Guanine deaminase in rat liver and mouse liver and brain

1. The guanine deaminase in rat liver supernatant preparations was resolved into two fractions, A and B, on DEAE-cellulose columns. The two differed in electrophoretic mobility and in various properties. The most noteworthy distinction between A and B components was that the enzyme A activity showed a sigmoid dependence on substrate concentration whereas the enzyme B showed classical Michaelis-Menten kinetics. The K(m) value of enzyme A for guanine was 5.3mum and that of enzyme B 20mum. 2. The entire guanine deaminase activity of mouse liver was contained in the 15000g supernatant of iso-osmotic homogenates. 3. A reinvestigation of the behaviour of rat brain 15000g supernatant guanine deaminase isoenzymes revealed that one enzyme had sigmoidal kinetics and the other enzyme showed a hyperbolic response. 4. Of the guanine deaminase in mouse brain iso-osmotic sucrose homogenate 80% was recovered in the 15000g supernatant and the rest from the particles. The supernatant guanine deaminase was resolvable into two fractions on DEAE-cellulose columns. One enzyme showed sigmoidal kinetics whereas the other showed a hyperbolic response to increasing substrate concentration; the K(m) values for the reaction with guanine were respectively 5 and 66mum. 5. The particulate fractions of mouse liver and brain were devoid of any overt inhibitory activity.     

Phospholipid-transfer proteins in human liver and primary liver carcinoma

Investigations have been carried out on phospholipid-transfer activity of the cytosol and the phospholipid composition of subcellular membranes from human liver and primary liver carcinoma. In both human liver and primary liver carcinoma cytosolic fractions, the transfer activity for phosphatidylcholine (PC), phosphatidylethanolamine (PE) and sphingomyelin has been observed for the first time. The transfer rate of PC and PE in normal human liver was almost equal, whereas sphingomyelin-transfer activity was much slower. In carcinoma cells, the transfer activity for PE and PC was significantly enhanced, while sphingomyelin transfer remained unchanged. Comparative investigations with HepG2 cultured cells have revealed a high PE-transfer activity in this cell line. Parallel with the phospholipid-transfer activity modifications in neoplasic cells, changes in the phospholipid composition of microsomes and mitochondria have been observed. The content of PC and PE in hepatocarcinoma cells was decreased in microsomes, while in the mitochondria it was increased. The possible role of the phospholipid-transfer proteins in the maintenance of membrane composition and structure is discussed.     

Low-molecular-weight chromium-binding substance from chicken liver and American alligator liver

Low-molecular-weight chromium-binding substance (LMWCr), also known as chromodulin, is a chromium-binding oligopeptide proposed to have a function in chromium transport and insulin signaling in mammals. In this work, LMWCr has been isolated and purified for the first time from non-mammalian sources: chicken and American alligator. Milligram quantities of the oligopeptide can be obtained from kilogram quantities of liver. The LMWCr's from both sources are asparatate- and glutamate-rich oligopeptides which possess multinuclear chromium assemblies. The composition and physical and spectroscopic properties of the avian and reptilian LMWCr's are extremely similar to those of their mammalian analogues, suggesting the multinuclear sites of the biomolecule from all three classes of animal possess very similar structures. The chicken and alligator oligopeptides may possess intrinsic phosphotyrosine phosphatase activity.     

Potential neural progenitor cells in fetal liver and regenerating liver

From unfractionated embryonic mice liver cells, appreciable amount of spherical bodies containing nestin-positive cells were generated in the presence of neuronal growth factors. Following cultivation on poly-d-lysine/laminin-coated slips, approximately 70% of the cells expressed neuronal markers, and 16% had long processes. Functional analysis of these long-process-bearing cells with the whole-cell patch clamp method showed an inward current in response to glutamate, GABA, and serotonin as the neuronal characteristics. Furthermore, regenerating liver in adult mice also contained nestin-positive cells to the same extent as fetal liver. Regenerating liver could have potential as a source of neural cells for autologous transplantation.     

Fetal and adult liver stem cells for liver regeneration and tissue engineering

For the development of innovative cell-based liver directed therapies, e.g. liver tissue engineering, the use of stem cells might be very attractive to overcome the limitation of donor liver tissue. Liver specific differentiation of embryonic, fetal or adult stem cells is currently under investigation. Different types of fetal liver (stem) cells during development were identified, and their advantageous growth potential and bipotential differentiation capacity were shown. However, ethical and legal issues have to be addressed before using fetal cells. Use of adult stem cells is clinically established, e.g. transplantation of hematopoietic stem cells. Other bone marrow derived liver stem cells might be mesenchymal stem cells (MSC). However, the transdifferentiation potential is still in question due to the observation of cellular fusion in several in vivo experiments. In vitro experiments revealed a crucial role of the environment (e.g. growth factors and extracellular matrix) for specific differentiation of stem cells. Co-cultured liver cells also seemed to be important for hepatic gene expression of MSC. For successful liver cell transplantation, a novel approach of tissue engineering by orthotopic transplantation of gel-immobilized cells could be promising, providing optimal environment for the injected cells. Moreover, an orthotopic tissue engineering approach using bipotential stem cells could lead to a repopulation of the recipients liver with healthy liver and biliary cells, thus providing both hepatic functions and biliary excretion. Future studies have to investigate, which stem cell and environmental conditions would be most suitable for the use of stem cells for liver regeneration or tissue engineering approaches.     

Isoprostanes and the liver

The liver has been central to our understanding of the physiology and biology of the F2-isoprostanes. The discovery of F2-IsoPs and the initial demonstration that they could be used to localize oxidative stress was first demonstrated in a rat model of oxidative liver injury (carbon tetrachloride), and the first demonstration that plasma concentrations are increased in a human disease was in patients with liver failure and the hepatorenal syndrome [J. Clin. Invest. 90 (6) (1992b) 2502; J. Lipid Mediat. 6 (1/3) (1993) 417]. This article will cover the measurement of F2-IsoPs as markers of lipid peroxidation in vivo in liver disease, and review their biological activity as mediators of disease.     

Apoptosis and liver diseases

Regulation of homeostasic balance between cell proliferation and cell death, called apoptosis, is essential for development and maintenance of multicellular organisms. Recent research into the molecular mechanisms of apoptosis has revealed that apoptosis is a genetically and evolutionarily conserved process that can become deranged when the components of the cellular apoptotic machinery are mutated, perturbated by viral gene products or present in inappropriated quantities. Analysis of the regulatory apoptotic pathways has led to a better understanding of the etiology and pathogenesis of many human diseases, notably cancers, infectious diseases or autoimmune diseases. Our understanding of the regulation of apoptosis in health and disease is far from complete and the use of understanding into new therapeutic modalities has only begun to be approached.     

Cysteinyl-leukotrienes and the liver

Leukotrienes are potent biological mediators implicated in an increasing number of disease processes. This review outlines the basic biology of leukotrienes and discusses recent developments in our understanding of the specific role of cysteinyl-leukotrienes (cLTs) in cholestasis, hepatic inflammation, portal hypertension, and the pathogenesis of the hepatorenal syndrome (HRS).     

Foods and liver health

Chronic liver damage is a worldwide common pathology, characterised by an inflammatory and fibrotic process that leads to a progressive evolution from chronic hepatitis to cirrhosis and hepatocellular carcinoma (HCC). A major role for fats and oxidative stress has been recently demonstrated in the pathogenesis of liver diseases. In the clinical practice, dietary recommendations in the management of chronic diseases often rely on denying patients certain foods, which results in a severe reduction of quality of life. In this paper a new perspective based on the development of Food intended for Specific Medical Purposes (FSMP) containing highly bioavailable antioxidant compounds or polyunsaturated-fatty acids, has been highlighted as a tool for preventive and curative medicine, to be associated to pharmacological treatments.     

Fatty liver and lipotoxicity

Fatty liver disease comprises a spectrum ranging from simple steatosis to steatohepatitis which can progress to liver cirrhosis and hepatocellular cancer. Hepatic lipotoxicity may ensue when the hepatic capacity to utilize, store and export fatty acids (FA) as triglycerides is overwhelmed. Additional mechanisms of hepatic lipotoxicity include abnormal FA oxidation with formation of reactive oxygen species, disturbances in cellular membrane FA and phospholipid composition, alterations of cholesterol content and ceramide signalling. Lipotoxicity is a key factor for the progression of fatty liver disease by inducing hepatocellular death, activating Kupffer cells and an inflammatory response, impairing hepatic insulin signalling resulting in insulin resistance, and activation of a fibrogenic response in hepatic stellate cells that can ultimately lead to cirrhosis. Therefore, the concept of hepatic lipotoxicity should be considered in future therapeutic concepts for fatty liver disease.     

补体系统及其在肝损伤再生中的作用

补体系统是机体免疫防御机制的重要组成部分,参与免疫识别和防御。近年来,系统研究发现补体除免疫调节外,还具有参与生殖发育、成骨、组织和器官再生等重要生理机能的作用。多项研究报道了补体活化和各种肝损伤／再生的关系,对此进行综述,以期促进对补体多样性功能的了解。     

清肝泄浊法治疗脂肪肝30例临床研究

目的:观察清肝泄浊法治疗脂肪肝的临床疗效.方法:治疗组30例口服自拟清肝泄浊汤,对照组24例口服复方益肝灵,观察肝功能及血脂变化.结果:治疗组30例,有效率90.0%,对照组30例,有效率36.6%.结论:清肝泄浊法治疗脂肪肝的疗效明显,适宜临床推广运用.     

Phosphorylation and inactivation of liver glycogen synthase by liver protein kinases

A rapid method for purifying glycogen synthase a from rat liver was developed and the enzyme was tested as a substrate for nine different protein kinases, six of which were isolated from rat liver. The enzyme was phosphorylated on a 17-kDa CNBr fragment to approximately 1 phosphate/87-kDa subunit by phosphorylase b kinase from muscle or liver with a decrease in the activity ratio (-Glc-6-P/+Glc-6-P) from 0.95 to 0.6. Calmodulin-dependent glycogen synthase kinase from rabbit liver produced a similar phosphorylation pattern, but a smaller activity change. The catalytic subunit of beef heart cAMP-dependent protein kinase incorporated greater than 1 phosphate/subunit initially into a 17-kDa CNBr peptide and then into a 27-30-kDa CNBr peptide, with an activity ratio decrease to 0.5. Glycogen synthase kinases 3, 4, and 5 and casein kinase 1 were purified from rat liver. Glycogen synthase kinase 3 rapidly phosphorylated liver glycogen synthase to 1.5 phosphate/subunit with incorporation of phosphate into 3 CNBr peptides and a decrease in the activity ratio to 0.3. Glycogen synthase kinase 4 produced a pattern of phosphorylation and inactivation of liver synthase which was very similar to that caused by phosphorylase b kinase. Glycogen synthase kinase 5 incorporated 1 phosphate/subunit into a 24-kDa CNBr peptide, but did not alter the activity of the synthase. Casein kinase 1 phosphorylated and inactivated liver synthase with incorporation of phosphate into a 24-kDa CNBr peptide. This kinase and glycogen synthase kinase 4 were more active against muscle glycogen synthase. Calcium-phospholipid-dependent protein kinase from brain phosphorylated liver and muscle glycogen synthase on 17- and 27-kDa CNBr peptides, respectively. However, there was no change in the activity ratio of either enzyme. The following conclusions are drawn. 1) Liver glycogen synthase a is subject to multiple site phosphorylation. 2) Phosphorylation of some sites does not per se control activity of the enzyme under the assay conditions used. 3) Liver contains most, if not all, of the protein kinases active on glycogen synthase previously identified in skeletal muscle.