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Half a century after their discovery:Structural insights into exonuclease and annealase proteins catalyzing recombineering

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Engineering Microbiology 2024年第1期4卷 39-54页

作者： Lucy J.Fitschen Timothy P.Newing Nikolas P.Johnston Charles E.Bell Gökhan Tolun School of chemistry and molecular Bioscience and Molecular HorizonsUniversity of WollongongWollongongNSWAustralia The ARC Training Centre for Cryo-electron Microscopy of Membrane Proteins University of WollongongWollongongNSWAustralia Faculty of Science University of Technology SydneyUltimoNSW 2007Australia department of biological chemistry and pharmacology The Ohio State University College of MedicineColumbusOH 43210

Recombineering is an essential tool for molecular biologists,allowing for the facile and efficient manipulation of bacterial genomes directly in cells without the need for costly and laborious in vitro manipulations involving restriction *** main workhorses behind recombineering are bacteriophage proteins that promote the single-strand annealing(SSA)homologous recombination pathway to repair double-stranded DNA *** there have been several reviews examining recombineering methods and applications,comparatively few have focused on the mechanisms of the proteins that are the key players in the SSA pathway:a 5′→3′exonuclease and a single-strand annealing protein(SSAP or“annealase”).This review dives into the structures and functions of the two SSA recombination systems that were the first to be developed for recombineering in ***:the RecET system from *** Rac prophage and the𝜆Red system from bacteriophageλ.By comparing the structures of the RecT and Red𝛽annealases,and the RecE and𝜆Exo exonucleases,we provide new insights into how the structures of these proteins dictate their *** the sequence conservation of the𝜆λExo and RecE exonucleases gives more profound insights into their critical functional ***,as recombineering accelerates and evolves in the laboratory,a better understanding of the mechanisms of the proteins behind this powerful technique will drive the development of improved and expanded capabilities in the future.

关键词： Recombineering Redβ λExo RecE RecT Protein structure Annealase Exonuclease

Nitrosamines crisis in pharmaceuticals-Insights on toxicological implications,root causes and risk assessment:A systematic review

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Journal of Pharmaceutical Analysis 2024年第5期14卷 639-652页

作者： Hemanth P.R.Vikram Tegginamath Pramod Kumar Gunjan Kumar Narasimha M.Beeraka Rajashree Deka Sheik Mohammed Suhail Sandeep Jat Namitha Bannimath Gayatiri Padmanabhan Ravandur S.Chandan Pramod Kumar Bannimath Gurupadayya department of Pharmaceutical chemistry JSS College of Pharmacy MysuruJSS Academy of Higher Education and Research(JSSAHER)Mysuru570015India Xenone Healthcare Pvt.Ltd. New Delhi110076India department of Pharmaceutics JSS College of Pharmacy MysuruJSS Academy of Higher Education and Research(JSSAHER)Mysuru570015India department of Human Anatomy I.M.Sechenov First Moscow State Medical University(Sechenov University)Moscow119991Russian Federation department of pharmacology Raghavendra Institute of Pharmaceutical Education and Research(RIPER)Ananthapuramu515721India Herman B.Wells Center for Pediatric Research Department of PediatricsIndiana University School of MedicineIndianapolisIN46202USA Animal Physiology and Biochemistry Laboratory Department of ZoologyGauhati UniversityGuwahati781014India department of pharmacology JSS College of Pharmacy MysuruJSS Academy of Higher Education and Research(JSSAHER)Mysuru570015India department of Pharmaceutical Analysis National Institute of Pharmaceutical Education and Research(NIPER)-GuwahatiChangsari781101India department of pharmacology University of GalwayGalwayH91 TK33Ireland

The presence of N-nitroso compounds,particularly N-nitrosamines,in pharmaceutical products has raised global safety concerns due to their significant genotoxic and mutagenic *** systematic review investigates their toxicity in active pharmaceutical ingredients(APIs),drug products,and pharmaceutical excipients,along with novel analytical strategies for detection,root cause analysis,reformulation strategies,and regulatory guidelines for *** review emphasizes the molecular toxicity of N-nitroso compounds,focusing on genotoxic,mutagenic,carcinogenic,and other physiological ***,it addresses the ongoing nitrosamine crisis,the development of nitrosamine-free products,and the importance of sensitive detection methods and precise risk *** comprehensive overview will aid molecular biologists,analytical scientists,formulation scientists in research and development sector,and researchers involved in management of nitrosamine-induced toxicity and promoting safer pharmaceutical products.

关键词： Carcinogenicity Genotoxicity Mutagenicity molecular toxicity Nitrosamine impurities Nitrosamine drug substance-related impurities

Neuroprotective effects of G9a inhibition through modulation of peroxisome-proliferator activator receptor gamma-dependent pathways by miR-128

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Neural Regeneration Research 2024年第11期19卷 2532-2542页

作者： Aina Bellver-Sanchis Pedro AAvila-López Iva Tic David Valle-García Marta Ribalta-Vilella Luis Labrador Deb Ranjan Banerjee Ana Guerrero Gemma Casadesus Coralie Poulard Mercè Pallàs Christian Grinán-Ferré department of pharmacology and Therapeutic chemistry Institut de Neurociències-Universitat de BarcelonaBarcelonaSpain department of Biochemistry and molecular Genetics Feinberg School of MedicineNorthwestern UniversityChicagoILUSA Institute of Biotechnology National Autonomous University of MexicoCuernavacaMexico department of pharmacology and Therapeutics Health Science Center-University of FloridaGainesvilleFLUSA department of chemistry National Institute of Technology DurgapurM G AvenueDurgapurWest BengalIndia Cancer Research Cancer Lyon Universitéde LyonLyonFrance Inserm U1052 Centre de Recherche en Cancérologie de LyonLyonFrance CNRS UMR5286 Centre de Recherche en Cancérlogie de LyonLyonFrance Centro de Investigación en Red Enfermedades Neurodegenerativas(CIBERNED)Instituto de Salud Carlos IIIMadridSpain Neuroimmunology department National Institute of Neurology and Neurosurgery“Manuel Velasco Suárez”Mexico CityMexico

Dysregulation of G9a,a histone-lysine N-methyltransferase,has been observed in Alzheimer’s disease and has been correlated with increased levels of chronic inflammation and oxidative ***,microRNAs are involved in many biological processes and diseases playing a key role in pathogenesis,especially in multifactorial diseases such as Alzheimer’s ***,our aim has been to provide partial insights into the interconnection between G9a,microRNAs,oxidative stress,and *** better understand the biology of G9a,we compared the global microRNA expression between senescence-accelerated mouse-prone 8(SAMP8)control mice and SAMP8 treated with G9a inhibitor *** found a downregulation of miR-128 after a G9a inhibition treatment,which interestingly binds to the 3′untranslated region(3′-UTR)of peroxisome-proliferator activator receptor γ(PPARG)***,Pparg gene expression levels were higher in the SAMP8 group treated with G9a inhibitor than in the SAMP8 control *** also observed modulation of oxidative stress responses might be mainly driven Pparg after G9a *** confirm these antioxidant effects,we treated primary neuron cell cultures with hydrogen peroxide as an oxidative *** this setting,treatment with G9a inhibitor increases both cell survival and antioxidant ***,up-regulation of PPARγby G9a inhibitor could also increase the expression of genes involved in DNA damage responses and *** addition,we also described that the PPARγ/AMPK axis partially explains the regulation of autophagy markers ***,PPARγ/GADD45αpotentially contributes to enhancing synaptic plasticity and neurogenesis after G9a ***,we propose that pharmacological inhibition of G9a leads to a neuroprotective effect that could be due,at least in part,by the modulation of PPARγ-dependent pathways by miR-128.

关键词： aging cognitive decline epigenetics G9a inhibition microRNAs miR-128 peroxisome-proliferator activator receptorγ(PPARγ) PPARG SAMP8

Modeling tuberous sclerosis complex with human induced pluripotent stem cells

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World Journal of pediatrics 2024年第3期20卷 208-218页

作者： Weibo Niu Benjamin Siciliano Zhexing Wen department of Psychiatry and Behavioral Sciences Emory University School of MedicineWhitehead Research Building 447615 Michael StreetAtlantaGA 30322USA The Graduate Program in molecular and Systems pharmacology Laney Graduate SchoolEmory UniversityAtlanta.GA 30322.USA department of Cell Biology Emory University School of MedicineWhitehead Research Building 447615 Michael StreetAtlantaGA30322USA department of Neurology Emory University School of MedicineWhitehead Research Building 447.615 Michael StreetAtlantaGA 30322USA

Background Tuberous sclerosis complex(TSC)is an autosomal dominant genetic disorder with a birth incidence of I:6000 in the United States that is characterized by the growth of non-cancerous tumors in multiple organ systems including the brain,kidneys,lungs,and ***,TSCis also associated with signicant neurological manifestations including epilepsy TSC-associated neuropsychiatric disorders,intellectual disabilities,and autism spectrum *** in the TSCI or TSC2 genes are well-established causes of TSC,which lead to TSC1/TSC2 deficiency in organs and hyper-activation of the mammalian target of rapamycin signaling *** models have been widely used to study the effect of TSCl/2 genes on the development and function of the *** considerable progress in understanding the molecular mechanisms underlying TSC in animal models,a human-specific model is urgently needed to investigate the effects of TSCl/2 mutations that are unique to human *** sources Literature reviews and research articles were published in PubMed-indexed *** Human-induced pluripotent stem cells(iPSCs),which capture risk alleles that are identical to their donors and have the capacity to differentiate into virtually any cell type in the human body,pave the way for the empirical study of previously inaccessible biological systems such as the developing human *** In this review,we present an overview of the recent progress in modeling TSC with human iPSC models,the existing limitations,and potential directions for future research.

关键词： Induced pluripotent stem cells Three-dimensional cultures Tuberous sclerosis complex Two-dimensional cultures

Metformin reduces testosterone level in type 2 diabetes mellitus patients and its therapy by traditional medicine

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Clinical Traditional Medicine and pharmacology 2024年第4期5卷 19-26页

作者： Gorika Tomar Mitushi Singh Isha Sharma Anuj Nautiyal Saurabh Aggarwal Subhash Chandra Havagiray R.Chitme Cassio Rocha Medeiros Micheline de Azevedo Lima Lucia Raquel de Lima Henrique Douglas Melo Coutinho Bhavna Kumar department of Pharmacy Practice School of Pharmaceutical SciencesSGRR UniversityPatel NagarDehradun 248001UttarakhandIndia department of Medicine Shri MahantIndiresh HospitalSGRR UniversityPatel NagarDehradun 248001UttarakhandIndia department of Pharmaceutical chemistry School of Pharmaceutical SciencesSGRR UniversityPatel NagarDehradun 248001UttarakhandIndia department of pharmacology and Toxicology Amity Institute of PharmacyAmity UniversitySector-125Noida 201313 UPGwaliorIndia CECAPE College Av.Padre Cícero3917-São JoséJuazeiro do Norte 63024-015CEBrazil department of molecular Biology Federal University of Paraiba-UFPBJoao Pessoa 58051-900ParaibaBrazil department of biological chemistry Regional University of Cariri-URCACrato 63105-000CEBrazil department of Pharmaceutics DIT UniversityDehradun 248009UttarakhandIndia

Background:Type 2 Diabetes Mellitus(T2DM)is a chronic metabolic disorder characterized by insulin resistance and ***,a commonly prescribed treatment for T2DM,is known for its effectiveness in improving glycemic *** studies suggest that metformin may also influence testosterone levels in male patients,a hormone crucial for male reproductive health,muscle mass maintenance,and overall quality of *** plants were traditionally used for influencing the testosterone level in the body which may be given as a therapy for the side–effects of metformin ***:To assess the impact of metformin treatment on testosterone levels in male patients with T2DM compared to non-diabetic males,and to evaluate the potential implications of traditional medical treatments in managing the consequences of reduced testosterone ***:This study is a prospective observational investigation conducted among a total of 60 patients,divided into two groups:30 males taking metformin as a hyperglycaemic agent with T2DM and 30 males without any diabetic *** study was be carried out in different outpatient department of Shri Mahant Indiresh Hospital,Patel Nagar,Dehradun to assess the level of testosterone in male patients with T2DM taking metformin and in non-Diabetic *** on these findings,potential traditional medicines methods for reducing testosterone levels were ***:It was demonstrated that male patients with T2DM undergoing metformin treatment exhibited significant reduction in testosterone serum levels compared to non-diabetic *** plants were examinedand,and clinical studies were conducted on *** was discovered that these plants have been effective in increasingtestosterone level,suggesting their potential use as therapy for ***:This study shows reduction in the testosterone levels,which may lead to low sex drives and induction of low testosterone induced erectile dy

关键词： Type 2 diabetes mellitus Testosterone Metformin Erectile dysfunction Hyperglycaemia

Supramolecular organizing centers(SMOCs) as signaling machines in innate immune activation

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Science China(Life Sciences) 2015年第11期58卷 1067-1072页

作者： QIAO Qi WU Hao department of biological chemistry and molecular pharmacology Harvard Medical School Program in Cellular and molecular Medicine Boston Children's Hospital

Innate immunity offers the first line of defense against infections and other types of danger such as tumorigenesis. Its discovery provides tremendous therapeutic opportunities for numerous human diseases. Delving into the structural basis of signal transduction by innate immune receptors, our lab has recently helped to establish the new paradigm in which innate immune receptors transduce ligand-binding signals through formation of higher-order assemblies containing intracellular adapters, signaling enzymes and their substrates. These large signalosome assemblies may be visible under light microscopy as punctate structures in the μm scale, connecting to the underlying molecular structures in the nm scale. They drive proximity-induced enzyme activation, and provide a mechanism for signaling amplification by nucleated polymerization. These supramolecular signaling complexes also open new questions on their cellular organization and mode of regulation, pose challenges to our methodology, and afford valuable implications in drug discovery against these medically important pathways.

关键词： innate immunity caspase-recruitment domain pyrin domain supramolecular organizing centers filament

HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth(Lamiaceae) against Candida strains

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Asian Pacific Journal of Tropical Biomedicine 2019年第3期9卷 123-128页

作者： Janaína Esmeraldo Rocha Henrique Douglas Melo Coutinho Cícero Roberto Nascimento Saraiva Antonia Thassya Lucas dos Santos Ant?nio Judson Targino Machado Jo?o Tavares Calixto Júnior Irwin Rose Alencar Menezes José Galberto Martins da Costa Jaime Ribeiro-Filho Aracelio Viana Colares Laboratory of Microbiology and Biology molecular Department of Biological Chemistry Regional University of Cariri Leao Sampaio University Center department of biological Sciences Regional University of Cariri Laboratory of pharmacology and molecular chemistry Department of Biological Chemistry Regional University of Cariri Laboratory of Natural Product Research Department of Biological Chemistry Regional University of Cariri

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration(MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction(DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥4096 μg/mL against Candida albicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC_(50) of 2.60 and 2.53 μg/mL respectively, demonstrating significant synergistic effects against Candida strains. The modulator activity of DHm might be due to the presence of caffeic acid. Moreover, DHm and caffeic acid did not cause significant hemolytic effects, indicating that they present low cytotoxicity. Conclusions: These data indicate that DHm potentiates the activity of the fluconazole, without enhancement of the toxicity, encouraging further toxicological, pharmacological and phytochemical studies to provide consistent evidence of the potential of this plant to be used in drug development.

关键词： Hyptis martiusii Caffeic acid Candida albicans Candida tropicalis Antifungal properties

利用中心原子价层轨道阐释软硬酸碱理论的原理与应用

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化学教育（中英文） 2024年第6期45卷 99-107页

作者：杨雪梅王志鹏张永臻马新雨 Chen Maggie 蒋振雄哈尔滨工业大学(深圳)理学院广东深圳518055 Division of Genetics Department of MedicineBrigham and Women’s HospitalDepartment of Biological Chemistry and Molecular PharmacologyHarvard Medical SchoolBostonMA02115USA department of chemistry Texas A&M UniversityCollege StationTX77840USA 清华大学化学系北京100084 澳门科技大学创新工程学院材料科学与工程系中国澳门999078 department of chemistry and Chemical Biology Harvard UniversityBostonMA02138USA department of Microbiology Miami UniversityOxfordOH45056USA Computational Biology and Bioinformatics Program Duke UniversityDurhamNC27703USA

软硬酸碱理论作为基础化学的重要理论,其在教学与教材中却存在定义不清、难以应用、适用范围不明、模棱两可等众多问题。借用前线轨道理论的观念,针对核心原子的外层轨道进行系统性讨论,并分类举例说明其适用范围与应用方式,以期对基础... 详细信息

软硬酸碱理论作为基础化学的重要理论,其在教学与教材中却存在定义不清、难以应用、适用范围不明、模棱两可等众多问题。借用前线轨道理论的观念,针对核心原子的外层轨道进行系统性讨论,并分类举例说明其适用范围与应用方式,以期对基础教学和相关科研有助。

关键词：软硬酸碱理论前线轨道理论亲核进攻电负性

Evolution of chemistry and selection technology for DNA-encoded library

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Acta Pharmaceutica Sinica B 2024年第2期14卷 492-516页

作者： Peixiang Ma Shuning Zhang Qianping Huang Yuang Gu Zhi Zhou Wei Hou Wei Yi Hongtao Xu Shanghai Key Laboratory of Orthopedic Implants Department of OrthopedicsShanghai Ninth People’s HospitalShanghai Jiao Tong UniversitySchool of MedicineShanghai 200025China Shanghai Institute for Advanced Immunochemical Studies ShanghaiTech UniversityShanghai 201210China Guangzhou Municipal and Guangdong Provincial Key Laboratory of molecular Target&Clinical pharmacology The NMPA and State Key Laboratory of Respiratory DiseaseGuangzhou Medical UniversityGuangzhou 511436China College of Pharmaceutical Science and Institute of Drug Development&Chemical Biology Zhejiang University of TechnologyHangzhou 310014China

DNA-encoded chemical library(DEL)links the power of amplifiable genetics and the nonself-replicating chemical phenotypes,generating a diverse chemical *** analogy with the biological world,the DEL world can evolve by using a chemical central dogma,wherein DNA replicates using the PCR reactions to amplify the genetic codes,DNA sequencing transcripts the genetic information,and DNA-compatible synthesis translates into chemical ***,DNA-compatible synthesis is the key to expanding the DEL chemical ***,the evolution-driven selection system pushes the chemicals to evolve under the selective pressure,i.e.,desired selection *** this perspective,we summarized recent advances in expanding DEL synthetic toolbox and panning strategies,which will shed light on the drug discovery harnessing in vitro evolution of chemicals via DEL.

关键词： DNA-encoded library Drug discovery High-throughput selection DNA-compatible synthesis In vitro evolution Chemical central dogma