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Homologous recombination mediates stable Fah gene integration and phenotypic correction in tyrosinaemia mouse-model

World Journal of Hepatology 2018年第2期10卷 277-286页

作者： Norman Junge Qinggong Yuan Thu Huong Vu Simon Krooss Christien Bednarski Asha Balakrishnan Toni Cathomen Michael P Manns Ulrich Baumann amar deep sharma Michael Ott Department of Pediatric Gastroenterology and Hepatology Hannover Medical School Department of Gastroenterology Hepatology and Endocrinology Hannover Medical School TWINCORE Centre for Experimental and Clinical Infection Research Medical Center University of Freiburg Institute for Cell and Gene Therapy Research Group Micro RNA in Liver Regeneration Cluster of Excellence REBIRTH Hannover Medical School

AIM To stably correct tyrosinaemia in proliferating livers of fumarylacetoacetate-hydrolase knockout(Fah-/-) mice by homologous-recombination-mediated targeted addition of the Fah *** C57 BL/6 Fah?exon5 mice served as an animal model for human tyrosinaemia type 1 in our study. The vector was created by amplifying human Fah c DNA including the TTR promoter from a lentivirus plasmid as described. The Fah expression cassette was flanked by homologous arms(620 bp and 749 bp long) of the Rosa26 gene locus. Mice were injected with 2.1 × 108 VP of this vector(r ***) via the tail vein. Mice in the control group were injected with 2.1 × 108 VP of a similar vector but missing the homologous arms(r ***). Primary hepatocytes from Fah-/-recipient mice, treated with our vectors, were isolated and 1 × 106 hepatocytes were transplanted into secondary Fah-/-recipient mice by injection into the spleen. Upon either vector application or hepatocyte transplantation NTBC treatment was stopped in recipient mice. RESULTS Here, we report successful HR-mediated genome editing by integration of a Fah gene expression cassette into the "safe harbour locus" Rosa26 by recombinant AAV8. Both groups of mice showed long-term survival, weight gain and FAH positive clusters as determined by immunohistochemistry analysis of liver sections in the absence of NTBC treatment. In the group of C57 BL/6 Fah?exon5 mice, which have been transplanted with hepatocytes from a mouse injected with r *** 156 d before, 6 out of 6 mice showed long-term survival, weight gain and FAH positive clusters without need for NTBC treatment. In contrast only 1 out 5 mice, who received hepatocytes from r *** treated mice, survived and showed few and smaller FAH positive clusters. These results demonstrate that homologous recombinationmediated Fah gene transfer corrects the phenotype in a mouse model of human tyrosinaemia type 1(Fah-/-mice) and

关键词： Gene therapy AAV8 Liver based metabolic disease Targeted integration ROSA26 Paediatric liver disease

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Cell atlas of CCl_(4)-induced progressive liver fibrosis reveals stage-specific responses

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Zoological Research 2023年第3期44卷 451-466页

作者： Peng-Cheng Guo Jing Zuo Ke-Ke Huang Guang-Yao Lai Xiao Zhang Juan An Jin-Xiu Li Li Li Liang Wu Yi-Ting Lin Dong-Ye Wang Jiang-Shan Xu Shi-Jie Hao Yang Wang Rong-Hai Li Wen Ma Yu-Mo Song Chang Liu Chuan-Yu Liu Zhen Dai Yan Xu amar deep sharma Michael Ott Qing Ou-Yang Feng Huo Rong Fan Yong-Yin Li Jin-Lin Hou Giacomo Volpe Long-Qi Liu Miguel A.Esteban Yi-Wei Lai State Key Laboratory for Zoonotic Diseases Key Laboratory for Zoonosis Research of Ministry of EducationInstitute of ZoonosisCollege of Veterinary MedicineJilin UniversityChangchunJilin 130062China BGI-Hangzhou HangzhouZhejiang 310012China BGI-Shenzhen ShenzhenGuangdong 518083China Key Laboratory of Biological Targeting Diagnosis Therapy and Rehabilitation of Guangdong Higher Education InstitutesFifth Affiliated Hospital of Guangzhou Medical UniversityGuangzhouGuangdong 510799China Laboratory of Integrative Biology Guangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhouGuangdong 510530China Joint School of Life Sciences Guangzhou Institutes of Biomedicine and Health and Guangzhou Medical UniversityGuangzhouGuangdong 510530China School of Life Sciences Division of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiAnhui 230026China College of Life Sciences University of Chinese Academy of SciencesBeijing 100049China Guangzhou Institutes of Biomedicine and Health Chinese Academy of SciencesGuangzhouGuangdong 510530China Biotherapy Centre Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouGuangdong 510630China Department of Gastroenterology Hepatology and EndocrinologyHannover Medical SchoolHannover 30625Germany Department of Hepatobiliary Surgery and Liver Transplant Center General Hospital of Southern Theater CommandGuangzhouGuangdong 510010China Department of Infectious Diseases Nanfang HospitalSouthern Medical UniversityGuangdong Provincial Key Laboratory of Viral Hepatitis ResearchGuangzhouGuangdong 510515China Hematology and Cell Therapy Unit IRCCS-Istituto Tumori‘Giovanni Paolo II’Bari 70124Italy Institute of Experimental Hematology Hannover Medical SchoolHannover 30625Germany

Chronic liver injury leads to progressive liver fibrosis and ultimately cirrhosis,a major cause of morbidity and mortality ***,there are currently no effective anti-fibrotic therapies available,especially for latestage patients,which is partly attributed to the major knowledge gap regarding liver cell heterogeneity and cellspecific responses in different fibrosis *** reveal the multicellular networks regulating mammalian liver fibrosis from mild to severe phenotypes,we generated a single-nucleus transcriptomic atlas encompassing 49919nuclei corresponding to all main liver cell types at different stages of murine carbon tetrachloride(CCl_(4))-induced progressive liver *** analysis distinguished the sequential responses to injury of hepatocytes,hepatic stellate cells and endothelial ***,we reconstructed the cell-cell interactions and gene regulatory networks implicated in these *** integrative analyses uncovered previously overlooked aspects of hepatocyte proliferation exhaustion and disrupted pericentral metabolic functions,dysfunction for clearance by apoptosis of activated hepatic stellate cells,accumulation of pro-fibrotic signals,and the switch from an anti-angiogenic to a pro-angiogenic program during CCl_(4)-induced progressive liver *** dataset thus constitutes a useful resource for understanding the molecular basis of progressive liver fibrosis using a relevant animal model.

关键词： Liver fibrosis Toxicity Single-cell and single-nucleus RNA-sequencing Hepatocytes Hepatic stellate cells Angiogenesis Cell-cell interactions Gene regulatory networks

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Effect of stacking sequence on physical, mechanical and tribological properties of glass-carbon hybrid composites

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Friction 2014年第4期2卷 354-364页

作者： Gaurav AGARWAL amar PATNAIK Rajesh kumar sharma Jyoti AGARWAL Department of Mechanical Engineering Invertis UniversityBareillyIndia Department of Mechanical Engineering National Institute of Technology(NIT)HamirpurIndia Department of Computer Engineering Rajshree Institute of Management&Technology(RIMT)BareillyIndia

A study was done to determine the effect of best stacking sequence(position and orientation)on the mechanical properties,friction and wear response of hybrid *** main purpose of the study was to determine the best available stacking sequence for which the physical,mechanical and three body abrasive wear rate is *** for mechanical properties(tensile strength,flexural strength,inter laminar shear strength,impact strength and hardness)was done on glass-carbon hybrid *** composites were then subjected to test for three body abrasive wear on a designed experimental *** different factors(sliding velocity,fabric weight percentage,normal load,sliding distance and abrasive particle size)varied in steps to evaluate the friction and three body abrasive wear response of the *** design of experiment using Taguchi’s orthogonal array and analysis of variance(ANOVA)was applied to find out minimum specific wear *** results revealed the significance in arrangement of two different layers of fabric(glass and carbon)and the difference in properties obtained by *** Taguchi experimental analysis it has been observed that three body abrasive wear rate is minimum at 72 cm/s sliding velocity,30 wt.%fabric content,80 N normal load,90 m sliding distance and 125μm abrasive particle *** scanning electron microscope(SEM)studies revealed the dynamics of three‐body abrasive wear and underlying micro‐mechanisms that result in the analysis of three body abrasive wear.

关键词： hybrid composite stacking sequence Taguchi’s method wear mechanism

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Enhanced Marathi Speech Recognition Facilitated by Grasshopper Optimisation-Based Recurrent Neural Network

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Computer Systems Science & Engineering 2022年第11期43卷 439-454页

作者： Ravindra Parshuram Bachate Ashok sharma amar Singh Ayman AAly Abdulaziz HAlghtani Dac-Nhuong Le School of Computer Science and Engineering Lovely Professional UniversityPunjab144001India Department of Computer Science and IT University of JammuJammu and Kashmir180006India School of Computer Applications Lovely Professional UniversityPunjab144001India Department of Mechanical Engineering College of EngineeringTaif UniversityTaif21944Saudi Arabia School of Computer Science Duy Tan UniversityDanang550000Vietnam Institute of Research and Development Duy Tan UniversityDanang550000Vietnam

Communication is a significant part of being human and living in the *** kinds of languages and their variations are there;thus,one person can speak any language and cannot effectively communicate with one who speaks that language in a different *** application fields such as education,mobility,smart systems,security,and health care systems utilize the speech or voice recognition models ***,various studies are focused on the Arabic or Asian and English languages by ignoring other significant languages like Marathi that leads to the broader research motivations in regional *** is necessary to understand the speech recognition field,in which the major concentrated stages are feature extraction and *** paper emphasis developing a Speech Recognition model for the Marathi language by optimizing Recurrent Neural Network(RNN).Here,the preprocessing of the input signal is performed by smoothing and median *** preprocessing the feature extraction is carried out using MFCC and Spectral features to get precise features from the input Marathi Speech *** optimized RNN classifier is used for speech recognition after completing the feature extraction task,where the optimization of hidden neurons in RNN is performed by the Grasshopper Optimization Algorithm(GOA).Finally,the comparison with the conventional techniques has shown that the proposed model outperforms most competing models on a benchmark dataset.

关键词： deep learning grasshopper optimization algorithm recurrent neural network speech recognition word error rate

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Plant Disease Diagnosis and Image Classification Using deep Learning

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Computers, Materials & Continua 2022年第5期71卷 2125-2140页

作者： Rahul sharma amar Singh Kavita N.Z.Jhanjhi Mehedi Masud Emad Sami Jaha Sahil Verma School of Computer Applications Lovely Professional University Jalandhar144411India Department of Computer Science and Engineering Chandigarh UniversityMohaliIndia Department of Computer Science and Engineering Tailor’s UniversityMalaysia Department of Computer Science College of Computer and Information TechnologyTaif UniversityTaif21944Saudi Arabia Department of Computer Science Faculty of Computer and Information TechnologyKing Abdulaziz UniversityJeddah21589Saudi Arabia

Indian agriculture is striving to achieve sustainable intensification,the system aiming to increase agricultural yield per unit area without harming natural resources and the *** farming employs technology to improve *** and accurate analysis and diagnosis of plant disease is very helpful in reducing plant diseases and improving plant health and food crop *** disease experts are not available in remote areas thus there is a requirement of automatic low-cost,approachable and reliable solutions to identify the plant diseases without the laboratory inspection and expert’s *** learning-based computer vision techniques like Convolutional Neural Network(CNN)and traditional machine learning-based image classification approaches are being applied to identify plant *** this paper,the CNN model is proposed for the classification of rice and potato plant leaf *** leaves are diagnosed with bacterial blight,blast,brown spot and tungro *** leaf images are classified into three classes:healthy leaves,early blight and late blight *** leaf dataset with 5932 images and 1500 potato leaf images are used in the *** proposed CNN model was able to learn hidden patterns from the raw images and classify rice images with 99.58%accuracy and potato leaves with 97.66%*** results demonstrate that the proposed CNN model performed better when compared with other machine learning image classifiers such as Support Vector Machine(SVM),K-Nearest Neighbors(KNN),Decision Tree and Random Forest.

关键词： Plant diseases detection CNN image classification deep learning in agriculture

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Ultrasonographic assessment of paediatric ocular emergencies： A tertiary eye hospital based observation

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World Journal of Emergency Medicine 2018年第4期9卷 272-275页

作者： amar Pujari deepa R Swamy Rashmi Singh Ritika Mukhija Rohan Chawla Pradccp sharma Dr. Rajendra Prasad Centre for Ophthalmic Sciences All India Institute of Medical Sciences New Delhi 110029 India

BACKGROUND: The purpose of this study is to assess the utility of ocular ultrasound B scan in the emergency at the fi rst point of care for detecting posterior segment and orbital pathologies in cases of paediatric ocular ***: A prospective observational study involving 122 paediatric patients presenting to eye emergency over a period of ninety days were assessed with ultrasonography for the posterior segment as well as orbital pathology whenever indicated. The ocular ultrasound was performed gently over closed ***: Posttraumatic globe injuries were the most common indication for posterior segment evaluation, which constituted 80(65.57%) eyes. Among these 52 patients had an anechoic posterior segment and 28 patients had variable findings such as vitreous haemorrhage(8.19%), retinal detachment(6.55%), choroidal detachment(4.91%), posteriorly dislocated clear lens(0.81%) and retained intraocular foreign body(5.73%). Non-traumatic cases constituted around 42(34.42%) eyes, which included corneal ulcer(7.37%), retinoblastoma(6.55%), endophthalmitis(4.91%), extraocular muscle cysticercosis(4.91%), orbital cellulitis(4.09%), periocular haemorrhage(2.45%), proptosis(1.63%), paediatric cataract(1.63%) and cryptophthalmos(0.81%). No adverse events of performing the ultrasound was ***: First point ultrasonography in paediatric ocular emergencies is a cheap, portable and an effective tool in the assertion of signifi cant posterior segment and orbital diseases.

关键词： Paediatric ocular emergencies Ultrasound B scan Posterior segment andorbital pathologies

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Liquefaction Potential Assessment of Guwahati City Using One Dimensional Ground Response Analysis

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Journal of Geoscience and Environment Protection 2020年第5期8卷 176-194页

作者： amar Farooq Siddique Binu sharma Department of Civil Engineering Assam Engineering College Guwahati India

Guwahati city which lies in the North Eastern region of India, falls in the highest seismic risk zonal level i.e. zone V in India. However, there are very few works on seismic hazard analysis of Guwahati soil considering the local site effects. The effect of large modifications in seismic waves that occur due to variation in soil properties near the surface of the earth is of great importance in geotechnical earthquake engineering. Seismic soil liquefaction, a soil seismic hazard, is evaluated in Guwahati city in terms of factor of safety against liquefaction along the soil profiles using ground response analysis. One dimensional ground response analysis has been conducted using equivalent linear and non linear method using the deepsoil software. The input motion of 2011 Sikkim earthquake (Mw = 6.9) having bedrock PGA of 0.152 g at 30 m depth is considered. A comparative study has been made of the equivalent linear and non linear analysis in terms of surface PGA (g), maximum strain (%), maximum stress ratio and liquefaction potential using soil profiles of Guwahati city. It has been observed that stiffer soil layer results in similar PGA from both the analysis however non linear analysis generally gives a lesser surface PGA than by equivalent linear analysis. Non linear analysis generally gives a higher strain range and a lower maximum stress ratio as compared to the equivalent linear method. A slightly higher factor of safety is obtained using non linear analysis than using equivalent linear analysis. A soil database of 200 bore holes was used for the study. Spatial distribution of soil liquefaction potential is presented in the form of GIS based maps of factor of safety values.

关键词： Ground Response Analysis Equivalent Linear Method Non Linear Method Soil Liquefaction

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Association of NFKB1 gene polymorphism (rs28362491) with levels of inflammatory biomarkers and susceptibility to diabetic nephropathy in Asian Indians

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World Journal of Diabetes 2017年第2期8卷 66-73页

作者： amar Gautam Stuti Gupta Mohit Mehndiratta Mohini sharma Kalpana Singh Om P Kalra Sunil Agarwal Jasvinder K Gambhir Department of Medicine University College of Medical Sciences (University of Delhi) and G.T.B. Hospital Molecular Diagnostic Laboratory Department of Biochemistry University College of Medical Sciences (University of Delhi) and G.T.B. Hospital

AIM To investigate the association of NFKB1 gene-94 ATTG insertion/deletion(rs28362491) polymorphism with inflammatory markers and risk of diabetic nephropathy in Asian *** A total of 300 subjects were recruited(100 each), normoglycemic,(NG); type 2 diabetes mellitus(T2DM) without any complications(DM) and T2 DM with diabetic nephropathy [DM-chronic renal disease(CRD)]. Analysis was carried out by polymerase chain reaction-restriction fragment length polymorphism and ELISA. Pearson's correlation, analysis of variance and logistic regression wereused for statistical *** The allelic frequencies of-94 ATTG insertion/deletion were 0.655/0.345(NG), 0.62/0.38(DM) and 0.775/0.225(DM-CRD). The-94 ATTG ins allele was associated with significantly increased levels of urinary monocyte chemoattractant protein-1(u MCP-1); u MCP-1(P = 0.026) and plasma tumor necrosis factor-alpha(TNF-α); TNF-α(P = 0.030) and almost doubled the risk of diabetic nephropathy(OR = 1.91, 95%CI: 1.080-3.386, P = 0.025).CONCLUSION-94 ATTG ins/ins polymorphism might be associated with increased risk of developing nephropathy in Asian Indian subjects with diabetes mellitus.

关键词： Diabetic nephropathy Inflammation NFKB1 -94 ATTG ins/del polymorphism Urinary monocyte chemoattractant protein-1 Tumor necrosis factor-alpha

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Loss of the BRCA1-PALB2 interaction accelerates p53-associated tumor development in mice

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Genes & Diseases 2022年第3期9卷 807-813页

作者： amar H.Mahdi Yanying Huo Ying Chen Pier Selenica Anchal sharma Elise Merritt Nicola Barnard Chang Chan Shridar Ganesan Jorge S.Reis-Filho Britta Weigelt Subhajyoti De Bing Xia Rutgers Cancer Institute of New Jersey New BrunswickNJ 08903USA Department of Radiation Oncology Rutgers Robert Wood Johnson Medical SchoolNew BrunswickNJ 08903USA Department of Pathology Memorial Sloan Kettering Cancer CenterNew YorkNY 10065USA Department of Pathology and Laboratory Medicine Rutgers Robert Wood Johnson Medical SchoolNew BrunswickNJ 08901USA

The BRCA1-PALB2-BRCA2 axis,or the BRCA pathway,plays key roles in genome stability maintenance and suppression of breast and several other *** to frequent p53 mutations in human BRCA1 breast cancers and mouse mammary tumors from Brcal,Brca2 and Palb2 conditional knockout models,it is often thought that p53 inactivation accelerates BRCA1/2 and PALB2-associated ***,we studied tumor development in mice with a mutation in Palb2 that disengages the PALB2-BRCA1 interaction in different Trp53 *** than mammary tumors,Palb2 and Trp53 compound mutant mice developed,with greatly reduced latencies,lymphomas and sarcomas that are typically associated with germline Trp53 *** exome sequencing failed to identify any significant differences in genomic features between the same tumor types of Trp53 single mutant and Palb2;Trp53 compound mutant *** results suggest that loss of the BRCA pathway accelerates p53-associated tumor development,possibly without altering the fundamental tumorigenic processes.

关键词： BRCA1 BRCA2 Osteosarcoma P53 PALB2 Thymic lymphoma

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