Biomedical subjects

E Verney

Publications and source records attributed to E Verney.

At least 37 records · Page 2Linked to original sources

PubMed1992

Effect of benzodiazepines on tryptophan binding to rat hepatic nuclei.

This study evaluates whether or not some of the benzodiazepines would influence the binding of L-tryptophan to rat hepatic nuclei or nuclear envelopes. Previous publications have indicated that binding of L-tryptophan to hepatic nuclear envelope proteins was saturable, stereospecific, and of high affinity. In this study, we investigated whether some of the benzodiazepines would influence L-tryptophan binding to rat hepatic nuclei or nuclear envelopes as assayed by in vitro L-(5-3H) tryptophan binding. Our results indicate that the addition of chlordiazepoxide, diazepam, prazepam, flurazepam, nordazepam, N-desalkylflurazepam, temazepam, oxazepam, lorazepam, or 4-chlorodiazepam has little influence on the L-(5-3H) tryptophan binding to hepatic nuclei in vitro. However, the addition of demoxepam, the N-desalkylated compound of chlordiazepoxide, caused marked competition with 3H-tryptophan binding to hepatic nuclei in vitro. When chlordiazepoxide (1 mg/100 g body weight) is administered intraperitoneally 20 min before killing, the isolated hepatic nuclei reveal decreased specific L-tryptophan binding compared to controls. Also, rats pretreated with chlordiazepoxide intraperitoneally before tube-feeding L-tryptophan revealed diminished tryptophan-induced hepatic nuclear RNA efflux and protein synthesis. Our results suggest that chlordiazepoxide, possibly by itself or through a metabolite, can act to affect hepatic nuclear binding of L-tryptophan and to inhibit the stimulatory effect of L-tryptophan on hepatic protein synthesis.

Animals↗

PubMed1990

Comparison of effects of L-tryptophan and a tryptophan analog, D,L-beta-(1-naphthyl)alanine, on processes relating to hepatic protein synthesis in rats.

Earlier studies reported that the administration of L-tryptophan increased polyribosomal aggregation, protein synthesis and levels of cytoplasmic poly(A) mRNA in rat liver. This study was concerned with the effects of an L-tryptophan analog, D,L-beta-(1-naphthyl)alanine, in comparison with those of L-tryptophan. Both D,L-beta-(1-naphthyl)alanine and L-tryptophan bound to the L-tryptophan receptor protein and increased poly(A)polymerase and nucleoside triphosphatase activities of hepatic nuclei. However, only L-tryptophan was associated with increases in the release of labeled nuclear RNA (in vitro), in protein synthesis, in polyribosomal aggregation and in glycosylation ([14C]glucosamine incorporation into proteins) of rat liver. These results indicate that although D,L-beta-(1-naphthyl)alanine affected hepatic nuclei (binding and enzyme levels), it did not stimulate nucleocytoplasmic translocation of mRNA and concomitant protein synthesis, as did L-tryptophan.

Alanine↗

PubMed1989

Effect of feeding a choline-deficient diet on the hepatic nuclear response to tryptophan in the rat.

In earlier studies the acute administration of tryptophan (TRP) to rats was reported to induce enhanced in vivo [14C]orotate-labeled hepatic nuclear RNA release in vitro. This change was considered to possibly be related to the induction of more and larger gamma-glutamyl transpeptidase-positive foci in the livers of rats treated with diethylnitrosamine and fed long-term elevated TRP in a choline-supplemented (CS) but not in a choline-deficient (CD) diet (comparisons with respective controls). In this study we investigated whether feeding a CD compared to a CS diet for 1 week would affect selected hepatic nuclear responses to TRP. Rats fed the CS but not the CD diet and tube-fed TRP 10 min before being killed revealed enhanced labeled hepatic nuclear RNA release in vitro. In all experiments, comparisons were made with the control groups (rats fed the CS or stock diet). When rats were fed elevated TRP (2%) in the diets (CS or CD) for 1 week, labeled hepatic nuclear RNA release was increased with the CS + TRP but not with the CD + TRP diet group. [3H]TRP binding to hepatic nuclei in vitro revealed no change in the CS + TRP group, decreased in the CD group, and markedly increased in the CD + TRP group in comparison with the control (CS) group. Hepatic nuclear nucleoside triphosphatase activity was increased only in the CS + TRP group while hepatic nuclear poly(A) polymerase activity was increased in the CS + TRP and in the CD +/- TRP groups. Serum cholesterol and triglycerides were decreased in the CD group and increased to control levels in the CD + TRP group.

Animals↗

PubMed1988

Nuclear envelope glycoprotein with poly(A) polymerase activity of rat liver: isolation, characterization, and immunohistochemical localization.

A protein with poly(A) polymerase activity has been identified and isolated from hepatic nuclear envelopes of rats to near homogeneity. The ability of the enzyme to bind to concanavalin A-agarose and to be eluted from the column with methyl alpha-D-mannopyranoside (0.2 M) as well as the inhibitory effects of alpha-mannosidase suggested that it was a glycoprotein. Poly(A) polymerase has an absolute requirement for a divalent cation, ATP, and an oligonucleotide primer. The enzyme activity with Mn2+ was about 20-fold higher than that with Mg2+. Several known inhibitors adversely affected poly(A) polymerase activity. The enzyme has a molecular weight of 64,000 when analyzed by polyacrylamide gel electrophoresis under denaturing conditions and has a sedimentation coefficient of 4.5 S. Immunohistochemical studies using polyclonal antibodies raised against the purified enzyme revealed that the antigen was localized in the nuclear membranes.

Adenosine Triphosphate↗

PubMed1988

Poly(A) polymerase activity in murine serum. Elevation in animals with proliferative changes.

Sera of normal rats contain polynucleotide adenylyltransferase [poly(A) polymerase] activity. The enzymatic activity has been optimized with regard to primer concentration, ion requirements, kinetics, and protein. Results based on inclusion of inhibitors in the assay system show that the enzyme is poly(A) polymerase. High levels of the enzymatic activity were prevalent in sera of (a) BUF/SimfBR rats bearing sc transplanted hepatomas; (b) Sprague-Dawley rats with hepatoma cells grown in ascites; (c) partially hepatectomized Sprague-Dawley rats; and (d) MRL/lpr mice, which are in a massive lymphoproliferative autoimmune state.

Animals↗

PubMed1988

Identification and immunohistochemical localization of a tryptophan binding protein in nuclear envelopes of rat liver.

A tryptophan binding protein which was identified by binding studies has been purified to apparent homogeneity from rat liver nuclear envelopes. Two affinity matrices, namely, concanavalin A-agarose and tryptophan-agarose, were utilized for purification of the binding protein. Findings with lectin affinity chromatography suggested that the binding entity was a glycoprotein since it could be eluted off the column with methyl alpha-D-mannopyranoside (0.2 M). Eluates from both columns, when electrophoresed separately (under denaturing conditions) on polyacrylamide gels, revealed the presence of a protein with an apparent molecular weight of approximately 33,000-34,000 which is the same as that observed when covalently bound (i.e., crosslinked) [3H]-tryptophan is analyzed on polyacrylamide gels under denaturing conditions and then autoradiographed. Polyclonal antibodies raised against the binding protein recognized polypeptides with molecular weights of 64,000 and 33,000-34,000 when analyzed by the Western blot technique, suggesting that the protein was probably a dimer. Immunohistochemical studies revealed that the antigen is localized in the nuclear membranes, thereby corroborating our biochemical premise that the binding protein was present in the nuclear envelopes.

Animals↗

PubMed1988

Effect of tryptophan on toxic cirrhosis induced by intermittent carbon tetrachloride intoxication in the rat.

The effects of the administration of tryptophan on toxic cirrhosis induced by intermittent carbon tetrachloride (CCl4) intoxication in the rat were investigated. Rats received CCl4 (0.45 ml/100 g body wt ip) twice weekly for 10-14 weeks. Tryptophan (30 mg/100 g body wt) by stomach tube was administered 1 hr before killing. Tryptophan improved hepatic polyribosomal aggregation and [14C]leucine incorporation into protein in vitro of control rats as well as long-term CCl4-treated rats that had developed toxic cirrhosis. However, the effects were more marked in control than in experimental rats. Tryptophan administration induced an increase in labeled nuclear RNA release in vitro and a decrease in labeled tryptophan binding to nuclear protein in vitro of livers of rats receiving long-term CCl4 and of control rats. The results indicate that the stimulatory effects of a single administration of tryptophan in toxic cirrhotic livers are similar to, but somewhat less than, those which occur in livers of normal, control rats.

Animals↗

PubMed1988

Effect of tryptophan on livers of pregnant and lactating rats and their fetuses and pups.

The effect of the administration of L-tryptophan on hepatic polyribosomes and protein synthesis in pregnant rats and their fetuses and in lactating rats and their pups was investigated. Pregnant rats tube-fed tryptophan 1 hr before killing revealed increased hepatic protein synthesis but essentially unmodified polyribosomal aggregation of maternal livers while no changes were observed in fetal livers in comparison to controls (water-treated). Lactating rats tube-fed tryptophan 1 hr before killing revealed increased polyribosomal aggregation and protein synthesis of the livers in comparison to controls. Pups of these mothers that received tryptophan intraperitoneally 1 hr before killing did not reveal a significant change in the hepatic polyribosomes or protein synthesis.

Animals↗

PubMed1987

Effect of tryptophan on isolated hepatocytes of rats.

The addition of tryptophan to adult rat hepatocyte cultures stimulated DNA synthesis. The increase in DNA synthesis as measured by 3H-thymidine incorporation into DNA was observed on treatment of the cultures with tryptophan for 48 h but also as short as for 6 h in comparison with control cultures. An increase was also apparent at 30 h which was maintained for up to 48 h post treatment with tryptophan. The increase in DNA synthesis by tryptophan cannot be attributed to cell injury or to increased DNA degradation. Of the degradative enzymes added after harvesting the hepatocytes, only DNase decreased incorporation of 3H-thymidine. The observed effect was specific for tryptophan since treatment with kynurenine, isoleucine, methionine or serine failed to show a significant effect. Pretreatment of cultured hepatocytes with hydroxyurea prevented the tryptophan stimulated increase in DNA synthesis suggesting that the latter was due to replicative and not to reparative DNA synthesis. Experiments performed with the addition of diethylnitrosamine also alluded to tryptophan's role in replicative DNA synthesis. The mechanism of tryptophan-induced DNA synthesis is discussed.

Animals↗

PubMed1987

Influence of level of dietary protein on tryptophan-induced promotional activity in induction of gamma-glutamyl transpeptidase-positive foci of rat liver.

The influence of varying the dietary protein content on the emergence of gamma-glutamyl transpeptidase (GGT)-positive foci in the livers of male rats fed elevated (2%) L-tryptophan (TRP) after being exposed to a hepatocarcinogen was investigated. Subtotal hepatectomies were performed, and 18 hr later the rats were treated with diethylnitrosamine (30 mg/kg). Ten days later four dietary groups were followed for 10 weeks: (1) control diet containing 21% protein (C); (2) control diet containing 5.3% protein (C-LP); (3) C + TRP; and (4) C-LP + TRP. Rats fed the C-LP diet developed heavier livers but fewer and smaller GGT + foci than did rats fed the C diet. Rats fed elevated TRP diets (C + TRP and C-LP + TRP) developed more and larger GGT + foci than did rats fed the regular control diets (C and C-LP), indicating that the promotional effect of elevated dietary TRP was similar at the two levels of dietary protein.

Animals↗

PubMed1986

Effects of varying fat content of a high tryptophan diet on the induction of gamma-glutamyltranspeptidase positive foci in the livers of rats treated with hepatocarcinogen.

The influence of varying the dietary fat content on the emergence of gamma-glutamyltranspeptidase (GGT)-positive foci in the livers of male rats fed elevated (2%) L-tryptophan (TRP) and exposed to a hepatocarcinogen was investigated. Subtotal hepatectomies were performed, and 18 h later the rats were treated with diethylnitrosamine (DEN) (30 mg/kg). Ten days later 4 dietary groups were followed for 10 weeks: (1) control diet containing 15% fat (C-HF); (2) control diet containing 5% fat (C-LF); (3) C-HF + TRP; (4) C-LF + TRP. Rats fed elevated TRP diets (C-HF + TRP and C-LF + TRP) similarly developed more and larger GGT+ foci than did rats fed the regular TRP diets (C-HF and C-LF), indicating that the promotional effect of elevated dietary TRP was not affected by the fat level (15% vs. 5%).

Animals↗

PubMed1986

The influence of hypertonic NaCl on nucleocytoplasmic translocation of RNA in the rat liver.

Hypertonic NaCl administered to rats or mice has been demonstrated to induce in the liver a rapid disaggregation of polyribosomes and inhibition of protein synthesis. This study was concerned with whether hypertonic NaCl would affect nucleocytoplasmic translocation of RNA in the livers of rats. The effect of tube-feeding a hypertonic (10.7%) NaCl solution (321 mg in 3 ml/100 g body wt) for 10 minutes on in vitro release of 14C-orotate-labeled nuclear RNA was assayed. Although the combination of nuclei and cytosols of livers of hypertonic NaCl-treated rats revealed diminished in vitro labeled nuclear RNA release in comparison with hepatic nuclei and cytosols of control (water-treated) rats, each of the two components varied in activity. Even though the overall effect was an inhibitory one, cytosols of livers of hypertonic NaCl-treated rats stimulated in vitro release of labeled nuclear RNA, whereas nuclei of livers of hypertonic NaCl-treated rats revealed diminished in vitro release of labeled nuclear RNA in comparison with cytosols and nuclei of livers of control rats. The stimulatory effect of the hepatic cytosols of the hypertonic NaCl-treated rats was essentially unaffected by pretreatment of the rats with puromycin or cycloheximide, but was abolished by pretreatment of the cytosols in vitro with alpha-mannosidase or beta-galactosidase. Passage of cytosols of control and experimental livers through concanavalin A-agarose columns concentrated the activities of the eluates in stimulating in vitro labeled nuclear RNA release. In vivo 14C-orotate labeling of hepatic nuclear RNA for 30 minutes was increased by hypertonic NaCl treatment in comparison with water treatment of control animals. In vivo 14C-glucosamine incorporation into hepatic proteins of nuclei and nuclear envelopes was increased in hypertonic NaCl-treated rats in comparison with controls. In vitro 3H-tryptophan binding to proteins (trichloracetic acid-precipitable) to cytosols of livers of hypertonic NaCl-treated rats was increased in comparison with binding of controls. The results suggest that the administration of hypertonic NaCl rapidly leads to a change in hepatic cytosol whereby the activity to stimulate in vitro labeled nuclear RNA release is enhanced. This occurs without new protein synthesis, and the effect is probably mediated through a glycoprotein. In contrast, the hepatic nuclei of the rats treated with hypertonic NaCl show a decreased ability to release in vitro labeled nuclear RNA, possibly because of the development of a nuclear lesion.

Animals↗

PubMed1986

Evidence for the role of glycosylation of proteins in the tryptophan-induced stimulation of nucleocytoplasmic translocation of mRNA in rat liver.

Tryptophan induces a rapid stimulation of polyribosomal aggregation and protein synthesis in the livers of rats or mice. This stimulatory effect of tryptophan correlates with increased levels of mRNA in the cytoplasm of the liver and is related to enhanced translocation of nuclear mRNA into the cytoplasm. In the present study the possible role of glycoproteins of the rat liver, particularly those in the nuclear membranes and in the cytosol, in relation to the effect of tryptophan on enhanced nucleocytoplasmic translocation of mRNA was investigated. In the standard treatment regimen, tryptophan (30 mg/100 gm of body weight) was administered by stomach tube 10 minutes before killing. The following results were noted in the liver: Administration of tryptophan increased [14C]glucosamine incorporation into proteins of the subcellular fractions, particularly those of the soluble, nucleus and nuclear membrane. In vivo treatment of rats with tryptophan increased the in vitro [14C]orotate-labeled nuclear RNA release and [3H]tryptophan binding to proteins of the nuclei or cytosols. Treatment of cytosol proteins with alpha-mannosidase or beta-galactosidase, but not with neuraminidase, inhibited these increases. After treatment of cytosol fractions on a concanavalin A agarose column, the effluents showed decreased activity and the eluates showed increased activity in in vitro nuclear RNA release and in tryptophan binding to proteins of the control and, particularly, of the tryptophan-treated animals. Administration of tryptophan increased the polyribosomal aggregation, the in vitro [14C]leucine incorporation into protein, and the in vitro nuclear RNA release by cytosol. However, tunicamycin pretreatment of rats prevented these increases due to tryptophan. The data of this study suggest that glycosylated proteins, one or more, are the active components whereby tryptophan acts upon the liver.

Animals↗

PubMed1985

Hepatic ornithine decarboxylase activity in the rat as influenced by retinyl acetate and ethanol or tryptophan.

Ethanol and tryptophan have been demonstrated earlier to induce a rapid stimulation of hepatic ornithine decarboxylase (ODC) activity in overnight-fasted rats. In this study the effect of the administration of retinyl acetate prior to administering ethanol or tryptophan was investigated. The levels of ODC activity in the livers of control and experimental rats were assayed in vitro by measuring the release of 14CO2 from DL-[1-14C]ornithine. Intraperitoneal administration of retinyl acetate (1 microgram/100 g body wt) 1 hr before tube feeding ethanol (0.75 g as a 50% solution/100 g body wt) or L-tryptophan (30 mg in 3 ml water/100 g body wt) and 3 hr before killing caused an enhanced stimulation of hepatic ODC activity compared to that when each agent was administered alone. In vitro [14C]leucine incorporation into protein using hepatic microsomes of tryptophan-treated rats with or without retinyl acetate was increased in comparison with that of controls while decreases were observed when using microsomes of ethanol-treated rats with or without retinyl acetate. Although retinyl acetate has been reported earlier to inhibit the stimulation of hepatic ODC activity due to a variety of agents, including some agents known as carcinogens or promoters, it did not act in this manner against the acute administration of ethanol or tryptophan.

Animals↗

PubMed1985

Protective effect of tryptophan and cysteine against carbon tetrachloride-induced liver injury.

The effects of the administration of tryptophan and/or cysteine on carbon tetrachloride (CCl4)-induced hepatic injury were investigated. Rats received CCl4 (1 ml/kg ip) followed 6 hr later by tryptophan (300 mg/kg) and/or cysteine (950 mg/kg) via stomach tube and rats were killed after 24 hr. Treatment with tryptophan, cysteine, or both reduced the degree of hepatic necrosis observed histologically. While CCl4 caused polyribosomal disaggregation and decreased [14C]leucine incorporation into liver proteins in vitro and in vivo, treatment with tryptophan, cysteine, or both caused a shift in polyribosomes toward heavier aggregation and protein synthesis was increased. Serum activities of lactic dehydrogenase (LDH), glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, and gamma-glutamyl-transpeptidase were markedly increased after CCl4 alone but after subsequent treatment with cysteine or with tryptophan and cysteine appreciable decreases occurred. Glutathione concentration decreased but total amount remained constant in the livers of CCl4-treated rats while subsequent treatment with cysteine alone or together with tryptophan elevated both levels of glutathione. Using isolated hepatocytes, CCl4 caused decreases in cell viability, in release of LDH, and in [14C]leucine incorporation into protein. Treatment with CCl4 and tryptophan and/or cysteine revealed that cysteine alone or with tryptophan improved cell viability and decreased LDH release of the cells, while tryptophan alone or with cysteine improved protein synthesis. Upon cytologic evaluation, the isolated hepatocytes revealed membrane distortions after CCl4 alone but these were less marked after CCl4 plus tryptophan, cysteine, or both (most improvement). Thus, tryptophan and cysteine act in a beneficial manner against CCl4-induced hepatic injury in the rat.

Animals↗

PubMed1985

Influence of dietary tryptophan on the induction of gamma-glutamyltranspeptidase-positive foci in the livers of rats treated with hepatocarcinogen.

The ingestion of an elevated level (2%) of L-tryptophan (TRP) in a purified diet was investigated to determine whether it would influence the induction of gamma-glutamyltranspeptidase (GGT)-positive foci in the livers of rats exposed to a hepatocarcinogen. Subtotal hepatectomies were performed, and 18 h later, the rats were given injections i.p. of diethylnitrosamine (30 mg/kg). Ten days later, groups of male rats were placed on choline-supplemented (CS), CS + TRP, choline-deficient (CD), or CD + TRP diets for 10 wk. In two separate experiments, the rats fed the CS + TRP diet or the CD diet developed more and larger GGT + foci than did rats fed the CS diet. Rats fed the CD + TRP diet revealed similar changes to those found in rats fed the CD diet. The liver weights of the rats fed the CD or the CD + TRP diet were greater than those of rats fed the CS or the CS + TRP diet. Hepatic GGT activity was somewhat elevated in rats fed the CS + TRP diet and markedly elevated in rats fed the CD or the CD + TRP diet. Hepatic ornithine decarboxylase activity was increased in rats fed the CD + TRP diet. The results suggest that increased dietary tryptophan has a promoting effect on liver carcinogenesis as measured by the induction of GGT + foci in the livers of rats exposed to diethylnitrosamine. A potentiating effect by tryptophan was not observed in the livers of rats fed a CD diet.

Amino Acids↗

PubMed1985

Chemical pathology of diamino acid deficiency: considerations in relation to lysinuric protein intolerance.

Young rats were force-fed a lysine + arginine-devoid diet or a complete diet for 3 days, and selected biochemical and morphologic studies were conducted. Rats force-fed the experimental diet in comparison with those force-fed the control diet for 3 days showed decreased body weight gain, hepatomegaly with periportal fatty liver, pancreatic and splenic atrophy, and enhanced 14C-leucine incorporation into hepatic proteins. Differences in the experimental animals were observed in the free amino acid levels of serum (decreased lysine, arginine, and ornithine) and liver (decreased ornithine), in blood chemistries (decreased levels of ammonia N2, uric acid, cholesterol, protein, albumin, alkaline phosphatase, LDH and SGOT) and in hematologic findings (leukocytopenia and thrombocytopenia after a morning feeding). The experimental findings in young rats force-fed the lysine + arginine-devoid diet were compared with those reported to develop in children with lysinuric protein intolerance (LPI), an autosomal recessive defect in diamino acid transport. Children with LPI as described by others reveal a number of similarities as well as a number of differences in comparison to the findings in the experimental animals. The comparison suggests that some of the pathological manifestations of LPI may be related to a deficiency of diamino acids but others must be due to different alterations in this complex human disease.

Amino Acids↗

PubMed1984

Effect of tryptophan on informosomal and polyribosome-associated messenger RNA in rat liver.

The possible role that informosomal mRNA may play in the stimulation of hepatic protein synthesis induced by the acute administration of tryptophan was investigated. L-tryptophan (30 mg/100 g body weight) tube-fed to rats induced an increase (20%) in the amount of polyribosome-associated poly(A+)mRNA in the liver within 1 hour. On the other hand, the amount of the informosomal poly(A+)- mRNA revealed no significant increase or decrease. The size of the increase in the polyribosome-associated poly(A+)mRNA pool was equal to the entire amount of informosomal poly(A+)mRNA present in the hepatic cells. Therefore it was concluded that the failure to detect a significant decrease in the size of the informosomal mRNA pool indicated that the increase in the polyribosome-associated poly(A+)mRNA must be due to a different mechanism, such as enhanced nuclear-to-cytoplasmic translocation of nuclear poly(A+)mRNA, which had been reported earlier. To determine whether any qualitative or quantitative changes occurred in the RNA sequences along with the increase in poly(A+)mRNA following tryptophan administration, DNA/RNA hybridization studies were conducted by using hepatic polyribosome-associated poly(A+)- mRNA from tryptophan-treated and control rats. Although qualitatively no new species of mRNA were detected in the mRNA from tryptophan-treated rats, kinetic analysis of the hybridization curves indicated that there was a shift or accumulation of hepatic poly(A+)mRNA belonging to the intermediate and possibly the high frequency classes of polyribosome-associated poly(A+)mRNA in the livers of the tryptophan-treated rats.

Animals↗