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Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Journal of Animal Science and Biotechnology 2015年第2期6卷 230-251页

作者： Nuradilla Mohamad-Fauzi Pablo J Ross Elizabeth A Maga james d murray department of Animal Science University of California Institute of Ocean and Earth Sciences University of Malaya department of Population Health and Reproduction University of California

Background： There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells （MSCs） have been isolated and characterized from various species, but are poorly characterized in goats. Results： Goat MSCs isolated from bone marrow （BM-MSCs） and adipose tissue （ASCs） have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express Cd90, Cd105, Cd73 but not Cd45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency （CFE）, yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection. Conclusions： Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

关键词： Adipose Bone marrow Characterization differentiation Goat Mesenchymal stem cells

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Hematologic and biochemical reference intervals for specific pathogen free 6-week-old Hampshire-Yorkshire crossbred pigs

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Journal of Animal Science and Biotechnology 2014年第3期5卷 286-290页

作者： Caitlin A Cooper Luis E Moraes james d murray Sean d Owens department of Animal Science University of California department of Population Health and Reproduction University of California department of Pathology Microbiology and ImmunologySchool of Veterinary MedicineUniversity of California One Shields AvenueDavisCA 95616USA

Background： Hematologic and biochemical reference intervals depend on many factors, including age. A review of the literature highlights the lack of reference intervals for 6-wk-old specific pathogen free （SPF） Hampshire-Yorkshire crossbred pigs. For translational research, 6-wk-old pigs represent an important animal model for both human juvenile colitis and diabetes mellitus type 2 given the similarities between the porcine and human gastrointestinal maturation process. The aim of this study was to determine reference intervals for hematological and biochemical parameters in healthy 6-wk-old crossbred pigs. Blood samples were collected from 66 clinically healthy Hampshire-Yorkshire pigs. The pigs were 6 wks old, represented both sexes, and were housed in a SPF facility. Automated hematological and biochemical analysis were performed using an AdVIA 120 Hematology System and a Cobas 6000 C501 Clinical Chemistry Analyzer. Results： Reference intervals were calculated using both parametric and nonparametric methods. The mean, median, minimum, and maximum values were calculated. Conclusion： As pigs are used more frequently as medical models of human disease, having reference intervals for commonly measured hematological and biochemical parameters in 6-wk-old pigs will be useful. The reference intervals calculated in this study will aid in the diagnosis and monitoring of both naturally occurring and experimentally induced disease. In comparison to published reference intervals for older non SPF pigs, notable differences in leukocyte populations, and in levels of sodium, potassium, glucose, protein, and alkaline phosphatase were observed.

关键词： Biochemical analytes Hematology Pigs Reference interval SPF

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Treating amyotrophic lateral sclerosis with allogeneic Schwann cell-derived exosomal vesicles: a case report

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Neural Regeneration Research 2025年第4期20卷 1207-1216页

作者： Pascal J.Goldschmidt-Clermont Aisha Khan George Jimsheleishvili Patricia Graham Adriana Brooks Risset Silvera Alexander J.P.Goldschmidt damien d.Pearse W.dalton dietrich Allan d.Levi james d.Guest Miller School of Medicine University of MiamiMiamiFLUSA Stem Cell Institute Miller School of MedicineUniversity of MiamiMiamiFLUSA The Miami Project to Cure Paralysis Miller School of MedicineUniversity of MiamiMiamiFLUSA Alzady International LLC MiamiFLUSA Neurological Surgery Miller School of MedicineUniversity of MiamiMiamiFLUSA

Schwann cells are essential for the maintenance and function of motor neurons,axonal networks,and the neuromuscular *** amyotrophic lateral sclerosis,where motor neuron function is progressively lost,Schwann cell function may also be ***,important signaling and potential trophic activities of Schwann cell-derived exosomal vesicles have been *** case report describes the treatment of a patient with advanced amyotrophic lateral sclerosis using serial intravenous infusions of allogeneic Schwann cell-derived exosomal vesicles,marking,to our knowledge,the first instance of such *** 81-year-old male patient presented with a 1.5-year history of rapidly progressive amyotrophic lateral *** initial diagnosis,the patient underwent a combination of generic riluzole,sodium phenylbutyrate for the treatment of amyotrophic lateral sclerosis,and *** patient volunteered to participate in an FdA-approved single-patient expanded access treatment and received weekly intravenous infusions of allogeneic Schwann cell-derived exosomal vesicles to potentially restore impaired Schwann cell and motor neuron *** confirmed that cultured Schwann cells obtained from the amyotrophic lateral sclerosis patient via sural nerve biopsy appeared impaired(senescent)and that exposure of the patient’s Schwann cells to allogeneic Schwann cell-derived exosomal vesicles,cultured expanded from a cadaver donor improved their growth capacity in *** a period of observation lasting 10 weeks,during which amyotrophic lateral sclerosis Functional Rating Scale-Revised and pulmonary function were regularly monitored,the patient received weekly consecutive infusions of 1.54×1012(×2),and then consecutive infusions of 7.5×1012(×6)allogeneic Schwann cell-derived exosomal vesicles diluted in 40 mL of dulbecco’s phosphate-buffered *** of the infusions were associated with adverse events such as infusion reactions(allergic or otherwise)or

关键词： allogeneic amyotrophic lateral sclerosis exosomes infusion neuromuscular junction Schwann cell

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Light controlled imaging probes for intracellular target recognition： photochromism as a tool to enhance sensing precision

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Science China Chemistry 2017年第12期60卷 1563-1564页

作者： Tony d. james department of Chemistry University of BathBath BA2 7AYUK

Chemosensors and imaging probes have been the focus of significant research interest over the past few decades. In part due to ease of preparation and simplicity in manipulation, fluorescent probes have been extensive... 详细信息

关键词： Light controlled imaging probes for intracellular target recognition photochromism enhance sensing precision

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Challenges of modeling solar disturbances' arrival times at the Earth

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Science China(Technological Sciences) 2008年第10期51卷 1580-1588页

作者： C. d. FRY Chin-Chun WU murray dRYER Center for Space Plasma and Aeronomic Research The University of Alabama in Huntsville Huntsville AL 35899 USA Exploration Physics International Inc. Huntsville AL 35806 USA Center for Space Plasma and Aeronomic Research The University of Alabama in Huntsville Huntsville AL 35899 USA Space Weather Laboratory NASA/Goddard Space Flight Center Greenbelt MD 20771 USA Center for Space Plasma and Aeronomic Research The University of Alabama in Huntsville Huntsville AL 35899 USA Exploration Physics International Inc. Huntsville AL 35806 USA Space Weather Prediction Center National Oceanic and Atmospheric Administration (NOAA) Boulder Colorado USA

In recent years remarkable advances have been made in the development of phys-ics based models of various parts of the solar-terrestrial system (see JASTP special issues, October, November 2004; February 2007). In this paper, we focus our dis-cussions in a specific region of the Sun to the Earth’s environment (i.e. 1 AU). It is well-known that geomagnetic storms are caused by solar eruptions. The conse-quences of these storms include particle acceleration, solar wind impact on the Earth’s magnetosphere and ionosphere, UV-EUV radiation effects on the lower at-mosphere, etc. One of the main challenges is to predict the arrival time at 1 AU of the solar disturbance. The prospects look good for an accurate, real-time forecast scheme built on the acquisition of solar, heliosphere and the near-Earth data and large-scale models. However, the accuracy of these models still needs improve-ment. We will discuss the present status of the models and challenges to improve the simulation models.

关键词： space weather――magnetohydrodynamic (MHd) Modeling MHd Sun-Earth connection MHd Sun-Corona

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Comprehensive Transcriptome Analysis of Auxin Responses in Arabidopsis

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Molecular Plant 2008年第2期1卷 321-337页

作者： Ivan A. Paponov Martina Paponovt William Teale Margit Menges Sohini Chakrabortee james A.H. murray Klaus Palme Faculty of Biology University of Freiburg Schanzlestrasse 1 79104 Freiburg Germany Institute of Biotechnology University of Cambridge Cambridge CB2 1QT United Kingdom

In plants, the hormone auxin shapes gene expression to regulate growth and development. despite the detailed characterization of auxin-inducible genes, a comprehensive overview of the temporal and spatial dynamics of auxinregulated gene expression is lacking. Here, we analyze transcriptome data from many publicly available Arabidopsis profiling experiments and assess tissue-specific gene expression both in response to auxin concentration and exposure time and in relation to other plant growth regulators. Our analysis shows that the primary response to auxin over a wide range of auxin application conditions and in specific tissues comprises almost exclusively the up-regulation of genes and iden- tifies the most robust auxin marker genes. Tissue-specific auxin responses correlate with differential expression of Aux/ IAA genes and the subsequent regulation of context- and sequence-specific patterns of gene expression. Changes in transcript levels were consistent with a distinct sequence of conjugation, increased transport capacity and down-regulation of biosynthesis in the temperance of high cellular auxin concentrations. Our data show that auxin regulates genes associated with the biosynthesis, catabolism and signaling pathways of other phytohormones. We present a transcriptional overview of the auxin response. Specific interactions between auxin and other phytohormones are highlighted, particularly the regulation of their metabolism. Our analysis provides a roadmap for auxin-dependent processes that underpins the concept of an ＇auxin code＇-a tissue-specific fingerprint of gene expression that initiates specific developmental processes.

关键词： Comprehensive Transcriptome Analysis Auxin Responses Arabidopsis

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Promises and paradoxes of regulatory T cells in inflammatory bowel disease

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World Journal of Gastroenterology 2015年第40期21卷 11236-11245页

作者： james d LordJ Translational Program Benaroya Research Institute at Virginia Mason Mailstop

Since their discovery two decades ago,Cd4+Cd25+Foxp3+ regulatory T cells(Tregs) have become the subject of intense investigation by immunologists. Unlike other T cells,which promote an immune response,Tregs actively inhibit inflammation when activated by their cognate antigen,thus raising hope that these cells could be engineered into a highly targeted,antigenspecific,immunosuppressant therapy. Although Tregs represent less than 10% of circulating Cd4+T cells,they have been shown to play an essential role in preventing or limiting inflammation in a variety of animal models and human diseases. In particular,spontaneous intestinal inflammation has been shown to occur in the absence of Tregs,suggesting that there may be a Treg defect central to the pathogenesis of human inflammatory bowel disease(IBd). However,over the past decade,multiple groups have reported no qualitative or quantitative deficits in Tregs from the intestines and blood of IBd patients to explain why these cells fail to regulate inflammation in Crohn's disease and ulcerative colitis. In this review,we will discuss the history of Tregs,what is known about them in IBd,and what progress and obstacles have been seen with efforts to employ them for therapeutic benefit.

关键词： Foxp3 Regulatory T cells Crohn’s disease Th17 Ulce

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Macrophage migration inhibitory factor gene polymorphisms in inflammatory bowel disease: An association study in New Zealand Caucasians and meta-analysis

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World Journal of Gastroenterology 2013年第39期19卷 6656-6664页

作者： james d Falvey Robert W Bentley Tony R Merriman Mark B Hampton murray L Barclay Richard B Gearry Rebecca L Roberts department of Medicine University of OtagoChristchurch 8011New Zealand department of Gastroenterology Christchurch HospitalChristchurch 8011New Zealand department of Biochemistry University of OtagoDunedin 9054New Zealand department of Pathology University of OtagoChristchurch 8011New Zealand department of Surgical Sciences Dunedin School of MedicineDunedin 9054New Zealand

AIM:To investigate the association of macrophage migration inhibitory factor(MIF)promoter polymorphisms with inflammatory bowel disease(IBd)***:One thousand and six New Zealand Caucasian cases and 540 Caucasian controls were genotyped for the MIF SNP-173G>C(rs755622)and the repeat polymorphism CATT5-8(rs5844572)using a predesigned TaqMan SNP assay and capillary electrophoresis,*** were analysed for single site and haplotype association with IBd risk and ***-analysis was employed,to assess cumulative evidence of association of MIF-173G>C with *** published genotype data for MIF-173G>C in IBd were identified using PubMed and subsequently searching the references of all PubMed-identified *** genotypes for MIF-173G>C were generated from the Wellcome Trust Case Control Consortium(and National Institute of diabetes and digestive and Kidney diseases).Separate meta-analyses were performed on Caucasian Crohn’s disease(Cd)(3863 patients,6031controls),Caucasian ulcerative colitis(UC)(1260 patients,1987 controls),and East Asian UC(416 patients and 789 controls)datasets using the Mantel-Haenszel *** New Zealand dataset had 93%power,and the meta-analyses had 100%power to detect an effect size of OR=1.40 atα=0.05,***:In our New Zealand dataset,single-site analysis found no evidence of association of MIF polymorphisms with overall risk of Cd,UC,and IBd or disease phenotype(all P values>0.05).Haplotype analysis found the CATT5/-173C haplotype occurred at a higher frequency in New Zealand controls compared to IBd patients(0.6 vs 0.01;P=0.03,OR=0.22;95%CI:0.05-0.99),but this association did not survive bonferroni ***-analysis of our New Zealand MIF-173G>C data with data from seven additional Caucasian datasets using a random effects model found no association of MIF polymorphisms with Cd,UC,or overall ***,meta-analysis of all published MIF-173G>C data from East Asian datasets(416UC patients

关键词： Crohn’s disease Ulcerative colitis Migration inhibitory factor rs755622 rs5844572 Genetic association study

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A new direction for Alzheimer's research

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Neural Regeneration Research 2018年第2期13卷 190-193页

作者： james d.Weinstein Marshall University School of Medicine Medical Center

despite decades of research, at present there is no curative therapy for Alzheimer＇s disease. Changes in the way new drugs are tested appear to be necessary. Three changes are presented here and will be discussed. The first change is that Alzheimer＇s disease must be considered a disease of four major pathological processes, not one. The four processes are： 1） vascular hy- poperfusion of the brain with associated mitochondrial dysfunction, 2） destructive protein inclusions, 3） uncontrolled oxidative stress, and 4） proinflammatory immune processes second- ary to microglial and astrocytic dysfunction in the brain. The second change recommended is to alter the standard cognitive measurement tools used to quantify mental decline in test patients. Specifically the dementia Severity Rating Scale （dSRS） should supersede Mini-Mental State Examination （MMSE） and other popular tests, and a measurement scale developed in research should be used to produce a linear and non-irregular baseline. Finally, accepting the concept that four etiologies cause Alzheimer＇s disease leads to the last necessary change, that new thera- pies must be employed directed against all four causes, likely as a combination. There are drugs ready to be employed in such a combinations which are available and used clinically for other purposes so can be used ＂offlabel＂ and one such combination is suggested.

关键词： Alzheimer's disease etiologies combination therapy cognitive testing failed research off-label use dementia Severity Rating Scale

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Engineering Nitrogenases for Synthetic Nitrogen Fixation:From Pathway Engineering to directed Evolution

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Biodesign Research 2023年第1期5卷 25-36页

作者： Emily M.Bennett james W.murray Mark Isalan department of Life Sciences Imperial College LondonLondon SW72AZUK.

Globally,agriculture depends on industrial nitrogen fertilizer to improve crop *** production consumes fossil fuels and contributes to environmental nitrogen pollution.A potential solution would be to harness nitrogenases—enzymes capable of converting atmospheric nitrogen N2 to NH3 in ambient *** is therefore a major goal of synthetic biology to engineer functional nitrogenases into crop plants,or bacteria that form symbiotic relationships with crops,to support growth and reduce dependence on industrially produced *** review paper highlights recent work toward understanding the functional requirements for nitrogenase expression and manipulating nitrogenase gene expression in heterologous hosts to improve activity and oxygen tolerance and potentially to engineer synthetic symbiotic relationships with plants.

关键词： crops synthetic Evolution

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