Esse trabalho teve como objetivo principal salientar o potencial da drenagem nasobiliar (DNB) como uma forma não cirúrgica de acesso à bile, utilizando como modelo uma téc- nica de DNB no estudo bacteriológico da bile em pesquisa. Para tal, foram estudados 17 pacientes portadores de coledo- colitíase submetidos eletivamente à colangiopancreatografia endoscópica retrógrada na Unidade de Endoscopia do Hospital de Clínicas de Porto Alegre. Foram realizadas DNB por até três dias com coletas seriadas de bile no momento do exame e a cada 24 horas, visando analisar os germes mais prevalentes e o perfil evolutivo da microbiota bacteriana. Correlacionou- se infecção biliar(IB), definida como 105 unidades formadoras de colônias (UFC)/ml de bile, com dados clínico-laboratori- ais obtidos dos prontuários (idade, sexo, presença ou não de febre, icterícia, leucocitose, elevação de fosfatase alca- lina, divertículos justapapilares, uso de antibióticos e co- lecistectomia prévia). A única intercorrência foi desconfor- to na retrofaringe em 28% dos casos. Foram tomadas medidas preventivas visando reduzir a contaminação do sistema. As enterobacteriácias (Klebsiella e E. coli) foram os germes mais encontrados. Ocorreu crescimento bacteriano em 71% dos casos na primeira coleta, embora 30% tivessem IB. Houve al- teração da microbiota biliar em 58% dos casos da primeira para a segunda coleta e em 81% dos casos desta para a terceira. Enquanto IB foi identificada em 30% dos casos na primeira coleta, esta atingiu 50% na segunda, 90% na terceira e 100% na última coleta, embora todos os pacientes tivessem evoluído satisfatóriamente. O perfil bacteriano qualitativo também se alterou, havendo predominância de Klebsiella e E. coli na primeira coleta, acréscimo de Streptococcus faecalis na segunda e apenas Pseudomonas na última. A associação entre IB e os dados clínico-laboratoriais não foi estatisticamente significativa. Concluiu-se que as enterobacteriácias Gram - foram os germes mais prevalentes nos pacientes com coledocolitíase, sendo que o perfil bacteriológico foi significativamente alterado com a DNB, embora sem implicação no quadro clínico. Além disto, não houve associação entre os dados clínico-laboratoriais estudados e a presença de IB. / The main purpose of this study was to evaluate the potential of nasobiliary drainage (NBD) as a non-surgical procedure to gain access to the bile, using as standard the description of the NBD technique for the bacteriological of bile analysis. NBD was performed in 17 patients with choledocolithiasis, all admitted for elective endoscopic retrograde cholangiopancreatography at the Endoscopy Unit of Hospital de Clínicas de Porto Alegre . NBD was kept in place for up to 3 days. Bile sampling was performed at the time of the procedure and every 24 hours, in order to identify the most prevalent bacteria, as well as the evolutive profile of the bacterial flora. Biliary infection ( BI ), as defined by the presence of at least 105 colony forming units ( CFU ) / ml of bile, was correlated to clinical-laboratorial data obtained from patient records ( age, sex, presence or absence of fever, jaundice, leucocitosis, raise of alkaline phosphatase, justapapillary diverticula, use of antibiotics and previous cholecystectomy ). The only complication observed was pharyngeal disconfort in 28% of the cases. Prophylactic measures were used to reduce contamination. Enteric organisms ( Klebsiella and E. coli ) were the most frequently found bacteria. Bacterial growth was identified in 71% of the cases in the first sampling, although only 30% of these had BI. The biliary flora changed in 58% of the cases from the first to the second sampling and a further 81% change was observed in third sampling. BI was found in 30% of the cases in the first sampling, while in 50, 90 and 100% in the second, third and last sampling, respectively. Notally, in spite of the latter results all patients had a favorable outcome. The qualitative bacterial profile also changed, showing the predominance of Klebsiella and E. coli in the first sampling, the appearence of Streptococcus faecalis in the second and only Pseudomonas in the last one. The correlation between BI and clinical-laboratorial data did not show statistical significance. It was concluded that the Gram - enterobacteriaceas were the most prevalent bacteria in the patients with choledocolithiasis under study.Futhermore, the bacteriological flora was significantly changed with NBD, however without clinical implications. Finally, there was no correlation between the clinical-laboratorial data studied and the presence of BI.

No description available.

Abstract Apolipoprotein E (apo E) is one of the key regulatory proteins in cholesterol and lipoprotein metabolism. The present research focuses on the role of apo E in gastrointestinal diseases. The polymorphism of apo E has been suggested to be associated with the cholesterol content in gallstones and the crystallization rate of gallbladder bile. The possible effect of apo E polymorphism on the susceptibility to gallstone disease at the population level was examined in comparison with the classical risk factors for gallstone disease. The data suggest that the apolipoprotein E2 isoform is a genetic factor that provides protection against gallstone disease in women. The alterations in plasma lipoprotein levels and bile acid metabolism observed in patients with colorectal adenoma and carcinoma may reflect a genetic background predisposing to tumors through altered lipid metabolism. To determine, whether the polymorphism of apo E is associated with proximal or distal colonic neoplasia, the apo E phenotype was determined in 135 patients with colorectal carcinoma, and 199 randomly selected control subjects. The frequency of the ε4 allele of apo E was low in the patients with proximal adenoma and those with carcinoma, respectively, compared with the control subjects. The patients with distal tumors showed no alteration in ε4 frequency. The data suggest that the ε4 allele of apo E provides protection against the development of adenoma and carcinoma of the proximal colon. The association of apo E polymorphism with tumors is not a generalized phenomenon as is shown by the lack of association with breast or prostate cancers. To further study the mechanisms by which apo E might affect colon cancer, the expression of apo E in human intestine and the localization of apo E in normal and malignant gastrointestinal tract was studied using immunohistochemistry and in situ hybridization. Both immunoreactive apo E protein and apo E mRNA were present throughout the stomach, small intestine and colon. The phagocytes of lamina propria were positive for apo E, but the number of positive cells and the staining intensity varied according to localization. Macrophages in the superficial lamina propria of normal colon were more strongly positive for apo E than those in the small intestine, where the most positively stained cells were dendritic cells and macrophages in the germinal centers of lymphoid follicles. In samples from colorectal carcinomas intensely positive macrophages surrounded the tumor area, suggesting that apo E might play a role in the proliferation of malignant cells. Apo E binds with very high affinity to heparin and proteoglycans and inhibits the proliferation of several cell types, but the antiproliferative mechanism of apo E is still largely unknown. The effects of apo E at the cellular levels were studied in cell culture experiments. The effect of recombinant human apo E3 on cell polarity and the distribution of β-catenin were examined in undifferentiated (G+) and differentiated (G+ reversed) HT29 human colon adenocarcinoma cell lines. In cultured undifferentiated HT29 cells, treatment with apo E improved cell polarity and translocated β-catenin from the cytoplasm to cell-cell adhesion sites. Apo E may thus modulate epithelial integrity and contribute to cell growth and malignant transformation.

The present study of Rana catesbeinana includes an analysis of the following throughout the course of post-embryonic development: the change in GI tract length and weight; the relative surfactant activity of the bile; the change in major bile pigments. The most rapid and profound morphological and physiological transitions occur during the period of metamorphic climax. Because no information relative to change in anurans is available during this period, particular emphasis is placed upon the examination of individuals during metamorphic climax.

A protease has been found in the bile of 11 mammalian species investigated. The protease, given the tentative name of cholelysin, has been studied intensively in Abattoir ox bile. It makes up less than 10% of the ox bile protein, and is a potent fibrinolysin, as well as being active against the substrates α-casein, and the synthetic esters of tyrosin (ATEE) and arginine (BAEE). It is inactive against trypsinogen, chymotrypsinogen and plasminogen. Isolation and purification of this protease from ox bile proved complex, and was finally achieved by an 8 step procedure which yielded a dry white powder, stable for 45 months (to date) at 4°C. Quality control of this procedure was effected by means o f a fibrin plate assay, using chymotrypsin as a reference standard: the dose response curves of cholelysin and chymotrypsin were closely similar on the fibrin plate, enabling cholelysin (units/l) to be substituted for chymotrypsin (mg/l), in equivalent diameters of fibrinolysis. Gradient elution by tris/NaCl from Whatman DE 32 produced four areas, or Peaks, of fibrinolytic activity of cholelysin, each with some differing characteristics against the various substrates. These complexities were not studied in detail, and a simplification of the procedure was discovered, using batch-wise elution with tris buffer. Thereafter, only the first peak was studied (Peak I). Studies on the inhibition of cholelysin were done using many known inhibitors, including serum of man, and 4 laboratory animals; serum of two patients with homozygous deficiency of α1-antitrypsin; α2-macroglobulin, soy bean trypsin inhibitor, and aprotonin. The serum studies were done with heated and unheated material; platelet-rich and platelet-poor plasma were also studied. Serum was fractionated by paper electrophoresis in an attempt to discover the globulin fraction containing the inhibitor. No inhibition was found in the α-globulin fractions, and inhibition was maximal in the inter-α- globulin and α-globulin fractions. ATEE-esterase activity of cholelysin was inhibited by serum as strongly as fibrinolytic activity. A limited series of studies of coagulation was done; cholelysin was only found to influence fibrinogen, being quite strongly fibrinogenolytic in vitro. A slight effect on ADP-induced aggregation of platelets was found at a low ADP concentration. Using a rat model originally devised for the study of the anticoagulant effect of ancrod, cholelysin was found to be weakly anticoagulant. The dose was low by comparison with ancrod, and the result approached, but did not reach, statistical significance. The split products of plasmin and cholelysin digestion of stabilised fibrin were studied by polyacrylamide gel electrophoresis (PAGE), and these were found to be entirely different. The kinetics of the reaction between cholelysin and fibrin were studied by means of a new technique (the composite cuvette method) and by this method it was shown that cholelysin made a two-phase attack on the fibrin molecule. The attack was studied at 2, 10, and 30 minutes following commencement of fibrinolysis, and the biphasic nature of the attack was proved by a sharp increase in the number of reaction products present between the 2 and 10-minute samples. The molecular weight of fibrin split products by plasmin were shown to agree with those found in published work. Finally, the molecular weight of cholelysin was estimated by PAGE and by column chromatography with and without SDS. It seems probable that the basic molecular weight is ~7 000, with a dimer of ~13 000 and a tetramer of ~ 28 000.

Thirty six consecutive patients with primary sclerosing cholangitis (PSC), 20 males median age 42 years, were studied in order to define prognostic variables and determine the influence of surgery on outcome. Presentation was usually with insidious cholestasis or recurrent cholangitis. Twenty six patients (72 per cent) had associated inflammatory bowel disease (ulcerative colitis 20, Crohn's disease 2, unclassified 4). Thirty two patients were followed prospectively for up to nine years. Twenty three remained either stable or had slowly progressive disease. Of the remaining nine patients, seven died (five from end-stage liver failure and two from cholangiocarcinoma) and two patients underwent liver transplantation. Actuarial survival at five years was 52 per cent. A raised serum bilirubin concentration was the only variable at presentation that independently predicted a poor outcome. Cholangiograms were available for detailed assessment in thirty PSC patients. Neither the extent of biliary involvement nor the presence of surgical correctable ("dominant") strictures in the extrahepatic ductal system were of prognostic importance. Six patients who developed obstructive jaundice associated with advanced liver disease underwent surgical drainage operations for dominant biliary strictures, but this did not seem to prevent progression of the disease. Two patients who progressed to end-stage liver disease went on to liver transplantation and were alive with functioning grafts at seven and fourteen months respectively. Nine patients with asymptomatic PSC were followed prospectively for up to twelve years. None of these patients developed overt liver disease but serum bilirubin levels became mildly elevated in two patients. It is concluded that symptomatic PSC is a progressive disease with a poor prognosis. Patients with advanced liver disease due to PSC should be considered directly for liver transplantation. In contrast, asymptomatic PSC patients may remain symptom-free for many years.

Listeria monocytogenes is an enteric pathogen that can replicate within bile, yet this capability differs between strains. This project analyzed whether the pathogenic potential of the strain affects the ability to resist bile. We tested this hypothesis by examining the effect of bile on the morphology of a virulent strain (EGD-e) and an avirulent strain (HCC23) under aerobic and anaerobic conditions. Our data showed that exposure to bile greatly impacted the growth of HCC23. Additionally, scanning electron microscopy and transmission electron microscopy analyses indicated that bile affects the cell envelope of EGD-e and HCC23 differently. Our results suggest that differences exist in the ability of EGD-e and HCC23 to survive and replicate in the presence of bile. We propose that the virulence capability of L. monocytogenes directly correlates to its ability to resist the detergent properties of bile.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Thesis (M.D.)--University of Adelaide, 1988. / Includes bibliographical references (leaves 171-211).

No description available.