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Fixation methods for electron microscopy of human and other liver

World Journal of Gastroenterology 2010年第23期16卷 2851-2866页

作者： Eddie Wisse Filip Braet Hans Duimel Celien Vreuls Ger Koek Steven WM Olde Damink Maartje AJ van den Broek Bart De Geest cees hc dejong Chise Tateno Peter Frederik Electron Microscope Unit University of Maastricht Australian Key Center for Microscopy and Microanalysis Sydney University Department of Internal Medicine University of Maastricht Department of Surgery University of Maastricht Center for Molecular and Vascular Biology University of Leuven PhoenixBio Co.

For an electron microscopic study of the liver,expertise and complicated,time-consuming processing of hepatic tissues and cells is *** interpretation of electron microscopy(EM) images requires knowledge of the liver fine structure and experience with the numerous artifacts in fixation,embedding,sectioning,contrast staining and microscopic ***,the aim of this paper is to present a detailed summary of different methods for the preparation of hepatic cells and tissue,for the purpose of preserving long-standing expertise and to encourage new investigators and clinicians to include EM studies of liver cells and tissue in their projects.

关键词： Liver Tissue fixation Perfusion Electron microscopy Biopsy

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Human small intestine is capable of restoring barrier function after short ischemic periods

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World Journal of Gastroenterology 2017年第48期23卷 8452-8464页

作者： Dirk HSM Schellekens Inca HR Hundscheid Claire AJI Leenarts Joep Grootjans Kaatje Lenaerts Wim A Buurman Cornelis hc dejong Joep PM Derikx Department of Surgery Maastricht University Medical Center NUTRIM School for Nutrition and Translational Research in MetabolismMaastricht University Department of Gastroenterology Academic Medical Center MHeNs School of Mental Health and Neuroscience Maastricht University Medical Center Pediatric Surgical Center of Amsterdam Emma Children’s Hospital Academic Medical Center and VU University Medical Center

AIM To assess intestinal barrier function during human intestinal ischemia and reperfusion(IR).METHODS In a human experimental model,6 cm of jejunum was selectively exposed to 30 min of ischemia(I) followed by 30 and 120 min of reperfusion(R). A sham procedure was also performed. Blood and tissue was sampled at all-time points. Functional barrier function was assessed using dual-sugar absorption tests with lactulose(L) and rhamnose(R). Plasma concentrations of citrulline,an amino acid described as marker for enterocyte function were measured as marker of metabolic enterocytes restoration. Damage to the epithelial lining was assessed by immunohistochemistry for tight junctions( TJs),by plasma marker for enterocytes damage(I-FABP) and analyzed by electron microscopy(EM) using lanthanum nitrate as an electrondense *** Plasma L/R ratio's were significantly increased after 30 min of ischemia(30 I) followed by 30 min of reperfusion(30 R) compared to control(0.75 ± 0.10 vs 0.20 ± 0.09,P < 0.05). At 120 min of reperfusion(120 R),ratio's normalized(0.17 ± 0.06) and were not significantly different from control. Plasma levels of I-FABP correlated with plasma L/R ratios measured at the same time points(correlation: 0.467,P < 0.01). TJs staining shows distortion of staining at 30 I. An intact lining of TJs was again observed at 30 I120 R. Electron microscopy analysis revealed disrupted TJs after 30 I with paracellular leakage of lanthanum nitrate,which restored after 30 I120 R. Furthermore,citrulline concentrations closely paralleled the histological perturbations during intestinal *** This study directly correlates histological data with intestinal permeability tests,revealing that the human gut has the ability of to withstand short episodes of ischemia,with morphological and functional recovery of the intestinal barrier within 120 min of reperfusion.

关键词： Intestinal ischemia-reperfusion Intestinal barrier function Intestinal permeability Human Dualsugar absorption test Tight Junctions Citrulline

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Life and death at the mucosal-luminal interface: New perspectives on human intestinal ischemia-reperfusion

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World Journal of Gastroenterology 2016年第9期22卷 2760-2770页

作者： Joep Grootjans Kaatje Lenaerts Wim A Buurman Cornelis hc dejong Joep PM Derikx Department of Surgery NUTRIM School of Nutrition and Translational Research in Metabolism Maastricht University Medical Center Brigham and Women’s Hospital Harvard Medical School NUTRIM School of Nutrition and Translational Research in Metabolism Maastricht University Medical Center

Intestinal ischemia is a frequently observed phenomenon. Morbidity and mortality rates are extraordinarily high and did not improve over the past decades. This is in part attributable to limited knowledge on the pathophysiology of intestinal ischemia-reperfusion(IR) in man, the paucity in preventive and/or therapeutic options and the lack of early diagnostic markers for intestinal ischemia. To improve our knowledge and solve clinically important questions regarding intestinal IR, we developed a human experimental intestinal IR model. With this model, we were able to gain insight into the mechanisms that allow the human gut to withstand short periods of IR without the development of severe inflammatory responses. The purpose of this review is to overview the most relevant recent advances in our understanding of the pathophysiology of human intestinal IR, as well as the(potential) future clinical implications.

关键词： Small intestinal ischemia Colon ischemia Epithelial lining Ischemia-reperfusion Human models

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