.
ESTHER : Bao_2022_J.Mol.Cell.Biol__
PubMedSearch : Bao_2022_J.Mol.Cell.Biol__
PubMedID: 36107452

Title : Identification of Compounds for Butyrylcholinesterase Inhibition - Li_2021_SLAS.Discov__24725552211030897
Author(s) : Li S , Li AJ , Travers J , Xu T , Sakamuru S , Klumpp-Thomas C , Huang R , Xia M
Ref : SLAS Discov , :24725552211030897 , 2021
Abstract : Butyrylcholinesterase (BChE) is a nonspecific cholinesterase enzyme that hydrolyzes choline-based esters. BChE plays a critical role in maintaining normal cholinergic function like acetylcholinesterase (AChE) through hydrolyzing acetylcholine (ACh). Selective BChE inhibition has been regarded as a viable therapeutic approach in Alzheimer's disease. As of now, a limited number of selective BChE inhibitors are available. To identify BChE inhibitors rapidly and efficiently, we have screened 8998 compounds from several annotated libraries against an enzyme-based BChE inhibition assay in a quantitative high-throughput screening (qHTS) format. From the primary screening, we identified a group of 125 compounds that were further confirmed to inhibit BChE activity, including previously reported BChE inhibitors (e.g., bambuterol and rivastigmine) and potential novel BChE inhibitors (e.g., pancuronium bromide and NNC 756), representing diverse structural classes. These BChE inhibitors were also tested for their selectivity by comparing their IC(50) values in BChE and AChE inhibition assays. The binding modes of these compounds were further studied using molecular docking analyses to identify the differences between the interactions of these BChE inhibitors within the active sites of AChE and BChE. Our qHTS approach allowed us to establish a robust and reliable process to screen large compound collections for potential BChE inhibitors.
ESTHER : Li_2021_SLAS.Discov__24725552211030897
PubMedSearch : Li_2021_SLAS.Discov__24725552211030897
PubMedID: 34269114

Title : Profiling the Tox21 Chemical Collection for Acetylcholinesterase Inhibition - Li_2021_Environ.Health.Perspect_129_47008
Author(s) : Li S , Zhao J , Huang R , Travers J , Klumpp-Thomas C , Yu W , MacKerell AD, Jr. , Sakamuru S , Ooka M , Xue F , Sipes NS , Hsieh JH , Ryan K , Simeonov A , Santillo MF , Xia M
Ref : Environmental Health Perspectives , 129 :47008 , 2021
Abstract : BACKGROUND: Inhibition of acetylcholinesterase (AChE), a biomarker of organophosphorous and carbamate exposure in environmental and occupational human health, has been commonly used to identify potential safety liabilities. So far, many environmental chemicals, including drug candidates, food additives, and industrial chemicals, have not been thoroughly evaluated for their inhibitory effects on AChE activity. AChE inhibitors can have therapeutic applications (e.g., tacrine and donepezil) or neurotoxic consequences (e.g., insecticides and nerve agents). OBJECTIVES: The objective of the current study was to identify environmental chemicals that inhibit AChE activity using in vitro and in silico models. METHODS: To identify AChE inhibitors rapidly and efficiently, we have screened the Toxicology in the 21st Century (Tox21) 10K compound library in a quantitative high-throughput screening (qHTS) platform by using the homogenous cell-based AChE inhibition assay and enzyme-based AChE inhibition assays (with or without microsomes). AChE inhibitors identified from the primary screening were further tested in monolayer or spheroid formed by SH-SY5Y and neural stem cell models. The inhibition and binding modes of these identified compounds were studied with time-dependent enzyme-based AChE inhibition assay and molecular docking, respectively. RESULTS: A group of known AChE inhibitors, such as donepezil, ambenonium dichloride, and tacrine hydrochloride, as well as many previously unreported AChE inhibitors, such as chelerythrine chloride and cilostazol, were identified in this study. Many of these compounds, such as pyrazophos, phosalone, and triazophos, needed metabolic activation. This study identified both reversible (e.g., donepezil and tacrine) and irreversible inhibitors (e.g., chlorpyrifos and bromophos-ethyl). Molecular docking analyses were performed to explain the relative inhibitory potency of selected compounds. CONCLUSIONS: Our tiered qHTS approach allowed us to generate a robust and reliable data set to evaluate large sets of environmental compounds for their AChE inhibitory activity.
ESTHER : Li_2021_Environ.Health.Perspect_129_47008
PubMedSearch : Li_2021_Environ.Health.Perspect_129_47008
PubMedID: 33844597

Title : Single-cell-resolved measurement of enzyme activity at the tissue level using drop-on-demand microkits - Huang_2021_Analyst__
Author(s) : Huang R , Jin R , Jiang D , Chen HY
Ref : Analyst , : , 2021
Abstract : Drop-on-demand microkits with a diameter of ~20 microm are used to measure the activity of acetylcholinesterase (AChE) in a brain slice with single-cell resolution. The relative standard deviation from 25 cellular regions reached 73.3% exhibiting the difference of enzyme activity in the brain slice. Therefore, this approach utilizing the well-established kits provides an alternative single-cell-resolved strategy for the elucidation of enzymatic heterogeneity at the tissue level.
ESTHER : Huang_2021_Analyst__
PubMedSearch : Huang_2021_Analyst__
PubMedID: 33427262

Title : Antioxidant and pancreatic lipase inhibitory effects of flavonoids from different citrus peel extracts: An in vitro study - Huang_2020_Food.Chem_326_126785
Author(s) : Huang R , Zhang Y , Shen S , Zhi Z , Cheng H , Chen S , Ye X
Ref : Food Chem , 326 :126785 , 2020
Abstract : Obesity and oxidative damage are two important risk factors associated closely with metabolic syndrome. Utilization of functional food ingredients is considered as a feasible way to tackle these challenges. In the present study, eight representative species of citrus peel extracts (CPEs) were evaluated and compared for their flavonoid profiles, antioxidant activities, and pancreatic lipase (PL) inhibitory capacities and mechanisms. Results indicated that hesperidin, naringin, neohesperidin, narirutin and eriocitrin were the five major flavonoids in CPEs, among which hesperidin was the main active PL inhibitor. Moreover, hesperidin could interact with PL by hydrogen bonds and van der Waals forces, and the interaction would not obviously change the secondary structure of PL. Overall, ponkan peel extract, having the strongest overall antioxidant activity, the highest content of hesperidin and total phenolic compounds among all tested CPEs, is a promising natural ingredient to scavenge free radicals and manage obesity.
ESTHER : Huang_2020_Food.Chem_326_126785
PubMedSearch : Huang_2020_Food.Chem_326_126785
PubMedID: 32438224

Title : Identification of Highly Selective Lipoprotein-Associated Phospholipase A2 (Lp-PLA2) Inhibitors by a Covalent Fragment-Based Approach - Huang_2020_J.Med.Chem_63_7052
Author(s) : Huang F , Hu H , Wang K , Peng C , Xu W , Zhang Y , Gao J , Liu Y , Zhou H , Huang R , Li M , Shen J , Xu Y
Ref : Journal of Medicinal Chemistry , 63 :7052 , 2020
Abstract : Covalent ligands are of great interest as therapeutic drugs or biochemical tools. Here, we reported the discovery of highly selective and irreversible inhibitors of lipoprotein-associated phospholipase A2 (Lp-PLA2) using a covalent fragment-based approach. The crystal structure of Lp-PLA2 in complex with a covalent fragment not only reveals the covalent reaction mechanism but also provides a good starting point to design compound 8, which has a more than 130,000-fold and 3900-fold increase in potency and selectivity, respectively, compared to those of the covalent fragment. Furthermore, fluorescent probes with high selectivity and sensitivity are developed to characterize Lp-PLA2 and its enzymatic activity in vitro or even in living cells in a way more convenient than immunoblotting tests or immunofluorescence imaging. Overall, we provide a paradigm for application of the covalent fragment-based strategy in covalent ligand discovery and the advantage of enol-cyclocarbamate as a new warhead in designing covalent inhibitors of serine hydrolases.
ESTHER : Huang_2020_J.Med.Chem_63_7052
PubMedSearch : Huang_2020_J.Med.Chem_63_7052
PubMedID: 32459096
Gene_locus related to this paper: human-PLA2G7

Title : Genomics-driven discovery of the biosynthetic gene cluster of maduramicin and its overproduction in Actinomadura sp. J1-007 - Liu_2020_J.Ind.Microbiol.Biotechnol_47_275
Author(s) : Liu R , Fang F , An Z , Huang R , Wang Y , Sun X , Fu S , Fu A , Deng Z , Liu T
Ref : J Ind Microbiol Biotechnol , 47 :275 , 2020
Abstract : Maduramicin is the most efficient and possesses the largest market share of all anti-coccidiosis polyether antibiotics (ionophore); however, its biosynthetic gene cluster (BGC) has yet to been identified, and the associated strains have not been genetically engineered. Herein, we performed whole-genome sequencing of a maduramicin-producing industrial strain of Actinomadura sp. J1-007 and identified its BGC. Additionally, we analyzed the identified BGCs in silico to predict the biosynthetic pathway of maduramicin. We then developed a conjugation method for the non-spore-forming Actinomadura sp. J1-007, consisting of a site-specific integration method for gene overexpression. The maduramicin titer increased by 30% to 7.16 g/L in shake-flask fermentation following overexpression of type II thioesterase MadTE that is the highest titer at present. Our findings provide insights into the biosynthetic mechanism of polyethers and provide a platform for the metabolic engineering of maduramicin-producing microorganisms for overproduction and development of maduramicin analogs in the future.
ESTHER : Liu_2020_J.Ind.Microbiol.Biotechnol_47_275
PubMedSearch : Liu_2020_J.Ind.Microbiol.Biotechnol_47_275
PubMedID: 31853778
Gene_locus related to this paper: 9actn-a0a6i4pr03

Title : Use of high-throughput enzyme-based assay with xenobiotic metabolic capability to evaluate the inhibition of acetylcholinesterase activity by organophosphorous pesticides - Li_2019_Toxicol.In.Vitro_56_93
Author(s) : Li S , Zhao J , Huang R , Santillo MF , Houck KA , Xia M
Ref : Toxicol In Vitro , 56 :93 , 2019
Abstract : The inhibition of acetylcholinesterase (AChE) has pharmaceutical applications as well as potential neurotoxic effects. The in vivo metabolites of some chemicals including organophosphorus pesticides can become more potent AChE inhibitors compared to their parental compounds. To account for the effects of biotransformation, we have developed and characterized a high-throughput screening method for identifying AChE inhibitors that become active or more potent following xenobiotic metabolism. In this study, an enzyme-based assay was developed in 1536-well plates using recombinant human AChE combined with human or rat liver microsomes. The AChE activity was measured by two methods with different readouts: colorimetric and fluorescent. The assay exhibited exceptional performance characteristics including large assay signal window, low well-to-well variability and high reproducibility. The performance of the assays with microsomes was characterized by testing a group of known AChE inhibitors including parent compounds and their metabolites. Large potency differences between the parent compounds and the metabolites were observed in the assay with microsome addition. Both assay readouts were required for maximal sensitivity. These results demonstrate that this platform is a promising method to profile large numbers of chemicals that require metabolic activation for inhibiting AChE activity.
ESTHER : Li_2019_Toxicol.In.Vitro_56_93
PubMedSearch : Li_2019_Toxicol.In.Vitro_56_93
PubMedID: 30625376

Title : The genome of broomcorn millet - Zou_2019_Nat.Commun_10_436
Author(s) : Zou C , Li L , Miki D , Li D , Tang Q , Xiao L , Rajput S , Deng P , Peng L , Jia W , Huang R , Zhang M , Sun Y , Hu J , Fu X , Schnable PS , Chang Y , Li F , Zhang H , Feng B , Zhu X , Liu R , Schnable JC , Zhu JK
Ref : Nat Commun , 10 :436 , 2019
Abstract : Broomcorn millet (Panicum miliaceum L.) is the most water-efficient cereal and one of the earliest domesticated plants. Here we report its high-quality, chromosome-scale genome assembly using a combination of short-read sequencing, single-molecule real-time sequencing, Hi-C, and a high-density genetic map. Phylogenetic analyses reveal two sets of homologous chromosomes that may have merged ~5.6 million years ago, both of which exhibit strong synteny with other grass species. Broomcorn millet contains 55,930 protein-coding genes and 339 microRNA genes. We find Paniceae-specific expansion in several subfamilies of the BTB (broad complex/tramtrack/bric-a-brac) subunit of ubiquitin E3 ligases, suggesting enhanced regulation of protein dynamics may have contributed to the evolution of broomcorn millet. In addition, we identify the coexistence of all three C4 subtypes of carbon fixation candidate genes. The genome sequence is a valuable resource for breeders and will provide the foundation for studying the exceptional stress tolerance as well as C4 biology.
ESTHER : Zou_2019_Nat.Commun_10_436
PubMedSearch : Zou_2019_Nat.Commun_10_436
PubMedID: 30683860
Gene_locus related to this paper: panmi-a0a3l6qvl9 , 9poal-a0a2s3hbt0 , panmi-a0a3l6sxg5 , 9poal-a0a2t7cdl4 , panmi-a0a3l6ta96 , panmi-a0a3l6qv47 , panmi-a0a3l6s688 , panmi-a0a3l6tph0

Title : Jia-Wei-Kai-Xin-San, an Herbal Medicine Formula, Ameliorates Cognitive Deficits via Modulating Metabolism of Beta Amyloid Protein and Neurotrophic Factors in Hippocampus of Abeta1-42 Induced Cognitive Deficit Mice - Zhu_2019_Front.Pharmacol_10_258
Author(s) : Zhu Y , Shi Y , Cao C , Han Z , Liu M , Qi M , Huang R , Zhu Z , Qian D , Duan JA
Ref : Front Pharmacol , 10 :258 , 2019
Abstract : Jia-Wei-Kai-Xin-San (JWKXS) is a Chinese medicine formula applied for treating morbid forgetfulness in ancient China. Today, this formula is frequently applied for Alzheimer's disease and vascular dementia (VD) in clinic. Here, we developed it as granules and aimed to evaluate its anti-AD effect on beta amyloid protein 1-42 (Abeta1-42) induced cognitive deficit mice and reveal the possible molecular mechanisms. Firstly, daily intra-gastric administration of chemically standardized of JWKXS granules for 7 days significantly ameliorated the cognitive deficit symptoms and inhibited cell apoptosis in hippocampus on Abeta1-42 injection mice. JWKXS granules significantly decreased Abeta level, increased superoxide dismutase activity and decreased malondialdehyde level in hippocampus of model mice. It also restored acetylcholine amounts, inhibited acetylcholinesterase activities and increased choline acetyltransferase activities. In addition, JWKXS granules enabled the transformation of precursors of NGF and BDNF into mature forms. Furthermore, JWKXS granules could regulate gene expressions related to Abeta production, transportation, degradation and neurotrophic factor transformation, which led to down-regulation of Abeta and up-regulation of NGF and BDNF. These findings suggested that JWKXS granules ameliorated cognitive deficit via decreasing Abeta levels, protecting neuron from oxidation damages and nourishing neuron, which could serve as alternative medicine for patients suffering from AD.
ESTHER : Zhu_2019_Front.Pharmacol_10_258
PubMedSearch : Zhu_2019_Front.Pharmacol_10_258
PubMedID: 30941041

Title : Dietary mulberry leaf powder affects growth performance, carcass traits and meat quality in finishing pigs - Liu_2019_J.Anim.Physiol.Anim.Nutr.(Berl)_103_1934
Author(s) : Liu Y , Li Y , Peng Y , He J , Xiao D , Chen C , Li F , Huang R , Yin Y
Ref : J Anim Physiol Anim Nutr (Berl) , 103 :1934 , 2019
Abstract : This study was conducted to evaluate the effect of mulberry leaves as an alternative source of protein on growth performance, carcass traits and meat quality in finishing pigs. A total of 180 Xiangcun Black pigs were randomly assigned to five treatment groups with six pens of six pigs per pen. The pigs were provided with a basal diet or a diet contained 3%, 6%, 9% or 12% of mulberry leaf powder during a 50-day experiment period. The results showed that dietary mulberry leaf powder had no negative effect on growth performance in Xiangcun Black pigs, except in the 12% mulberry group, where final body weight and average daily gain decreased (p < .05) and feed to gain ratio of the pigs increased (p < .05). Dietary mulberry inclusion decreased (quadratic, p < .05) the back fat thickness, fibre mean cross-sectional area (CSA) in the longissimus dorsi (LD) muscle and mRNA expression levels of myosin heavy chain (MyHC) IIb in LD and biceps femoris (BF) muscles, while increased (linear or quadratic, p < .05) the plasma concentration of albumin, levels of crude protein (CP), inosine monophosphate (IMP) and several amino acids in muscle tissues. When compared with the other groups, the 9% mulberry diet increased (p < .05) loin-eye area and contents of CP and IMP in muscles, while decreased (p < .05) plasma activity of cholinesterase and concentrations of uric acid and urea. The 6% mulberry diet had the lowest fibre mean CSA and shear force and increased total fibre number of the LD muscle, when compared with the other groups. These results suggest that including mulberry in the diet at <12% is an effective feed crop to improve meat quality and the chemical composition of muscle without negatively affecting growth performance.
ESTHER : Liu_2019_J.Anim.Physiol.Anim.Nutr.(Berl)_103_1934
PubMedSearch : Liu_2019_J.Anim.Physiol.Anim.Nutr.(Berl)_103_1934
PubMedID: 31478262

Title : Identification of acetylcholinesterase inhibitors using homogenous cell-based assays in quantitative high-throughput screening platforms - Li_2017_Biotechnol.J_12_
Author(s) : Li S , Huang R , Solomon S , Liu Y , Zhao B , Santillo MF , Xia M
Ref : Biotechnol J , 12 : , 2017
Abstract : Acetylcholinesterase (AChE) is an enzyme responsible for metabolism of acetylcholine, a neurotransmitter associated with muscle movement, cognition, and other neurobiological processes. Inhibition of AChE activity can serve as a therapeutic mechanism, but also cause adverse health effects and neurotoxicity. In order to efficiently identify AChE inhibitors from large compound libraries, homogenous cell-based assays in high-throughput screening platforms are needed. In this study, a fluorescent method using Amplex Red (10-acetyl-3,7-dihydroxyphenoxazine) and the Ellman absorbance method were both developed in a homogenous format using a human neuroblastoma cell line (SH-SY5Y). An enzyme-based assay using Amplex Red was also optimized and used to confirm the potential inhibitors. These three assays were used to screen 1368 compounds, which included a library of pharmacologically active compounds (LOPAC) and 88 additional compounds from the Tox21 program, at multiple concentrations in a quantitative high-throughput screening (qHTS) format. All three assays exhibited exceptional performance characteristics including assay signal quality, precision, and reproducibility. A group of inhibitors were identified from this study, including known (e.g. physostigmine and neostigmine bromide) and potential novel AChE inhibitors (e.g. chelerythrine chloride and cilostazol). These results demonstrate that this platform is a promising means to profile large numbers of chemicals that inhibit AChE activity.
ESTHER : Li_2017_Biotechnol.J_12_
PubMedSearch : Li_2017_Biotechnol.J_12_
PubMedID: 28294544

Title : Increased permeability of the blood-brain barrier and Alzheimer's disease-like alterations in slit-2 transgenic mice - Li_2015_J.Alzheimers.Dis_43_535
Author(s) : Li JC , Han L , Wen YX , Yang YX , Li S , Li XS , Zhao CJ , Wang TY , Chen H , Liu Y , Qi CL , He XD , Gu QL , Ye YX , Zhang Y , Huang R , Wu YE , He RR , Kurihara H , Song XY , Cao L , Wang LJ
Ref : J Alzheimers Dis , 43 :535 , 2015
Abstract : Alzheimer's disease (AD) is a progressive neurological disorder that primarily affects memory, and its prevalence is rising. Increasing evidence suggests that dysfunction of the blood-brain barrier (BBB) may be involved in AD and other neurodegenerative diseases. Herein, we report that the permeability of the BBB is increased and that AD-like alterations are present in Slit-2 overexpressing transgenic mice. We found that behavioral change and the corresponding molecular diagnostic markers of AD, such as hippocampal neuron apoptosis, amyloid-beta (Abeta) protein deposition, and acetylcholinesterase expression, were increased in the Slit-2 transgenic mice. Moreover, the endothelial cells were dysfunctional, the size of the lateral ventricle cavity increased, and the permeability of the BBB increased. Additionally, there was an increased serum level of glutamate indicating that the BBB is related to AD. Finally, histopathological analysis of other organs in the Slit-2 overexpressing mice did not show any marked abnormalities. These findings demonstrate that Slit2 overexpression may be responsible for AD-like alterations and the increased BBB permeability in these mice. Our study provides a potential novel mechanism for the development of AD.
ESTHER : Li_2015_J.Alzheimers.Dis_43_535
PubMedSearch : Li_2015_J.Alzheimers.Dis_43_535
PubMedID: 25114073

Title : Pratensein attenuates Abeta-induced cognitive deficits in rats: Enhancement of synaptic plasticity and cholinergic function - Wei_2015_Fitoterapia_101C_208
Author(s) : Wei L , Lv S , Huang Q , Wei J , Zhang S , Huang R , Lu Z , Lin X
Ref : Fitoterapia , 101C :208 , 2015
Abstract : An isoflavone was isolated from Trifolium pratense using bioassay-guided screening. The structure of this natural compound was elucidated based on its spectral data, and it was identified as pratensein. The protective effect of pratensein was evaluated using a cognitive impairment model induced by injecting amyloid beta (1-42) (Abeta1-42) into the bilateral hippocampus of rats. The results showed that pratensein treatment significantly protected against Abeta1-42-induced cognitive impairments, as evidenced by the improvement in learning and memory and the attenuation of neuronal degeneration and apoptosis in hippocampus. Analysis of the potential mechanisms of action showed that pratensein significantly decreased inflammatory indicators such as MDA, NO, nNOS, IL-1beta and TNF-alpha. Pratensein markedly decreased the content and deposition of beta-amyloid peptide through regulating the expressions of Abeta-related genes including APP, BACE1, CatB, NEP and IDE. Moreover, pratensein significantly increased the expressions of synapse plasticity-related proteins, i.e., PSD-95, p-NMDAR1, p-CaMKII, p-PKACbeta, PKCgamma, p-CREB and BDNF. In addition, pratensein significantly decreased the activity of cholinesterase, then subsequently elevated the level of acetylcholine. In summary, our study indicated that pratensein may have a likely protective effect against Alzheimer's disease (AD) via improving synaptic plasticity and increasing cholinesterase activity.
ESTHER : Wei_2015_Fitoterapia_101C_208
PubMedSearch : Wei_2015_Fitoterapia_101C_208
PubMedID: 25665942

Title : Encapsulation of enzyme via one-step template-free formation of stable organic-inorganic capsules: A simple and efficient method for immobilizing enzyme with high activity and recyclability - Huang_2015_Biotechnol.Bioeng_112_1092
Author(s) : Huang R , Wu M , Goldman MJ , Li Z
Ref : Biotechnol Bioeng , 112 :1092 , 2015
Abstract : Enzyme encapsulation is a simple, gentle, and general method for immobilizing enzyme, but it often suffers from one or more problems regarding enzyme loading efficiency, enzyme leakage, mechanical stability, and recyclability. Here we report a novel, simple, and efficient method for enzyme encapsulation to overcome these problems by forming stable organic-inorganic hybrid capsules. A new, facile, one-step, and template-free synthesis of organic-inorganic capsules in aqueous phase were developed based on PEI-induced simultaneous interfacial self-assembly of Fmoc-FF and polycondensation of silicate. Addition of an aqueous solution of Fmoc-FF and sodium silicate into an aqueous solution of PEI gave a new class of organic-inorganic hybrid capsules (FPSi) with multi-layered structure in high yield. The capsules are mechanically stable due to the incorporation of inorganic silica. Direct encapsulation of enzyme such as epoxide hydrolase SpEH and BSA along with the formation of the organic-inorganic capsules gave high yield of enzyme-containing capsules ( approximately 1.2 mm in diameter), >90% enzyme loading efficiency, high specific enzyme loading (158 mg protein g(-1) carrier), and low enzyme leakage (<3% after 48 h incubation). FPSi-SpEH capsules catalyzed the hydrolysis of cyclohexene oxide to give (1R, 2R)-cyclohexane-1,2-diol in high yield and concentration, with high specific activity (6.94 U mg(-1) protein) and the same high enantioselectivity as the free enzyme. The immobilized SpEH demonstrated also excellent operational stability and recyclability: retaining 87% productivity after 20 cycles with a total reaction time of 80 h. The new enzyme encapsulation method is efficient, practical, and also better than other reported encapsulation methods. Biotechnol. Bioeng. 2015;112: 1092-1101. (c) 2015 Wiley Periodicals, Inc.
ESTHER : Huang_2015_Biotechnol.Bioeng_112_1092
PubMedSearch : Huang_2015_Biotechnol.Bioeng_112_1092
PubMedID: 25580912

Title : Self-assembly of amphiphilic janus particles into monolayer capsules for enhanced enzyme catalysis in organic media - Cao_2015_ACS.Appl.Mater.Interfaces_7_465
Author(s) : Cao W , Huang R , Qi W , Su R , He Z
Ref : ACS Appl Mater Interfaces , 7 :465 , 2015
Abstract : Encapsulation of enzymes during the creation of an emulsion is a simple and efficient route for enhancing enzyme catalysis in organic media. Herein, we report a capsule with a shell comprising a monolayer of silica Janus particles (JPs) (referred to as a monolayer capsule) and a Pickering emulsion for the encapsulation of enzyme molecules for catalysis purposes in organic media using amphiphilic silica JPs as building blocks. We demonstrate that the JP capsules had a monolayer shell consisting of closely packed silica JPs (270 nm). The capsules were on average 5-50 mum in diameter. The stability of the JP capsules (Pickering emulsion) was investigated with the use of homogeneous silica nanoparticles as a control. The results show that the emulsion stabilized via amphiphilic silica JPs presented no obvious changes in physical appearance after 15 days, indicating the high stability of the emulsions and JP capsules. Furthermore, the lipase from Candida sp. was chosen as a model enzyme for encapsulation within the JP capsules during their formation. The catalytic performance of lipase was evaluated according to the esterification of 1-hexanol with hexanoic acid. It was found that the specific activity of the encapsulated enzymes (28.7 U mL(-1)) was more than 5.6 times higher than that of free enzymes in a biphasic system (5.1 U mL(-1)). The enzyme activity was further increased by varying the volume ratio of water to oil and the JPs loadings. The enzyme-loaded capsule also exhibited high stability during the reaction process and good recyclability. In particular, the jellification of agarose in the JP capsules further enhanced their operating stability. We believe that the monolayer structure of the JP capsules, together with their high stability, rendered the capsules to be ideal enzyme carriers and microreactors for enzyme catalysis in organic media because they created a large interfacial area and had low mass transfer resistance through the monolayer shell.
ESTHER : Cao_2015_ACS.Appl.Mater.Interfaces_7_465
PubMedSearch : Cao_2015_ACS.Appl.Mater.Interfaces_7_465
PubMedID: 25478712

Title : Plasmonic ELISA for the ultrasensitive detection of Treponema pallidum - Nie_2014_Biosens.Bioelectron_58C_314
Author(s) : Nie XM , Huang R , Dong CX , Tang LJ , Gui R , Jiang JH
Ref : Biosensors & Bioelectronics , 58C :314 , 2014
Abstract : In this report, we have developed a plasmonic ELISA strategy for the detection of syphilis. Plasmonic ELISA is an enzyme-linked immunoassay combined with enzyme-mediated surface plasmon resonance (SPR) of gold nanoparticles (AuNPs). Immune response of the Treponema pallidum (T. pallidum) antibodies triggers the acetylcholinesterase-catalyzed hydrolysis of acetylthiocholine to produce abundant thiocholine. The positive charged thiol, in turn, alters the surface charge distribution the AuNPs and leads to the agglomeration of the AuNPs. The induced strong localized SPR effect of the agglomerate AuNPs can, thus, allow the quantitative assay of T. pallidum antibodies due to the remarkable color and absorption spectral response changes of the reaction system. The plasmonic ELISA exhibited a quasilinear response to the logarithmic T. pallidum antibody concentrations in the range of 1pg/mL-10ng/mL with a detection limit of 0.98pg/mL. Such a low detection limit was 1000-fold improvements in sensitivity over a conventional ELISA. The results of plasmonic ELISA in syphilis assays of serum specimens from 60 patients agreed with those obtained using a conventional ELISA method. The plasmonic ELISA has characteristics (analyte specific, cost-effective, ease of automatic, low limit of detection) that provide potential for diagnosis and therapeutic monitoring of syphilis.
ESTHER : Nie_2014_Biosens.Bioelectron_58C_314
PubMedSearch : Nie_2014_Biosens.Bioelectron_58C_314
PubMedID: 24662060

Title : Protective effect of Millettia pulchra polysaccharide on cognitive impairment induced by d-galactose in mice - Lin_2014_Carbohydr.Polym_101_533
Author(s) : Lin X , Huang Z , Chen X , Rong Y , Zhang S , Jiao Y , Huang Q , Huang R
Ref : Carbohydr Polym , 101 :533 , 2014
Abstract : A polysaccharide (PMP) was isolated from Millettia pulchra and purified by DEAE-cellulose and Sephadex G-75 chromatography. The results showed that PMP was composed of d-glucose and d-arabinose in a molar ratio of 90.79% and 9.21%, with an average molecular weight of about 14,301Da. Furthermore, the effect of PMP on cognitive impairment induced by d-galactose in mice was evaluated. Treatment with PMP significantly reversed d-galactose-induced learning and memory impairments, as measured by behavioral tests. One of the potential mechanisms of this action was to reduce oxidative stress and suppress inflammatory responses. Furthermore, our results also showed that PMP markedly reduced the content and deposition of beta-amyloid peptide, improved the dysfunction of synaptic plasticity, increased the levels of acetylcholine, but decreased cholinesterase activity. These results suggest that PMP exerts an effective protection against d-galactose-induced cognitive impairment, and PMP may be a major bioactive ingredient in M. pulchra.
ESTHER : Lin_2014_Carbohydr.Polym_101_533
PubMedSearch : Lin_2014_Carbohydr.Polym_101_533
PubMedID: 24299809

Title : Identification of differentially expressed proteins and phosphorylated proteins in rice seedlings in response to strigolactone treatment - Chen_2014_PLoS.One_9_e93947
Author(s) : Chen F , Jiang L , Zheng J , Huang R , Wang H , Hong Z , Huang Y
Ref : PLoS ONE , 9 :e93947 , 2014
Abstract : Strigolactones (SLs) are recently identified plant hormones that inhibit shoot branching and control various aspects of plant growth, development and interaction with parasites. Previous studies have shown that plant D10 protein is a carotenoid cleavage dioxygenase that functions in SL biosynthesis. In this work, we used an allelic SL-deficient d10 mutant XJC of rice (Oryza sativa L. spp. indica) to investigate proteins that were responsive to SL treatment. When grown in darkness, d10 mutant seedlings exhibited elongated mesocotyl that could be rescued by exogenous application of SLs. Soluble protein extracts were prepared from d10 mutant seedlings grown in darkness in the presence of GR24, a synthetic SL analog. Soluble proteins were separated on two-dimensional gels and subjected to proteomic analysis. Proteins that were expressed differentially and phosphoproteins whose phosphorylation status changed in response to GR24 treatment were identified. Eight proteins were found to be induced or down-regulated by GR24, and a different set of 8 phosphoproteins were shown to change their phosphorylation intensities in the dark-grown d10 seedlings in response to GR24 treatment. Analysis of these proteins revealed that they are important enzymes of the carbohydrate and amino acid metabolic pathways and key components of the cellular energy generation machinery. These proteins may represent potential targets of the SL signaling pathway. This study provides new insight into the complex and negative regulatory mechanism by which SLs control shoot branching and plant development.
ESTHER : Chen_2014_PLoS.One_9_e93947
PubMedSearch : Chen_2014_PLoS.One_9_e93947
PubMedID: 24699514

Title : Tumebacillus flagellatus sp. nov., an alpha-amylase\/pullulanase-producing bacterium isolated from cassava wastewater - Wang_2013_Int.J.Syst.Evol.Microbiol_63_3138
Author(s) : Wang Q , Xie N , Qin Y , Shen N , Zhu J , Mi H , Huang R
Ref : Int J Syst Evol Microbiol , 63 :3138 , 2013
Abstract : A novel alpha-amylase/pullulanase-producing bacterium, designated strain GST4(T), was isolated from samples collected from the wastewater of a cassava starch factory in Nanning, Guangxi Autonomous Region, southern China. Cells of strain GST4(T) were rod-shaped bacilli containing ellipsoidal terminal spores and found to be Gram-reaction-positive, aerobic, motile, oxidase-positive, catalase-negative and formed light yellow colonies on agar plates. Strain GST4(T) was able to grow at pH 4.5-8.5 (optimum at pH 5.5), temperatures ranging from 20 to 42 degrees C (optimum at 37 degrees C) and salt concentrations of 0-1% (w/v) NaCl (optimum at 0.5%, w/v) on R2A medium. Strain GST4(T) grew heterotrophically on complex carbon substrates and chemolithoautotrophically on inorganic sulfur compounds, as demonstrated by growth on sodium thiosulfate and sulfite as sole electron donors. It can reduce nitrate and nitrite. Strain GST4(T) contained iso-C(15:0) and anteiso-C(15:0) as the major cellular fatty acids and menaquinone 7 (MK-7) as the major respiratory quinone. The cell-wall peptidoglycan was of type A1gamma. The genomic DNA G+C content of strain GST4(T) was 53.7 mol%. Physiological and chemotaxonomic characteristics combined with phylogenetic analysis based on 16S rRNA gene sequences revealed that strain GST4(T) was a member of the genus Tumebacillus and most closely related to Tumebacillus permanentifrigoris DSM 18773(T) and Tumebacillus ginsengisoli DSM 18389(T) with 97.3 and 94.5% sequence similarity, respectively. The DNA-DNA relatedness values between strain GST4(T) and T. permanentifrigoris DSM 18773(T), and strain GST4(T) and T. ginsengisoli DSM 18389(T) were 44.0 and 60.4%, respectively. The new isolate differed from those species of the genus Tumebacillus in that it has peritrichous flagella for motility. Based on the evidence obtained from this study, strain GST4(T) represents a novel species of the genus Tumebacillus, for which the name Tumebacillus flagellatus sp. nov. is proposed. The type strain is GST4(T) ( =CGMCC 1.12170(T) =DSM 25748(T)).
ESTHER : Wang_2013_Int.J.Syst.Evol.Microbiol_63_3138
PubMedSearch : Wang_2013_Int.J.Syst.Evol.Microbiol_63_3138
PubMedID: 23435245
Gene_locus related to this paper: 9bacl-a0a074lri5 , 9bacl-a0a074ltm6

Title : The oyster genome reveals stress adaptation and complexity of shell formation - Zhang_2012_Nature_490_49
Author(s) : Zhang G , Fang X , Guo X , Li L , Luo R , Xu F , Yang P , Zhang L , Wang X , Qi H , Xiong Z , Que H , Xie Y , Holland PW , Paps J , Zhu Y , Wu F , Chen Y , Wang J , Peng C , Meng J , Yang L , Liu J , Wen B , Zhang N , Huang Z , Zhu Q , Feng Y , Mount A , Hedgecock D , Xu Z , Liu Y , Domazet-Loso T , Du Y , Sun X , Zhang S , Liu B , Cheng P , Jiang X , Li J , Fan D , Wang W , Fu W , Wang T , Wang B , Zhang J , Peng Z , Li Y , Li N , Chen M , He Y , Tan F , Song X , Zheng Q , Huang R , Yang H , Du X , Chen L , Yang M , Gaffney PM , Wang S , Luo L , She Z , Ming Y , Huang W , Huang B , Zhang Y , Qu T , Ni P , Miao G , Wang Q , Steinberg CE , Wang H , Qian L , Liu X , Yin Y
Ref : Nature , 490 :49 , 2012
Abstract : The Pacific oyster Crassostrea gigas belongs to one of the most species-rich but genomically poorly explored phyla, the Mollusca. Here we report the sequencing and assembly of the oyster genome using short reads and a fosmid-pooling strategy, along with transcriptomes of development and stress response and the proteome of the shell. The oyster genome is highly polymorphic and rich in repetitive sequences, with some transposable elements still actively shaping variation. Transcriptome studies reveal an extensive set of genes responding to environmental stress. The expansion of genes coding for heat shock protein 70 and inhibitors of apoptosis is probably central to the oyster's adaptation to sessile life in the highly stressful intertidal zone. Our analyses also show that shell formation in molluscs is more complex than currently understood and involves extensive participation of cells and their exosomes. The oyster genome sequence fills a void in our understanding of the Lophotrochozoa.
ESTHER : Zhang_2012_Nature_490_49
PubMedSearch : Zhang_2012_Nature_490_49
PubMedID: 22992520
Gene_locus related to this paper: cragi-k1qzk7 , cragi-k1rad0 , cragi-k1p6v9 , cragi-k1pa46 , cragi-k1pga2 , cragi-k1pp63 , cragi-k1pwa8 , cragi-k1q0b1.1 , cragi-k1q0b1.2 , cragi-k1q1h2 , cragi-k1q2z6 , cragi-k1qaj8 , cragi-k1qaw5 , cragi-k1qhl5 , cragi-k1qly1 , cragi-k1qqb1.1 , cragi-k1qqb1.2 , cragi-k1qs61 , cragi-k1qs99 , cragi-k1qwl6 , cragi-k1r068 , cragi-k1r0n3.1 , cragi-k1r0n3.2 , cragi-k1r0r4 , cragi-k1r1i9 , cragi-k1r8q9 , cragi-k1rgi1 , cragi-k1rig4 , cragi-k1s0a7.1 , cragi-k1s0a7.2 , cragi-k1s0a7.3 , cragi-k1q6q0 , cragi-k1rru1 , cragi-k1qfi4 , cragi-k1qvm5 , cragi-k1qq58 , cragi-k1qdc0 , cragi-k1r754 , cragi-k1pje5 , cragi-k1qca6 , cragi-k1qdt5 , cragi-k1qkz7 , cragi-k1rgd2 , cragi-k1puh6 , cragi-k1raz4 , cragi-k1qqj4 , cragi-k1rbs1

Title : No association of five candidate genetic variants with amyotrophic lateral sclerosis in a Chinese population - Chen_2012_Neurobiol.Aging_33_2721 e3
Author(s) : Chen Y , Zeng Y , Huang R , Yang Y , Chen K , Song W , Zhao B , Li J , Yuan L , Shang HF
Ref : Neurobiology of Aging , 33 :2721 e3 , 2012
Abstract : Recently, 5 single nucleotide polymorphisms (SNPs), rs2306677 in the inositol 1,4,5-triphosphate receptor 2 gene (ITPR2), rs1541160 in the kinesin-association protein 3 gene (KIFAP3), rs6690993 and rs6700125 in the FLJ10986 gene, and rs10260404 in the dipeptidyl-peptidase 6 gene (DPP6) have been reported to be associated with the risk of developing sporadic amyotrophic lateral sclerosis (SALS) in Caucasian populations. However, this association is not consistent among different studies and yet to be tested in Chinese SALS patients. We examined the above SNPs in a large cohort consisting of 395 SALS patients and 288 controls from Southwest China. Our results suggest that these SNPs are unlikely to be a common cause of SALS in Chinese populations.
ESTHER : Chen_2012_Neurobiol.Aging_33_2721 e3
PubMedSearch : Chen_2012_Neurobiol.Aging_33_2721 e3
PubMedID: 22795786

Title : Identification and characterization of a novel thermostable pyrethroid-hydrolyzing enzyme isolated through metagenomic approach - Fan_2012_Microb.Cell.Fact_11_33
Author(s) : Fan X , Liu X , Huang R , Liu Y
Ref : Microb Cell Fact , 11 :33 , 2012
Abstract : BACKGROUND: Pyrethroid pesticides are broad-spectrum pest control agents in agricultural production. Both agricultural and residential usage is continuing to grow, leading to the development of insecticide resistance in the pest and toxic effects on a number of nontarget organisms. Thus, it is necessary to hunt suitable enzymes including hydrolases for degrading pesticide residues, which is an efficient "green" solution to biodegrade polluting chemicals. Although many pyrethroid esterases have consistently been purified and characterized from various resources including metagenomes and organisms, the thermostable pyrethroid esterases have not been reported up to the present.
RESULTS: In this study, we identified a novel pyrethroid-hydrolyzing enzyme Sys410 belonging to familyV esterases/lipases with activity-based functional screening from Turban Basin metagenomic library. Sys410 contained 280 amino acids with a predicted molecular mass (Mr) of 30.8 kDa and was overexpressed in Escherichia coli BL21 (DE3) in soluble form. The optimum pH and temperature of the recombinant Sys410 were 6.5 and 55 degrees C, respectively. The enzyme was stable in the pH range of 4.5-8.5 and at temperatures below 50 degrees C. The activity of Sys410 decreased a little when stored at 4 degrees C for 10 weeks, and the residual activity reached 94.1%. Even after incubation at 25 degrees C for 10 weeks, it kept 68.3% of its activity. The recombinant Sys410 could hydrolyze a wide range of rho-nitrophenyl esters, but its best substrate is rho-nitrophenyl acetate with the highest activity (772.9 U/mg). The enzyme efficiently degraded cyhalothrin, cypermethrin, sumicidin, and deltamethrin under assay conditions of 37 degrees C for 15 min, with exceeding 95% hydrolysis rate. CONCLUSION: This is the first report to construct metagenomic libraries from Turban Basin to obtain the thermostable pyrethroid-hydrolyzing enzyme. The recombinant Sys410 with broad substrate specificities and high activity was the most thermostable one of the pyrethroid-hydrolyzing esterases studied before, which made it an ideal candidate for the detoxification of pyrethroids.
ESTHER : Fan_2012_Microb.Cell.Fact_11_33
PubMedSearch : Fan_2012_Microb.Cell.Fact_11_33
PubMedID: 22409882
Gene_locus related to this paper: 9bact-Sys410

Title : Genesis of muscle fiber-type diversity during mouse embryogenesis relies on Six1 and Six4 gene expression - Richard_2011_Dev.Biol_359_303
Author(s) : Richard AF , Demignon J , Sakakibara I , Pujol J , Favier M , Strochlic L , Le Grand F , Sgarioto N , Guernec A , Schmitt A , Cagnard N , Huang R , Legay C , Guillet-Deniau I , Maire P
Ref : Developmental Biology , 359 :303 , 2011
Abstract : Adult skeletal muscles in vertebrates are composed of different types of myofibers endowed with distinct metabolic and contraction speed properties. Genesis of this fiber-type heterogeneity during development remains poorly known, at least in mammals. Six1 and Six4 homeoproteins of the Six/sine oculis family are expressed throughout muscle development in mice, and Six1 protein is enriched in the nuclei of adult fast-twitch myofibers. Furthermore, Six1/Six4 proteins are known to control the early activation of fast-type muscle genes in myocytes present in the mouse somitic myotome. Using double Six1:Six4 mutants (SixdKO) to dissect in vivo the genesis of muscle fiber-type heterogeneity, we analyzed here the phenotype of the dorsal/epaxial muscles remaining in SixdKO. We show by electron microscopy analysis that the absence of these homeoproteins precludes normal sarcomeric organization of the myofiber leading to a dystrophic aspect, and by immunohistochemistry experiments a deficiency in synaptogenesis. Affymetrix transcriptome analysis of the muscles remaining in E18.5 SixdKO identifies a major role for these homeoproteins in the control of genes that are specifically activated in the adult fast/glycolytic myofibers, particularly those controlling Ca(2+) homeostasis. Absence of Six1 and Six4 leads to the development of dorsal myofibers lacking expression of fast-type muscle genes, and mainly expressing a slow-type muscle program. The absence of restriction of the slow-type program during the fetal period in SixdKO back muscles is associated with a decreased HDAC4 protein level, and subcellular relocalization of the transcription repressor Sox6. Six genes thus behave as essential global regulators of muscle gene expression, as well as a central switch to drive the skeletal muscle fast phenotype during fetal development.
ESTHER : Richard_2011_Dev.Biol_359_303
PubMedSearch : Richard_2011_Dev.Biol_359_303
PubMedID: 21884692

Title : Codon optimization, expression and enzymatic comparison of Rhizopus oryzae lipases pro-ROL and m-ROL in Pichia pastoris - Yang_2011_Sheng.Wu.Gong.Cheng.Xue.Bao_27_1780
Author(s) : Yang J , Yan X , Huang R , Zhang B
Ref : Sheng Wu Gong Cheng Xue Bao , 27 :1780 , 2011
Abstract : Rhizopus oryzae lipase (ROL) is not only a biocatalyst used in a broad range of biotechnological fields, but also a model to investigate the function of intramolecular chaperone in the post-translational processing of lipase. In this study, we cloned and expressed the mature lipase gene (m-ROL) containing the pre-sequence (pro-ROL) of R. oryzae HU3005 in Pichia pastoris GS115 and characterized their enzymatic activities. m-ROL exhibited higher hydrolysis activity towards middle-chain substrates (C10 and C12) at pH 9.0, whereas pro-ROL preferred short-chain substrates (C4) and displayed maximal activity at pH 8.0. Moreover, pro-ROL possessed better thermal stability than m-ROL. This enzymatic discrepancy between m-ROL and p-ROL may be due to the pre-sequence that affects the folding and conformation of the mature lipase domain. To improve the expression level of m-ROL in P. pastoris, overlap extension PCR was conducted to substitute eight less-frequently used codons of m-ROL with frequently used codons. After methanol-induced expression for 72 h, the activity and protein content of the codon optimized m-ROL reached 132.7 U/mL and 50.4 mg/L, while the activity of the parental m-ROL and pro-ROL are 28.7 U/mL and 14.4 mg/L, 29.6 U/mL and 14.1 mg/L, respectively.
ESTHER : Yang_2011_Sheng.Wu.Gong.Cheng.Xue.Bao_27_1780
PubMedSearch : Yang_2011_Sheng.Wu.Gong.Cheng.Xue.Bao_27_1780
PubMedID: 22506419

Title : A systematic review and meta-analysis of the relationship between lipoprotein lipase Asn291Ser variant and diseases - Hu_2006_J.Lipid.Res_47_1908
Author(s) : Hu Y , Liu W , Huang R , Zhang X
Ref : J Lipid Res , 47 :1908 , 2006
Abstract : This systematic review attempted to summarize the associations between the Asn291Ser variant in the lipoprotein lipase (LPL) gene and dyslipidemia, the risk of type 2 diabetes mellitus (T2DM), and coronary heart disease (CHD). In addition, the relationships between the Asn291Ser variant and other metabolic diseases such as obesity and high blood pressure were also investigated in this systematic review. We systematically reviewed the literature by means of a meta-analysis. Twenty-one articles, including 19,246 white subjects, were selected for this meta-analysis. The summary standardized mean difference (SMD) of plasma triglyceride (TG) for carriers compared with noncarriers of the Asn291Ser variant was 3.23 (P < 0.00001). The summary SMD of plasma HDL-cholsterol (HDL-C) for carriers compared with noncarriers of the Asn291Ser variant was -3.42 (P < 0.0001). The summary SMD of the association of the Asn291Ser variant with plasma TG increased with increasing age and weight gain. Significant interactions between the LPL Asn291Ser variant and fasting glucose, T2DM, and CHD were seen (P = 0.02, 0.04, and 0.01, respectively). No significant interactions were seen between the LPL Asn291Ser variant and body mass index, waist-hip ratio, and blood pressure (P > 0.05). This meta-analysis indicates that the Asn291Ser variant in the LPL gene is a risk factor for dyslipidemia, characterized by hypertriglyceridemia and low HDL-C levels. And the Asn291Ser variant in the LPL gene predisposes to more severe dyslipidemia with increasing age and weight gain. Also, this meta-analysis shows that the LPL Asn291Ser variant is associated with CHD and T2DM.
ESTHER : Hu_2006_J.Lipid.Res_47_1908
PubMedSearch : Hu_2006_J.Lipid.Res_47_1908
PubMedID: 16741292

Title : The mechanism of ageing of phosphonylated acetylcholinesterase - Sun_1979_J.Med.Chem_22_1306
Author(s) : Sun M , Chang Z , Shau M , Huang R , Chou T
Ref : Journal of Medicinal Chemistry , 22 :1306 , 1979
Abstract : 1. The extent of potential reactivation of organophosphate-inhibited acetylcholinesterase decreases with time, a phenomenon called ageing. Ageing is due to dealkylation of the alkoxyl group of the residue bound to the enzyme. The rate of ageing is proportional to the electron-donating capacity of the alkyl group. 2. The ageing of phosphophonylated cholinesterase cal also be demonstrated using a phrenic nerve-diaphragm preparation. The same relationship between the rate of ageing and the structure of the alkyl group was observed. 3. Ageing occurs much faster in electrically stimulated preparations than in resting preparations. This may be due to production of a more acidic environment for the enzyme at the active centre by the products of hydrolysis of the acetylcholine released by stimulation.
ESTHER : Sun_1979_J.Med.Chem_22_1306
PubMedSearch : Sun_1979_J.Med.Chem_22_1306
PubMedID: 510296