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Culture conditions for equine bone marrow mesenchymal stem cells and expression of key transcription factors during their differentiation into osteoblasts

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Journal of Animal Science and Biotechnology 2014年第2期5卷 163-172页

作者： elizabeth r a glynn Alfredo Sanchez Londono Steven A Zinn Thomas A Hoagl Kristen E Govoni Department of Animal Science University of Connecticut3636 Horsebarn Road Ext.Unit 4040StorrsCT 06269-4040USA Department of Environmental and Population Health Tufts University North GraftonMA 01536USA

Background： The use of equine bone marrow mesenchymal stem cells （BMSC） is a novel method to improve fracture healing in horses. However, additional research is needed to identify optimal culture conditions and to determine the mechanisms involved in regulating BMSC differentiation into osteoblasts. The objectives of the experiments were to determine： 1） if autologous or commercial serum is better for proliferation and differentiation of equine BMSC into osteoblasts, and 2） the expression of key transcription factors during the differentiation of equine BMSC into osteoblasts. Equine BMSC were isolated from the sterna of 3 horses, treated with purchased fetal bovine serum （FBS） or autologous horse serum （HS）, and cell proliferation determined. To induce osteoblast differentiation, cells were incubated with L-ascorbic acid-2-phosphate and glycerol-2-phosphate in the presence or absence of human bone morphogenetic protein2 （BMP2）, dexamethasone （DEX）, or combination of the two. Alkaline phosphatase （ALP） activity, a marker of osteoblast differentiation, was determined by ELISA. Total rNA was isolated from differentiating BMSC between d 0 to 18 to determine expression of runt-reloted tronscrJption foctor2 （runx2）, osterix （Osx）, and T-box3 （Tbx3）. Data were analyzed by ANOVA. results： relative to control, FBS and HS increased cell number （133 ± 5 and 116 ± 5%, respectively; P 〈 0.001） and 5-bromo- 2＇-deoxyuridine （BrdU） incorporation （167 ± 6 and 120 ± 6%, respectively; P 〈 0.001）. Treatment with DEX increased ALP activity compared with control （1,638 ± 38%; P 〈 0.001）. In the absence and presence of Dex, BMP-2 did not alter ALP activity （P 〉 0.8）. runt-reloted transcription foctor2 expression increased 3-fold （P 〈 0.001） by d 6 of culture. Osterix expression increased 94old （P 〈 0.05） by d 18 of culture. Expression of Tbx3 increased 1.8-fold at d 3 （P 〈 0.01）; however expression was reduced 4-fold at d 18 （P 〈 0.01）. Conclusions： Dexamethasone, but not BMP-2, is required for differentiation of equine BMSC into osteo

关键词： Bone marrow mesenchymal stem cells Cell culture Equine Osteoblasts Transcription factors

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Hepatitis E virus: Western Cape, South Africa

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World Journal of Gastroenterology 2016年第44期22卷 9853-9859页

作者： richie G Madden Sebastian Wallace Mark Sonderup Stephen Korsman Tawanda Chivese Bronwyn Gavine Aniefiok Edem roxy Govender Nathan English Christy Kaiyamo Odelia Lutchman Annemiek A van der Eijk Suzan D Pas glynn W Webb Joanne Palmer elizabeth Goddard Sean Wasserman Harry r Dalton C Wendy Spearman royal Cornwall Hospital Trust and European Centre for Environment and Human Health University of Exeter Division of Hepatology Department of Medicine Faculty of Health Sciences University of Cape Town and Groote Schuur Hospital National Health Laboratory Service Groote Schuur Hospital Cape Town and Division of Medical Virology University of Cape Town Faculty of Health Sciences Department of Viroscience Erasmus Medical Centre Medical Center Rotterdam Department of Paediatrics Faculty of Health Sciences University of Cape Town and Red Cross Children’s Hospital Division of Infectious Diseases and HIV Medicine Department of Medicine Faculty of Health Sciences University of Cape Town and Groote Schuur Hospital

AIM To conduct a prospective assessment of anti-hepatitis E virus(HEV) Ig G seroprevalence in the Western Cape Province of South Africa in conjunction with evaluating risk factors for exposure.METHODS Consenting participants attending clinics and wards of Groote Schuur, red Cross Children's Hospital and their affiliated teaching hospitals in Cape Town, South Africa, were sampled. Healthy adults attending blood donor clinics were also recruited. Patients with known liver disease were excluded and all major ethnic/race groups were included to broadly represent local demographics. relevant demographic data was captured at the time of sampling using an interviewer-administered confidential questionnaire. Human immunodeficiency virus(HIV) status was self-disclosed. HEV Ig G testing was performed using the Wantai? assay.rESULTS HEV is endemic in the region with a seroprevalence of 27.9%(n = 324/1161) 95%CI: 25.3%-30.5%(21.9% when age-adjusted) with no significant differences between ethnic groups or HIV status. Seroprevalence in children is low but rapidly increases in early adulthood. With univariate analysis, age ≥ 30 years old, pork and bacon/ham consumption suggested risk. In the multivariate analysis, the highest risk factor for HEV Ig G seropositivity(Or = 7.679, 95%CI: 5.38-10.96, P rs or older followed by pork consumption(Or = 2.052, 95%CI: 1.39-3.03, P recent clinical case demonstrates that HEV genotype 3 may be currently circulating in the Western Cape.CONCLUSION Hepatitis E seroprevalence was considerably higher than previously thought suggesting that hepatitis E warrants consideration in any patient presenting with an unexplained hepatitis in the Western Cape, irrespective of travel history, age or ethnicity.

关键词： Hepatitis E Seroprevalence South Africa Pork consumption Genotype

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