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Application of three-dimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey （Macaca mulatta）

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Asian Journal of Andrology 2015年第6期17卷 972-980,I0009页

作者： Mahmoud Huleihel Seyedmehdi Nourashrafeddin Tony M Plant the shraga segal department of microbiology Immunology and Genetics Faculty of Health Sciences Ben-Gurion University of the Negev Beer-Sheva Israel department of Obsteirics Gynecology and Reproductive Sciences Univeisity of Pittsburgh School of Medicine Magee-Womens Research InstitutePittsburgh PA 15213 USA

In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules （including those generated by the SSC niche） and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system （SACS） and a 3D methylcellulose culture system （MCS）. It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we （1） provide a brief overview of the differences in spermatogenesis in rodents and primates, （2） summarize data related to attempts to generate sperm in vitro, （3） report for the first time formation of colonies/clusters of cells and differentiation of meiotic （expression of CREM-1） and postmeiotic （expression of acrosin） germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and （4） indicate research needed to optimize 3D systems for in vitro primate spermatogenesis and for possible future application to man.

关键词： in vitrospermatogenesis methylcellulose culture system monkey primates soft agar culture system three-dimensional culture system

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Differentiation of murine male germ cells to spermatozoa n a soft agar culture system

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Asian Journal of Andrology 2012年第2期14卷 285-293页

作者： Mahmoud Abu Elhija Eitan Lunenfeld Stefan Schlatt Mahmoud Huleihel the shraga segal department of microbiology and Immunology Faculty of Health Sciences Ben-Gurion University of the Negev Beer-Sheva 84105 Israel department of Obstetrics and Gynecology Soroka University Medical Center and Faculty of Health Sciences Ben-Gurion University of the Negev Beer-Sheva 84105 Israel Centre of Reproductive Medicine and Andrology of the University of Menster Menster 84149 Germany

Establishment of an in vitro system that allows the development of testicular germ cells to sperm will be valuable for studies of spermatogenesis and future treatments for male infertility. In the present study, we developed in vitro culture conditions using three-dimensional agar culture system （SACS）, which has the capacity to induce testicular germ cells to reach the final stages of spermatogenesis, including spermatozoa generation. Seminiferous tubules from testes of 7-day-old mice were enzymatically dissociated, and intratubular cells were cultured in the upper layer of the SACS in RPMI medium supplemented with fetal calf serum （FCS）. The lower layer of the SACS contained only RPMI medium supplemented with FCS. Colonies in the upper layer were isolated after 14 and 28 days of culture and were classified according to their size. Immunofluorescence and real-time PCR were used to analyse specific markers expressed in undifferentiated and differentiated spermatogonia （Vasa, Dazl, OCT-4, C-Kit, GFR- a-l, CD9 and a-6-integrin）, meiotic cells （LDH, Crem-1 and Boule） and post-meiotic cells （Protamine-1, Acrosin and SP-IO）. Our results reveal that it is possible to induce mouse testicular pre-meiotic germ cell expansion and induce their differentiation to spermatozoa in SACS. The spermatozoa showed normal morphology and contained acrosomes. Thus, our results demonstrate that SACS could be used as a novel in vitro system for the maturation of pre-meiotic mouse germ cells to post-meiotic stages and morphologically-normal spermatozoa.

关键词： acrosome agar in vitro culture meiosis spermatogenesis spermatogonia spermatozoa testis

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Targeting the overexpressed mitochondrial protein VDAC1 in a mouse model of Alzheimer’s disease protects against mitochondrial dysfunction and mitigates brain pathology

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Translational Neurodegeneration 2022年第1期11卷 1-30页

作者： Ankit Verma Anna Shteinfer-Kuzmine Nikita Kamenetsky Srinivas Pittala Avijit Paul Edna Nahon Crystal Alberto Ouro Vered Chalifa-Caspi Swaroop Kumar Pandey Alon Monsonego Noga Vardi Shira Knafo Varda Shoshan-Barmatz department of Life Sciences Ben-Gurion University of the Negev84105 Beer-ShevaIsrael National Institute for Biotechnology in the Negev Ben-Gurion University of the Negev84105 Beer-ShevaIsrael Achva Academic College 79804 ShikmimIsrael department of Physiology Faculty of Health SciencesBen-Gurion University of the Negev84105 Beer-ShevaIsrael Ilse Katz Institute for Nanoscale Science and Technology Ben-Gurion University of the Negev84105 Beer-ShevaIsrael the shraga segal department of microbiology Immunologyand GeneticsFaculty of Health SciencesBen-Gurion University of the Negev84105 Beer-ShevaIsrael NeuroAging Group(NEURAL) Clinical Neurosciences Research Laboratory(LINC)Health Research Institute of Santiago de Compostela(IDIS)15706 Santiago de CompostelaSpain.

Background Alzheimer’s disease(AD)exhibits mitochondrial dysfunctions associated with dysregulated metabolism,brain inflammation,synaptic loss,and neuronal cell death.As a key protein serving as the mitochondrial gatekeeper,the voltage-dependent anion channel-1(VDAC1)that controls metabolism and Ca2+homeostasis is positioned at a convergence point for various cell survival and death signals.Here,we targeted VDAC1 with VBIT-4,a newly developed inhibitor of VDAC1 that prevents its pro-apoptotic activity,and mitochondria dysfunction.Methods To address the multiple pathways involved in AD,neuronal cultures and a 5×FAD mouse model of AD were treated with VBIT-4.We addressed multiple topics related to the disease and its molecular mechanisms using immunoblotting,immunofluorescence,q-RT-PCR,3-D structural analysis and several behavioral tests.Results In neuronal cultures,amyloid-beta(Aβ)-induced VDAC1 and p53 overexpression and apoptotic cell death were prevented by VBIT-4.Using an AD-like 5×FAD mouse model,we showed that VDAC1 was overexpressed in neurons surrounding Aβplaques,but not in astrocytes and microglia,and this was associated with neuronal cell death.VBIT-4 prevented the associated pathophysiological changes including neuronal cell death,neuroinflammation,and neuro-metabolic dysfunctions.VBIT-4 also switched astrocytes and microglia from being pro-inflammatory/neurotoxic to neuroprotective phenotype.Moreover,VBIT-4 prevented cognitive decline in the 5×FAD mice as evaluated using several behavioral assessments of cognitive function.Interestingly,VBIT-4 protected against AD pathology,with no significant change in phosphorylated Tau and only a slight decrease in Aβ-plaque load.Conclusions The study suggests that mitochondrial dysfunction with its gatekeeper VDAC1 is a promising target for AD therapeutic intervention,and VBIT-4 is a promising drug candidate for AD treatment.

关键词： Alzheimer’s disease Metabolism Mitochondria Neuroinflammation VDAC1

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Stellettin B renders glioblastoma vulnerable to poly (ADPribose) polymerase inhibitors via suppressing homologydirected repair

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Signal Transduction and Targeted Therapy 2023年第4期8卷 1388-1391页

作者： Xin Peng Yingying Wang Shaolu Zhang Zhennan Tao Yuxiang Dai Francois X.Claret Moshe Elkabets Hou-Wen Lin Zhe-Sheng Chen Dexin Kong Tianjin Key Laboratory of Technologies Enabling Development of Clinical Therapeutics and Diagnostics School of PharmacyTianjin Medical UniversityTianjin 300070China Key Laboratory of Immune Microenvironment and Diseases(Ministry of Education) Tianjin Medical UniversityTianjin 300070China department of Systems Biology The University of Texas MD Anderson Cancer CenterHoustonTX 77030USA department of Neurosurgery the Affiliated Drum Tower HospitalSchool of MedicineNanjing UniversityNanjing 210008China the shraga segal department of microbiology Immunology and GeneticsFaculty of Health SciencesBen-Gurion University of the NegevBeer-Sheva 84105Israel Research Center for Marine Drugs State Key Laboratory of Oncogenes and Related GenesDepartment of PharmacyRenji HospitalSchool of MedicineShanghai Jiaotong UniversityShanghai 200127China department of Pharmaceutical Sciences College of Pharmacy and Health SciencesSt.John’s UniversityQueensNY 11439USA

Dear Editor,Glioblastoma(GBM)is one of the most fatal brain tumors.Current first-line post-surgery regimens for GBM including radiotherapy and temozolomide(TMZ)chemotherapy show very limited efficacy.1,2 Novel therapeutic approaches for GBM patients are urgently needed.Natural products are important sources for drug discovery,especially in the field of cancer treatment.3 We previously isolated stellettin B(STELB)(Fig.1a)from marine sponge(Jaspis stellifera)and reported the remarkable and specific anticancer activities.Recently,a series of stellettins has been totally synthesized and the core chemical structure has been indicated.4 However,the specific mechanism and its role in regulating tumor biology remain largely unknown.

关键词： chemotherapy surgery render

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Cell transplantation therapy using pluripotent stem cells

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World Journal of Immunology 2013年第2期3卷 15-17页

作者： Noah Isakov the shraga segal department of microbiology Immunology and Genetics Faculty of Health Sciences and the Cancer Research Center Ben Gurion University of the NegevBeer Sheva 84105 Israel

The 2012 Nobel Prize in Physiology or Medicine was awarded jointly to Sir John B Gurdon and Shinya Yamanaka "for the discovery that mature cells can be reprogrammed to become pluripotent". Professor John B Gordon who pioneered the field of somatic cell nuclear transfer was the first to show that a nucleus of a mature cell can be transplanted into an enucleated egg and give rise to a living organism. His pioneering "cloning" technique paved the way for genome reprogramming and has led to subsequent cloning of differentanimal species. Professor Shinya Yamanaka revolutionized the filed of stem cell production by showing that the introduction of four selected genes into cells transform them into induced pluripotent stem cells that resemble embryonic stem cells and serve as promising cells for future regenerative medicine.

关键词：免疫医学英文文摘

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PICOT promotes T lymphocyte proliferation by down-regulating cyclin D2 expression

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World Journal of Immunology 2020年第1期10卷 1-12页

作者： Pinakin Pandya Noah Isakov the shraga segal department of microbiology Immunology and GeneticsFaculty of Health Sciences and the Cancer Research CenterBen Gurion University of the NegevBeer Sheva 84105Israel department of Computational and System biology UPMC Hillman Cancer CenterUniversity of PittsburghSchool of MedicinePittsburghPA 15232United States

The mammalian protein kinase C-interacting cousin of thioredoxin(PICOT;also termed glutaredoxin 3)is a multi-domain monothiol glutaredoxin that is involved in a wide variety of signaling pathways and biological processes.PICOT is required for normal and transformed cell growth and is critical for embryonic development.Recent studies in T lymphocytes demonstrated that PICOT can translocate to the nucleus and interact with embryonic ectoderm development,a polycomb group protein and a core component of the polycomb repressive complex 2,which contributes to the maintenance of transcriptional repression and chromatin remodeling.Furthermore,PICOT was found to interact with chromatin-bound embryonic ectoderm development and alter the extent of histone 3 lysine 27 trimethylation at the promoter region of selected polycomb repressive complex 2 target genes.PICOT knockdown in Jurkat T cells led to increased histone 3 lysine 27 trimethylation at the promoter region of CCND2,a cell cycle-regulating gene which encodes the cyclin D2 protein.As a result,the expression levels of CCND2 mRNA and protein levels were reduced,concomitantly with inhibition of the cell growth rate.Analysis of multiple data sets from the Cancer Genome Atlas revealed that a high expression of PICOT correlated with a low expression of CCND2 in a large number of human cancers.In addition,this parameter correlated with poor patient survival,suggesting that the ratio between PICOT/CCND2 mRNA levels might serve as a predictor of patient survival in selected types of human cancer.

关键词： Protein kinase C-interacting cousin of thioredoxin Glutaredoxin 3 Cyclin D2 Histone methylation T lymphocyte Histone 3 lysine 27 trimethylation

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Glycans to improve efficacy and solubility of protein aggregation inhibitors

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Neural Regeneration Research 2021年第11期16卷 2215-2216页

作者： Ashim Paul Daniel segal Elsa Zacco department of Molecular microbiology and Biotechnology Shmunis School of Biomedicine and Cancer ResearchTel Aviv UniversityRamat AvivTel Aviv 6997801Israel RNA Central Lab Center for Human TechnologiesIstituto Italiano di Tecnologia16152 GenovaItaly

Misfolding and subsequent aberrant selfassembly of certain proteins into toxic amyloid deposits are hallmarks of various diseases,most notably neurodegenerative disorders such as Alzheimer’s disease(AD)and Parkinson’s disease(Chiti and Dobson,2017).Aromatic residues in amyloidogenic proteins have been shown to be key factors in protein oligomerization and fibrilization,mostly driven byπ-πinteractions.Together with aromaticity,post-translational modifications can greatly affect a protein’s solubility and conformation and,as a consequence,its propensity to aggregate.

关键词： solubility aggregation mostly

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Author Correction to: SARS-CoV-2 interaction with Siglec-1 mediates trans-infection by dendritic cells

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Cellular & Molecular Immunology 2022年第8期19卷 965-966页

作者： Daniel Perez-Zsolt Jordana Muñoz-Basagoiti Jordi Rodon Marc Elosua-Bayes Dàlia Raïch-Regué Cristina Risco Martin Sachse Maria Pino Sanjeev Gumber Mirko Paiardini Jakub Chojnacki Itziar Erkizia Xabier Muñiz-Trabudua Ester Ballana Eva Riveira-Muñoz Marc Noguera-Julian Roger Paredes Benjamin Trinité Ferran Tarrés-Freixas Ignacio Blanco Victor Guallar Jorge Carrillo JuliàBlanco Amalio Telenti Holger Heyn Joaquim segalés Bonaventura Clotet Javier Martinez-Picado Júlia Vergara-Alert Nuria Izquierdo-Useros IrsiCaixa AIDS Research Institute 08916 Badalona Spain Germans Trias i Pujol Research Institute (IGTP) Can Ruti Campus 08916 Badalona Spain University of Vic—Central University of Catalonia (UVic-UCC) Vic 08500 Spain Catalan Institution for Research and Advanced Studies (ICREA) 08010 Barcelona Spain IRTA Centre de Recerca en Sanitat Animal (CReSA IRTA-UAB) Campus de la UAB 08193 Bellaterra (Cerdanyola del Vallès) Spain CNAG-CRG Centre for Genomic Regulation (CRG) Barcelona Institute of Science and Technology (BIST) 08028 Barcelona Spain Universitat Pompeu Fabra (UPF) Barcelona Spain Centro Nacional de Biotecnología CSIC 28049 Madrid Spain Division of microbiology and Immunology Yerkes National Primate Research Center Emory University Atlanta GA USA department of Pathology and Laboratory Medicine School of Medicine Emory University Atlanta GA USA Division of Pathology Yerkes National Primate Research Center Emory University Atlanta GA USA Barcelona Supercomputing Center (BSC) 08034 Barcelona Spain department of Integrative Structural and Computational Biology Scripps Research La Jolla CA 92037 USA UAB CReSA (IRTA-UAB) Campus de la UAB 08193 Bellaterra (Cerdanyola del Vallès) Spain Departament de Sanitat i Anatomia Animals Facultat de Veterinària UAB 08193 Bellaterra (Cerdanyola del Vallès) Spain

Correction to:Cellular&Molecular Immunology https://***/10.1038/s41423-021-00794-6,published online 15 November 2021 In the version of this correspondence initially published,one of the SARS-CoV-2 variants used in Fig.1B,which was originally described in the article as the SARS-CoV-2 variant‘B.1.***.248.2 Gamma’,is actually the‘P.2 Zeta’SARS-CoV-2 variant of interest.The GISAID accession ID EPI_ISL_1831696 provided is correct。

关键词： interaction media initially

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SARS-CoV-2 interaction with Siglec-1 mediates trans-infection by dendritic cells

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Cellular & Molecular Immunology 2021年第12期18卷 2676-2678页

作者： Daniel Perez-Zsolt Jordana Muñoz-Basagoiti Jordi Rodon Marc Elosua-Bayes Dàlia Raïch-Regué Cristina Risco Martin Sachse Maria Pino Sanjeev Gumber Mirko Paiardini Jakub Chojnacki Itziar Erkizia Xabier Muñiz-Trabudua Ester Ballana Eva Riveira-Muñoz Marc Noguera-Julian Roger Paredes Benjamin Trinité Ferran Tarrés-Freixas Ignacio Blanco Victor Guallar Jorge Carrillo JuliàBlanco Amalio Telenti Holger Heyn Joaquim segalés Bonaventura Clotet Javier Martinez-Picado Júlia Vergara-Alert Nuria Izquierdo-Useros IrsiCaixa AIDS Research Institute 089BadalonaSpain IRTA Centre de Recerca en Sanitat Animal(CReSAIRTA-UAB)Campus de la UAB08193Bellaterra(Cerdanyola del Vallès)Spain CNAG-CRG Centre for Genomic Regulation(CRGBarcelona Institute of Science and Technology(BIST)08028BarcelonaSpain Centro Nacional de Biotecnología CSIC28049MadridSpain Division of microbiology and Immunology Yerkes National Primate Research CenterEmory UniversityAtlantaGAUSA department of Pathology and Laboratory Medicine School of MedicineEmory UniversityAtlantaGeorgiaUSA Division of Pathology Yerkes National Primate Research CenterEmory UniversityAtlantaGeorgiaUSA Germans Trias i Pujol Research Institute(IGTP) Can Ruti Campus089BadalonaSpain Barcelona Supercomputing Center(BSC) 08034BarcelonaSpain Catalan Institution for Research and Advanced Studies(ICREA) 08010BarcelonaSpain University of Vic-Central University of Catalonia(UVic-UCC) Vic08500Spain department of Integrative Structural and Computational Biology Scripps ResearchLa JollaCA92037USA Universitat Pompeu Fabra(UPF) BarcelonaSpain UAB CReSA(IRTA-UAB)Campus de la UAB08193Bellaterra(Cerdanyola del Vallès)Spain Departament de Sanitat i Anatomia Animals Facultat de VeterinàriaUAB08193Bellaterra(Cerdanyola del Vallès)Spain

Antigen-presenting cells(APCs)may be resistant to SARS-CoV-2 infection but still contribute to viral pathogenesis.Lectins such as sialic acid-binding Ig-like lectin 1(Siglec-1/CD169)mediate the attachment of viruses to APCs.Here,we show that APCs effectively capture SARS-CoV-2 within compartments via recognition of Siglec-1.This receptor interacts with sialylated gangliosides on membranes of SARS-CoV-2 variants.

关键词： infection Antigen media

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Detection and Characterization of β-Lactamase Encoding Genes in Carbapenem Non-Susceptible Gram-Negative Bacteria and Susceptibility of Isolates to Ceftazidime-Avibactam at a New York City Community Hospital

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Open Journal of Medical microbiology 2016年第4期6卷 150-157页

作者： Carl Urban Rita Colon-Urban Vincent J. LaBombardi Noriel Mariano Nishant Prasad Geeti Dhillon Marina Guralnik Sorana segal-Maurer The Dr. James J. Rahal Jr. Division of Infectious Diseases Department of Medicine New York-Presbyterian/Queens Flushing NY USA The Dr. James J. Rahal Jr. Division of Infectious Diseases Department of Medicine NewYork-Presbyterian/Queens Flushing NY USA department of Biological Sciences SUNY Old Westbury Old Westbury New York NY USA department of Pathology NewYork-Presbyterian/Queens Flushing NY USA

A surveillance study was undertaken to identify prominent β-lactamase encoding genes in 131 carbapenem non-susceptible gram-negative clinical isolates at a New York City community hospital. KPC carbapenemases were detected in 89% of Enterobacteriaceae as well as additional TEM, SHV, and CTX-M class A enzymes. OXA-23 and OXA-24 were the prevalent class D carbapenemases identified in Acinetobacter species. One OXA-23 in M. morganii and one OXA-48 in K. pneumoniae were also identified. Among class C β-lactamases CMY, ACT/MIR, DHA, and FOX were detected. The in vitro activity of ceftazidime-avibactam by E-test methodology was tested with minimal inhibitory concentrations (MIC) of ≤3 μg/ml for 97.8% of all Enterobacteriaceae, MIC50/90 of 16/>256 μg/ml for carbapenem non-susceptible Acinetobacter, and 3/6 μg/ml for carbapenem non-susceptible Pseudomonas aeruginosa. Periodic surveillance of isolates to characterize current and emerging β-lactamase genotypes present in local isolates may help identify outbreak situations, provide assistance to infection control and antibiotic stewardship programs, and potentially improve patient outcomes.

关键词： Carbapenem Non-Susceptible Check-MDR CT103 XL Microarray β-Lactamase Detection Resistance Mechanisms Ceftazidime-Avibactam

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