Effect of lysosomal loading with Triton WR 1339 on the distribution of albumin-14C in the liver of rats with chronic hepatitis

The mechanism of the protective effect of the lysosomotropic detergent Triton WR 1339 in chronic hepatitis was examined. Assuming that the improvement in the condition is connected with potentiation of the heterophagous function of the lysosomes, the intensity of uptake of albumin-¹⁴C by the liver and its subcellular distribution in the liver of rats were studied during administration of the detergent to animals with chronic carbon tetrachloride hepatitis. Preliminary injection of the detergent did not affect the intensity of uptake of albumin-¹⁴C, but subsequent injection of Triton WR 1339 into rats with toxic hepatitis reduced the protein uptake to values obtained in intact rats. In chronic hepatitis albumin-¹⁴C is concentrated in the lysosomal fraction. After injection of Triton WR 1339 into the poisoned animals the peaks of labeled protein and lysosomal enzymes did not coincide. The selective role of lysosomes of the Kupffer cells of the liver in producing the more rapid recovery of the liver from chronic hepatitis is examined.Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 7, pp. 816–819, July, 1976.     

Effect of loading the liver lysosomes with Triton WR 1339 on the development of chronic toxic hepatitis

Preliminary administration of Triton WR 1339 has a favorable effect on the course of chronic toxic hepatitis. The zones of necrosis are reduced, the development of connective tissue is delayed, and liver function is improved. The liver lysosomes of animals poisoned with CCl₄ after preliminary administration of the detergent are more stable when exposed to harmful procedures in vitro.Central Research Laboratory, Novosibirsk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR, D. D. Yablokov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 79, No. 1, pp. 21–23, January, 1975.     

Studies on the mechanism of enhancement of carbon tetrachloride hepatotoxicity by Triton WR 1339.

Pretreatment of rats with Triton WR 1339 significantly enhanced the intensity of CC14-induced liver necrosis. Previous workers suggested that this effect might be due to enhancement by Triton WR 1339 of cellular degradative processes. This pretreatment, however, also enhanced the intensity of covalent binding of [14C]CC14 metabolites to microsomal protein at 3 or 6 h, but not 1 h after its administration. This effect is not due to changes of microsomal P-450 content or increased activity of mixed-function oxygenase-metabolizing drugs like pentobarbital. Pretreatment with Triton WR 1339 also partially increased CC14-induced peroxidation of microsomal lipids at 1, 3 or 6 h after administration of the hepatotoxin. Liver concentrations of CC14 in Triton WR 1339-treated rats were significantly higher at 3 or 6 h but not at 1, 10 or 24 after its i.p. administration. Triton WR 1339 treatment decreased the body temperature of the rats and further intensified the decrease produced by CC14. Results suggest that, in addition to possible effects of Triton WR 1339 administration on liver-cell degradative processes, there are other actions of this detergent on CC14 activation and lipid peroxidation which might play a role in the heightened response of the liver of CC14-induced injury.     

Changes in rat liver lysosomes during stimulation of regeneration in the injured organ by triton WR-1339

The effect of a single dose of the lysosomotropic agent Triton WR-1339 on the properties of the liver lysosomes of rats with chronic toxic hepatitis and on the course of the pathological process was investigated. The compound was shown to promote the more rapid restoration of liver structure and function. The possible mechanism of the beneficial effect of Triton WR-1339 is discussed.Central Research Laboratory, Novosibirsk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 8, pp. 230–232, August, 1978.     

Action of sodium aurothiomalate in acute toxic hepatitis

The effect of the lysosomotropic compound sodium aurothiomalate on the character of the liver damage and changes in the lysosomes in acute toxic hepatitis was studied. After the combined action of this compound and CCl₄ the extent of spread of necrosis was reduced and the area of zones with degenerative changes in the parenchymatous cells of the liver was increased. The degree of solubilization of acid RNase was reduced. Nonsedimented -galactosidase and cathepsin D activity was the same as in CCl₄ hepatitis.Central Scientific-Research Laboratory, Novosibirsk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 5, pp. 539–542, May, 1977.     

Liver sinusoidal cell ultrastructure in partially hepatectomized rats

Two-thirds of the liver was removed from male Wistar rats weighing 160–200 g. In response to the operation phasic changes occurred in the structure of the lysosomal apparatus in the Kupffer cells. The number of primary lysosomes in the Kupffer cells was increased 2.5 h after partial hepatectomy, and the size and polymorphism of the lysosomes were increased after 9 h. At the peak of mitotic activity of the hepatocytes (30 h after partial hepatectomy) mainly secondary lysosomes were identified in the Kupffer cells, but 48 h after the operation, on the other hand, their number was reduced, although young forms of primary lysosomes appeared. Manifestations of fatty infiltration predominated in the endothelial cells and reached a maximum at the peak of mitosis of the hepatocytes. The results are evidence that ultrastructural changes in the sinusoidal cells depend on the phases of reparative regeneration of the liver.Laboratory of Pathophysiology, Department of General Pathology, Institute of Clinical and Experimental Medicine, Siberian Branch, Academy of Medical Sciences of the USSR, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 12, pp. 729–733, December, 1979.     

Structural and functional changes in lysosomes of regional lymph nodes and liver of dogs with toxic hepatitis

Structural and functional changes in lysosomes of cells in the regional lymph nodes and liver were investigated in the course of toxic hepatitis caused by single or repeated administration of CCl₄ to dogs. Total activity of the lymph node lysosomal enzymes studied (-galactosidase, acid RNase, cathepsin D) exceeded the corresponding activity in the liver of intact dogs, and this reflects the barrier or protective function of the organ. With the development of acute toxic hepatitis a sharp increase was found in acid RNase and cathepsin D activity. At the times of testing (8 and 30 days) the parameters studied had not returned to normal. Meanwhile the mass of the regional lymph nodes and the relative number of macrophages and neutrophils in their sinuses were increased. The increase in lysosomal enzyme activity in the regional lymph nodes following liver damage is connected with the increased functional load on the lymph nodes, which are involved in the hydrolysis of biopolymers brought to the regional lymph nodes with the lymph flow.Department of Normal Anatomy and Central Research Laboratory, Novosibirsk Medical Institute. Translated from Byulleten' Éxperimental'noi Biologii i Meditsiny, Vol. 86, No. 10, pp. 428–431, October, 1978.     

The Effects of Delta Sleep-Inducing Peptide on the Intensity of Lipid Peroxidation and Xanthine Oxidase Activity in Rat Tissues during Cold Stress

Exogenous delta sleep-inducing peptide given i.p. to intact rats at a dose of 12 g/100 g decreased the levels of diene conjugates and Schiff bases in liver and brain tissues and had no effect on xanthine oxidase activity in these tissues. Cold stress was accompanied by increases in xanthine oxidase activity in rat liver and brain, with a consequent accumulation of diene conjugates and Schiff bases, as compared with intact animals. Preliminary administration of delta sleep-inducing peptide before three days of cold stress led to decreases in xanthine oxidase activity and lipid peroxidation products in the liver and brain, as compared with values in stressed rats. The protective effect of delta sleep-inducing peptide in stress is discussed.     

Fractional Composition of Blood Serum Lipoproteins in Mice and Rats with Triton WR 1339-Induced Lipemia

We compared fractional composition of blood serum lipoproteins (LP) in female ICR mice and Wistar rats induced by single administration of a nonionic detergent Triton WR 1339 in doses of 300 and 500 mg/kg. Lipemia in animals of both species was characterized by a sharp increase in the concentration of cholesterol and, particularly, of triglycerides in blood serum lipoproteins by the 24th hour after administration of the detergent. We revealed a significant increase in the concentrations of atherogenic VLDL cholesterol (due to VLDL2), intermediate density lipoproteins, and LDL. These changes were more pronounced in rats. The model of lipemia can be used to study the role of fractional composition of lipoproteins and, particularly, of triglycerides in the pathogenesis of atherosclerosis. Moreover, this model holds much promise for evaluation of the efficiency of hypolipidemic drugs (statins and fibrates) in normalizing the increased level of atherogenic cholesterol of VLDL and LDL.     

Status of the progeny of rats treated with platinum-containing cytostatics

Wistar rat pups from rats treated with first- and second-generation platinum-containing cytostatics (platidiam, carboplatin) 1, 3, and 6 months before mating with intact partners had similar disorders. The severity of these disorders depended in many cases on the chemical structure of the drug and sex of the parent treated with the cytostatic. The severity of toxic effects in the progeny of intact females mated with cytostatic-treated males decreased with prolongation of the period elapsing between the treatment and mating. Carboplatin produced a more potent toxic effect on the reproductive function of rats compared to platidiam.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 9, pp. 302–305, September, 2004     

Aggravation of injury to liver lysosomes of rats with CCl4 hepatitis by preliminary prolonged administration of chlorpromazine

Preliminary prolonged administration of chlorpromazine (5 mg/kg for three weeks) aggravated the injury to liver lysosomes of rats with acute CCl₄ hepatitis. Similar marked changes were observed in lysosomes sedimented with heavy and light mitochondrial fractions.Central Scientific-Research Laboratory and Department of Psychiatry, Novosibirsk Medical Institute. (Presented by Academician, of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 1, pp. 38–41, January, 1977.     

Activity of lysosomal enzymes in the bile and serum of mice with intrahepatic cholestasis

Lysosomal enzyme activity in the bile and blood serum was compared in mice with experimental intrahepatic cholestasis induced by α-naphthyl isothiocyanate and Triton WR 1339. Triton WR 1339 increases the synthesis of cholesterol (fatty acid precursor) in liver cells. The development of intrahepatic cholestasis was confirmed by the increase in activities of alkaline phosphatase and γ-glutamyltransferase in blood serum. Administration of Triton WR 1339 in a dose of 100 mg/100 g was followed by a 10-fold increase in β-galactosidase activity (hepatocyte lysosomal enzyme) in the bile, but not in the serum of mice. β-Galactosidase activity significantly increased in the bile, but decreased in the serum of mice after treatment with α-naphthyl isothiocyanate in a dose of 200 mg/kg. Our results indicate that intrahepatic cholestasis is manifested in increased secretion of lysosomal glycosidases into the bile. Bile components can aggravate damage to liver cells by affecting the processes of hepatocyte apoptosis and necrosis. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 496–499, May, 2008     

Porphyrobilinogen biosynthesis from δ-aminolevulinic acid by the viscera of albino rats

Ability to synthesize porphyrobilinogen (PBG) from -aminolevulinic acid (ALA) was determined in homogenates of tissues of the lungs, heart, liver, kidneys, spleen, pancreas, and small intestine of 77 albino rats. All these organs were found to be able to synthesize PBG. Highest ALA dehydratase activity was found in the liver tissue, followed in descending order by the kidneys, lungs, pancreas, small intestine, heart, and spleen. On the addition of a lead solution to the synthesizing system a significant decrease in enzyme activity was observed in the liver tissue, but in kidney tissue its activity was unchanged. On the addition of lead and D-penicillamine simultaneously no changes were found in the toxic effect of lead.M. F. Vladimirskii Moscow Regional Clinical Research Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR S. S. Debov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 12, pp. 687–689, December, 1978.     

Effect of Transitory Ischemia on Liver Lysosomal Apparatus in Rats with Different Resistance to Hypoxia

We studied the state of lysosomal apparatus and pro- and antioxidant activity in the liver of rats with different resistance to hypoxia during postischemic recovery. Under normal conditions the lysosomal apparatus did not differ in highly and low resistant animals. During ischemia and reperfusion the damage to hepatic lysosomal membranes in rats highly resistant to hypoxia was less pronounced than in low resistant animals. These differences also concerned labilization of lysosomes during exposure to damaging factors (hypotonia and Triton X-100). The rats highly resistant to hypoxia differed from low resistant animals by higher stability of lysosomal membranes, lower prooxidant activity (malonic dialdehyde content), and higher tissue concentration of -tocopherol during reperfusion.     

Copper supplementation in the rat: Preliminary observations on the clinical,hematological and histopathological profile

Complete toxicological examinations in female rats fed an anti-inflammatory 200 ppm copper-containing diet for 30 and 58 days were performed.Copper was found to accumulate in liver, kidneys and paws; however, at the hematological, clinical chemical and histopathological levels, this treatment did not seem able to induce any toxic accumulation phenomenon in the examined rats.     

Effects of aldosterone on lipid metabolism and renal oxygen consumption in the rat

Summary The plasma level of free fatty acids (FFA) in adrenalectomized rats increases by 50% after treatment with aldosterone (2 g/100 g rat).Lipolytic activity in peripheral fat tissue is lowered after adrenalectomy and doubles after in vivo administration of aldosterone to adrenalectomized rats (measured as free fatty acid release in vitro from epididymal fat tissue).Lypolysis of adipose tissue stimulated by the in vitro presence of ACTH also increases after in vivo administration of aldosterone.Incorporation of intravenously administered label from U¹⁴C-palmitate into total extractable lipid of renal tissue is augmented 3 h after aldosterone administration to adrenalectomized rats, while no increase of the radioactivity is observed in total lipid from liver tissue. Treatment with aldosterone does not affect the total lipid content of kidney or liver in adrenalectomized rats.The oxygen consumption rate of kidney cortex slices with lactate, -hydroxybuterate or acetoacetate as substrates is lowered after in vivo administration of aldosterone to adrenalectomized rats. With succinate, however, the respiratory rate of kidney slices increases after aldosterone treatment of adrenalectomized rats, the ouabain-sensitive respiration being more affected than the ouabain-insensitive respiration. An interpretation of the O₂ consumption data implicating competition of lipid metabolism for CoA-SH is discussed.     

Myelin glycosphingolipid/cholesterol-enriched microdomains selectively sequester the non-compact myelin proteins CNP and MOG

Plasma membranes are complex arrays of protein and lipid subdomains. Detergent-insoluble, glycosphingolipid/cholesterol-enriched micro-domains (DIGCEMs) have been implicated in protein sorting and/or as sites for signaling cascades in the plasma membrane. We previously identified the presence of DIGCEMs in oligodendrocytes in culture and purified myelin and characterized a novel DIGCEM-associated tetraspan protein, MVP17/rMAL (Kim et al. (1995) Journal of Neuroscience Research 42, 413–422). We have now analyzed the association of known myelin proteins with DIGCEMs in order to provide a better understanding of their roles during myelin biogenesis. We used four well-established criteria to identify myelin DIGCEM-associated proteins: insolubility in a non-ionic detergent Triton X-100 at low temperature (4°C), flotation of the insoluble complexes to low density fractions in sucrose gradients, and TX-100 solubilization at 37°C, or at 4°C following treatment with the cholesterol-binding detergent saponin. We demonstrate that these proteins fall into four distinct groups. Although all tested proteins could be floated to a low-density fraction, proteolipid protein (PLP), myelin basic protein (MBP) and myelin associated glycoprotein (MAG) were solubilized by the detergent extraction, and connexin32 (Cx32) and oligodendrocyte-specific protein (OSP) met only some of the criteria for DIGCEMs. Only the non-compact myelin proteins 2,3-cyclic nucleotide 3-phosphodiesterase (CNP) and myelin/oligodendrocyte glycoprotein (MOG) satisfied all four criteria for DIGCEM-associated proteins. Significantly, only 40% of CNP and MOG were selectively associated with DIGCEMs. This suggests that they may have both non-active soluble, and functionally active DIGCEM-associated, forms in the membrane, consistent with current views that DIGCEMs provide platforms for bringing together and activating components of the signal transduction apparatus. We therefore propose that CNP and MOG may have unique roles among the major myelin proteins in signaling pathways mediated by lipid-protein microdomains formed in myelin.     

DNA content and size of cell nuclei in the regenerating rat liver

The DNA content in single nuclei and the size of the nuclei were investigated in the intact and regenerating rat liver from 18 h to 21 days after partial hepatectomy. The results of the measurements show that the mean DNA content per nucleus in the intact rat liver is 6.5 pg, and that most nuclei are about equal in size to the diploid nucleus (42.5 ²). DNA synthesis began in the regenerating liver before 18 h after the operation. By 24 h the DNA content in most nuclei of the experimental animals was twice that in the intact rats. This shows that the first wave of synthesis involved 85–90% of the liver cells. After mitosis, which in most cells took place before 36 h after partial hepatectomy weaker waves of DNA synthesis followed, after approximately 42 and 60 h.Department of Normal Anatomy and Department of General Biology, P. J. Safarik University, Kosice, Czechoslovakia. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 8, pp. 369–371, March, 1977.     

Involvement of lysosomes in the uptake of macromolecular material by bloodstream forms of Trypanosoma brucei

To investigate whether the lysosomes of Trypanosoma brucei are capable of uptake of macromolecules after internalization by the cell, we used Triton WR-1339, a non-digestible macromolecular compound, which is known to cause a marked decrease in the density of hepatic lysosomes due to massive intralysosomal storage. Intraperitoneal administration of 0.4 g/kg Triton WR-1339 to rats infected with T. brucei led to the development of a large vacuole in the trypanosomes between nucleus and kinetoplast within 22 h. Higher doses (2 g/kg) led to the disappearance of the trypanosomes from the blood and resulted in permanent cures (greater than 100 days). Lysosomes isolated from the trypanosomes of animals treated with a sub-curative dose showed a decrease in equilibrium density of 0.03 g/cm3 in sucrose gradients. These lysosomes were partly damaged as evidenced by a reduction in latency and an increase in the non-sedimentable part of lysosomal enzymes. We conclude that acid proteinase and alpha-mannosidase-containing organelles of T. brucei take up exogenous macromolecules and must therefore be considered as true lysosomes and that Triton WR-1339 acts in T. brucei as a true lysosomotropic drug. Its trypanocidal action probably results from an interference with lysosomal function.