Effects of alcohol and acetaldehyde on metabolism and function of neurotransmitter systems in cerebral cortical neurons in primary culture.

Effects of alcohol (ethanol) and acetaldehyde (AcAl) on the metabolism and function of gamma-amino-butyric acid (GABA)ergic and cholinergic systems were investigated using mouse cerebral cortical neurons in primary culture. Exposure to alcohol in vitro had no significant effects on the content of neuroactive amino acids as well as the activities of glutamic acid decarboxylase (GAD), GABA-transaminase (GABA-T), choline acetyltransferase (CAT) and acetylcholinesterase (AChE). In contrast, AcAl showed remarkable reductions of neuroactive amino acids content, and of CAT and AChE activities, but induced no alteration in the activities of GAD and GABA-T.

Alcohol, acetaldehyde and salsolinol-induced alterations in functions of cerebral GABA/benzodiazepine receptor complex.

Effects of alcohol (ethanol) and acetaldehyde on the metabolism and function of primary cultured GABAergic neurons and that of salsolinol, a condensation product of acetaldehyde with dopamine, on cerebral GABA/benzodiazepine (BZP) receptor complex were investigated. In vitro addition of acetaldehyde induced a significant reduction not only on the content of GABA but also on the basal and GABA-activated [3H]flunitrazepam ([3H]FLN) bindings in primary cultured neurons. In contrast, alcohol exhibited only suppressive effects on [3H]FLN binding as well as on the enhancement of [3H]FLN binding by GABA.

Transcellular transport of fluorescein dextran through an arterial endothelial cell monolayer.

Transcellular transport of fluorescein dextran (FD) of various molecular weights (4K, 10K, 20K, 70K and 150K daltons) through porcine arterial endothelial cells cultured on a type I collagen gel supported by a dacron sheet was studied and compared with the transport of low density lipoprotein labeled with rhodamine B (RB-LDL) described previously (Hashida et al., Cell Struct. Funct.

Stress and immune responses. II. Identification of stress-sensitive cells in murine spleen cells.

The influences of restraint stress on the functions of T cells, B cells and adherent cells in antibody responses were investigated. Antibody response against sheep red blood cells (SRBC), a T cell-dependent antigen, in cultured splenocytes from restrained mice was reduced to about 40-50% of that from the control mice. Addition of normal T cells to these cultures, however, restored the suppressed response.

The effects of basic substances and acidic ionophores on the digestion of exogenous and endogenous proteins in mouse peritoneal macrophages.

Basic substances and acidic ionophores that increase the lysosomal pH in cultured macrophages (Ohkuma, S., and B. Poole, 1978, Proc.

Transcellular transport of lipoprotein through arterial endothelial cells in monolayer culture.

To study the mechanism of lipoprotein transport through arterial endothelial cells, porcine endothelial cells were cultured on gelated type I collagen supported by a dacron sheet, and the transport of low density lipoprotein (LDL) labeled with rhodamine B isothiocyanate (RB-LDL) through the cells was measured. Light and scanning electron microscopy showed that the cells on the gel were confluent. There was little RB-LDL transport through the endothelial monolayer at 0 degrees C.

Peroxidized lipids isolated by HPLC from atherosclerotic aorta.

Conjugated dienes of fatty acids were detected in cholesterol esters in lipid droplets prepared from atherosclerotic aorta of WHHL rabbits. Two peroxidized fatty acids were recovered by reverse phase HPLC. These peroxidized fatty acids showed maximum absorption at 233 nm, suggesting that they were peroxidized lipids containing cis-trans conjugated diene, and one of them was shown to be 13-hydroxy octadecadienoic acid.

Thioridazine enhances lysosomal accumulation of epidermal growth factor and toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin.

Thioridazine, a phenothiazine calmodulin inhibitor, aggravated the cytotoxic effect of a conjugate (EGF-PE) of epidermal growth factor (EGF) coupled with Pseudomonas exotoxin against cultured HeLa cells. Other phenothiazine calmodulin inhibitors, trifluoperazine and chlorpromazine, also intensified the cytotoxic effect of EGF-PE, whereas N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W7) had no such effect. By using iodinated epidermal growth factor ( [125I]EGF), the effect of thioridazine on intracellular transport of EGF was examined.

Similarity of lysosomal H+-ATPase to mitochondrial F0F1-ATPase in sensitivity to anions and drugs as revealed by solubilization and reconstitution.

Lysosomal H+-translocating ATPase (H+-ATPase) was solubilized with lysophosphatidylcholine and reconstituted into liposomes (Moriyama, Y., Takano, T. and Ohkuma, S.

Development of taurine biosynthesizing system in cerebral cortical neurons in primary culture.

Developmental patterns of taurine biosynthesizing system were investigated using primary cultured neurons prepared from the neopallium of 15-day-old fetal mice by a trypsin treatment in comparison with those in cerebral cortices obtained from age-matched fetal and neonatal mice. The morphological observations by phase contrast and scanning electron micrographies indicated that the cells in primary culture used in the present study possessed typical features of neurons. In addition, the immunohistochemical studies using the antibody to glial fibrillary acidic protein (GFAP), a specific marker for astroglia, revealed that the contamination of astroglias was negligible.

Studies on fine structure and location of lipids in quick-freeze replicas of atherosclerotic aorta of WHHL rabbits.

The fine structure of intracellular and extracellular lipids in the atherosclerotic aorta of Watanabe-heritable hyperlipidemic (WHHL) rabbits was demonstrated by a quick-freeze etching technique. Many lipid droplets, with and without a membrane, were observed in the foam cells. Membrane-free droplets were observed as onion-like structure with a concentric lamellar structure surrounded by 10 nm filaments.

Monoclonal antibodies recognizing lipid-laden cells and extracellular regions with lipid-deposits in atherosclerotic aorta.

Monoclonal antibodies against lipid-laden cells and against extracellular regions of lipid-deposits in atherosclerotic aorta were prepared. Mice were immunized with a delipidated homogenate of atherosclerotic aorta of Watanabe-heritable hyperlipidemic rabbits. Hybridomas were obtained by fusion and cultured in hypoxanthine, aminopterin and thymidine selection medium.

Effect of phospholipids on the hydrolysis of cholesterol oleate liquid crystals by lysosomal acid lipase.

Cholesterol oleate liquid crystals were prepared in vitro as a model of lipid droplets accumulated in the atheromatous aorta. The hydrolysis of cholesterol oleate crystals by lysosomal acid lipase was examined in the presence and absence of various phospholipids. Phosphatidylserine markedly stimulated the hydrolysis of cholesterol oleate liquid crystals (20 times the basal value); it increased the Vmax value about 15 times and decreased the Km value to 1/20 times the basal value.

Positional specificity of lysosomal acid lipase purified from rabbit liver.

The hydrolysis of tri-, di-, and monooleoylglycerols by highly purified lysosomal acid lipase from rabbit liver (Imanaka, T., et al. (1984) J.

Regulatory role of cysteine dioxygenase in cerebral biosynthesis of taurine. Analysis using cerebellum from 3-acetylpyridine-treated rat.

The effect of 3-acetylpyridine (3-AP) administration on the biosynthesis of taurine in the rat brain has been studied. Treatment with 3-AP induced a significant decrease in the cerebellar contents of taurine and its metabolic precursors, cysteine sulfinic acid (CSA) and cysteic acid (CA), as well as a selective degeneration of climbing fibers in the molecular layer of the cerebellum. It was found that the activity of cerebral cysteine dioxygenase, the enzyme catalyzing the formation of CSA from cysteine, consisted of two systems with low and high Km values.

Subcellular localization of acid lipase in rat liver.

The intracellular localization of acid lipase was examined by several subcellular fractionation techniques. Upon differential centrifugation, acid lipase was detected mainly in the lysosomal fraction. When linear sucrose density gradient centrifugation was used, the mean density of acid lipase was similar to that of lysosomal marker enzymes.

Preparation of blood components with saline-adenine-glucose-phosphate-maltose quadruple-pack system.

A saline-adenine-glucose-phosphate-maltose (SAGP-maltose) quadruple-pack system was devised to improve the conventional method of preparing blood components. One of the features of this method is that high-speed centrifugation was used as the initial spin in the two-step centrifugation of whole blood. This feature permitted the removal of the buffy coat from red cells at the same time the platelets were isolated.

Hepatic venocclusive disease associated with the consumption of pyrrolizidine-containing dietary supplements.

Venocclusive disease, a form of Budd-Chiari syndrome, was diagnosed in a 49-yr-old woman. The patient had portal hypertension associated with obliteration of the smaller hepatic venules. A liver biopsy specimen showed centrilobular necrosis and congestion.

Effect of esterastin, an acid lipase inhibitor, on the free and esterified cholesterol contents of cultured aortic smooth muscle cells treated with LDL and cholesterol ester liquid crystals.

The effects of esterastin, an acid lipase inhibitor, on the free and esterified cholesterol contents of cultured smooth muscle cells from pig aorta were examined. The post-nuclear supernatant fraction of the cell homogenate showed maximum acid cholesterol esterase activity at pH 4.5, and 50% of this activity was inhibited by 0.

[Cellular responses and the H+ pump in the vacuolar system].

Intracellular acidic compartments are now recognized to be ubiquitous in the vacuolar system of eukaryotic cells, where the unique ATP-dependent H+ pump (s) functions to maintain their internal low pH. Furthermore, various basic substances and acidic ionophores have been found to raise the internal pH of these compartments to neutrality. These chemicals, on the other hand, have been found to affect various biological reactions in multicellular organisms, including those involved in intercellular communication.

Biliary excretion of novel pneumotoxic metabolites of the pyrrolizidine alkaloid, monocrotaline.

Perfusion of the pyrrolizidine alkaloid, monocrotaline, through the isolated rat liver resulted in the appearance of Ehrlich-positive metabolites in both the perfusate and bile. Livers from male rats released greater quantities of metabolite into both bile and perfusate. Metabolite release was stimulated by pretreatment with phenobarbital.