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R C Lin

Publications and source records attributed to R C Lin.

At least 37 records · Page 2Linked to original sources
PubMed1989

Further studies on the 37 kD liver protein-acetaldehyde adduct that forms in vivo during chronic alcohol ingestion.

We have previously reported the detection of a 37 kD liver protein-acetaldehyde adduct in rats fed alcohol chronically with the AIN'76 diet. It was surprising that only one liver protein-acetaldehyde adduct was found. In this report, we have tried to detect additional protein-acetaldehyde adducts by electroimmunotransblot with rabbit anti-hemocyanin-acetaldehyde adduct IgG and to further characterize the 37 kD liver protein-acetaldehyde adduct. Sensitivity of electroimmunotransblot increased 10- to 20-fold when alkaline phosphatase-linked antibody was used in place of horseradish peroxidase, but only one protein-acetaldehyde adduct band was detected in liver. Feeding rats the Lieber-DeCarli alcohol diet also did not produce more protein-acetaldehyde adduct bands in electroimmunotransblot. Addition of cyanamide, an aldehyde dehydrogenase inhibitor, to the AIN'76 alcohol diet greatly increased the intensity of the 37-kD protein-acetaldehyde adduct band on electroimmunotransblot but did not produce other bands. The 37 kD protein-acetaldehyde adduct decayed in vivo with a half-life of 4 days when alcohol was removed from the diet. The 37 kD protein-acetaldehyde adduct in liver is cytosolic. Its interaction with anti-hemocyanin-acetaldehyde adduct IgG was blocked by polylysine-acetaldehyde adduct and polytyrosine-acetaldehyde adduct. It could be removed by immunosorption with anti-hemocyanin-acetaldehyde adduct IgG-bound immunoresin. When immunoblotted with anti-alcohol dehydrogenase and anti-aldehyde dehydrogenase antibodies, the alcohol dehydrogenase and aldehyde dehydrogenase bands in liver of alcohol-fed rats showed identical intensities before and after immunosorption.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetaldehyde↗
PubMed1988

Effects of hormones on apolipoprotein secretion in cultured rat hepatocytes.

When rat hepatocytes were cultured in serum-free and hormone-free Waymouth's medium, secretion rates of apolipoproteins (apo) AI and AIV were stable for two days, while the secretion rate of apo E decreased only 20% on the second day. Effects of insulin and dexamethasone on hepatic secretion of apo AI, apo E, and apo AIV were studied in primary culture of rat liver cells within two days. Adding insulin alone and dexamethasone alone, 1 mumol/L each, to cultured hepatocytes for 20 hours had little effect on the amounts of apo AI secreted by the cells. But when the treatment with either insulin or dexamethasone was prolonged for 44 hours, apo AI secretion by treated cells was increased 2.0-fold and 1.4-fold over that by control cells, respectively. If both hormones were added together, secretion of apo AI was synergistically increased 1.5-fold and 7-fold after 20 and 44 hours incubation, respectively. The optimal concentrations of both hormones for the synergistic effect were 0.1 mumol/L. Insulin alone did not affect, while dexamethasone alone slightly suppressed, apo E secretion by hepatocytes. However, when hepatocytes had been incubated with both hormones, a 70% increase in the release of apo E into the culture medium was also observed after 20 hours. Insulin caused a two-fold increase in cellular apo E in hepatocytes. The insulin-mediated cellular accumulation of apo E could be enhanced only very slightly by dexamethasone, but was completely blocked by glucagon.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Detection of a protein-acetaldehyde adduct in the liver of rats fed alcohol chronically.

We report here the formation in vivo of a protein-acetaldehyde adduct (protein-AA) in liver when rats were fed alcohol chronically. This chemically modified protein was demonstrated by electroimmunotransblot technique and with rabbit polyclonal antibodies that recognize acetaldehyde adduct as an epitope (i.e., both anti-hemocyanin-AA IgG and anti-myoglobin-AA IgG). It has a molecular weight of 37,000. It can be detected in the liver of rats fed the alcohol-containing American Institute of Nutrition 1976 liquid diet for only 1 wk. Since the protein profiles of soluble hepatic proteins from alcohol-fed and control rats were identical on SDS-PAGE, the peroxidase-positive band demonstrated by electroimmunotransblot was most likely not a new protein synthesized de novo. Borohydride reduction was not necessary to stabilize this protein-AA. Intraperitoneal injections of ethanol (2 g/kg body wt) at 8-h intervals to rats over a 24-h period did not produce any detectable protein-AA in the liver. Incubation of the liver homogenate from a control liver with acetaldehyde without sodium cyanoborohydride for 4 h also failed to generate any protein-AA. Therefore, the formation of the 37-kD protein-AA in vivo reported here is dependent on chronic alcohol consumption.

Acetaldehyde↗
PubMed1987

Differing effects of arginine deficiency on the urea cycle enzymes of rat liver, cultured hepatocytes and hepatoma cells.

We have confirmed that arginine-deficient diets increase the liver activities (units per 100 g) of the first four arginine biosynthetic enzymes of the urea cycle in Wistar rats, but not the activity of arginase. In contrast, rat liver cells cultured in monolayers for 48, 72 or 96 h in arginine-free L-15 or minimum essential medium showed no changes in carbamoyl-phosphate synthase (EC 6.3.4.16), ornithine transcarbamylase (EC 2.1.3.3), argininosuccinate synthase (EC 6.3.4.5), argininosuccinase (EC 4.3.2.1) or arginase (EC 3.5.3.1) activities. The arginine content of the cells grown on deficient medium was 36% of that of cells grown on 2.9 mM arginine-sufficient L-15, yet the urea excretion rate into the medium was reduced to 7% of the rate in control cells and the excretion of orotic acid was 400% of that in control cells. A Morris rat hepatoma cell line, 7800C1, which maintains activities of all five urea cycle enzymes, showed no consistent increases in the activities of the first four enzymes when the arginine in the medium was varied between 0 and 2 mM. Thus, in spite of severe arginine deficiency, cultured rat liver cells and hepatoma cells do not show the derepression-like response seen by other investigators when nonliver cells were cultured in arginine-deficient media. The difference between in vivo and in vitro effects of arginine deficiency on urea cycle activities remains unexplained.

Ammonium Chloride↗
PubMed1986

Quantification of apolipoproteins in rat serum and in cultured rat hepatocytes by enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay (ELISA) has been developed to measure apolipoproteins in rat serum. Nondelipidated whole serum was heat-treated at 52 degrees C for 3 h in phosphate-buffered saline containing 0.1% Tween-20 before assay. Monospecific rabbit anti-rat apolipoprotein antibodies were added to 96-well polystyrene microtiter plates which had been coated with purified rat serum apolipoproteins or unknown samples. After incubation and washing, goat anti-rabbit serum antibodies conjugated with horseradish peroxidase were added to the plates and incubated. The bound peroxidase activity was assayed after further washing. Serum apolipoprotein concentrations were calculated by comparison against purified standards that were assayed simultaneously with the unknown samples. The intraassay coefficients of variation for apolipoprotein AI, E, and AIV (Apo AI, E, and AIV) were 2.3, 4.4, and 5.3%, and interassay coefficients of variation were 6.1, 5.5, and 7.9%, respectively. The ELISA assay is sensitive to nanogram quantities of rat serum apolipoproteins and the results agree well with those measured by densitometry. The serum concentrations of Apo AI, E, and AIV of a normal fed rat were found to be 504 +/- 8, 413 +/- 20, and 262 +/- 20 micrograms/ml, respectively. When cultured as monolayers in Waymouth's medium for 1 day, rat hepatocytes secreted Apo AI, E, and AIV at rates of 2.51, 61.8, and 48.9 ng protein/mg cell protein/h.

Animals↗
PubMed1985

Serum cholesterol, lecithin-cholesterol acyltransferase, and hepatic hydroxymethylglutaryl coenzyme A reductase activities of lean and obese Zucker rats.

Serum cholesterol concentrations, lecithin-cholesterol acyltransferase (LCAT), and hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activities of lean and obese Zucker rats were compared. The excess serum cholesterol of the female obese rat is found to be mainly free cholesterol associated with very low-density lipoproteins, whereas that of the male obese rat is carried as cholesterol esters associated with high-density lipoproteins. The high level of serum free cholesterol in the female obese rat is not due to a deficiency in lecithin-cholesterol acyltransferase activity. This enzyme activity is found to be elevated in the male obese rat. Hepatic HMG-CoA reductase activity declines as rats mature; this observation is most apparent in obese male rats. Lean rats exhibit the normal diurnal rhythm, but mature obese rats show little diurnal variation in HMG-CoA reductase activity. Obese female rats maintain high reductase activities, but the activities of obese male rats remain low at all times. Starvation suppresses liver HMG-CoA reductase and serum cholesterol in both lean and obese female rats. Thus, an increase in hepatic cholesterol synthesis may contribute to hypercholesterolemia in the obese female Zucker rat. On the other hand, factors such as nonhepatic synthesis or a decreased cholesterol catabolism may play more important roles in maintaining high serum cholesterol in the obese male Zucker rat.

Animals↗
PubMed1984

Secretion of the newly synthesized cholesterol by rat hepatocytes in primary culture.

We used monolayer cultured rat hepatocytes as an experimental model to study the secretion of the newly synthesized cholesterol by the liver. Cellular cholesterol was labeled by exposing cultured hepatocytes to [14C]acetate prior to the study of secretion. Secretion of the newly synthesized cholesterol was measured by extracting cholesterol in the culture medium and assaying for the radioactivity of [14C]cholesterol. We found that: (a) cultured hepatocytes could secrete newly synthesized cholesterol in serum-free medium; (b) secreted [14C]cholesterol was bound to macromolecule(s) and the secretion rate was not affected by cycloheximide for up to 5 h; (c) serum added to the culture medium greatly enhanced hepatic cholesterol secretion; (d) serum high-density lipoproteins were most effective, lipoprotein-deficient serum (d greater than 1.21) less effective in stimulating cholesterol secretion, whereas low-density and very-low-density lipoproteins had little effect; (e) when the serum-free culture medium was fractionated by ultracentrifugation, a major portion of the secreted [14C]cholesterol was found in the high-density lipoprotein fraction; (f) part of the medium [14C]cholesterol also turned up in the high-density lipoprotein fraction when lipoprotein-deficient serum was added as the acceptor; (g) secreted [14C]cholesterol was found only in free form, although some of the cellular [14C]cholesterol was found as esters.

Animals↗
PubMed1982

Effect of dexamethasone on 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity and cholesterol synthesis in rat liver.

Dexamethasone increases reductase activity in cultured liver cells after a lag period of 2 h. The increases of activity are linear from 10(-9) to 10(-5) M dexamethasone, the maximum responses ranging from 2- to 4-fold. The increased reductase activity after dexamethasone treatment is not due to a change of the state of phosphorylation/dephosphorylation of the enzyme nor to an increase of cytosolic activating factor(s) for the reductase. Cholesterol synthesis, measured by incorporation of either [14C]acetate or 3H2O, increases 3-fold after dexamethasone (10(-6) M) treatment, as does the hydroxymethylglutaryl-CoA reductase activity, confirming that this enzyme is rate-controlling for cholesterol synthesis in cultured liver cells as it is in vivo. Dexamethasone (10 micrograms/100 g rat), given after onset of the light cycle, increases reductase activity over control rats at the nadir of the circadian cycle of this enzyme. When given after onset of the dark cycle, dexamethasone does not increase reductase activity over controls at the peak of their circadian cycle. Thus, physiologic doses of glucocorticoids partially reverse the decline in reductase activity due to the circadian rhythm.

Acetates↗
PubMed1981

Induction of urea cycle enzymes of rat liver by amino acids.

To determine which amino acids in a high casein diet are responsible for induction of the five urea cycle enzyme activities in rat liver, we tube-fed 21 L-amino acids singly to rats over 2 days at maximum doses which did not cause toxicity. The results were compared with the 1.3- to 1.9-fold increases (units/100 g rat) obtained by tube-feeding 2 g N/kg for 2 days as casein hydrolysate. Ala (2 g N/kg), Gly /2 g N/kg), Met (0.2--0.4 g N/kg) and Cys (0.4 g N/kg) were the only amino acids which increased all five activities. Moreover, Met. Ala, Gly and casein hydrolysate in these doses increased immuno-precipitable arginase as much as they increased its activity. A combination of Met, Ala and Gly (2 g N/kg) increased all five activities more than 2 g N/kg of casein hydrolysate. Met (0.05 g N/kg) + Ala (0.08 g N/kg) + Gly (0.1 g N/kg), the amounts of these contained in 2 g N/kg of casein, increased all five enzymes in 2 days as much as this dose of casein hydrolysate. Met (0.06 g N/kg) alone increased all five activities (units/100 g rat) 1.2 to 1.4-fold over controls by increasing g liver/100 g rat. Ammonium citrate or acetate tube-feedings over 8 days at 2 g N/kg increased only AS. The keto-acid of alanine, pyruvate, or the alpha-hydroxy acid of methionine did not increase any enzyme whereas the same molar dose of their amino acids increased all five activities. Thus three amino acids of casein, Ala, Gly and especially Met, account for the enzyme adaptation of the urea cycle on a high casein diet.

Alanine↗
PubMed1981

pH Determination in acidified foods: collaborative study.

A proposed method for determining pH of acidified foods has been developed and subjected to collaborative study. The method appears to be both accurate and precise. Five samples consisting of pimientos, marinated pimientos, 2 pH buffer solutions, and chocolate syrup were sent to each of 12 collaborators along with a copy of the method. Two of the collaborators were FDA District laboratories while the remainder were representatives from industry, universities, and state health agencies. Many different types of pH meters and combinations of electrodes were used by the collaborators. The tabulated results from the collaborators are presented. The method has been adopted official first action.

Food Analysis↗
PubMed1978

Induction of urea cycle enzymes of rat liver by glucagon.

All five urea cycle enzymes of rat liver increased in activity 48 h after subcutaneous administration of crystalline zinc glucagon to male rats and remained elevated after 7 days of continuous glucagon infusion. The maximum ratios of enzyme activities over those of controls were 2.0 for carbamyl phosphate synthetase, 1.3 for ornithine transcarbamylase, 2.7 for argininosuccinate synthetase, 3.2 for argininosuccinase, and 2.2 for arginase. Actinomycin D or puromycin prevented these responses to glucagon. The increase in arginase activity after zinc glucagon treatment was matched by an increase in immunoprecipitable enzyme. All five enzymes were induced by physiological plasma levels of glucagon. Tube feeding of casein hydrolysate for 2 days increased all five enzyme activities 1.5- to 2.2-fold and resulted in plasma glucagon levels similar to those required for induction by exogenous glucagon. Thus, glucagon is an inducer of the entire urea cycle in rat liver and plays a role in the induction of the cycle by protein feeding.

Animals↗
PubMed1969

Lysergic acid diethylamide: role in conversion of plasma tryptophan to brain serotonin (5-hydroxytryptamine).

Injections of D-lysergic acid diethylamide decrease the turnover rate of 5-hydroxytryptamine of rat brain, as measured from the conversion of (14)C-tryptophan into (14)C-5-hydroxytryptamine. The 2-bromolysergic acid diethylamide given in doses fivefold greater than those of lysergic acid diethylamide fails to change the rate of (14)C-tryptophan conversion into (14)C-5-hydroxytryptamine. The effect of D-lysergic acid diethylamide is discussed with regard to its action on brain serotonergic neurons and its psychotomimetic effects.

Animals↗