bromochloroacetic-acid and Hernia--Inguinal

Epithelial-mesenchymal transition (EMT) describes rapid changes in cellular phenotype. During EMT, epithelial cells down-modulate cell-cell adhesion, alter polarity, reorganize cytoskeleton, become isolated, motile, and resistant to anoikis. Epithelial breakdown and epithelial cell migration are the key processes involved in the obliteration of processus vaginalis. The great majority of abnormalities are because of nonobliteration or incomplete fusion of PV. We aimed to analyze the quantitative changes of epithelial genes in adult/child patients and their controls to examine differences of the genesis of these hernias. We also aimed to investigate the potential epigenetic causes of indirect inguinal hernia in adult patients.. Ten adult, ten child indirect inguinal hernia sacs and ten adult, ten child parietal peritonea were used. Hernial sac samples were obtained from indirect inguinal hernia surgeries. Peritonea of adult patients who underwent open cholecystectomy for cholelithiasis via subcostal incision were included in the study as the healthy control groups. Ages of the children were selected to be between 0 and 5 whereas the age of adults was chosen as 35-55, respectively. Total RNA isolation and cDNA synthesis were made from hernia sacs and peritoneum samples. Relative Keratin 1, Keratin 15, Filaggrin2 and STAT3 expressions were analyzed via qPCR. Indirect inguinal hernia sac cells were seeded and grown in vitro. Child diseased cells were employed in immunocytochemistry (ICC) analysis for Cytokeratin 15, Filaggrin2 and Bcl-2. Adult indirect inguinal hernia cells were examined for H3 modifications through ICC.. In child indirect inguinal hernia, Keratin expressions were found higher than their controls. They were meaningfully lower than the healthy group in adult indirect inguinal hernia. Keratin 15, Keratin 1 and Filaggrin2 expressions were all correlating since they are members of related pathways. STAT3 expressions were opposite to Keratin and Filaggrin expressions suggesting that adult cells might have a switch to the mesenchymal state from the epithelial state. Adult indirect inguinal hernia samples have switched to the mesenchymal state whereas child indirect inguinal hernia samples have shown lack of transition.. Irregular changes of EMT associated genes act in the progression of indirect inguinal hernia. Hence, the information on the epigenetic regulation of EMT in patients with primary inguinal hernia can aid to comprehend the pathogenesis in adults and infers new therapeutic approaches for this disease.. Prognosis study.. Level 2.

Topics: Adult; Case-Control Studies; Child, Preschool; Epigenesis, Genetic; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Filaggrin Proteins; Hernia, Inguinal; Humans; Immunohistochemistry; Infant; Infant, Newborn; Inguinal Canal; Keratins; Male; Middle Aged; Peritoneum

To evaluate the utility of peritoneal pathologic samples, unrelated to peritoneal dialysis (PD) treatment, for the study of peritoneal fibrosis and inflammation.. Comparative morphologic and immunohistochemical study of peritoneal pathologic samples unrelated to PD with peritoneal biopsies from PD patients with special emphasis on the expression of myofibroblastic and epithelial-to-mesenchymal transition markers.. Regarding morphology, PD-related simple fibrosis was less cellular, with greater stromal hyalinization, determining a homogeneous, hypocellular aspect of the submesothelium. In contrast, non-PD fibrosis was more cellular with an extracellular matrix showing a dense and fibrillar quality with wide bundles of collagen. Hylinazing vasculopathy was only present in PD samples. Myofibroblastic differentiation and epithelial-to-mesenchymal transition were common findings in all situations of peritoneal fibrosis. Calponin and calretinin are useful cellular markers to study such fibrogenic mechanisms and correlate with other well-known markers such as a -SMA and cytokeratins. Their expression was much more intense in those samples showing acute inflammation (peritonitis).. Non-PD models of peritoneal fibrosis seem very useful to evaluate important features of human peritoneal pathology such us fibrogenesis, and inflammation. Fibrogenic events such as myofibroblastic differentiation and epithelial-to-mesenchymal transition are evident in these tissue samples allowing us to use them as an accessible source for in vivo and ex vivo studies. Both events show their maximal expression in situations of acute inflammation supporting the important role that peritonitis episodes play in the progression of fibrosis.

Topics: Actins; Biomarkers; Biopsy; Calbindin 2; Calcium-Binding Proteins; Calponins; Case-Control Studies; Cell Differentiation; Edema; Epithelium; Fibrin; Fibroblasts; Fibrosis; Hernia, Inguinal; Humans; Hyalin; Keratins; Microfilament Proteins; Neutrophils; Peritoneum; S100 Calcium Binding Protein G; Sclerosis; Tissue Adhesions

Recent evidence has suggested that calcitonin gene-related peptide (CGRP), which is released from the genitofemoral nerve, may trigger fusion of the patent processus vaginalis in children with inguinal hernia. The purpose of this study was to determine whether CGRP triggers the release of mesenchymal factors leading to subsequent fusion of the processus vaginalis.. The response of cultured epithelial cells derived from the patent processus vaginalis was analysed by a novel in vitro culture system. Epithelial cells lining fresh hernial sacs (removed at inguinal herniotomy) were detached enzymatically and cultured for 72 hours on Micropore filters, in the presence of either 100 ng/mL hepatocyte growth factor (HGF), 7.4 x 10(-6) mol/L CGRP (amino acids 1 to 37), 7.4 x 10(-6) mol/L CGRP (8 to 37) antagonist, 10% fetal calf serum (FCS), or serum-free medium (SFM) alone. Transformation from an epithelial cell morphology to motile mesenchymal fibroblast-like cells was assessed by an average migration score (AMS), ranging from 0 with no sign of migration, to 3 with greater than 75% of cells migrating. Confocal microscopy was used to record changes in expression of epithelial (cytokeratin) and mesenchymal (vimentin) markers, as well as actin. Beta-catenin also was examined because it is part of the molecular complex that links cadherins to actin resulting in cell-cell adhesion.. Epithelial and mesenchymal markers underwent either down-regulation or up-regulation as epithelial cell sheets broke apart and individual cells commenced migration. The AMS after 72 hours of culture was 0.22 with SFM (control); with FCS the score was 1.4 (P < .01). The AMS score with CGRP (1 to 37) was 0.55 (P = .165) and with its analogue, CGRP (8 to 37), which is a competitive inhibitor, 0.67 (P = .309). Neither was significant. HGF caused a significant increase in the AMS to 1.56 (P = .01).. Both HGF and FCS (which contains various undefined peptides and growth factors) produced transformation of hernial sac epithelial cells, whereas CGRP and its inactive analogue did not. CGRP receptors are localised to mesenchymal fibroblasts within the processus vaginalis connective tissue, suggesting that CGRP could act indirectly via HGF, which, in turn, promotes fusion of the processus vaginalis. In the future, a nonsurgical treatment of inguinal herniae in children might be possible by the local administration of agents which promote fusion.

Topics: Actins; beta Catenin; Calcitonin Gene-Related Peptide; Cell Movement; Cells, Cultured; Child; Cytoskeletal Proteins; Epithelial Cells; Female; Hepatocyte Growth Factor; Hernia, Inguinal; Humans; Immunohistochemistry; Keratins; Male; Trans-Activators; Vimentin