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Impact of Leucine 278 Residue on Fatty Acid Length Specificity of <i>Candida antarctica</i>Lipase B

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Advances in Microbiology 2015年第7期5卷 493-499页

作者： Fanghua Wang Shulin Hou Qian Wang Pu Wang Jinsong Liu Bo Yang Yonghua Wang College of Light Industry and Food Sciences South China University of Technology Guangzhou China State Key Laboratory of Respiratory Disease Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China School of Bioscience and Bioengineering South China University of Technology Guangzhou China

Structural analysis of Candida antarctica lipase B (CALB) indicates that side chain of leucine at 278 site lies above the entrance of the catalytic pocket, which prognosticates its potential role on substrate Specificity of the enzyme. To verify this presumption, shortened side chain of glycine or proline was rational designed and mutants were constructed by site-directed mutagenesis method. The colorimetric assay using p-nitrophenyl esters of fatty acids with various chain-lengths was used to study the substrate preference of lipases. Results indicated that L278G or L278P mutations both induced the drift of substrate Specificity of CALB from p-nitrophenyl caprylate (pNP-C8) to longer carbon chain length of p-nitrophenyl caprate (pNP-C10). Meanwhile, Vmax value of two mutants to pNP-C10 was both higher than that of wild-type. Docking results also indicated that shortened side chain of glycine or proline residues substitution at this site could get rid of the space block present above the catalytic pocket, and made longer chain substrate (pNP-C10) enter into the catalytic pocket easier. The modulation of Specificity observed allowed for building substrate binding model and opened new possibilities for designing ligand specific lipases.

关键词： Candida antarctica Lipase B (CALB) Site-Directed Mutagenesis Substrate Specificity Stereospecific Blockade

Rapid Internal Control Reference Recombinase-Aided Amplification Assays for EBV and CMV Detection

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Biomedical and Environmental Sciences 2021年第8期34卷 650-655页

作者： GAO Yuan TIE Yan Qing ZHAO Lin Qing TAN He DING Nan DING Ya Xin GUO Qi ZHANG Rui Qing WANG Jin Rong CHEN Zi Wei FAN Guo Hao SHEN Xin Xin FENG Zhi Shan MA Xue Jun Hebei Medical University Shijiazhuang 050031HebeiChina NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Hebei General Hospital Shijiazhuang 050070HebeiChina Laboratory of Virology Beijing Key Laboratory of Etiology of Viral Diseases in ChildrenCapital Institute of PediatricsBeijing 100020China The Third Xiangya Hospital of Central South University Changsha 410013HunanChina Center for Biosafety Mega-Science Chinese Academy of SciencesWuhan 430071Hubei China

Epstein-Barr virus(EBV)and cytomegalovirus(CMV),two of the most prevalent human herpesviruses,cause a wide spectrum of diseases and symptoms and are associated with serious health problem.In this study,we developed an internal control reference recombinase-aided amplification(ICR-RAA)assay for the rapid detection of EBV and CMV within 30 min.The assay had a sensitivity of 5 and 1 copies/test for EBV and CMV,respectively,with no cross reaction with other pathogens.In comparison with those of the commercial quantitative polymerase chain reaction(qPCR),the sensitivity of the EBV and CMV ICR-RAAs using extracted DNA was 93.33%and 84.84%,respectively;the Specificity was 98.75%and 100.00%,respectively;and the Kappa values were 0.930 and 0.892(PSpecificity was 100.00%。

关键词： Specificity Internal EBV

sTWEAK is a marker of early haematoma growth and leukoaraiosis in intracerebral haemorrhage

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Stroke & Vascular Neurology 2021年第4期6卷 528-535,I0011,I0012页

作者： Andrés da Silva-Candal Iria López-Dequidt Manuel Rodriguez-Yañez PauloÁvila-Gómez JoséManuel Pumar JoséCastillo Tomás Sobrino Francisco Campos Ramón Iglesias-Rey Pablo Hervella Clinical Neurosciences Research Laboratory(LINC) Health Research Institute of Santiago de Compostela(IDIS)Santiago de CompostelaSpain Department of Neurology Hospital Clínico UniversitarioSantiago de CompostelaSpain Department of Neuroradiology Hospital Clínico UniversitarioSantiago de CompostelaSpain

Objective To study the association between early growth of haematoma with biomarkers of endothelial dysfunction such as leukoaraiosis(LA)and the soluble tumour necrosis factor-like weak inducer of apoptosis(sTWEAK)in patients with intracerebral haemorrhage(ICH).Methods This is a retrospective observational study of patients with nontraumatic ICH.Clinical and biochemical parameters were analysed.sTWEAK levels were measured by ELISA.LA was analysed in the hemisphere without haemorrhage to avoid interference with the acute injury.The main endpoint was the haematoma growth evaluated by the difference in volume between the second and the initial neuroimage.Poor functional outcome,defined as a modified Rankin Scale>2 at 3 months,was considered as secondary endpoint.Receiver operating characteristic curve analysis was performed to stablish the best cut-off for sTWEAK levels associated with haematoma growth.Results We included 653 patients with ICH in our analysis(71.1±11.9 years,44%women).Haematoma growth was observed in 188 patients(28.8%).sTWEAK levels≥5600 pg/mL predicted ICH growth with a sensitivity of 84%and a Specificity of 87%.sTWEAK levels≥5600 pg/mL and the presence of LA were associated with haematoma growth(OR:42.46;(CI 95%22.67 to 79.52)and OR:2.73(CI 95%1.39 to 5.34),respectively).Also,the presence of LA(OR:4.31(CI 95%2.89 to 6.42))and the interaction between ICH growth and sTWEAK(OR:2.23(CI 95%1.40 to 3.55))were associated with poor functional outcome at 3 months.Conclusion sTWEAKs,together with the presence and grade of LA,are biomarkers able to predict ICH growth and poor functional outcome in patients with ICH.

关键词： TWEAK intracerebral Specificity

Recombinase Polymerase Amplification Based Rapid Detection of Aroma Gene in Rice

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Rice science 2023年第2期30卷 96-99,I0010-I0013页

作者： Amrita BANERJEE S.BHARTI J.KUMAR P.SAR PRIYAMEDHA N.P.MANDAL S.SARKAR Somnath ROY Central Rainfed Upland Rice Research Station Indian Council of Agricultural Research(ICAR)-National Rice Research InstituteHazaribagJharkhand 825301India Department of Biotechnology Vinoba Bhave UniversityHazaribagJharkhand 825301India Crop Improvement Division ICAR-National Rice Research InstituteCuttackOdisha 753006India

Recombinase polymerase amplification(RPA) has been emerged as an alternative to PCR due to its high Specificity and sensitivity in diagnostics of animal and plant pathogens. In this study, an RPA protocol was develope... 详细信息

关键词： Specificity Rec emerged

Three Novel Real-Time RT-PCR Assays for Detection of COVID-19 Virus

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China CDC weekly 2020年第25期2卷 453-457页

作者： Peihua Niu Roujian Lu Li Zhao Huijuan Wang Baoying Huang Fei Ye Wenling Wang Wenjie Tan Key Laboratory of Biosafety National Health and Family Planning CommissionNational Institute for Viral Disease Control and PreventionChina CDCBeijingChina

What is already known on this topic?A novel human coronavirus,known as SARS-CoV-2 or 2019-nCoV,is the causative agent of the coronavirus disease 2019(COVID-19).We have released the primers and probes of real-time reverse transcription polymerase chain reaction(rRT-PCR)assays for the laboratory detection of COVID-19 infection.What is added by this report?Here we provide detailed technical data and evaluate the performance of three novel rRT-PCR assays targeting the ORF1ab,N,and E genes for detection of COVID-19 infection.The application of rRT-PCR assays among four types of specimens(alveolar lavage,sputum,throat swabs,and stool)from patients with COVID-19 indicated that the mean viral loads detected in sputum were higher than other specimens.What are the implications for public health practice?These rRT-PCR assays reported here could be used for laboratory diagnosis of COVID-19 infection with high sensitivity,Specificity,and applicability.Sputum rather than throat swabs and stool should be a priority for specimen collection for laboratory detection of COVID-19.

关键词： diagnosis Specificity stool

Personalized neoantigen pulsed dendritic cell vaccine for advanced lung cancer

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Signal Transduction and Targeted Therapy 2021年第2期6卷 519-530页

作者： Zhenyu Ding Qing Li Rui Zhang Li Xie Yang shu Song Gao Peipei Wang Xiaoqing Su Yun Qin Yuelan Wang Juemin Fang Zhongzheng Zhu Xuyang Xia Guochao Wei Hui Wang Hong Qian Xianling Guo Zhibo Gao Yu Wang Yuquan Wei Qing Xu Heng Xu Li Yang State Key Laboratory of Biotherapy and Cancer Center West China HospltalSIchuan Universityand Collaborative Innovation Center tor BlotherapyChengdu 610041China Department of Biotherapy Cancer CenterWest China HospitalSichuan UniversityChengduChina Department of Oncology Cancer CenterShanghai Tenth People's HospitalTongil UniversityShanghai 200072China Department of Radiology West China Hospitalsichuan UniversityChengduSichuanChina YuceBio Technology Co LtdShenzhenChina

Neoantigens are considered to be ultimate target of tumor immunotherapy due to their high tumor Specificity and immunogenicity.Dendritic cell(DCs)vaccines based on neoantigens have exciting effects in treatment of some malignant tumors and are a promising therapeutic modality.Lung cancer is a lethal disease with the highest morbidity and mortality rate in the world.Despite the rapid development of targeted therapy and immune checkpoint inhibitors for lung cancer in recent years,their efficacy is still unsatisfactory overall.Therefore,there is an urgent unmet clinical need for lung cancer treatment.Here,we attempted to treat lung cancer using a personalized neoantigen peptide-pulsed autologous DC vaccine and conducted a single-arm,2 medical centers,pilot study initiated by the investigator(ChiCTR-ONC 16009100,NCT02956551).The patients enrolled were patients with heavily treated metastatic lung cancer.Candidate neoantigens were derived from whole-exome sequencing and RNA sequencing of fresh biopsy tissues as well as bioinformatics analysis.A total of 12 patients were enrolled in this study.A total of 85 vaccine treatments were administered with a median value of 5 doses/person(range:3-14 doses/person).In total,12-30 peptide-based neoantigens were selected for each patient.AIl treatment-related adverse events were grade 1-2 and there were no delays in dosing due to toxic effects.The objective effectiveness rate was 25%;the disease control rate was 75%;the median progression-free survival was 5.5 months and the median overall survival was 7.9 months.This study provides new evidence for neoantigen vaccine therapy and new therapeutic opportunities for lung cancer treatment.

关键词： vaccine doses Specificity

Innovation focus in 2023

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The Innovation 2024年第1期5卷 1-2页

作者： [Anonymous] The Innovation Editorial Team China

Review of scientific breakthroughs in 2023,from neuromodulation and brainmachine interfaces to AI,superconductivity,and space exploration by The Innovation’s editorial team(Figure 1).NEUROMODULATION LEADS TO BREAKTHROUGHS Neuromodulation aims to modulate the activity of specific neurons through physical,chemical,or biological means,thereby modifying related perception,action,or even reversing disease states.Among the currently available neuromodulation techniques,chemogenetics has emerged as a rising star in recent years,offering advantages such as cell-type and pathway Specificity,less invasiveness,and increased flexibility.

关键词： Specificity Figure breakthrough

Gamma oscillations and their role in orchestrating balance and communication following stroke

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Neural Regeneration Research 2025年第2期20卷 477-478页

作者： Montana Samantzis Cong Wang Matilde Balbi Queensland Brain Institute The University of QueenslandBrisbaneQLDAustralia Engineering Research Centre of Traditional Chinese Medicine Intelligent Rehabilitation Ministry of EducationShanghaiChina

Induced brain oscillations in the gamma range have recently garnered attention due to their reported neuroprotective effects in the treatment of Alzheimer’s disease.This method differs from pharmacological approaches by tapping into the neuronal population dynamics that underlie the homeostatic processes in the brain that are crucial for the recovery of function.Recently,induced gamma-range oscillations have been used to improve cerebral blood flow,motor function,and synaptic plasticity in a mouse model of focal stroke,highlighting the broad potential of recruiting intrinsic recovery processes for the treatment of neurological conditions.Addressing open questions,such as the frequency Specificity of the benefits,will shed light on the intrinsic processes involved and allow clinicians to optimize recovery after stroke.

关键词： protective Specificity Alzheimer

Pentatricopeptide repeat protein PHOTOSYSTEM I BIOGENESIS FACTOR2 is required for splicing of ycf3

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Journal of Integrative Plant Biology 2020年第11期62卷 1741-1761页

作者： Xuemei Wang Zhipan Yang Yi Zhang Wen Zhou Aihong Zhang Congming Lu Key Laboratory of Ph otobiology Institute of BotanyChinese Academy of SciencesBeijing 100093China College of Life Sciences U niversity of C hinese Academy of SciencesBeijing 100049China State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTaian 271018China

To gain a better understanding of the molec-ular mechanisms of photosystem!(PSI)biogenesis,wecharacterized the Arabidopsis thaliana photosystem l bio-genesis factor 2(pbf2)mutant,which lacks PSl complex.PBF2 encodes a P-class pentatricopeptide repeat(PPR)protein.In the pbf2 mutants,we observed a striking de-crease in the transcript level of only one gene,thechloroplast gene ycf3,which is essential for PSl assembly.Further analysis of ycf3 transcripts showed that PBF2 isspecifically required for the splicing of ycf3 intron 1.Computational prediction of binding sequences and elec-trophoretic mobility shift assays reveal that PBF2 specifi-cally binds to a sequence in ycf3 intron 1.Moreover,we found that PBF2 interacted with two general factors forgroup ll intron splicing CHLOROPLAST RNA SPLICING2-AsSOCIATED FACTOR1(CAF1)and CAF2,and facilitated theassociation of these two factors with ycf3 intron 1.Ourresults suggest that PBF2 is specifically required for thesplicing of ycf3 intron 1 through cooperating with CAF1and CAF2.Our results also suggest that additional proteinsare required to contribute to the Specificity of CAF-dependent group ll intron splicing.

关键词： complex. striking Specificity