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Multifaceted nature of membrane microdomains in colorectal cancer

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World Journal of Gastroenterology 2011年第6期17卷 681-690页

作者： Kristina A Jahn filip braet Australian Centre for Microscopy and Microanalysis The University of Sydney

Membrane microdomains or lipid rafts are known to be highly dynamic and to act as selective signal transduction mediators that facilitate interactions between the cell's external and internal *** rafts play an important mediating role in the biology of cancer:they have been found in almost all existing experimental cancer models,including colorectal cancer (CRC),and play key regulatory roles in cell migration,metastasis,cell survival and tumor *** paper explores the current state of knowledge in this field by highlighting some of the pioneering and recent lipid raft studies performed on different CRC cell lines and human tissue *** this literature review,it becomes clear that membrane microdomains appear to be implicated in all key intracellular signaling pathways for lipid metabolism,drug resistance,cell adhesion,cell death,cell proliferation and many other processes in *** signal transduction pathways seem to originate directly from those peculiar lipid islands,thereby orchestrating the colon cancer cells' state and *** confirmed by recent animal and preclinical studies in different CRC models,continuing to unravel the structure and function of lipid rafts-including their associated complex signaling pathways-will likely bring us one step closer to better monitoring and treating of colon cancer patients.

关键词： Actin Caveolae Cytoskeleton Combined imaging Detergent-resistant membranes Drug targeting Electron microscopy Lipid domains Membrane rafts Prognosis Staging Tomography Lipid-mediated therapy

Unlocking the ultrastructure of colorectal cancer cells in vitro using selective staining

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World Journal of Gastroenterology 2010年第22期16卷 2743-2753页

作者： Joanna M Biazik Kristina A Jahn filip braet Australian Key Centre for Microscopy and Microanalysis Madsen Building F09 The University of Sydney

AIM: To characterise differences between three widely used colorectal cancer cell lines using ultrastructural selective staining for glycogen to determine variation in metastatic properties. METHODS: Transmission electron microscopy was used in this investigation to help identify intracellular structures and morphological features which are precursors of tumor invasion. In addition to morphological markers, we used selective staining of glycogen as a marker for neoplastic cellular proliferation and determined whether levels of glycogen change between the three different cell lines. RESULTS: Ultrastructural analysis revealed morphological differences between the cell lines, as well as differentiation into two sub-populations within each cell line. Caco-2 cells contained large glycogen deposits as well as showing the most obvious morphological changes between the two sub-populations. SW480 cells also contained large glycogen stores as well as deep cellular protrusions when grown on porous filter membranes. HT-29 cells had trace amounts of glycogen stores with few cellular projections into the filter pores and no tight junction ***: Morphology indicative of metastatic properties coincided with larger glycogen deposits, providing strong evidence for the use of selective staining to determine the neoplastic properties of cells.

关键词： Cancer Colorectal Electron microscopy Glycogen Potassium ferrocyanide

Influence of kavain on hepatic ultrastructure

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World Journal of Gastroenterology 2008年第4期14卷 541-546页

作者： Shuang Fu Emine Korkmaz filip braet Quan Ngo Iqbal Ramzan Faculty of Pharmacy The Universityof Sydney NSW 2006 Australia Australian Key Centre forMicroscopy and Microanalysis Electron Microscopy Unit TheUniversity of Sydney NSW 2006 Australia Faculty of Medicine The University of Sydney NSW2006 Australia

AIM: To investigate whether the major kavalactone kavain imposes adverse effects on the liver ultrastructure and function by affecting vascular and microvascular architecture and altering hepatocellular morphology. METHODS: Kavain solution (10 μg/mL or 43.5 μmol/L) was perfused for 2 h in isolated rat livers. After standard fi xation and tissue preparation,the samples were examined by scanning electron microscopy (SEM),transmission electron microscopy (TEM),and light microscopy (LM). RESULTS: LM,SEM,and TEM examinations indicated kavain-treated rat livers (n = 4) displayed severe vascular and endothelial damage compared to control livers (n = 4). CONCLUSION: The data so far support the hypothesis that kavain induces adverse effects on liver; additional investigations with other kavalactones and their effects on liver are urgently needed.

关键词： Kava Kavalactones Kavain Liver Electronmicroscopy

Hydrodynamics based transfection in normal and fibrotic rats

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World Journal of Gastroenterology 2006年第38期12卷 6149-6155页

作者： Rita Yeikilis Shunit Gal Natalia Kopeiko Melia Paizi Mark Pines filip braet Gadi Spira Department of Anatomy and Cell Biology The Bruce Rappaport Faculty of Medicine Rappaport Family Institute for Research in the Medical Sciences Technion Haifa 31096 Israel Department of Experimental Surgery The Bruce Rappaport Faculty of Medicine Rappaport Family Institute for Research in the Medical Sciences Technion Haifa 31096 Israel Institute of Animal Sciences Agricultural Research Organization the Volcani Center Bet Dagan 50250 Israel Australian Key Centre for Microscopy and Microanalysis The University of Sydney Sydney NSW 2006 Australia

AIM： Hydrodynamics based transfection （HBT）, the injection of a large volume of naked plasmid DNA in a short time is a relatively simple, efficient and safe method for in vivo transfection of liver cells. Though used for quite some time, the mechanism of gene transfection has not yet been elucidated. METHODS： A lucJferase encoding plasmid was injected using the hydrodynamics based procedure into normal and thioacetamide-induced fibrotic Sprague Dawley rats. Scanning and transmission electron microscopy images were taken. The consequence of a dual injection of Ringer solution and luciferase pDNA was followed. Halofuginone, an anti collagen type I inhibitor was used to reduce ECM load in fibrotic rats prior to the hydrodynamic injection. RESULTS： Large endothelial gaps formed as soon as 10＇ following hydrodynamic injection; these gradually returned to normal 10 d post injection. Hydrodynamic administration of Ringer 10 or 30 m prior to moderate injection of plasmid did not result in efficient transfection suggesting that endothelial gaps by themselves are not sufficient for gene expression. Gene transfection following hydrodynamic injection in thioacetamide induced fibrotic rats was diminished coinciding with the level of fibrosis. Halofuginone, a specific collagen type I inhibitor, alleviated this effect. CONCLUSION： The hydrodynamic pressure formed following HBT results in the formation of large endothelial gaps. These gaps, though important in the transfer of DNA molecules from the blood to the space of Disse are not enough to provide the appropriate conditions for hepatocyte transfection. Hydrodynamics based injection is applicable in fibrotic rats provided that ECM load is reduced.

关键词： Gene transfection Fibrosis In vivo transfection Fibrotic Hepatic Endothelial lining Endothelium Sinusoidal Fenestrae Space of disse Extracellular matrix

The hepatic sinusoidal endothelial lining and colorectal liver metastases

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World Journal of Gastroenterology 2007年第6期13卷 821-825页

作者： filip braet Keissuke Nagatsuma Masaya Saito Lilian Soon Eddie Wisse Tomokazu Matsuura Australian Key Centre for Microscopy and Microanalysis Electron Microscopy Unit The University of Sydney NSW 2006 Australia Department of Pathology The Jikei University School of Medicine Tokyo Japan Division of Gastroenterology and Hepatology Department of Internal Medicine The Jikei University School of Medicine Tokyo Japan Department of Laboratory Medicine The Jikei University School of Medicine Tokyo Japan

Colorectal cancer (CRC) is a common malignant disease and the severe nature of cases in men and women who develop colorectal cancer makes this an important socio-economic health issue. Major challenges such as understanding and modeling colorectal cancer pathways rely on our understanding of simple models such as outlined in this paper. We discuss that the development of novel standardized approaches of multidimensional (correlative) biomolecular microscopy methods facilitates the collection of (sub) cellular tissue information in the early onset of colorectal liver metastasis and that this approach will be crucial in designing new effective strategies for CRC treatment. The application of X-ray micro-computed tomography and its potential in correlative imaging of the liver vasculature will be discussed.

关键词： Apoptosis Australia Correlative microscopy Endothelial cells Hepatic metastasis Colorectal cancer CC531 Gaps Interferon gamma Kupffer cells Natural killer cells Nitric oxide Macrophages Modeling Phagocytosis Plugging Pit cells Stellate cells X-ray micro-computed tomography

Reconstruction of liver organoid using a bioreactor

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World Journal of Gastroenterology 2006年第12期12卷 1881-1888页

作者： Masaya Saito Tomokazu Matsuura Takahiro Masaki Tetsuro Suzuki Haruka Maehashi Keiko Shimizu Yoshiaki Hataba Tohru Iwahori filip braet Division of Gastroenterology and Hepatology Department of Internal MedicineThe Jikei University School of MedicineTokyoJapan Department of Laboratory Medicine The Jikei University School of MedicineTokyoJapan Department First Department of Biochemistry The Jikei University School of MedicineTokyoJapan DNA Medical Institute The Jikei University School of MedicineTokyoJapan Fifth Division of Blood Purification Department of SurgeryTokyo Medical UniversityTokyoJapan Australian Key Center for Microscopy & Microanalysis Electron Microscope UnitThe University of SydneyNSW 2006 Australia

AIM： To develop the effective technology for reconstruction of a liver organ in vitro using a bio-artificial liver. METHODS： We previously reported that a radial-flow bioreactor （RFB） could provide a three-dimensional highdensity culture system. We presently reconstructed the liver organoid using a functional human hepatocellular carcinoma cell line （FLC-5） as hepatocytes together with mouse immortalized sinusoidal endothelial cell （SEC） line M1 and mouse immortalized hepatic stellate cell （HSC） line A7 as non parenchymal cells in the RFB. Two × 10^7 FLC-5 cells were incubated in the RFB. After 5 d, 2 × 10^7 A7 cells were added in a similar manner followed by another addition of 10^7 M1 cells 5 d later. After three days of perfusion, some cellulose beads with the adherent cells were harvested. The last incubation period included perfusion with 200 nmol/L swinholide A for 2 h and then the remaining cellulose beads along with adherent cells were harvested from the RFB. The cell morphology was observed by transmission electron microscopy （TEM） and scanning electron microscopy （SEM）. To assess hepatocyte function, we compared mRNA expression for urea cycle enzymes as well as albumin synthesis by FLC-5 in monolayer cultures compared to those of single-type cultures and cocultures in the RFB. RESULTS： By transmission electron microscopy, FLC-5, M1, and A7 were arranged in relation to the perfusion side in a liver-like organization. Structures resembling bile canaliculi were seen between FCL-5 cells. Scanning electron microscopy demonstrated fenestrae on SEC surfaces. The number of vesiculo-vacuolar organelles （WO） and fenestrae increased when we introduced the actin-binding agent swinholide-A in the RFB for 2h. With respect to liver function, urea was found in the medium, and expression of mRNAs encoding arginosuccinate synthetase and arginase increased when the three cell types were cocultured in the RFB. However, albumin synthesis decreased. CONCLUSION： Co-culture

关键词： Liver organoid Organ reconstruction Bioartificial liver Coculture Liver sinusoidal endothelial cell Hepatocytes Fenestrae Vesiculo vacuolar organelles Radial flow bioreactor

Fixation methods for electron microscopy of human and other liver

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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

Electronic Properties of Immobilised DNA-Based Molecules at the Single-Molecule Level

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Electronic Properties of Immobilised DNA-Based Molecules at ...

On the cell biology of pit cells,the liver-specific NK cells

第五届全国化学生物学学术会议

作者： Wenrong Yang~(1*) Pall Thordarson~2 filip braet~1 J. Justin Gooding~3 1 Australian Key Centre for Microscopy and Microanalysis(AKCMM),Electron Microscopy Unit,The University of Sydney,Australia 2 School of Chemistry,the University of Sydney,Australia ~3School of Chemistry,The University of New South Wales,Australia

＜正＞Using molecules as electronic components is a challenging new research direction in the science and technology of nanometre-scale *** requires understanding the electrical behaviour and properties of single molecules embedded between metal *** how electrons flow through single molecules is important in several areas such as:fabricating nanoelectronic devices and sensors;and specially rationalizing electron transfer（ET）in biological *** the later case, the nature of electrical transport along the DNA duplex string is important in our understanding of DNA damage in vivo and in exploring the possible use of DNA in molecular electronics and direct detection of DNA *** this project,we have beeing investigated the ET properties of

关键词： Single Molecules DNA Bioelectronic devices

来源： cnki会议评论

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World Journal of Gastroenterology 2000年第1期6卷 1-11页

作者： Luo DZ Vermijlen D Ahishali B Triantis V Plakoutsi G braet F Vanderkerken K Wisse E Laboratory for Cell Biology and Histology Free University of Brussels(VUB)Brussels-JetteBelgium Department of Pathology Guangxi Medical University NanningChina Laboratory for Cell Biology and Histology Free University of Brussels(VUB)Brussels-JetteBelgium Department of Hematology and Immunology Free University of Brussels(VUB)Brussels-JetteBelgium

INTRODUCTION Natural killer (NK) cells are functionally defined by their ability to kill certain tumor cells and virus-infected cells without prior

关键词： Subject headings pit cells hepatic natural killer cells large granular lymphocytes