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Laser capture microdissection and genetic analysis of carbon-labeled Kupffer cells

World Journal of Gastroenterology 2009年第14期15卷 1708-1718页

作者： Stephan Gehring edmond Sabo maryann e san martin elizabeth M Dickson Chao-Wen Cheng Stephen H Gregory Department of Medicine Rhode Island Hospital and The Warren Alpert Medical School of Brown University Providence RI 02903 United States Department of Pathology Rhode Island Hospital and The Warren Alpert Medical School of Brown University Providence RI 02903 United States zentrum fur Kinder- und Jugendmedizin der Universitaitsmedizin der Johannes Gutenberg-Universitait 55101 Mainz Germany

AIM： To develop a method of labeling and microdissecting mouse Kupffer cells within an extraordinarily short period of time using laser capture microdissection （LCM）. MeTHODS： Tissues are complex structures comprised of a heterogeneous population of interconnected cells. LCM offers a method of isolating a single cell type from specific regions of a tissue section. LCM is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCM prior to mRNA isolation can be elaborate, thereby subjecting the RNA to considerable degradation. Kupffer cell labeling is achieved by injecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model. ReSULTS： mRNA extracted from the microdissected cell population displayed marked increases in colonystimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. Gene expression by Kupffer ceils derived from bile-duct-ligated, versus sham-operated, mice was compared. Microarray analysis revealed a significant （2.5-fold, q value 〈 10） change in 493 genes. Based on this fold-change and a standardized PubMed search, 10 genes were identified that were relevant to the ability of Kupffer cells to suppress liver injury. CONCLUSION; The methodology outlined herein provides an approach to isolating high quality RNA from Kupffer cells, without altering the tissue integrity.

关键词： Kupffer cells India ink Laser capturemicrodissection Bile duct ligation DNA microarray

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A novel regulatory region controls IgH locus transcription and switch recombination to a subset of isotypes

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Cellular & Molecular Immunology 2019年第11期16卷 887-889页

作者： Rocío Amoretti-Villa Mélanie Rogier Isabelle Robert Vincent Heyer Bernardo Reina-san-martin Institut de Génétique et de Biologie Moléculaire et Cellulaire(IGBMC) IllkirchFrance Institut National de la santéet de la Recherche Médicale(INSeRM) U1258IllkirchFrance Centre National de la Recherche Scientifique(CNRS) UMR7104IllkirchFrance Universitéde Strasbourg IllkirchFrance

Class switch recombination(CSR)occurs at the IgH locus and replaces the immunoglobulin(Ig)isotype expressed from IgM to IgG,Ige or IgA,endowing the B cell receptor with novel effector *** is triggered by activation-induced cytidine deaminase(AID),1 an enzyme that deaminates cytosines to uracils in single-stranded DNA exposed by *** distinct antibody isotypes are encoded in the IgH locus in individual transcription units composed of a cytokine-inducible promoter,an intronic exon,and a switch region(Sx),followed by the exons encoding the constant region(Cx)(Fig.S1a).During CSR,the choice of recombination to a particular isotype is determined by the stimulation-dependent activation of specific promoters,triggering the generation of noncoding germline transcripts(GLTs).2 Thus far,the transcriptional regulation of the IgH locus is known to be controlled by the eμenhancer,located downstream of the variable region and upstream of the donor switch region(Sμ),and the 3′regulatory region(3′RR)super-enhancer located downstream of Cα.

关键词： IgH stimulation replace

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Mediator contributes to IgH locus VDJ rearrangements by promoting usage of most distal V segments

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Cellular & Molecular Immunology 2020年第4期17卷 407-409页

作者： Iman Dalloul Zeinab Dalloul sandrine Le Noir François Boyer Bernardo Reina-san-martin Jeanne Cook-Moreau Michel Cogné UMR CNRS 7276 INSERM U1262Universite de LimogesLimoges 87025France INSeRM U1258 UMR CNRS 7104Institut de Genetique et de Biologie Moleculaire et CellulaireUniversitéde StrasbourgIllkirch 67400France

INTRODUCTION The immunoglobulin heavy(IgH)chain locus includes a large cluster of V,D and J segments spanning thousands of kilobases in mammals and its expression implies long-range 3D-regulation of various gene modifications.1 V(D)J recombination occurs during early B-cell ontogeny.2 It is regulated by multiple transcription factors and cis-regulatory elements,and most notably by the intronic 5′eμelement located in the JH to Cμintron.3 The intergenic control region 1(IGCR1)located between the VH and D gene clusters inhibits rearrangement of the most DH-proximal VH gene segments,while promoting usage of distal VH gene segments.4 Other downstream elements of the locus,flanking its 3′end might have more subtle roles:while the large 3′regulatory region(3′RR)shows strong internal synergies2 and behaves as a superenhancer in mature B *** deletion showed no influence on the VDJ repertoire and rather increased transcription of unrearranged V segments.3–5 Insulators binding CTCF farther downstream of the 3′RR were shown,however,to promote usage of distal VH segments.6 Altogether,there are strong indications that elements involved in long-distance regulation of transcription might also influence VDJ ***-frame splicing of VDJ exons is also needed for full Ig expression.7 Given the functional importance of gene architecture and long-range interactions for the IgH locus physiology,we explored the dependence of the VDJ repertoire on Med1,a known actor of long-range enhancer interactions.

关键词： B cells mediator VDJ repertoire

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effects of absolute fasting on reproduction and survival of the invasive apple snail Pomacea canaliculata in its native range

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Current Zoology 2016年第4期62卷 369-375页

作者： Nicolas e. TAMBURI Pablo R. martin INBIOSUR (UNS-CONICeT) Universidad Nacional del Sur Departamento de Biologla Bioquimica y Farmacia San Juan 670 8000 Bahia Blanca Argentina

A South American freshwater gastropod, the apple snail Pomacea canaliculata, has become a driver of ecosystemic changes in wetlands and an important rice pest after its introduction to various parts of the world, mainly Asia. The objective of this study was to study the effect of an abrupt interruption in food availability in the short term （up to 4 weeks） and long term （up to 8 months） on survival and re- productive activity. The main results indicate that short-term fasting mainly affects the survival of males, but only when they are raised together with females, probably due to a greater mate-searching activity that increases mortality in the individuals with lower reserves. The number of copulating snails or egg-laying females shows an abrupt drop when fasting and a rapid recovery after the food supply is restored. The strategy of discontinuing reproductive activity prioritizes energy conservation for the survival of the females. Interpopulation variation in resistance to starvation was observed in adults, which can be explained to some extent by the food availability that they experienced in their natural environment. No interpopulational differences in survival were seen in hatchlings. The mean maximum values of survival under starvation were 52.6 days in hatchlings and the 3.3% of adults survive over than 200 days, which may be a relevant trait in dispersal and establishment in new habitats.

关键词： GASTROPODA POMACeA canaliculata WeTLANDS STARVATION eNeRGY conservation

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Spectrum and visual outcomes of Vogt-Koyanagi-Harada disease in Argentina

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International Journal of Ophthalmology(english edition) 2017年第1期10卷 98-102页

作者： Veronica e.Giordano Ariel Schlaen martin J.Guzman-sanchez Cristobal Couto Department of Ophthalmology Inflammatory Eye Diseases ServiceHospital de Clinicas Jose de San MartinBuenos Aires C1120AARArgentina Department of Ophthalmology Oculoplastic ServiceAssociation for the Prevention of Blindness in MexicoVicente Garcia Torres 46San Lucas 04030CoyoacanMexico CityMexico

AIM： To review all cases of Vogt-Koyanagi-Harada （VKH） disease in an Inflammatory eye Disease Service in Argentina and to describe the clinical profile and outcomes of ***： The records from patients with VKH disease between January 1980 and December 2008 were retrospectively analyzed for clinical profile, complications, and treatment. Patients were classified according to their initial treatment in group 1： high corticosteroid dose [≥1 mg/（kg·d）] within 2wk of illness onset; group 2： high corticosteroid dose, 2 to 4wk of onset and group 3： patients who received the high dose after 1mo of illness onset, patients who received lower oral doses than 1 mg/（kg·d） without regarding the time of beginning of the ***： A total of 210 eyes of 105 patients were included. The mean age at presentation was 32.6±13y （range： 10-74y）, and 86.7% were female. The mean duration of follow up was 144±96.6mo. Patients in the group 1 had significantly higher visual acuity than the other groups （P〈0.0001）, none had （loss of, or no） light perception at the end of follow up, whereas 24.7% patients in group 3 ended in light perception （P〈0.004）.CONCLUSION： Patients with early high dose corticosteroid treatment have better visual acuity and fewer complications. Proper timing in referral and treatment is critical for better visual outcome in VKH disease.

关键词： Vogt-Koyanagi-Harada disease Argentine population clinical spectrum treatment

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Late Protective effects of the Anticalmodulin Drug Fluphenazine on Carbon Tetrachloride-induced Liver Necrosis

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Biomedical and environmental Sciences 1995年第3期8卷 218-225页

作者： e. C. De FeRReYRA A. S. BeRNACCHI M. F. san martin G. D. CASTRO AND J. A. CASTRO (Centro de Investigaciones Toxicologicas (CeITOX)-CITeFA/CONICeT,Zufriategui 4380, (1603) Villa Martelli, Buenos Aires, Angentina) Centro de Investigaciones Toxicológicas (CeITOX)-CITeFA/CONICeT Zufriategui Buenos Aires Argentina.

Fluphenazine (FP) treatment (50mg/kg bw, ip in saline) 30 min before or 6 or 10 h after CCl4 administration (1 ml/kg ip in olive oil) significantly prevented the liver necrosis produced by the hepatotoxin at 24 h. FP had enhancing effects on the covalent binding of CCl4 reactive metabolites to cellular constituents and on CCl4 induced lipid *** lowered bOdy temperature of the CCl4-poisoned animals during the 24 h observation period. The obtained results are compatible but do not prove the hypothesis that calmodulin (CaM) had participation in late occurring events preceding necrosis. FP lowering action on body temperature, however, might also play a role in the effects of this drug on the onset of CCl4 induced liver necrosis. FP levels in liver tissue as determined by gas chromatography-mass spectrometry evidenced the presence of the drug in amounts suffi cient to inhibit CaM and that suggests that not all preventive effects of FP are due to its indirect actions on the central nervous system via decreased body temperature

关键词： ab Late Protective effects of the Anticalmodulin Drug Fluphenazine on Carbon Tetrachloride-induced Liver Necrosis

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Induction of Defensive Responses in <i>eucalyptus globulus</i>(Labill) Plants, against <i>Ctenarytaina eucalypti</i>(Maskell) (Hemiptera: Psyllidae)

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American Journal of Plant Sciences 2012年第5期3卷 589-595页

作者： Christian Troncoso Jose Becerra Claudia Perez Victor Hernandez Aurelio san martin Manuel sanchez-Olate Darcy Rios Centro de Biotecnologia Laboratorio de Cultivo de Tejidos Vegetales Universidad de Concepcion Concepcion Chile Departamento de Botanica Facultad de Ciencias Naturales y Oceanograficas Universidad de Concepcion Concepcion Chile Departamento de Quimica Facultad de Ciencias Universidad de Chile Santiago Chile Facultad de Ciencias Forestales Universidad de Concepcion Concepcion Chile

This study evaluated the expression of defense compounds from the secondary metabolism of eucalyptus globulus plants, subjected to direct and indirect stimuli by the insect Ctenarytaina eucalypti (blue gum Psyllid). Results showed that defense responses were activated in plants in all tested cases. Were detected and identified thirty-two compounds in the leaves of treated plants, of which five compounds differed with the control, and all are part of the chemical defenses from the plants, three of them were oxygenated monoterpenes (borneol, exo-2-hydroxy cineole and thymol), a aromatic carboxylic acid (benzoic acid) and a quinone (6-acethyl-flaviolin). The plants induced by volatile compounds and by indirect entomological manner, showed its capability to synthesize defensive compounds without a wound that promotes these responses. Were also found some constitutive secondary metabolites over expressed in the different inductions compared with the control.

关键词： Chemical Defense e. globules Secondary Metabolites Terpenes

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Glassy carbon microelectrodes minimize induced voltages,mechanical vibrations,and artifacts in magnetic resonance imaging

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Microsystems & Nanoengineering 2019年第1期5卷 89-99页

作者： Surabhi Nimbalkar erwin Fuhrer Pedro Silva Tri Nguyen martin Sereno Sam Kassegne Jan Korvink MeMS Research Lab Department of Mechanical EngineeringCollege of EngineeringSan Diego State University5500 Campanile DriveSan DiegoCA 92182USA NSF-eRC Center for Neurotechnology(CNT) SeattleWAUSA Institute of Microstructure Technology–Karlsruhe Institute of Technology Hermann-von-Helmholtz-Platz76344 Eggenstein-LeopoldshafenGermany Magnetic Resonance Imaging Lab San Diego State UniversitySan DiegoCA 92182USA

The recent introduction of glassy carbon(GC)microstructures supported on flexible polymeric substrates has motivated the adoption of GC in a variety of implantable and wearable *** probes such as electrocorticography and penetrating shanks with GC microelectrode arrays used for neural signal recording and electrical stimulation are among the first beneficiaries of this *** the expected proliferation of these neural probes and potential clinical adoption,the magnetic resonance imaging(MRI)compatibility of GC microstructures needs to be established to help validate this potential in clinical ***,we present GC microelectrodes and microstructures—fabricated through the carbon micro-electro-mechanical systems process and supported on flexible polymeric substrates—and carry out experimental measurements of induced vibrations,eddy currents,and *** induced vibration,induced voltage,and MRI experiments and finite element modeling,we compared the performances of these GC microelectrodes against those of conventional thin-film platinum(Pt)microelectrodes and established that GC microelectrodes demonstrate superior magnetic resonance compatibility over standard metal thin-film ***,we demonstrated that GC microelectrodes experienced no considerable vibration deflection amplitudes and minimal induced currents,while Pt microelectrodes had significantly larger *** also showed that because of their low magnetic susceptibility and lower conductivity,the GC microelectrodes caused almost no susceptibility shift artifacts and no eddy-current-induced artifacts compared to Pt *** together,the experimental,theoretical,and finite element modeling establish that GC microelectrodes exhibit significant MRI compatibility,hence demonstrating clear clinical advantages over current conventional thin-film materials,further opening avenues for wider adoption of GC microelectrodes in chronic clinical app

关键词： magnetic compatibility resonance

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effect of Sn on synthesis of nanocrystalline Ti-based alloy with fcc structure

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Transactions of Nonferrous Metals Society of China 2020年第8期30卷 2119-2131页

作者： C.AGUILAR e.PIO A.MeDINA C.PARRA R.MANGALARAJA P.martin I.ALFONSO K.TeLLO Departamento de Ingeniería Metalúrgica y de Materiales Departamento de Física(+)Universidad Técnica Federico Santa MaríaValparaísoChile Instituto de Investigación en Metalurgia y Materiales Universidad Michoacana de San Nicolás de HidalgoMoreliaMéxico Laboratorio de Cerámicos Avanzados y Nanotecnologia Departamento de Ingeniería de MaterialesUniversidad de ConcepciónConcepciónChile Instituto de Investigaciones en Materiales Universidad Nacional Autónoma de MéxicoCampus Morelia UNAMMoreliaMéxico

The effect of the amount of Sn on the formation of fcc phase in Ti-13 Ta-x Sn(x=3,6,9 and 12,at.%)alloys was *** alloys were synthesized by mechanical alloying using a planetary mill,jar and balls of stabilized *** Rietveld refinement,it was found that the obtained fcc phase has crystallite size smaller than 10 nm and microstrain larger than *** conditions are required to form an fcc phase in Ti-based *** all samples,the microstructure of the fcc phase consists of equiaxial crystallites with sizes smaller than 10 *** largest presence of fcc phase in the studied Ti alloy was found with 6 at.%Sn,because this alloy exhibits the largest microstrain(1.5×10-2)and crystallite size of 6.5 *** data reveal that a solid solution and an amorphous phase were formed during *** necessary conditions to promote the formation of solid solution and amorphous phases were determined using thermodynamic *** the amount of Sn increases,the energy required to form an amorphous phase varies from approximately 10 to approximately-5 k J/mol for 3 and 12 at.%Sn,*** thermodynamic calculations are in agreement with XRD patterns analysis and HRTeM results.

关键词： Ti-based alloy phase transformation fcc phase Rietveld analysis mechanical alloying

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Annual variation of adult survival of a south-temperate House Wren population in Argentina

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Avian Research 2024年第2期15卷 229-237页

作者： Gustavo J.Fernandez Mariana e.Carro Paulo e.Llambías Grupo de Investigacion en ecofisiología de Fauna Silvestre INIBIOMA-CONICETAsentamiento Universitario San Martín de los Andes(AUSMA)Universidad Nacional Del ComahuePasaje de la Paz 235(8370)San Martín de los AndesNeuquenArgentina Grupo de Investigacion de Biología de Aves IADIZA CCT-MendozaUniversidad Nacional de CuyoAvenida RuizLeal s/n Parque General San Martin5500MendozaArgentina

Identifying factors affecting the survival of individuals is essential for understanding the evolution of life-history traits and population *** numerous studies on this subject in north-temperate environments,there is a lack of equivalent studies at similar latitudes in the ***,we used a 14-year dataset of capture,banding,and resighting to estimate the annual variation in the apparent adult survival probability of a south-temperate population of House Wrens(Troglodytes aedon bonariae).We evaluated temporal variation in sur-vival and the effect of environmental(climatic)and demographic variables(adult abundance,total number of fledglings produced during each breeding season)on survival *** found that the probability of adult survival decreased as the abundance of breeding adults *** density-dependent effect could be related to the resident lifestyle of southern House Wrens,which could determine an intense competition for territories and resources that ultimately would affect their survival.

关键词： Annual variation CJS Density-dependence Survival Troglodytes aedon bonariae

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