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Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA

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Journal of Pharmaceutical Analysis 2023年第9期13卷 1013-1023页

作者： Huiyi Yang Meng Qi Qiyi He Sung Hee Hwang Jun Yang Mark McCoy Christophe Morisseau Suqing Zhao Bruce D.Hammock department of Pharmaceutical Engineering School of Biomedical and Pharmaceutical SciencesGuangdong University of TechnologyGuangzhouChina department of entomology and Nematology and ucd comprehensive cancer center University of CaliforniaDavisCA 95616USA Langfang Normal University LangfangHebei065000China

To ensure proper dosage of a drug,analytical quantification of it in biofluid is *** chromatography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and ***,it requires expensive instrumentation and is not appropriate for bedside *** soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human *** optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and *** assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate *** addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory *** results indicate that nanobody-based ELISA methods can efficiently analyze drug like *** methods could be easily implemented by the bedside,in the field in remote areas or in veterinary *** work illustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and *** of nanobody based pharmaceutical assays are discussed.

关键词： Nanobody Immunoassay Soluble epoxide hydrolase inhibitors Metabolites Small molecules

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Synergistic effects of Pleistocene geological and climatic events on complex phylogeographic history of widespread sympatric species of Megaloptera in East Asia

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Zoological Research 2024年第5期45卷 1131-1146页

作者： Ai-Li Lin Ming-Ming Zou Li-Jun Cao Fumio Hayashi Ding Yang Xing-Yue Liu department of entomology China Agricultural UniversityBeijing 100193China Sanya Institute of China Agricultural University SanyaHainan 572025China International Joint Laboratory of Taxonomy and Systematic Evolution of Insecta Henan Institute of Science and TechnologyXinxiangHenan 453003China Institute of Plant and Environmental Protection Beijing Academy of Agriculture and Forestry SciencesBeijing 100097China department of Biology Tokyo Metropolitan UniversityHachioji-shiTokyo 192-0397Japan

Unraveling the phylogeographic histories of species remains a key endeavor for comprehending the evolutionary processes contributing to the rich biodiversity and high endemism found in East *** this study,we explored the phylogeographic patterns and demographic histories of three endemic fishfly and dobsonfly species(Neochauliodes formosanus,Protohermes costalis,and Neoneuromus orientalis)belonging to the holometabolan order *** species,which share a broad and largely overlapping distribution,were analyzed using comprehensive mitogenomic *** findings revealed a consistent influence of vicariance on the population isolation of *** and *** between Hainan,Taiwan,and the East Asian mainland during the early Pleistocene,potentially hindering subsequent colonization of the later diverged *** to these ***,we unveiled the dual function of the major mountain ranges in East Asia,serving both as barriers and conduits,in shaping the population structure of all three ***,we demonstrated that these co-distributed species originated from Southwest,Southern,and eastern Central China,respectively,then subsequently migrated along multi-directional routes,leading to their sympatric distribution on the East Asian ***,our results highlighted the significance of Pleistocene land bridges along the eastern coast of East Asia in facilitating the dispersal of mountain-dwelling insects with low dispersal ***,this study provides novel insight into the synergistic impact of Pleistocene geological and climatic events in shaping the diversity and distribution of aquatic insects in East Asia.

关键词： Comparative phylogeography Island biogeography Pleistocene glaciation Corydalidae East Asia

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Structure-guided discovery of potent and oral soluble epoxide hydrolase inhibitors for the treatment of neuropathic pain

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Acta Pharmaceutica Sinica B 2022年第3期12卷 1377-1389页

作者： Fangyu Du Ruolin Cao Lu Chen Jianwen Sun Yajie Shi Yang Fu Bruce D.Hammock Zhonghui Zheng Zhongbo Liu Guoliang Chen Key Laboratory of Structure-Based Drug Design&Discovery of Ministry of Education School of Pharmaceutical EngineeringShenyang Pharmaceutical UniversityShenyang 110016China department of entomology and Nematology and UC Davis comprehensive cancer center University of California DavisDavisCA 95616USA Shandong Xinhua Pharmaceutical Co. Ltd.Zibo 2550S6China School of Pharmacy Shenyang Pharmaceutical UniversityShenyang 110016China

Soluble epoxide hydrolase(sEH) is related to arachidonic acid cascade and is over-expressed in a variety of diseases, making sEH an attractive target for the treatment of pain as well as inflammatory-related diseases. A new series of memantyl urea derivatives as potent sEH inhibitors was obtained using our previous reported compound 4 as lead compound. A preferential modification of piperidinyl to 3-carbamoyl piperidinyl was identified for this series via structure-based rational drug design. Compound A20 exhibited moderate percentage plasma protein binding(88.6%) and better metabolic stability in vitro. After oral administration, the bioavailability of A20 was 28.6%. Acute toxicity test showed that A20 was well tolerated and there was no adverse event encountered at dose of 6.0 g/kg. Inhibitor A20 also displayed robust analgesic effect in vivo and dose-dependently attenuated neuropathic pain in rat model induced by spared nerve injury, which was better than gabapentin and sEH inhibitor(±)-EC-5026. In one word, the oral administration of A20 significantly alleviated pain and improved the health status of the rats, demonstrating that A20 was a promising candidate to be further evaluated for the treatment of neuropathic pain.

关键词： Soluble epoxide hydrolase Analgesia Synthesis Neuropathic pain Inhibitor

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Nanobody-based bispecific T-cell engager (Nb-BiTE): a new platform for enhanced T-cell immunotherapy

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Signal Transduction and Targeted Therapy 2023年第10期8卷 4444-4447页

作者： Xiao-mei Yang Xuan-dong Lin Wei Shi Shen-xia Xie Xia-ning Huang Shi-hua Yin Xiao-bing Jiang Bruce D.Hammock Zhi Ping Xu Xiao-ling Lu Guangxi Key Laboratory of Nanobody Research Guangxi Nanobody Engineering Research CenterCollege of StomatologyHospital of StomatologyLaboratory Animal CenterSchool of Basic Medical SciencePharmaceutical CollegeGuangxi Medical UniversityNanning CityGuangxi 530021P.R.China department of Neurosurgery Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan CityHubei 430022P.R.China department of entomology and Nematology UCD Comprehensive Cancer CenterUniversity of CaliforniaDavisCA 95616USA Australian Institute for Bioengineering and Nanotechnology The University of QueenslandBrisbaneQLD 4072Australia Institute of Biomedical Health Technology and Engineering and Institute of Systems and Physical Biology Shenzhen Bay LaboratoryShenzhen 518107P.R.China

A novel bispecific T-cell engager(BiTE)has been developed as an efficient immunotherapeutic molecule specifically bringing the T-cell and the tumor target together for enhanced *** general BiTE construct consists of two single-chain variable antibody fragments(scFvs)targeting a tumor-associated antigen(TAA)and a T-cell marker in *** binding of BiTEs to tumor antigens induces immediate T-cells’cytotoxicity against tumor cells without involving any typical costimulatory signals,specific TCR and MHC recognition.

关键词： immunotherapy tumor cytotoxicity

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Generation of Dual functional Nanobody-Nanoluciferase Fusion and its potential in Bioluminescence Enzyme Immunoassay for trace Glypican-3 in Serum

Generation of Dual functional Nanobody-Nanoluciferase Fusion...

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第十四届全国免疫学学术大会

作者： Siliang Duan Sheng Yu Zhenfeng Li Jingzhang Li Shimei Zhao Hongjing Liu Shanguang Wu Xiongjie Bi Dongyang Li Jiexian Dong Bruce D. Hammock Medical College Guangxi University of Science and Technology The Second Clinical Medical College of Guangxi University of Science and Technology department of entomology and Nematology and ucd comprehensive cancer center University of California Liuzhou People's Hospital The First Affiliated Hospital of Guangxi University of Science and Technology

Glypican-3(GPC3) is a serological biomarker for the diagnosis of Hepatocellular carcinoma(HCC), but it is a challenging task to develop a bioassay for determination of the trace GPC3 in serum. In this study, Bioluminescense immunoassay based on bifunctional nanobody-nanoluciferase fusion was developed with the ultra-sensitive feature to achieve this goal. First, nanobodies special against GPC3 binder as biological recognition element were generated by immunization and phage display technology. Second, The best clone GPN2 was fused with nanoluciferase as a dual-functional immunoreagent to establish an ultra-sensitive bioluminescence enzyme immunoassay(BLEIA), which is 30 and 5 times more sensitive than the traditional colorimetric assay and fluorescent assay, respectively. The cross-reactivity analysis of BLEIA showed that there was no cross-reactivity with HCC related tumor markers AFP, CEA, CA19-9 and GPC1/GPC2. The limit of detection(LOD) of developed BLEIA was 1.5 ng/mL, which assured its application in the diagnosis of GPC3 in 94 serum samples. This study indicates that BLEIA based on nanobody-nanoluciferase fusion could be used as a useful tool for the diagnosis of HCC patients.

关键词： Anti-GPC3 binder Nanobody/nanoluciferase fusion Bioluminescence enzyme immunoassay Diagnosis for HCC

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对硫磷纳米抗体筛选及分子识别机制研究

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分析化学 2019年第9期47卷 1419-1428页

作者：张玉琪张瑾如王锋李家冬司睿吕丽珊沈玉栋王弘 HAMMOCK Bruce D 孙远明华南农业大学食品学院广东省食品质量与安全重点实验室畜禽产品精准加工与安全控制技术国家地方联合工程研究中心(广东) 广州安诺科技股份有限公司 department of entomology and ucd comprehensive cancer center University of CaliforniaDavis

对硫磷是一种广谱的有机磷酸酯类杀虫、杀螨剂,毒性极强,可对环境和人类健康造成严重危害。尽管我国已禁止使用对硫磷,但仍存在违禁使用现象,加强对其监测十分必要。纳米抗体是已知最小的与抗原结合的片段（15 kDa）,来源于骆驼科动物... 详细信息

对硫磷是一种广谱的有机磷酸酯类杀虫、杀螨剂,毒性极强,可对环境和人类健康造成严重危害。尽管我国已禁止使用对硫磷,但仍存在违禁使用现象,加强对其监测十分必要。纳米抗体是已知最小的与抗原结合的片段（15 kDa）,来源于骆驼科动物体内重链抗体的可变区,具有稳定性强、灵敏度高、易表达、水溶性好等特性。本研究基于噬菌体展示技术构建了抗对硫磷纳米抗体库,其库容为2.41×10;cfu/mL,经抗原-抗体固相亲和筛选及噬菌体酶联免疫法（Phage ELISA）鉴定,获得5种抗对硫磷纳米抗体VHH1、VHH6、VHH13、VHH21和VHH24。以结合能力最强的VHH1为基础,采用同源建模和分子对接技术探讨VHH1与对硫磷分子的识别机制,发现VHH1与对硫磷存在氢键和疏水间分子作用力,关键氨基酸是Ser60、Lys104、Phe105、Gly106和Arg107。本研究为抗体的进化改造和半抗原的设计提供了新思路。

关键词：纳米抗体对硫磷噬菌体展示分子模拟

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A highly sensitive bio-barcode immunoassay for multi-residue detection of organophosphate pesticides based on fluorescence antiquenching

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Journal of Pharmaceutical Analysis 2022年第4期12卷 637-644页

作者： Lingyuan Xu Xiuyuan Zhang A.M.Abd El-Aty Yuanshang Wang Zhen Cao Huiyan Jia J.-Pablo Salvador Ahmet Hacimuftuoglu Xueyan Cui Yudan Zhang Kun Wang Yongxin She Fen Jin Lufei Zheng Baima Pujia Jing Wang Maojun Jin Bruce D.Hammock Institute of Quality Standard and Testing Technology for Agro-Products Chinese Academy of Agricultural SciencesBeijing100081China department of Pharmacology Faculty of Veterinary MedicineCairo UniversityGiza12211Egypt department of Medical Pharmacology Medical FacultyAtaturk UniversityErzurum25240Türkiye Institute of Livestock and Poultry Ningbo Academy of Agricultural SciencesNingboZhejiang315040China Nanobiotechnology for Diagnostics Group Instituto de Química Avanzada de CatalunaIQAC-CSICBarcelona08034Spain CIBER de Bioingeniería Biomateriales y Nanomedicina(CIBER-BBN)Madrid28029Spain Inspection and Testing center of Agricultural and Livestock Products of Tibet Department of Agriculture and Rural Affairs of Tibet Autonomous RegionLhasa850000China department of entomology&Nematology and the UC Davis comprehensive cancer center University of CaliforniaDavisCA95616USA

Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and *** nanoparticles were functionalized with monoclonal antibodies against the tested *** oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal *** detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the *** resulting fluorescence signal enables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concentration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,*** mean recovery ranged between 80.3% and 110.8% with relative standard deviations of 7.3%-17.6%,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.

关键词： Organophosphate pesticides Fluorescence anti-quenching Gold nanoparticles DNA-RNA hybridization DNA oligonucleotides Ribonuclease H

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Comparative analysis of peptidoglycan recognition proteins in endoparasitoid wasp Microplitis mediator

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Insect Science 2017年第1期24卷 2-16页

作者： Rui-Juan Wang Zhe Lin Hong Jiang Jiancheng Li Tusar T. Saha Ziyun Lu Zhiqiang Lu Zhen Zou department of entomology College of Plant Protection Northwest A&F University Yangling Shaanxi State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of Zoology Chinese Academy of Sciences Beijing Key Laboratory of Integrated Pest Management on Crops in Nor/hem Region of North China Ministry of Agriculture IPM Center of Hebei Province Institute of Plant Protection Hebei Academy of Agriculture and Forestry Sciences Baoding China department of entomology University of California Riverside CA USA

Peptidoglycan recognition proteins （PGRPs） are a family of innate immune receptors that specifically recognize peptidoglycans （PGNs） on the surface of a number of pathogens. Here, we have identified and characterized six PGRPs from endoparasitoid wasp, Microplitis mediator （MmePGRPs）. To understand the roles of PGRPs in parasitoid wasps, we analyzed their evolutionary relationship and orthology, expression profiles during different developmental stages, and transcriptional expression following infection with Gram-positive and -negative bacteria and a fungus. MmePGRP-S1 was significantly induced in response to pathogenic infection. This prompted us to evaluate the effects of RNA interference mediated gene specific knockdown ofMmePGRP-S1. The knockdown of MmePGRP-S1 （iMmePGRP-S1） dramatically affected wasps＇ survival following challenge by Micrococcus luteus, indicating the involvement of this particular PGRP in immune responses against Gram-positive bacteria. This action is likely to be mediated by the Toll pathway, but the mechanism remains to be determined. MmePGRP-S 1 does not play a significant role in anti-fungal immunity as indicated by the survival rate of iMmePGRP-S wasps. This study provides a comprehensive characterization of PGRPs in the economically important hymenopteran species M. mediator.

关键词： endoparasitoid wasp insect immunity microbial infection Microplitis mediator peptidoglycan recognition protein

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Elucidation of the MicroRNA Transcriptome in Western Corn Rootworm Reveals Its Dynamic and Evolutionary Complexity

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Genomics, Proteomics & Bioinformatics 2021年第5期19卷 800-814页

作者： Xiaozeng Yang Elane Fishilevich Marcelo A.German Premchand Gandra Robert E.McEwan André Billion Eileen Knorr Andreas Vilcinskas Kenneth E.Narva Beijing Agro-biotechnology Research center Beijing Academy of Agriculture and Forestry SciencesBeijing 100097China Corteva Agriscience Agriculture Division of DowDuPontIndianapolisIN 46268USA University of Nebraska-Lincoln Department of EntomologyLincolnNE 68583USA Fraunhofer Institute for Molecular Biology and Applied Ecology Department of BioresourcesGiessen 35394Germany

Diabrotica virgifera virgifera(western corn rootworm,WCR)is one of the most destructive agricultural insect pests in North *** is highly adaptive to environmental stimuli and crop protection ***,little is known about the underlying genetic basis of WCR behavior and *** specifically,the involvement of small RNAs(s RNAs),especially micro RNAs(mi RNAs),a class of endogenous small non-coding RNAs that regulate various biological processes,has not been examined,and the datasets of putative s RNA sequences have not previously been generated for *** achieve a comprehensive collection of s RNA transcriptomes in WCR,we constructed,sequenced,and analyzed s RNA libraries from different life stages of WCR and northern corn rootworm(NCR),and identified 101 conserved precursor mi RNAs(pre-mi RNAs)in WCR and other *** also identified 277 corn rootworm specific pre-mi *** analyses of s RNA populations in WCR revealed that its s RNA transcriptome,which includes PIWI-interacting RNAs(pi RNAs)and mi RNAs,undergoes a dynamic change throughout insect *** analysis of mi RNA datasets from model species reveals that a large pool of species-specific mi RNAs exists in corn rootworm;these are potentially evolutionarily *** of WCR mi RNA clusters to other insect species highlight conserved mi RNA-regulated processes that are common to *** Analysis of RNA Ends(PARE)also uncovered potential mi RNA-guided cleavage sites in ***,this study provides a new resource for studying the s RNA transcriptome and mi RNA-mediated gene regulation in WCR and other Coleopteran insects.

关键词： Western corn rootworm Small RNA MicroRNA Parallel Analysis of RNA Ends PIWI-interacting RNA

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New insights into the systematics of Entomobryoidea(Collembola: Entomobryomorpha): first instar chaetotaxy, homology and classification

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Zoological Systematics 2019年第4期44卷 249-278页

作者： Feng Zhang Bruno CBellini Felipe NSoto-Adames department of entomology College of Plant ProtectionNanjing Agricultural UniversityNanjing 210095China department of Botany and Zoology Biosciences CenterFederal University of Rio Grande do NorteNatalBrazil Division of Plant Industry Florida Department of Agriculture and Consumer ServicesGainesvilleFL 32608USA

The largest superfamily Entomobryoidea is one of the key groups in ***,incorrect recognition of chaetotaxic homology within Entomobryoidea severely impedes the accurate species comparison of adults and phylogenetic *** classification of the superfamily at suprageneric level is disputable in the light of recent *** homology of tergal chaetotaxy was traced and revised based on 38 species of first instar and partial early *** phylogenetic reconstructions were reconstructed mainly relying on first instar characters using both parsimony and likelihoodbased *** selection and several rogue taxa impacted on resolution and support of otherwise well-supported *** published molecular phylogeny,a revised classification of three families and nine subfamilies was presented:Orchesellidae,Entomobryidae and *** includes all basal taxa having a short fourth abdominal *** and Microfalculidae taxa were sunk into *** Paronellidae was divided into two subfamilies:Paronellinae(Paronellini+Cyphoderini+Bromacanthini)and Salininae(Cremastocephalini+Callyntrurini).Microfalculidae(Microfalcula)was closer to Salina and Akabosia and thus transferred into *** study erected a new classification framework for Entomobryoidea based upon comprehensive *** homologization across a wide range provided a standardized,comparable,powerful tool for taxonomy.

关键词： Phylogeny chaetotaxic homologization Orchesellidae Entomobryidae Paronellidae.

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