Browsing by Department "UCT/MRC Liver Research Centre"

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    Albumin metabolism following partial hepatectomy in the rat
    (1980) Lloyd, Elwyn Allden; Saunders, Stuart J
    In the work reported here the author set out to investigate five aspects of albumin metabolism following partial hepatectomy in the rat. 1. The plasma albumin levels following partial hepatectomy. 2. The effect of partial hepatectomy on the albumin synthesis rate. 3. The effect of supplementary amino acids on the albumin synthesis rate following partial hepatectomy. 4. The effect of hydrocortisone on the albumin synthesis rate following partial hepatectomy. 5. The effect of partial hepatectomy on the albumin catabolic rate.
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    The Budd-Chiari syndrome : a study of diagnosis, haemodynamics and treatment
    (1968) Clain, David Jocelyn
    Symptomatic occlusion of the hepatic veins is a rare condition caused by tumour or thrombus arising either locally or by extension from the inferior vena cava. It is usually called the Budd-Chiari syndrome. The etiology remains unknown in over two-thirds of the patients. Its rarity and interest has led to a large number of individual case reports. 322 instances of symptomatic hepatic vein occlusion have been reported, of which 184 are single case publications. There are only six series of more than five cases (Nishikawa, 1910; Corinini and Oberson. 1937; Palnar, 1954; Parker, 1959; Gibson, 1960; Safouh and Shehata, 1965) and these have been largely drawn from autopsy records, although Palmer (1954) described seven patients seen during life. The clinical and pathological features of hepatic vein occlusion have been described in a number of papers (Hess, 1905; Thompson and Turnbull, 1912; Armstrong and Carnes, 1944; Kelsey and Comfort, 1945; Thompson, 1947; Parker, 1959; Gibson, 1960) during the one hundred and twenty years since the publication of Budd's treatise. However, accurate diagnosis has generally relied on autopsy, and detailed investigations have seldom been performed. Consequently, little is known of the roentgenographic and haemodynamic features. The diagnosis of liver disease has been revolutionized by such special techniques as percutaneous liver biopsy, portal pressure measurements, isotope scanning and selective arteriography and venography. This study describes six patients with the Budd-Chiari syndrome in whom these methods have been applied to establish the diagnosis, to ascertain the underlying cause and to assess the possibility of surgical intervention. Special attention has been given to hepatic venography and hepatography. The vascular pattern in the Budd-Chiari syndrome has been compared with that in normals and in patients with other diseases of the liver. Diagnostic features have been determined and an attempt made to evaluate compensatory changes in the lymphatic drainage and venous blood supply following hepatic vein obstruction. Alterations in portal dynamics have also been recorded. The clinical course has been followed and the effect of treatment assessed in each patient. Finally, the literature has been reviewed with particular reference to the diagnosis and treatment of hepatic vein thrombosis. The studies reported in this thesis were carried out during the tenure of a Research Fellowship in the Royal Free Hospital School of Medicine, and they were supported by a grant from the William Shepherd Bequest to the Royal Free Hospital. The special radiological procedures, haemodynamic studies, isotope investigations and laboratory work were personally performed with the exception of the scintillation scans, coeliac axis arteriograms and the other individual tests acknowledged overleaf.
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    CTLA4 gene polymorphisms in autoimmune hepatitis (AIH) : gene and clinical disease correlations in South African patients
    (2007) Marais, Surita; Hairwadzi, H N; Meissner, P N; Corrigall, A V; Spearman, W
    Also available online. Includes bibliographical references (leaves 76-85).
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    The fumonisin B₁-fed rat as a model for liver injury, oval ('progenitor') cell proliferation, and carcinogenesis
    (1999) Lemmer, Eric Richard; Hall, Pauline; Shephard, Enid; Cruse, Peter
    Fumonisin B₁ (FB₁‚ ) is a carcinogenic mycotoxin produced by the fimgus Fusarium moniliforme in maize, and is hepatotoxic and hepatocarcinogenic in rats. The goal of this dissertation was to characterise the FB₁-fed rat as a model for liver injury and carcinogenesis, and to examine the role of oval ('progenitor') cells during these processes. Male Fischer 344 rats were fed FB₁ 250 mg/kg diet for five weeks, and this basic feeding regimen was modified in individual experiments. Short-term feeding of FB₁ caused a severe 'toxic' hepatitis, apoptosis and regeneration of hepatocytes, fibrosis, proliferation of OV-6 positive oval cells, and formation of GST pi positive hepatic foci and nodules. Oval cells were noted inside some of the hepatic nodules. There were marked increases in the expression of mRNA transcripts for mature TGF-β1 and c-myc in livers of FB₁-fed animals. The overexpression of TGF-β1 by hepatocytes may be responsible for the prominent apoptosis and fibrosis seen with liver injury due to FB₁. Increased expression of c-myc and TGF-β1 may cooperate during FB₁-induced promotion of liver tumours, possibly by providing an environment that selects for the growth of TGFβ1-resistant transformed liver cells. In rats given FB₁ in the presence of dietary iron overload, FB₁ augmented iron-induced lipid peroxidation in the liver. However, dietary iron loading appeared to protect against the cancer-promoting properties of FB₁, possibly due to a stimulatory effect on hepatocyte regeneration. Long-term feeding of FB₁ caused fibrosis and regenerative nodules, dysplastic hepatic nodules, cholangiofibrotic lesions, intraductal cholangiocarcinomas, and a hepatocellular carcinoma. 2-Acetylaminofluorene enhanced the effects of FB₁ in the liver, presumably by blocking hepatocyte regeneration in response to FB₁ toxicity. Proliferating oval cells were found inside/adjacent to GST pi positive lesions, dysplastic nodules, and cholangiofibrotic lesions, suggesting that oval cells may be involved in FBI-induced hepato- and cholangiocarcinogenesis in the liver. Furthermore, the OV-6 antigen was expressed by proliferating oval cells and bile ductules, hepatic nodules, cholangiofibrotic lesions, and cystic lesions, indicating that all of these cells may have a common ('stem') cell of origin. In conclusion, the FB₁-fed rat is a promising model for the study of liver injury, oval ('progenitor') cell proliferation, and carcinogenesis.
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    Human glutathione S-transferases : characterization, tissue distribution and kinetic studies
    (1988) Corrigall, Anne Vint; Kirsch, Ralph E
    In this study the purification of human basic and near-neutral liver, and human basic and acidic lung glutathione S-transferases (GSH S-T) was undertaken. Purification of the basic and near-neutral GSH S-T was achieved using a combination of affinity chromatography, chromatofocusing and immunoaffinity chromatography. Affinity and ion exchange chromatography were employed in the purification of the basic and acidic lung forms. The purified proteins had similar physicochemical characteristics to the GSH S-T purified by others. The binding of 1-chloro-2,4-dinitrobenzene (CDNB) to the 3 classes of human GSH S-T, viz. basic, near-neutral and acidic and the effects of such binding, if any, were examined. Human acidic lung GSH S-T is irreversibly inactivated by CDNB in the absence of the co-substrate glutathione (GSH). The time-dependent inactivation is pseudo-first order and demonstrates saturation kinetics, suggesting that inactivation occurs from an EI complex. GSH protects the enzyme against CDNB inactivation. In contrast, the basic and near-neutral GSH S-T are not significantly inactivated by CDNB. Incubation with [¹⁴C]-CDNB indicated covalent binding to all 3 classes of GSH S-T. When the basic and acidic GSH S-T were incubated with [¹⁴C]-CDNB and GSH, cleaved with cyanogen bromide, and chromatographed by HPLC, a single peptide fraction was found to be labelled in both classes. Incubation in the absence of GSH yielded 1 and 2 additional labelled peptide fractions for the basic and acidic transferases, respectively. These results suggest that while CDNB arylates all 3 classes of human GSH S-T, only the acidic GSH S-T possesses a specific GSH-sensitive CDNB binding site, which when occupied leads to time-dependent inactivation of the enzyme. The tissue distribution and localization of the 3 classes of human GSH S-T in normal and tumour tissue was examined. Antibodies to representatives of the 3 classes were raised in rabbits, and radial immunodiffusion employed to quantitate their concentrations in the cytosol of 18 organs from 9 individuals. The data provide the first direct, quantitative evidence for the inter-individual and inter-organ variation suggested by earlier workers. The absence of the near-neutral GSH S-T in 5 of the 9 individuals studied confirms an earlier suggestion of a "null" allele for this transferase. Basic and acidic GSH S-T (apart from in a single liver), were always present. Near-neutral GSH S-T, when present, were found in all tissues examined. The marked inter-organ and inter-individual variation observed in this study may explain individual and organ susceptibility to drugs, toxins and carcinogens. The immunohistochemical localization of the 3 classes of GSH S-T reveals important differences in their localization, and may provide insight into their functions in various organs and tissues.
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    The identification, purification and characterization of the fetal rat liver glutathione S-transferase isoenzyme YcYfetus
    (1988) Scott, Trevor Robert; Kirsch, Ralph E; Folb, Peter I
    This study has examined the expression of the glutathione S-transferases (GSH S-T) in fetal rat livers in order to provide more information about the role played by this important group of enzymes in the fetus. The study commenced with an examination of the subunit composition of adult and fetal rat liver GSH S-T using affinity chromatography followed by polyacrylamide gel electrophoresis in sodium dodecyl sulphate. Adult livers contained four major GSH S-T subunits. An additional and previously unidentified subunit was detected in fetal livers. This subunit, which differed from that found in rat placenta, had a Mᵣ of approximately 25 500. Densitometric measurements suggest that the newly detected subunit accounts for as much as 26% of the GSH S-T in fetal livers. The novel fetal isoenzyme comprising this subunit was purified using a combination of affinity chromatography, carboxymethyl-cellulose column chromatography and chromatofocusing. The six major basic rat liver GSH S-T were purified for reference and comparative purposes. The fetal isoenzyme is composed of two non-identical subunits, namely, subunit Yc (Mᵣ 28 000) and the fetal subunit referred to as 'Yfetus'· The enzyme which I have termed GSH S-transferase Yc Y fetus has an isoelectric point of approximately 8.65 and has GSH S-T activity towards a number of substrates. Significantly, the fetal isoenzyme has one of the highest glutathione peroxidase activities yet described for the purified rat liver GSH S-T towards the model substrate, cumene hydroperoxide. Kinetic studies reveal that the fetal isoenzyme has a catalytic efficiency for the peroxide substrate which is four fold higher than that of the adult rat liver isoenzyme, GSH S-T YcYc. The in vitro effect of the GSH S-T substrate and teratogen, acrolein, on this fetal isoenzyme was investigated and compared with acrolein's effect on some of the adult rat liver GSH S-T isoenzymes in the standard 1-chloro-2,4-dinitrobenzene assay. Surprisingly, acrolein was identified as a non-competitive inhibitor of the GSH S-T. Exposure to acrolein in various guises could therefore result in inhibition of the fetal isoenzyme and its subsequent failure in inhibiting lipid peroxidation. Inhibitor studies were performed to look at the effect of acrolein, as well as other substrate and non-substrate ligands, on the glutathione peroxidase activity of GSH S-T YcY fetus and YcYc. The glutathione peroxidase activity of the fetal isoenzyme was far less susceptible to acrolein inhibition than the YcYc isoenzyme and the fetal isoenzyme was found to retain significant glutathione peroxidase activity despite saturating concentrations of non-substrate ligand. This study suggests that the fetal isoenzyme serves a specific function in protecting fetuses against the possible teratogenic effects of organic peroxides.
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    A neutral protease of the neutrophil surface : role in the proteolysis of C-reactive protein and fibrinogen
    (1995) Kelly, Sharon Lesley; Shephard, Enid
    Both of the acute phase reactants, C-reactive protein and fibrinogen, as well as neutrophils have been shown to accumulate at sites of tissue injury or inflammation. The association of C-reactive protein with neutrophils and the concomitant degradation of this ligand by a phorbol 12-myristate 13 acetateactivatable membrane-associated neutral protease has been shown in previous studies. Degradation of C-reactive protein by the neutrophil protease was shown to result in peptides with an ability to modulate various immune functions of the neutrophil. The aim of this study has been to investigate specific characteristics of the protease, with respect to cellular distribution and molecular size. The ability of this neutrophil membrane-associated protease to degrade the acute phase protein, fibrinogen was investigated. The mechanism of degradation of both C-reactive protein and fibrinogen during their association with the neutrophil was also examined. The neutrophil protease, capable of degrading C-reactive protein, was also associated with the cytoskeleton and was proposed to be a submembrane protease localised at sites of attachment of the membrane with the cytoskeleton. The protease was found to have a molecular mass of approximately 600 kDa which, on sodium dodecyl sulphate polyacrylamide gel electrophoresis, separated into four bands which migrated to molecular mass values of 209 kDa, 316 kDa, 398 kDa and 501 kDa. This protease also possessed fibrinogenolytic activity. The fibrinogen degradation products generated by this neutrophil membrane-associated protease were distinct from the products generated by the fibrinogenolytic systems of plasmin, human neutrophil elastase and neutrophil lysosomal enzymes and were unclottable through cleavage of the Aα chain from the N-terminus and the Bβ and γ chains from the C-terminus. N-terminal cleavage of the Aα chain by the neutrophil membrane-associated protease generated the Aα1-21 peptide, previously regarded as a unique consequence of elastase activity. Degradation of C-reactive protein and fibrinogen occurred as a result of their interaction with the neutrophil near to the CD11c integrin receptor. This interaction resulted in the egress of proteolytic activity into the extracellular medium. The fibrinogen products generated outside the cell associated with the neutrophil via the β₂ integrin receptors and the IgG Fc receptor. The interaction of the Creactive protein degradation products with the neutrophil could not be determined. Both C-reactive protein and fibrinogen are degraded by non-stimulated neutrophils but activation with phorbol 12- myristate 13 acetate resulted in maximum degradation This upregulation of activity was achieved through activation of H7 and trifluoperazine inhibitable cellular kinases and changes in microfilament assembly. The generation of non-clottable fibrinogen together with possible modulation of neutrophil receptormediated functions by the fibringen degradation products as well as the knowledge that the neutrophil protease generates C-reactive protein peptides with immunomodulatory activity implicates this neutrophil membrane-associated protease in the modulation of various inflammatory processes.
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    Pig liver perfusion : a role in hepatic assist?
    (1972) Hickman, Rosemary
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    Proteases of the neutrophil membrane represent an alternative fibrinolytic pathway to that mediated by plasmin
    (1996) Adams, Susan Ann; Shephard, Enid
    The cellular components of the blood, which become associated with fibrin through specific cellular adhesive processes, play a significant role in the breakdown of fibrin. Fibrinolysis by elastase and cathepsin G, enzymes present within the azurophilic granules of the neutrophil, has previously been shown. Recent studies have demonstrated neutrophil-mediated fibrinogenolysis by a membrane-associated protease which suggests that proteases connected with the neutrophil membrane might also be capable of clot dissolution. Investigations showed that neutrophil-mediated clot lysis was effected by a membrane-associated serine protease that can be dissociated by SDS-PAGE to bands that migrate to apparent molecular weights of 501 kDa, 398 kDa, 316 kDa, 245 kDa and 209 kDa. This degradation was distinct from that produced by plasmin, neutrophil lysosomal enzymes and purified human neutrophil elastase and enhanced the action of plasmin in clot solubilization. Preincubation of neutrophils with monoclonal antibodies directed against the CD 11 c/CD 18 integrin was able to significantly inhibit neutrophil membrane-dependent fibrinolytic activity. Upregulation of enzyme activity occurred following association of fibrin substrate with the cell membrane and was dependent on the activation of cellular kinases, in particular protein kinase C. Fibrin products generated by neutrophil membrane proteolytic activity were found to possess biological activity. The low molecular weight peptides effected substantial inhibition of thrombin-induced platelet aggregation while the presence of the higher molecular weight material could partially overcome platelet-induced resistance to plasmic lysis. No modulation of platelet-mediated fibrin clot retraction was observed using these same fibrin products. Neutrophil lysosomal enzyme activity was shown to further degrade the end products of plasmic fibrin degradation into low molecular weight material, followed by reassembly of higher molecular weight products in a process dependent on calcium and factor XIII. The reformed products have a similar molecular weight to those produced by plasmic lysis of fibrin, as well as a putative crosslinked site. However, the isoelectric point of these reformed products indicates they are distinctly different from plasmin-derived fibrin products. These reassembled products were recognized by a monoclonal antibody raised against D-dimer. Processing by neutrophils of the end products of plasmic fibrin degradation may have the potential for modulating the immune response as well as compromising the predictive value of tests measuring D-dimer, used as a laboratory marker of a number of thromboembolic disorders encountered in clinical practice.
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    The relative roles of portal hypertension and of cirrhosis in the pathogenesis of pulmonary lesions associated with chronic liver disease
    (1987) O'Brien, John A
    There have been numerous reports of cardiovascular and pulmonary abnormalities in patients with cirrhosis and portal hypertension. The role of portal hypertension in the pathogenesis of pulmonary abnormalities in patients with liver disease has not been defined. The present study was therefore undertaken to clarify this. Pulmonary function, including exercise testing, was evaluated in two groups of patients, 11 with portal hypertension due to cirrhosis and 10 with extrahepatic portal vein thrombosis and normal liver histology. Carbon monoxide gas transfer (TLCOsb) was less than 75% of predicted values in four patients from each group. One patient from each group had clinical and catheter confirmed evidence of pulmonary hypertension. Abnormal cardiorespiratory responses to exercise occurred in three patients in the extrahepatic group. Two had associated low TLCOsb and one developed arterial desaturation on exercise. A similar pattern was seen in three patients with cirrhosis. All had low TLCOsb and one developed arterial desaturation during exercise. In the cirrhotic group however three additional patients showed reduction in Pa02 unassociated with elevated heart rate response on exercise. There was no significant correlation with the presence of autoimmune antibodies which appear to be a secondary phenomenon. Our results suggest that pulmonary hypertension is linked to the presence of portal hypertension. Reduction in arterial P02, appears to occur only in patients with liver disease, presumably on the basis of intrapulmonary shunting.
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