Дисертації з теми "Mesothelial"
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Lin, Judy Li-Wen. "Mechanisms of mesothelial tissue lubrication." Thesis, Massachusetts Institute of Technology, 2006. http://hdl.handle.net/1721.1/36249.
Повний текст джерела"June 2006."
Includes bibliographical references (p. 63-64).
In the pleural space, sliding between the lung and chest wall induces shear stress that could damage the delicate mesothelial cells covering the tissue surfaces. Normally, the pleural space, which is filled with fluid, is able to sustain continuous shear loading throughout its lifetime. To understand the mechanisms in preventing frictional damage on mesothelial tissue, we conducted experiments using abdominal tissue excised from a rat. We allowed the tissue to slide against a glass surface, and measured the fluid thickness and shear force between them. We also studied independent variables such as location on the tissue, applied normal load, sliding velocity and direction to determine their effects on mesothelial tissue lubrication. Both thickening and thinning of the fluid layer were observed during sliding. The fluid thickness was found to change with sliding velocity and direction, but invariant with location on tissue surface. In tribological experiments, shear force decreased with increasing velocity until it reached a minimum value varying with different tissue samples. Normal load had a strong effect on shear force, but not on friction coefficient.
(cont.) Overall, the friction curves had similar shape as described by the mixed/elasto-hydrodynamic regions of the Stribeck curve. Results were consistent within each tissue sample, but varied among samples. The dependency on velocity and direction suggested elasto-hydrodynamic lubrication. Taken together, we conclude that elasto-hydrodynamic lubrication is likely to be an important lubrication mechanism for mesothelial tissue sliding in the pleural region. Our findings support the existence of a continuous fluid layer separating the pleural surfaces. The fluid pressure gradient generated by surface roughness redistributes fluid from thick to thin regions preventing surface contact.
by Judy Li-Wen Lin.
S.M.
Davidowitz, Rachel Alexis. "Mechanisms Governing Mesothelial Clearance by Ovarian Cancer Spheroids." Thesis, Harvard University, 2013. http://dissertations.umi.com/gsas.harvard:10719.
Повний текст джерелаDixit, Radhika Nagaraj. "The contribution of mesothelial cells to lung development." Thesis, Boston University, 2013. https://hdl.handle.net/2144/12825.
Повний текст джерелаMesothelium-derived progenitors have been demonstrated to contribute to differentiated mesenchymal components of the heart, liver, and gut during organogenesis. The precise contribution of the mesothelium to lung development, however, has not been fully clarified and the key signals regulating mesothelial cell entry have not been identified. To rigorously address this issue, we employed mice with an inducible Cre expressed from the Wilm's tumor-1 (WT1) locus for high fidelity lineage tracing after confirming that Cre-recombinase was mesothelial-specific and faithfully recapitulated endogenous WT1 gene expression. We visualized WT1+ mesothelial cell entry into the fetal lung by live imaging and identified their progenies in subpopulations of bronchial smooth muscle cells, vascular smooth muscle cells, and desmin+ fibroblasts by lineage tagging. In view of the role of Sonic Hedgehog (Hh) signaling in regulating mesenchymal cell differentiation and epithelial-mesenchymal transition, we hypothesized that this pathway regulates events associated with migration of mesothelial cells into the developing lung. To examine for this, we first used two independent reporter mice to show that Hh signaling is active within the lung mesothelium at time points coinciding with the appearance of mesothelium-derived cells in the lung parenchyma. Using loss-of-function assays in organ cultures, and targeted mesothelial-restricted loss-of hedgehog function mice, we demonstrated that mesothelial cell movement into the lung requires the direct action of Hh signaling. In order to examine whether WT1 interacts with Hh pathway, we conducted ChIP assays on fetal lung mesothelial cells, and found that WT1 directly binds and regulates promoter elements of downstream targets of Hh pathway. Consistent with this observation, Hh pathway gene expression was down-regulated in isolated WT1 deficient fetal lung mesothelial cells. Taken together, these findings lend further support to a paradigm in which mesothelial cells are an important source of progenitors for mesenchymal structures. Our findings also reveal a role for Hh pathway in the early events associated with mesothelial cell entry and indicate that WT1 likely acts upstream of Hh signaling.
Gulyás, Miklós. "Mesothelial differentiation, mesothelioma and tumor markers in serous cavities /." Stockholm, 2003. http://diss.kib.ki.se/2003/91-7349-566-2/.
Повний текст джерелаYung, S. S. Y. "Characterization of proteoglycans synthesized by human peritoneal mesothelial cells." Thesis, Swansea University, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.636722.
Повний текст джерелаDauleh, S. "Characterising mesothelial cell cultures derived from the murine omentum." Thesis, University of Liverpool, 2016. http://livrepository.liverpool.ac.uk/3004240/.
Повний текст джерелаSwain, William Alexander. "Cell signalling pathways in mesothelial cells treated with mineral fibres." Thesis, University of Leicester, 2002. http://hdl.handle.net/2381/30767.
Повний текст джерелаMohamed, Moinuddin Mohammed. "Characterisation of peritoneal calcification in encapsulating peritoneal sclerosis." Thesis, University of Manchester, 2017. https://www.research.manchester.ac.uk/portal/en/theses/characterisation-of-peritoneal-calcification-in-encapsulating-peritoneal-sclerosis(003593cd-01f0-4e7b-a22b-d216451f6a93).html.
Повний текст джерелаMedcalf, James Frederick. "The role of mesothelial cell biology in peritoneal fibrosis on CAPD." Thesis, University of Leicester, 2001. http://hdl.handle.net/2381/29376.
Повний текст джерелаSun, Xiaojuan. "Studies on mesothelial differentiation : prognostic and therapeutic approaches to malignant mesothelioma /." Stockholm, 2005. http://diss.kib.ki.se/2005/91-7140-513-5/.
Повний текст джерелаThompson, Joyce K. "The Role of Inflammasomes in Asbestos-Induced Mesothelial to Fibroblastic Transition." ScholarWorks @ UVM, 2017. http://scholarworks.uvm.edu/graddis/691.
Повний текст джерелаWu, Xuan. "Role of peritoneal mesothelial cells and the inflammatory response in peritoneal fibrosis." Thesis, University of Edinburgh, 2011. http://hdl.handle.net/1842/5937.
Повний текст джерелаGardner, M. J. "Adhesion molecules in the interactions of ovarian tumour cells and mesothelial cells." Thesis, University of Newcastle Upon Tyne, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.336754.
Повний текст джерелаWard, K. L. "Analysing the angiogenic potential of mesothelial cells in vitro and in vivo." Thesis, University of Liverpool, 2017. http://livrepository.liverpool.ac.uk/3008101/.
Повний текст джерелаCrosby, Lynn Marie. "Studies on Mechanisms of Potassium Bromate-Induced Mesothelial Carcinogenesis in the Male F344 Rat." NCSU, 2000. http://www.lib.ncsu.edu/theses/available/etd-20000509-143301.
Повний текст джерелаPotassium bromate (KBrO3) is a drinking water disinfection by-product and may represent a health hazard if found to be carcinogenic for humans, as was determined in rats (DeAngelo et al., 1998; Hayashi et al., 1986; Kurokawa, 1985; Kurokawa et al., 1985; Kurokawa et al., 1986a; Kurokawa et al., 1982; Kurokawa et al., 1983a; Kurokawa et al., 1987a; Kurokawa et al., 1983b; Kurokawa et al., 1986b; Ohno et al., 1982; Onodera et al., 1985). The purpose of this research was to determine the mechanism of toxicity. The peritoneal mesothelium is a (rat) target organ of potassium bromate carcinogenicity, but has not been studied due to the inherent difficulties of working with this one cell layer-thick tissue. Data from a two-year bioassay were used to more precisely map the location of origin, revealing that these tumors in the male F344 rat originate on the mesorchium of the tunica vaginalis testis, the mesosplenium, or at a point in between. An in vitro cell culture system was developed to study the mechanism of toxicity. It was demonstrated that KBrO3 caused cell cycle arrest and markedly increased the number of apoptotic cells. KBrO3 is a powerful oxidant and glutathione (GSH) is the major cellular antioxidant. Therefore, GSH-related responses were studied revealing mesothelial cells contained substantially less GSH than a human hepatocellular carcinoma cell line (Hep-G2). Studies employing GSH ester or N-acetyl cysteine (a GSH precursor) pre-treatment demonstrated abatement of toxicity in mesothelial, but not Hep-G2, cells. Experiments carried out to determine the chemical or enzymatic nature of the reaction between GSH and KBrO3 revealed differences between the reaction kinetics of the unbuffered and buffered chemical and cell-free/cell lysate reactions, probably due to reaction pH. Both chemical and cellular reactions exhibited a similar first step reaction between GSH and KBrO3; thus, enzyme participation is probably not required. Experiments using diethylmaleate, which depletes GSH by a reaction involving KBrO3, showed that GSH depletion greatly enhanced KBrO3 toxicity, indicating GSH glutathione S-transferase, buthionine sulfoximine (which prevents synthesis of GSH) and was protective. This does not support the hypothesis that the reaction of KBrO3 and GSH itself produces a radical/reactive species that oxidatively damages lipids, proteins and DNA. Rather, depletion of GSH likely precedes oxidative damage. Gene expression studies demonstrated that peritoneal mesothelial cells displayed expression changes in a discrete set of genes, including oxidative stress-responsive genes, after treatment with KBrO3 for four or 12 hours. Mesothelial cells severely damaged by five days KBrO3 treatment recovered from complete cell cycle arrest after four weeks and exhibited explosive growth, focus formation and altered morphology. The redox imbalance created by GSH depletion appears to mediate increased expression of known oxidative stress responsive genes (e.g., HO-1,GADD45, GADD153, QR), activation of transcription factors (AP-1 and NFkB) and down-regulation of cell cycle initiating cyclins (and up-regulation of the CDK inhibitor p21waf1/cip1) in KBrO3-mediated toxicity. These alterations may permit cell survival, as observed after severe toxicity, and may be accompanied by transforming mutations or clastogenic changes. Taken together, these data suggest that mesothelial cells represent a population susceptible to KBrO3-mediated toxicity in vitro, and suggest that tissue susceptibility in vivo plays a role in the nascence of mesotheliomas in the male F344 rat.
Crosby, Lynn M. "Studies on mechanisms of potassium bromate-induced mesothelial carcinogenesis in the male F344 rat." Raleigh, NC : North Carolina State University, 2000. http://www.lib.ncsu.edu/etd/public/etd-5032149410021291/etd.pdf.
Повний текст джерелаNagai, Hirotaka. "Diameter and rigidity of multiwalled carbon nanotubes are critical factors in mesothelial injury and carcinogenesis." Kyoto University, 2012. http://hdl.handle.net/2433/157431.
Повний текст джерелаBussadori, Giulio. "Mesothelial Signature in Mesenchymal Stem/Stromal cells derived from HighGrade Serous Ovarian Cancer marks their Identity." Doctoral thesis, Università degli studi di Trieste, 2015. http://hdl.handle.net/10077/11100.
Повний текст джерелаMesenchymal Stem/Stromal Cells (MSCs) are the precursors of various cell types that compose both normal and cancer tissue microenvironments. In order to support the widely diversified parenchymal cells and tissue organization, MSCs are characterized by a large degree of heterogeneity, although available analyses of molecular and transcriptional data do not provide clear evidence. Moreover a wealth of studies has demonstrated a significant role of the microenvironment and MSCs in tumor growth. In the course of the years MSCs were isolated by different groups from different tissues, both healthy and cancerous. The laboratory in which I conducted my PhD project was able to purify MSCs from different healthy tissues (N-MSCs) and, using an adapted protocol, from High-Grade Serous Ovarian Carcinomas (HG-SOC-MSCs). It was possible to shown that these cells do not possess gross chromosomal aberrations and are not tumorigenic in vivo. To better characterize these cells, an integrative bioinformatics analysis was conducted using the deep-CAGE-derived expression profiles obtained from HG-SOC-MSCs, N-MSCs and the FANTOM5 large comprehensive primary cells and tissues dataset. When compared to the other cells and tissues, HG-SOC-MSCs showed a correlation with mesothelial cells and cells hypothesized to have a mesothelial origin, such as smooth muscle cells and fibroblasts. It is known that mesothelial cells in culture can alternate between epithelioid and fibroblastoid morphologies and express high levels of either keratin or vimentin or both depending on their state of growth and the presence of EGF. When cultivated in the absence of EGF, which is known to induce a morphological switch in mesothelial cells, HG-SOC-MSCs switch from a fibroblast-like to an epithelial-like shape. N-MSCs in the same culture conditions, instead, do not change morphology. Moreover, in absence of EGF, HG-SOC-MSCs but not N-MSCs raise the levels of Keratin 7 both in protein and in mRNA. Starting from the list of up-regulated genes in HG-SOC-MSCs compared to N-MSCs a list of mesothelial-related genes was generated. This mesothelial-related gene list was compared to high-throughput gene expression datasets of MSCs derived from other tissues. Such analysis revealed that the mesothelial-related signature is specific to HG-SOC-MSCs. Moreover, Kaplan-Meier survival analysis conducted on a comprehensive SOC microarray dataset showed that patients with higher levels of the mesothelial-related gene signature displayed shorter progression-free survival time. Such correlation was rather specific for HG-SOC given that its performance was either statistically non-significant in the case of lung cancer or correlated with good prognosis in the case of breast cancer. Altogether, the study allowed us to assign a specific identity for MSCs derived from high-grade serous ovarian cancer. We demonstrated a cell-type specific transcriptional activity associated with HG-SOC-MSCs, which identifies them compared to N-MSCs from other districts and position them close to primary mesothelial and mesothelail-derived cells within the FANTOM5 dataset.
XXV Ciclo
1985
Sone, Michihiko. "Mesothelial Cell Proliferation in the Scala Tympani: a Reaction to the Rupture of the Round Window Membrane." 名古屋大学医学部, 1998. http://hdl.handle.net/2237/6200.
Повний текст джерела陳曉瑞 and Xiaorui Chen. "Effects of high glucose, peritoneal dialysis fluid and heparin on proteoglycan synthesis in human peritoneal mesothelial cell." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31242960.
Повний текст джерелаChen, Xiaorui. "Effects of high glucose, peritoneal dialysis fluid and heparin on proteoglycan synthesis in human peritoneal mesothelial cell /." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B23457387.
Повний текст джерелаTOYOKUNI, SHINYA, NORIHIKO KOHYAMA, NOBUAKI MISAWA, HIROTAKA NAGAI, YUE WANG, AKIHIRO SAKAI, SHAN HWU CHEW, YASUMASA OKAZAKI, and DILINUER AIERKEN. "Rat model demonstrates a high risk of tremolite but a low risk of anthophyllite for mesothelial carcinogenesis." Nagoya University School of Medicine, 2014. http://hdl.handle.net/2237/19493.
Повний текст джерелаGuo, Hong, and 郭紅. "Effects of anti-DNA antibodies on pleural mesothelial cells: in vitro studies to explore thepathogenetic mechanism of pulmonary lupus." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2003. http://hub.hku.hk/bib/B26631945.
Повний текст джерелаpublished_or_final_version
abstract
toc
Medicine
Master
Master of Philosophy
Rosso, L. P. A. "RUOLO DEL GEL PIASTRINICO DA SANGUE PLACENTARE NEI PROCESSI DI RIPARAZIONE PLEURICA." Doctoral thesis, Università degli Studi di Milano, 2013. http://hdl.handle.net/2434/217629.
Повний текст джерелаOBJECTIVES: Prolonged air leak is the major cause of morbidity after pulmonary resection. In this study we tested an innovative approach based on the use of the platelet gel derived from human umbilical cord blood in repairing pleural damage through in vitro and in vivo experimental approaches. METHODS: The in vitro model scratch assay was performed to test the tissue repair capability mediated by platelet gel compared to the standard culture conditions using human primary mesothelial cells. In vivo the animal model consisted of an iatrogenic injury on the left lung surface. Fifty-four Wistar rats were divided into a treated group with cord blood platelet gel (n=31) and a control group (n=23). After the damage, the cord blood platelet gel was placed on the injured area only in treated animals. Rats were sacrificed to evaluate and study histological changes, and possible presence of pleural adhesions and infections. In addition, changes in the soluble inflammatory factor pattern were tested using a multiplex proteome array. RESULTS: In vitro, cord blood platelet gel repaired the damage of mesothelial cells in a shorter time in comparison with the control (24 vs 35 hours, respectively). In vivo, the formation of new mesothelial tissue was already visible at 45+1 hours in treated group vs 130+2.5 hours in control group; complete recovery was obtained respectively 75+1 hours compared to 160+6 hours. There was direct evidence of pleural adhesions in 43% of treated compared to 17% of controls. The gel was not associated with the development of any complications. Interestingly, some crucial soluble factors involved in inflammation were significantly reduced in the treated animals. CONCLUSIONS: Cord blood platelet gel significantly reduces the time to repair the pleural damage. In addition, it positively stimulates the development of pleural adhesions, particularly useful in the management of prolonged air leaks. We hypothesize that platelet gel play at least a double role releasing cytokines and growth factors that support the faster tissue repair and consequently reduce the inflammation site.
Misri, Ripen. "Molecular imaging of mesothelin expressing cancers." Thesis, University of British Columbia, 2010. http://hdl.handle.net/2429/30461.
Повний текст джерелаThattamparambil, Pravin. "Telomerase-Expression in reaktiven und neoplastischen mesothelialen Läsionen." [S.l.] : [s.n.], 2004. http://deposit.ddb.de/cgi-bin/dokserv?idn=975790242.
Повний текст джерелаBaxter, Katherine Elizabeth. "Developing an Oncolytic Prime-Boost Vaccine Targeting the Tumour Associated Antigen Mesothelin for the Treatment of Pancreatic Cancer." Thesis, Université d'Ottawa / University of Ottawa, 2020. http://hdl.handle.net/10393/40024.
Повний текст джерелаYoung, Vicky Jane. "The role of the peritoneum and transforming growth factor β in the aetiology of endometriosis". Thesis, University of Edinburgh, 2015. http://hdl.handle.net/1842/21099.
Повний текст джерелаBetz, Georg. "Pulmonales Blastom Abgrenzung vom malignen diffusen Mesotheliom und pulmonalen Adenokarzinom /." [S.l.] : [s.n.], 2003. http://deposit.ddb.de/cgi-bin/dokserv?idn=970850182.
Повний текст джерелаDobra, Katalin. "Malignant mesothelioma: an experimental study with emphasis on proteoglycans in mesothelias cell growth and differentiation /." Stockholm, 2002. http://diss.kib.ki.se/2002/91-7349-168-3/.
Повний текст джерелаFegan, Kenneth Scott. "Study of inflammatory signalling in epithelial ovarian cancer and the normal human mesothelium." Thesis, University of Edinburgh, 2010. http://hdl.handle.net/1842/4819.
Повний текст джерелаSantos, Camila Ramos dos. "Estudos estruturais das proteinas Q4DV70 de Trypanosoma cruzi e mesotelina de Homo sapiens." [s.n.], 2009. http://repositorio.unicamp.br/jspui/handle/REPOSIP/314349.
Повний текст джерелаTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
Made available in DSpace on 2018-08-14T15:40:05Z (GMT). No. of bitstreams: 1 Santos_CamilaRamosdos_D.pdf: 7751754 bytes, checksum: 779a2794a6dc213431c4264c43cc0af6 (MD5) Previous issue date: 2009
Resumo: Neste trabalho realizamos estudos estruturais com duas proteínas, a Q4DV70 de Trypanosoma cruzi e a mesotelina de Homo sapiens. O objetivo foi contribuir para a compreensão da função dessas proteínas, as quais possivelmente são importantes para a doença de Chagas e o câncer, respectivamente. A proteína Q4DV70 estava anotada no genoma de T. cruzi como hipotética conservada. Em nosso estudo, a proteína foi pela primeira vez detectada em amostras do parasita. Essa expressão ocorre na fase epimastigota, mas não na fase tripomastigota metacíclica, indicando que a proteína pode ter uma função importante no ciclo de vida do T. cruzi. A estrutura cristalográfica da Q4DV70 foi resolvida por substituição molecular e refinada com dados até 1,5 Å de resolução. Ela apresenta enovelamento tiorredoxina, formado por uma folha ß de 5 fitas cercada por duas hélices a de cada lado. As proteínas que apresentam maior identidade seqüencial e superposição estrutural com Q4DV70 são as tiorredoxinas e PDIs, oxidorredutases de pontes dissulfeto. Porém, as duas cisteínas do sítio ativo dessas proteínas estão substituídas por serinas em Q4DV70, o que impossibilita a função de formação, redução ou isomerização de pontes dissulfeto. Diversas tiorredoxina-like apresentam atividade chaperona independente da função oxidorredutase. Essa função está relacionada a regiões hidrofóbicas na superfície e/ou depende da presença de outro domínio. Q4DV70 é monomérica, composta apenas pelo domínio tiorredoxinalike, e não apresenta regiões hidrofóbicas em sua superfície. Além disso, não demonstrou capacidade de aumentar o enovelamento da GAPDH de T. cruzi. Esses resultados indicam que Q4DV70 não apresenta atividade chaperona. Mesotelina é uma proteína expressa em mesotélio normal e em diversos tipos de câncer, como mesotelioma, câncer de ovário e de pâncreas e leucemia mielóide aguda. Ela é considerada um marcador diagnóstico para esses cânceres e tem sido alvo para o desenvolvimento de drogas anti-tumor. Além disso, ela interage com MUC16, uma proteína presente na superfície de células de câncer de ovário. Apesar da reconhecida importância e uso da mesotelina, pouco se sabe sobre sua estrutura e função. A proteína foi purificada em condições desnaturantes e submetida ao re-enovelamento por diálise. Experimentos de dicroísmo circular, fluorescência e proteólise limitada comprovaram que a mesotelina foi corretamente re-enovelada. Essa amostra foi submetida a ensaios de cristalização, os quais resultaram em cristais que difrataram a baixa resolução. A estrutura de baixa resolução da mesotelina foi calculada a partir de dados de espalhamento de raios X a baixo ângulo e mostra que sua forma é alongada e curvada. O espectro de dicroísmo circular da mesotelina é típico de proteínas ricas em hélices a. Recentemente foi proposto que a estrutura da mesotelina é formada por uma estrutura em super-hélice, composta por repetições do tipo ARM, as quais apresentam 3 hélices a cada uma. Nossos resultados experimentais indicam que esse modelo teórico está correto. Proteólise limitada com quimotripsina resultou em um domínio estável de 20 kDa na região N-terminal da proteína. Esse domínio contém os 64 resíduos descritos como possível sítio de ligação a MUC16 e deve ser formado pelas 4 primeiras repetições do tipo ARM, de acordo com o modelo publicado.
Abstract: In this work we carried out studies with two proteins, Q4DV70 from Trypanosoma cruzi and mesothelin from Homo sapiens. The aim was to help the understanding of the function of these proteins which possibly are important to Chagas disease and cancer, respectively. Q4DV70 protein was annotated in T. cruzi genome as a conserved hypothetical protein. In our studies, the protein was detected for the first time in parasite samples. Its expression occurs in the epimastigote form but not in the metacyclic trypomastigote form indicating that Q4DV70 is important for the pathogen life cycle. Q4DV70 crystal structure was solved by molecular replacement and refined with data to 1.5 Å maximum resolution. It shows a thioredoxin fold, formed by a five stranded ß-sheet flanked by two a-helixes in each side. The proteins more sequentially identical and better structurally superposed to Q4DV70 are thioredoxins and protein disulfide isomerases, which are disulfide oxidoreductases. However, the cysteine residues from CXXC motif of the active site are replaced by serines in Q4DV70, what prevents the function of formation, reduction and isomerization of disulfide bonds. Different thioredoxin-like proteins show chaperone activity independent of oxidoreductase function. This function is related to hydrophobic regions on the surface and/or is dependent of another domain. Q4DV70 is monomeric, composed only by thioredoxin-like domain and does not show hydrophobic regions on its surface. Moreover the protein does not increase the refolding of GAPDH from T. cruzi. These results indicate that Q4DV70 does not present chaperone activity. Mesothelin is a protein expressed in normal mesothelium and in different types of cancer, such as mesothelioma, ovarian cancer, acute myeloid leukemia and cancer of pancreas. It is considered a diagnostic marker for these cancers and it has been used in the development of antitumor drugs. Besides that, it binds to MUC16, a protein present on the surface of ovarian cancer cells. In spite of its recognized significance and use in cancer, little is known about its structure and function. The protein was purified under denaturing conditions and submitted to refolding by dialysis. Circular dichroism, fluorescence and limited proteolysis experiments confirmed that mesothelin was correctly refolded. This sample was submitted to crystallization trials that resulted in crystals which diffracted at low resolution. The low resolution structure of mesothelin was calculated from small angle X-ray scattering data and it shows an elongated and curved shape. The circular dichroism spectrum for mesothelin is typical of proteins that are rich in a-helixes. Recently it was proposed that mesothelin has a superhelix structure made by ARM repeats which are composed by 3 a-helixes each one. Our experimental results indicate that this theoretical model is correct. Limited proteolysis with chymotrypsin resulted in a stable domain of 20 kDa in the Nterminal region of the protein. This domain has the 64 residues described as the possible binding site for MUC16 and should be composed by the first four ARM repeats according to the model.
Doutorado
Bioquimica
Doutor em Biologia Funcional e Molecular
OHAN, JEANNY. "Etude des cellules mesotheliales humaines et porcines comme cibles pour des applications cliniques : ensemencement des protheses vasculaires - therapie genique." Paris 7, 2001. http://www.theses.fr/2001PA077106.
Повний текст джерелаCUNHA, M. N. PINTO DA. "IMMUNOPROFILE IN EFFUSION CYTOLOGY." Doctoral thesis, Università degli Studi di Milano, 2010. http://hdl.handle.net/2434/150058.
Повний текст джерелаROUGIER, JEAN-PHILIPPE. "Effets des dialysats et des cytokines sur le renouvellement de la matrice extracellulaire par les cellules mesotheliales peritoneales humaines : etude in vitro." Paris 6, 1997. http://www.theses.fr/1997PA066717.
Повний текст джерелаAsgarov, Kamal. "Development of more precise and efficient antibodies for cancer targeting : membrane associated form specific anti-mesothelin antibodies and CAR as an example." Thesis, Besançon, 2016. http://www.theses.fr/2016BESA3013/document.
Повний текст джерелаAntibody based immune treatment is a promising component of cancer therapy. To date there are more than 30 approved monoclonal antibodies for cancer therapy. More than 350 antibodies are also in different phases of clinical development. Mesothelin is one of the most promising targets for immunotherapy. It is present at relatively low levels in mesothelial cells of the pleura, peritoneum and pericardium of healthy individuals, but is highly expressed in a number of different cancers, including mesotheliomas, stomach cancers, squamous cell carcinomas, as well as prostate, pancreatic, lung, and ovarian cancers. Mesothelin is a glycosylphosphatidylinositol (GPI)-linked glycoprotein synthesized as a 69 kDa precursor and proteolytically processed into a 30 kDa NH2-terminal secreted form (formerly referred to as Megakaryocyte Potentiating Factor (MPF)) and a 40 kDa membrane-bound form. Besides that it can be cleaved by a protease leading to the production of a soluble, shedded, form of mesothelin. It has already been shown that this soluble form of mesothelin acts as a ligand and neutralizes the mesothelin targeting therapeutic antibodies. Therefore antibodies could not reach cancer cells and remained inefficient. In our work we decided to develop discriminating antibodies specific to a membrane associated form so as to overcome the antagonism produced by soluble forms of mesothelin. To this aim we used a novel method of mouse immunization, in which we first tolerized the mouse with soluble mesothelin before immunization with mesothelin expressing cells. By using phage display technology we obtained nearly 150 mesothelin recognizing clones in 34 VH-CDR3 families, among which we identified only 2 families that bind membrane mesothelin with high affinity and do not recognize any other soluble form of mesothelin. Here we suggest that this Fab can be effective candidates to be used for mesothelin expressing cancer therapy being allowed to pass through the soluble mesothelin barrier. To show their efficacy for therapeutic use we constructed a CAR with the sc-Fv of a membrane-form discriminating antibody
Yegles, Michel. "Cytotoxicite et segregations anormales de chromosomes produites sur des cultures de cellules mesotheliales pleurales de rat par des fibres minerales. Importance des caracteristiques dimensionnelles des fibres." Paris 7, 1994. http://www.theses.fr/1994PA077196.
Повний текст джерелаTurini, Marc. "Développement d'anticorps bispécifiques de lama pour le traitement de cancers du sein réfractaires à l'action du trastuzumab." Thesis, Aix-Marseille, 2014. http://www.theses.fr/2014AIXM4021.
Повний текст джерелаTrastuzumab is established as standard of care for the treatment of HER2high breast cancers. However, in addition to Fc-related limitations inherent to IgG antibodies, trastuzumab is inefficient to treat low- (triple-negative) or moderate-HER2-overexpressing (hormone-receptor-positive) breast cancers. Based on the unique structural and functional properties of llama single domain antibodies (sdAbs), we report the design of two Fab-like bispecific antibodies targeted to HER2 (HER2bsFab) and mesothelin (MesobsFab), an antigen overexpressed in several human tumors, including triple-negative breast cancers. The two bsFabs display a unique, specific and high affinity for FcγRIII. As a consequence, they do not bind the FcγRIIB inhibitor receptor and bypass competition with endogenous IgGs. HER2bsFab mediated ADCC at picomolar concentration against HER2high as well as HER2moderate cell lines. In vivo HER2bsFab potently inhibited HER2high tumor growth and more importantly, exhibited a net superiority over trastuzumab at inhibiting HER2moderate tumor growth. Moreover, FcγRIIIA-engagement by HER2bsFab was independent of FcγRIIIA-158 polymorphism and induced a stronger NK cells activation in response to target cell recognition. Such findings led us to investigating the efficacy of bsFabs in a context of low-HER2-overexpression displays by triple-negative breast cancers. In vitro characterization showed that both HER2bsFab and MesobsFab trigger efficient lysis of two different triple-negative breast cancer cell lines. Altogether, these findings would enable the treatment of a broader population of patients than that eligible with current HER2-targeted therapies
Ney, Britta Joanna Berenike [Verfasser], and Annette [Akademischer Betreuer] Staebler. "Das Expressionsmuster von CA-125 und Mesothelin in Primärtumor und Metastasen des high-grade serösen Ovarialkarzinoms. - Über den Einfluss dieser tumorassoziierten Antigene und tumor-infiltrierenden Lymphozyten auf die Prognose. / Britta Joanna Berenike Ney ; Betreuer: Annette Staebler." Tübingen : Universitätsbibliothek Tübingen, 2020. http://d-nb.info/1204422095/34.
Повний текст джерелаHasanova, Reyhan. "Identification de nouveaux biomarqueurs des cancers colorectaux." Thesis, Bourgogne Franche-Comté, 2019. http://www.theses.fr/2019UBFCE017.
Повний текст джерелаColorectal cancer (CRC) is the third most commonly diagnosed cancer after lung cancer, and prostate cancer. The early prognostic estimation could reduce the mortality and the implementation of new biomarkers is getting indispensable. In this study we investigated two molecules for better characterization and prognosis of CRC: Angiopietin2 (Ang2) and Mesothelin (Msln). We first analyzed transcriptomic and clinicopathological data of primary tumors collected from online repositories. 1617 transcriptomes had been produced by microarrays and were downloaded from NCBI Gene Expression Omnibus (GEO) and additionally 573 RNAseq transcriptomic data were collected from The Cancer Genome Atlas (TCGA) repository. Plasma ELISA assays for Ang2 and Msln were then performed in 20 healthy donors and in 51 CRC patients from a local cohort in order to validate those proteins as potential biomarkers. Ang2 survival analyses performed on transcriptomic data showed that high intratumoral Ang2 expression was associated with poor overall survival (OS) and relapse-free survival (RFS) in CRC patients with localized stages. As expected, Ang2 expression was associated with stroma infiltration and angiogenesis but multivariate analyses showed that Ang2 was an independent factor for OS after adjusting it with all available variables (including Consensus Molecular Subtypes of CRC). The plasma measurement of Ang2 showed that the metastatic CRC (mCRC) patients had higher levels than the healthy donors and the progression free survival (PFS) was lower in patients with higher Ang2. High intratumoral MSLN expression in CRC transcriptomes was associated with poor OS and RFS. It was also an independent factor for OS and RFS. Pathway analyses revealed that Msln expression was associated with tumor exosome pathway and cell invasion. ELISA measurements of Msln levels showed that the patients with colorectal cancer had significantly higher Msln levels than healthy donors. Furthermore, the CRC patients with increased levels of Msln had poor OS than the patients with low Msln levels. Taken together, these results suggest that both Ang2 and Msln are independent prognostic biomarkers for CRC. We now aim to stratify risk using both Ang2 and Msln to improve the treatment of CRC patients
yun, Liu shin, and 劉心雲. "Mitochondria in high glucose induced human peritoneal mesothelial cell apoptosis." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/00745300800019609859.
Повний текст джерела臺北醫學大學
醫學技術學系
94
In Taiwan, there are at present more than forty thousand patients with end-stage renal disease (ESRD) under regular dialysis therapy. Kidney disease is the seventh of ten leading causes of mortality in our country.The incidence rate of ESRD in Taiwan has gained the second place in the world. Continuous ambulatory peritoneal dialysis (CAPD) is a convenient therqpy as one kind of renal replacement therapy which interferes less of daily activity in ESRD patients. Peritoneal fibrosis (PF) is one of the most common complications in peritoneal dialysis (PD). It has been found that bio-incompatible PD solutions bearing characteristics of high glucose (HG), hyperosmolality, and low pH value may leads to the development of PF. The glucose concentration of PD solutions in Taiwan is 1.5%(83.8mM), 2.5%(138mM), or 4.25%(236mM). After long-term exposure of HG condition, human peritoneal mesothelial cells (HPMCs) will result in changes of peritoneal structure and function. Histological evaluations had demonstrated that HPMCs apoptosis and accumulation of extracellular matrix (ECM) are main pathogenesis of PF. However, the mechanisms of HG-induced apoptosis of HPMCs and the role of mitochondria as well as reactive oxygen species (ROS) in PF remains undetermined. This research is aims to evaluate effects of HG in apoptosis of HPMCs. We also investigated the efficacy of mitochondria oxidative phosphorylation and ROS generation in HPMCs under HG condition. It was found that HG induced HPMC apoptosis, cytochrome c release, PARP cleavage, activations of caspase-9, caspase-3, and an up-regulated gene expression of type I collagen with 138mM and 236mM glucose treatment. It was also demonstrated that these detrimental effects of HG in RMC caould be significantly debased apoptosis 50% by 1mM l-N-acetylcystein (L-NAC), debased apoptosis 60% by 1μM rotenone or debased apoptosis 12%by 1μM (CCCP) carbonyl cyanide m-chlorophenylhydrazone supplemented. Induced apoptosis and ECM accumulation were also revealed in rat mesangial cell cuture with 35mM glucose treatment. Our work may have great clinical implications in management of diabetic nephropathy and provide pharmacological information for prevention of PF in long-term PD patients.
Liu, Yo-Min, and 劉佑民. "Decoy Receptor 3 (DcR3) Protects Transdifferentiated Human Mesothelial Cells from Apoptosis." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/35289121632713501411.
Повний текст джерела國立陽明大學
生理學研究所
96
Background. Myofibroblasts are known to be important in the pathogenesis of peritoneal fibrosis by promoting cytokine release, extracellular matrix formation and angiogenesis. Fas-mediated apoptosis plays a vital role in the regulation of myofibroblast clearance, while decoy receptor 3 (DcR3) is a soluble receptor that competitively binds to Fas ligand (FasL). In the present study, we sought to investigate the protective effects of DcR3 in the Fas-mediated apoptosis of human peritoneal mesothelial cells (HPMCs) , in both its native and transdifferentiated states. Methods. HPMCs and human peritoneal fibroblasts (HPFs) were isolated from human omentum tissue, and transdifferented HPMCs (tPMCs) were initiated by induction with 3 ng/ml of transforming growth factor β1 (TGF-β1) for 7 days. Morphological differences and the expression of e-cadherin, vimentin, cytokeratins, and α-smooth muscle actin were observed. Production of DcR3 was induced by TNFα and subsequently measured using ELISA and reverse transcription PCR of DcR3 mRNA. Fas receptor expression was characterized by immunofluorescent cytometry and RT-PCR. Apoptosis was induced by sensitizing the cells with cycloheximide (CHX) and then treatment with FasL 100 ng/ml, and incremental doses of human recombinant DcR3-Fc (rhDcR3-Fc) were added to evaluate their effects on FasL-induced apoptosis. Apoptosis was assessed by FACS measurement of TUNEL, PI staining and immunoblotting for the active form of caspase 3. Results. HPMCs and HPFs exhibit a negligible constitutive production of DcR3 and low production even after induction with TNFα, while tPMCs show a constitutive expression of DcR3 that was significantly increased after induction with TNFα. Fas receptor expression was significantly down-regulated in tPMCs. The percentage of apoptotic tPMCs were not significantly increased with FasL treatment alone, but pretreatment with CHX sensitized cells to FasL-induced apoptosis. HPMCs exhibited a dramatic increase in apoptosis percentage after induction with CHX/FasL, while apoptosis in tPMCs and HPFs were less pronounced. The addition of rhDcR3-Fc inhibits FasL-induced cell death in all three cell lines at a concentration of 100ng/ml. Conclusion. tPMCs up-regulate DcR3 expression and down-regulate Fas receptor expression. tPMCs are also more resistant to FasL-mediated apoptosis, and the presence of DcR3 in culture medium protects HPMCs, tPMCs and HPFs cells against FasL-induced apoptosis. tPMCs have previously been implicated as a protagonist in peritoneal fibrosis, and their higher DcR3 secretion and lower Fas expression may play an important role in the balance between healing and fibrosis after peritoneal inflammation.
Liaw, Yuang-Shuang, and 廖永祥. "Regulation of Intracellular pH and Ion Transporters in Human Pleural Mesothelial Cell." Thesis, 1998. http://ndltd.ncl.edu.tw/handle/30998346568803536248.
Повний текст джерела國立臺灣大學
臨床醫學研究所
87
Pleural mesothelial cells (PMCs), which cover the surface of the parietal and visceral pleura, play important roles to reconstruct and restore the normal pleural function after pleural injury. Although infectious, inflammatory and neoplastic diseases frequently involved the pH change of pleural effusion, the regulatory mechanisms have not yet completely illustrated in a variety of pleural disorders. We hypothesized the pH of pleural effusion might be regulated by the transmembrane transporters in the pleural lining cells, i.e., pleural mesothelial cells. We established the methods to isolate and identify the cultured pleural mesothelial cells from pleural effusions and pleural tissues. Morphologically, the cells formed a homogeneous cell population, and polygonal when confluent. Cultured PMCs co-expressed cytokeratin and vimentin, which is a characteristic of epithelial and mesodermal origin, respectively. Then we provided a model to study intracellular pH (pHi) regulation in PMCs, pleural effusion formation, and pleural injury and carcinogenesis. Responding extrinsic or intrinsic stimuli, PMCs can produce a broad spectrum of extracellular matrix, including at least three collagen types (I, III, IV), elastin, fibronectin and laminin. The extracellular matrix could be the ligands, which interact with integrins, a family of cell adhesion molecules, in PMCs. Integrins are heterodimers, * and * subu-nits, which mediate cell- to- cell or extracellular connections. Interactions of ligands and integrins mediated cell morphological change, cellular attachment and detachment, activation Na-H exchanger, then induced intracellular alkalization. By using flow cytometry, we found the a2, a3, a5, b1, b3, and avb3 subunits were uniformly expressed at higher (>70%) positive cells for integrin staining on cultured human PMCs. The expression a1 subunit was intermediate (30-70%), and a4 and a6 subunits were low (<30%). The immunofluorescent stain also showed similar findings. The expression of integrins on Met-5A cells showed that a3, a5, a6, and b1 subunits were high, a1 subunit, intermediate and a2, a4, b3 and avb3 subunits, low. The patterns of immnunohistochemical staining of integrin expression on pleural tissues were generally and consistently similar to the results of the flow cytometry in the cultured human PMCs, except b3. To further investigate the regulation of pHi in PMCs, we studied the intrinsic buffering power of H+ ions (bi) and the pHi regulatory systems by using a pH-sensitive fluorescent probe, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) with microspectrofluorimetry. We found: (1) that at the resting pHi, the bi was low and increased as the pHi decreased; (2) that the pHi recovery was largely inhibited either with Na-free medium or nominally HCO3 free medium containing ethyl-isopropyl amiloride (EIPA); (3) a 4-4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS)-sensitive, Na/HCO3-dependent, but Cl-independent acid extrusion mechanism in CO2/HCO3 buffer; and (4) that in the same buffer, a DIDS-sensitive but Na-independent alkalosis was induced by intracellular Cl depletion. We therefore conclude that at least three membrane pHi regulators are involved in regulating the pHi in PMCs, these being the EIPA-sensitive Na-H exchanger; a novel electroneutral, DIDS-sensitive Na-HCO3 cotransporter; and the DIDS-sensitive Cl-HCO3 exchanger. Furthermore, under physiologic conditions, the Na-HCO3 cotransporter plays a more important role in extrusion of excess intracellular H+ ions than does the Na-H exchanger. EGF (epidermal growth factor) has been shown to stimulate cell proliferation in various cell types, including rat mesothelial cells. EGF also stimulates the Na-H exchanger, leading to enhanced cell proliferation. In human PMCs, the intracellular signaling mechanism mediating the EGF-induced stimulation of the Na-H exchanger has not yet been identified. Using a pH-sensitive fluorescent probe, BCECF with microspectrofluorimetry, to measure changes in the intracellular pH (pHi), we have found that: (1) EGF and 12-O-tetradecanoyl-phorbol-13-acetate (TPA, a phorbol ester) both stimulate the ethyl-isopropyl amiloride (EIPA)-sensitive Na+-H+ exchanger; (2) the TPA-induced alkalosis can be blocked by protein kinase C (PKC) inhibitors (chelerythrine and staurosporine) or by PKC down-regulation, indicating that PKC activation is involved in the stimulation of the Na+-H+ exchanger. However, the TPA-induced alkalosis is not blocked by tyrosine kinase inhibitors; (3) the stimulatory effect of EGF on the Na-H exchanger acts via stimulation of receptor tyrosine kinase activity, since it is inhibited by tyrosine kinase inhibitors (genistein, lavendustin A and herbimycin A), but, in addition, involves PKC activation, since the EGF-induced alkalosis is blocked by PKC inhibitors. (4) cell proliferation is both significantly enhanced by addition of EGF and the changes in pHi resulting from the change of extracellular pH (pHo). Taken together our observations indicate that the stimulatory effect on the Na-H exchanger with addition of EGF is mainly caused by PKC activation. The proliferative ability induced by EGF in human PMCs, however, is at least partly controlled by the changes in pHi which are mainly regulated by the Na- H exchanger. Based on these studies, we established a model to investigate the cellular function and physiology of human PMCs. The expression of integrins was first reported in PMCs. We also illustrated the regulation of pHi, through ion transporters and intrinsic buffering capacity. Further, we found the EGF-stimulated Na-H exchanger in PMCs via PKC pathway. This information provides new modalities for treatment of pleural injury and pleural carcinogenesis. We will investigate further the relationship of the effects and the mechanisms of fibrinolytics and pleurodesis agents with the integrins and ion transporters in PMCs.
Chen, Jinn-Yang, and 陳進陽. "The implication of peritoneal mesothelial cell apoptosis in peritonitis of peritoneal dialysis patients." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/09114279108536729942.
Повний текст джерела國立陽明大學
臨床醫學研究所
91
Background: Peritoneal dialysis (PD) is a widely used renal replacement therapy modality for end-stage renal disease patient. Patients receiving PD have higher technical failure rate than that of hemodialysis patients. Repeated peritonitis and ultrafiltration failure after long-term use of bio-incompatible peritoneal dialysate are two main reasons of technical failure. Peritonitis in PD patients is characterized by prolonged repair and was believed to cause peritoneum damage due to chronic inflammation. However, peritoneal mesothelial cells have good proliferating ability. Therefore, what is the reason of prolonged repair of peritoneum in PD peritonitis? We try to investigate the significance of human peritoneal mesothelial cells (HPMC) apoptosis during peritonitis and the role of nitric oxide, Fas-Fas ligand system in the process of peritoneal mesothelial cells apoptosis. Methods: We use cytokines (Tumor necrosis factor-a (TNF-α) 、Interleukin-1b (IL-1b)、Interferon-g (IFN-γ) 及lipopolysaccharide (LPS)) singly or in various combinations to stimulate HPMC and scrutinize the expression of nitric oxide synthase type II (iNOS) by western blot, reverse transcriptase polymerase chain reaction and Griess method. HPMC apoptosis was investigated by propidium iodide staining and TUNEL method. We also studied the expression of Fas in HPMC by flow cytometry and use anti-Fas antibody (clone CH11) to stimulate cytokine-primed HPMC. Apoptosis of HPMC was demonstrated by propidium iodide, TUNEL method and M30 cytodeath antibody staining. The quantity of apoptosis was studied by flow cytometry. The activities of caspase-3, caspase-8 and Bcl-2 were investigated by western blot. The apoptotic HPMC were co-incubated with adhered macrophages to see if macrophages could engulf apoptotic HPMCs. The peritoneal dialysate effluents of PD peritonitis patients were studied for the presence of apoptotic HPMCs. Results: Our results showed that iNOS is inducible after concomitant stimulation of TNF-a (5ng/ml) and IFN-g (5ng/ml). However, endogenous production of nitric oxide did not lead to apoptosis of HPMC. On the other hand, pre-incubation of HPMC with TNF-a (5ng/ml) and then stimulation with anti-Fas antibody (clone CH11) cause apoptosis of HPMC. Activation of both caspase-8 and caspase-3 were noted during the apoptotic process. The addition of caspase inhibitor could prevent the occurrence of apoptotic process. The apoptotic HPMC could be engulfed by macrophages. We found increased mesothelial cell apoptosis in peritoneal dialysate effluent during recovery phase of peritonitis. Conclusions: Our results reveal that bio-incompatible peritoneal dialysate probably aggravate the magnitude of HPMC apoptosis or affect the removal of apoptotic HPMC by macrophages during peritonitis. Failure to clear apoptotic HPMC might prolong peritoneal inflammation and therefore hamper the repair of peritoneum. The apoptosis of HPMC is related to the regulation of inflammation and subsequent repair of peritoneum during peritonitis.
RANIERI, DANILO. "Role of the mesothelial microenvironment in the peritoneal dissemination of gastric and colorectal cancers." Doctoral thesis, 2012. http://hdl.handle.net/11573/917634.
Повний текст джерелаHsiao, Hui-Hsu, and 蕭惠旭. "N-acetylcysteine prevents conventional peritoneal dialysate fluid - induced oxidative stress in human peritoneal mesothelial cells." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/98872248144017365362.
Повний текст джерела高雄醫學大學
醫學研究所
96
Introduction and aims: Chronic peritoneal dialysis is one of the major therapies for patients with end stage renal disorders. Dialysates with bio-incompatibility may lead to peritoneal mesothelial cell injury, fibrosis and subsequent poor dialysis efficacy. The aim of this study investigates the toxic effects of conventional glucose-based dialysates in primary human peritoneal mesothelial cells (HPMCs) and the possible roles of amtioxidants in HPMC during conventional dialysate exposure. Methods:HPMCs were collected from peritoneal effluent of patients receiving peritoneal dialysis (PD). Conventional 1.5% dextrose peritoneal dialysate was utilized. HPMCs were exposed to conventional peritoneal dialysate with 1.5% dextrose for different time course and then the survival rate of HPMCs was evaluated by MTT assay. Curcumin(CUR)and N-acetylcysteine (NAC) were utilized as antioxidants. ROS accumulation, such as intracellular hydrogen peroxide, and superoxide anion in HPMCs were detected with flow cytometry by using intracellular probes, H2DCFDA and dHE, respectively,. Protein expression of HSP72 and HSP27 were detected by Western blots analysis. Antioxidant enzymes, such as catalase and superoxide dimutase (SOD), and glutathione were detected by ELSA reader. Result:Present study shows that conventional 1.5% dextrose dialysate exposure resulted in significant decrease of cell survival in a time dependent manner. NAC treatment contributes to decrease the conventional dialysate-induced cell death. However, curcumin can not offer the protective effects. Moreover, HSP72 is induced in HPMCs after dialysate exposure. NAC treatment decreases HSP72 induction. The intensity of DCF green fluorescence and the EB red fluorescence are increased, and shifting to right showed in histogram after conventional dialysate exposure. The accumulation of intracellular hydrogen peroxide and superoxide anion are in an exposure time-dependent manner. However, NAC treatment contributes to decrease dialysate induced-ROS accumulation. Activity of SOD and catalase is not different in every group. However, the reduced glutathione is significant decrease after dialysate exposure and NAC treatment contributes to preserve the expression of reduced glutathione in HPMCs. Conclusion: ROS accumulation during convention peritoneal dialysate exposure is an important cause of leading cell damage. NAC treatment contributes to protect the HPMCs from conventional dialysates-induced cellular damage by preserving the reduced glutathione to decrease ROS accumulation and oxidative stresses. HSP72 could be used as a stress marker.
Li, Chia-Ling, and 李佳玲. "Effects of Different Peritoneal Dialysis Fluids on Intracellular pH Regulation in Human Peritoneal Mesothelial Cells." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/57113378249320888344.
Повний текст джерела國防醫學院
藥理學研究所
96
Introduction Intracellular pH (pHi) changes were found to influence many cellular functions. To date, the pHi regulators include Na+/H+ exchanger (NHE), Na+/HCO3- symporter (NHS), lactate-/H+ symporter (LHS), Cl-/OH- exchanger (CHE) and Cl-/HCO3- exchanger (AE) in the mammalian cells. Then, peritoneal dialysis (PD) is one method of treating chronic uremic patients. Available 4 commercial peritoneal dialysis fluids (PDFs), Dianeal, Extraneal, Nutrineal and Physioneal, have various but dramatic different on pH values (pH is 5.2, 5.2, 6.3 and 7.4 respectively). Objects The aims of the study used the human peritoneal mesothelial cells (HPMCs) to: (1.) investigate the underlying mechanisms for its effects on pHi regulation (NHE and NHS) and NHE isoforms; (2.) explore the effects of commercial PDFs and different pHo solutions on pHi; (3.) investigate the LHS, the effects of 40 mM lactate and different buffered solutions on LHS activity. Materials and Methods 1.HPMCs were cultured directly from peritoneal tissue and identified by immunocytochemistry. 2.The measurement of pHi was detected by microspectrofluorimetry method with BCECF-AM. We recorded fluorescence ratio (R490/440), which represented the change of pHi. 3.Immunocyto/histochemistry were used to identify NHE-1, NHE-2 and NHE-3 in HPMCs or peritoneal biopsy tissues. Results 1.We have successfully cultured human peritoneal mesothelial primary cells from tissue and set-up a reproduceable model for routine use. 2.In HEPES-buffered solution, removal of extracellular Na+ or adding HOE 694 (a specific NHE inhibitor) alone can totally inhibit the recovery from NH4Cl-induced intracellular acidosis, which prove the existence of NHE. 3.In bicarbonate-buffered solution, either HOE 694 or DIDS (a specific NHS inhibitor) alone only partially inhibited the recovery from NH4Cl-induced intracellular acidosis, while removal of Na+ or adding together HOE 694 and DIDS totally inhibited the recovery. This demonstrate the existence of NHS. 4.Both in HEPES- and bicarbonate-buffered solution, all 4 different PDFs (pH 5.2, 5.2, 6.3, 7.4, respectively) significantly changed HPMCs pHi. 5.Both in HEPES- and bicarbonate-buffered solution, we demonstrated that, in the HPMCs, pHi is increased linearly as pHo rising from 5.2 to 8.0. 6.In sodium citrate solution, CHC (a LHS inhibitor) inhibited pHi recovery from lactate-induced intracellular acidosis, which proves the existence of LHS. Lactate-induced intracellular acidosis kinetice was not affected by HOE 694 or 40 mM lactate, either in bicarbonate- or HEPES-buffered solutions. Conclusions 1.We have demonstrated for the first time that, two Na+-dependent acid-extruders (NHE and NHS) and LHS are functionally existed in the HPMCs. NHE-1 and NHE-3 were identified in HPMCs. 2.Commercial PDFs used in clinics significantly affect HPMCs pHi, which suggest the space for the improvement or developing new PDFs. 3.We have demonstrated that pHi is functional linearly as pHo. In bicarbonate- buffered solution (pH 5.2~7.4), it was best buffered solution for HPMCs.
Lin, Shih-Pi, and 林士弼. "The effects of amino acid-based peritoneal dialysate on peritoneal dialytic patient and peritoneal mesothelial cell." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/84921090443579795412.
Повний текст джерела高雄醫學大學
醫學研究所碩士班
93
Traditional peritoneal dialysis solutions are bioincompatible resulting from high glucose concentration, high osmolality, low pH, and glucose degradation products(GDP). Nutrineal○R is 1.1% amino acid-based peritoneal dialysis solution and is designed to replace daily losses of amino acids and proteins during peritoneal dialysis. Its pH value is 6.5, closer to physiological value. And there is no GDPs in Nutrineal○R . It may be more biocompatible. L-Methionine, 85 mg/dl in Nutrineal®, is one of the components. However, the absorption of methionine may increase the synthesis of a well-recognized independent risk factor for cardiovascular diseases, homocysteine (Hcy). Poor appetite was reported after use of Nutrineal®. In this study, we observed the changes of serum biochemistry, serum homocysteine and leptin, Kt/V and nPCR, CA125 and PICP of peritoneal effluent, and the response of human peritoneal mesothelial cell after treating with various peritoneal dialysis solution. Methods: Seventeen adults CAPD patients (11M/6F) who have received CAPD for at least 6 months were enrolled. They all received daily four-exchange CAPD treatment and Nutrineal® was used as once daily substitute, given at the first or second exchange. Blood samples and peritoneal effluents were collected every month. Serum albumin, total protein, BUN, creatinine, homocysteine, and leptin were measured. Kt/V , nPCR, and body mass index (BMI) were calculated. And we checked the CA125 and PICP of peritoneal effluent. Human peritoneal mesothelial cells were cultured from peritoneal effluent and then were treated with various peritoneal dialysis solutions. LDH, heat shock protein 70, and CA125 were measured. Results: Serum BUN and homocysteine increased significantly(p=0.007, p=0.003). Other serum data did not change. Body weight and BMI increased significantly(p = 0.0344, p = 0.037); but there was significant change in serum leptin. Peritoneal effluent CA125 increased significantly(p=0.045) and PICP did not change. There were a similar pattern in increase of LDH, heat shock protein 70, and CA125 measured from cultured mesothelial cell treated by various solutions. The increase by Nutrineal○R was lower; the increase by 4.25% Dianeal○R and Extraneal○R were higher. Conclusion: Nutrineal○R is more biocompatible to peritoneal membrane, but there was a disadvantage in increases of BUN and homocysteine after use of Nutrineal○R .Body weight and BMI were increased after use of Nutrineal○R . CA125 from human peritoneal mesothelial cells was increased after stimulated by various solutions. Further studies will be needed.
CHEN, SHIH-I., and 陳世宜. "The Role of Prostaglandins in Mesothelial Regeneration and Collagen Gene Expression: Relation to Hepatocyte Growth Factor." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/82274501455165208257.
Повний текст джерела國立臺灣大學
臨床醫學研究所
91
The patients undertaking peritoneal dialysis are inclined to have peritonitis and further, denudation of the mesothelial cell monolayer. The recovery from the said requires regeneration of the mesothelial cells (including adhesion, migration and proliferation) to keep the integrity of the peritoneum. This is highly related to the degree of achievement of prevention in peritoneal fibrosis and the maintenance of peritoneal function. It’s known that the occurrence of peritonitis would stimulate the production of prostaglandin (PG), however, the role it plays is not yet clear. Based on some reports, PG can promote migration of the mesangial cells and the endothelial cells though there is no answer that is there similar effect on the mesothelial cells. Moreover, PG also helps the proliferation of gastric epithelial cells through the increasing production of hepatocyte growth factor (HGF). But the PG’s effect on the mesothelial cells is not yet studied. PG also works to prevent the production of extracellular matrix in the mesangial cells. Is there similar effect on the mesothelial cells still needs to clarified. Hence, our study is designed to ask (1) whether PG can work to help migration, adhesion and proliferation in the mesothelial cell as well. And if the promotion of proliferation is proved, this effect is related to the incremental of HGF or not; (2) PG can also function in prohibiting the expression of collagen gene of the mesothelial cell or not; and (3) the clinical use of NSAID or COX-2 inhibitor does harm to the peritoneal repair. Our study evaluated adhesion of the mesothelial cell by trypsin assay first. Secondly, migration and proliferation of the cells would be evaluated with cell culture techniques by establishing a simulation model of damaging mesothelial cell in vitro. And the expression of collagen gene is measured by Northern blot analysis. Test the mesothelial cells with various dosage of PGE2、PGI2、IL-1β、indomethacin and COX-2 inhibitor (NS398) to verify their influences on the mesothelial cell’s adhesion, migration, proliferation and expression of collagen gene. And further check there is IL-1β → PG → HGF Pathway existing in the mesothelial cells or not by the enzymeimmunoassay analysis. The results showed both PGE2 and PGI2 can promote the mesothelial cell’s adhesion, as well as the IL-1β. PGE2 does not stimulate migration of mesothelial cells. However, PGI2 has the opposite trend on migration though there is no statistic significance. IL-1β can increase the secretion of PGE2 which peaks at the 16 hour of treatment and decades at the following hours. Nevertheless, both PGE2 and PGI2 have no effect on the secretion of HGF. The Northern blot analysis reveals PGE2 and PGI2 can suppress the expression of collagen gene, and COX-2 inhibitor and NSAID can block the suppression effect of PG in the stimulation of IL-1β. This study proves part of our hypothesis that both PGE2 and PGI2 can promote adhesion and suppress the collagen gene expression of the mesothelial cells. Therefore, we know that the clinical use of NSAID and COX-2 inhibitor may be harmful to the integrity of peritoneum, especially during the course of peritonitis when COX-2 is highly activated and the repair of peritoneum needs the function of PG.
Yi-JerLee and 李宜哲. "The effect of dwelling time on epithelial-to-mesenchymal transition of mesothelial cells during peritoneal dialysis." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/19732257154659647236.
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