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PubMed1981

Effect of casein and starch infusion in the large intestine on nitrogen metabolism of growing swine.

Eight crossbred female pigs (40 kg) with cannulae placed in the terminal ileum were used to evaluate the effect of infusions of casein starch and casein plus starch on organic matter fermentation and microbial protein synthesis in the large intestine, and their effect on urinary urea and orotic acid excretion, and on nitrogen (N) retention. Infused casein and starch were both totally digested. Nitrogen retention was increased (P greater than 0.05) when casein was infused. Starch infusion resulted in an increase (P greater than 0.05) in fecal N in the form of total protein (amino acids). The high correlation (P greater than 0.01) between fecal total protein and RNA indicates that the increase in fecal N resulted from an increase in microbial protein synthesis. About 5.2 g of bacterial protein was synthesized per 100 g of cornstarch fermented in the large intestine. Casein infusion increase (P greater than 0.05) total urinary N. Differences between treatments for urinary N were entirely because of changes in urinary urea. Urinary ammonia and unaccounted N were not affected by treatments. Urinary orotic acid was a good indicator of the urea cycle activity because of its hig correlation (P greater than 0.0) with urinary urea. Plasma urea N concentration was increased (P greater than 0.05) only when casein plus starch was infused.

Animals↗
PubMed2000

Indicators of L-arginine metabolism and cardiovascular risk factors--a cross-sectional study in healthy middle-aged men.

This study examines the relationship between traditional risk factors of coronary artery disease and indicators involved in the metabolism of L-arginine (plasma and urine L-arginine, plasma L-citrulline, serum creatinine and urine orotic acid). Our study population consisted of 40 healthy male volunteers aged between 35 and 55 years. We found an inverse association between serum creatinine and blood pressure, between plasma L-citrulline and blood pressure, as well as between urine L-arginine and blood pressure. We also found a positive association between plasma LDL-cholesterol and urine L-arginine and a negative correlation between plasma L-arginine and LDL-cholesterol. Orotic acid measured from urine was not associated with any of the indicators of L-arginine metabolism. Our results indicate that L-arginine metabolism is of profound significance for cardiovascular health. However, our study does not answer questions relating to causality. Further studies are needed to clarify the causal relationship between cardiovascular risk factors, especially elevated blood pressure and high LDL-cholesterol, and indicators of L-arginine metabolism.

Adult↗
PubMed1973

Thymidine kinase and deoxyribonucleic acid metabolism in growing and regenerating livers from rats on controlled feeding schedules.

Rats accustomed to eating during the first 8h of a daily 12h dark period re-established about 80% of intact liver weight, protein and DNA within 4 days following partial hepatectomy; further increases were not observed. Liver thymidine kinase activity and thymidine incorporation into liver DNA exhibited marked daily oscillations during liver regeneration. Maximum values were observed near the end of the dark period both in intact growing rats and in rats partially hepatectomized 2h before the end of the dark period. The time of day of surgery affected thymidine kinase activity and thymidine incorporation into DNA at specific times following partial hepatectomy. This seriously affects the interpretation of reports of experiments where the time of day of killing has been held constant and time of surgery varied. Highly significant correlation coefficients were observed for thymidine incorporation before killing versus thymidine kinase activity at time of killing and for thymidine versus orotic acid incorporation into DNA of livers from rats partially hepatectomized 2h before the end of the dark period and killed at 12h intervals. Thymidylate phosphatase activity returned to the normal amount at a rate similar to that for liver protein. Thymidylate phosphatase did not affect the validity of the thymidine kinase assay. The relationship of [(14)C]orotic acid to [(3)H]thymidine incorporation into liver DNA varied with the time of day, with the age of the rat and during the regeneration of the liver.

Age Factors↗
PubMed1966

Ribonucleic acid synthesis during the early action of thyroid hormones.

1. The effect on RNA synthesis in rat liver of thyroidectomy and the administration of thyroid hormone, especially during its physiological latent period, was studied by determining: (a) the activity of DNA-dependent RNA polymerase in isolated nuclei; (b) the rate of synthesis of nuclear and cytoplasmic RNA in vivo; (c) polyribosomal sedimentation profiles; (d) the response of microsomes and ribonucleoprotein particles to polyuridylic acid; (e) the effect of inhibitors of RNA and protein synthesis on the biological activity of hormones. 2. The DNA-dependent RNA-polymerase activity of isolated rat-liver nuclei was lowered by thyroidectomy and stimulated by the administration of tri-iodo-l-thyronine or l-thyroxine (2-25mug./100g. body wt.) to both normal and thyroidectomized rats. In thyroidectomized rats, the activity of the Mg(2+)-activated RNA-polymerase reaction (for which the product is mainly ribosomal type of RNA) was stimulated at 10-12hr. after a single injection of tri-iodothyronine, reaching a peak value of 60-90% stimulation at 45hr. after hormone administration. The Mn(2+)/ammonium sulphate-activated RNA-polymerase reaction (for which the RNA product is more DNA-like) was not affected for 24hr. after hormone administration but stimulated by 30-40% at 45hr. The response of both RNA-polymerase reactions to the hormone in vivo paralleled the physiological response but the enzyme was not stimulated by the addition in vitro of the hormone to isolated nuclei. 3. Within 3-4hr. after tri-iodothyronine administration to thyroidectomized rats, the specific activity of rapidly labelled nuclear RNA, after a 10min. pulse of [6-(14)C]orotic acid, was 30-40% greater than the control values, the stimulation reaching 100 and 200% at 11 and 16hr. respectively after hormone administration. Longer exposures to [6-(14)C]orotic acid and [(32)P]phosphate showed that the hormone accelerated the synthesis of mitochondrial, microsomal (or ribosomal) and soluble RNA. The greater part of the labelled nuclear RNA was of the ribosomal type. The hormone-induced increases in the incorporation of radioactive precursors into RNA were not preceded, but followed, by enhanced uptake of the precursor. There was no change, per g. of liver, of DNA, nuclear RNA or soluble RNA, but there was a 40-60% increase in the amount of ribosomal RNA between 35 and 45hr. after a single injection of tri-iodothyronine to thyroidectomized rats. 4. Coinciding with the increase in ribosomal RNA after hormone administration was an increase in the average size and amount of polyribosomes. The newly formed ribonucleoprotein particles, or messenger RNA attached to them, or both, were more firmly bound to microsomal membranes after hormone treatment. 5. Polyuridylic acid caused a bigger stimulation of incorporation of [(14)C]phenyl-alanine by ribonucleoprotein particles, but not by microsomes, from thyroidectomized rats as compared with preparations from normal animals. The response of ribonucleoprotein particles to polyuridylic acid was lowered after tri-iodothyronine treatment of thyroidectomized rats. 6. Actinomycin D, 5-fluorouracil, puromycin and cycloheximide caused a 70-100% inhibition of the stimulatory effect of l-thyroxine and tri-iodo-l-thyronine on basal metabolic rate and growth rate in both normal and thyroidectomized animals. Administration of actinomycin D also abolished the stimulation of RNA polymerase by tri-iodothyronine. 7. It is concluded that regulation of nuclear and ribosomal RNA synthesis is an essential step leading to the biological action of thyroid hormones and that the formation of new ribosomes is an important aspect of the control of cytoplasmic protein synthesis by these hormones.

Animals↗
PubMed1978

Turnover of messenger RNA in transplantable hepatomas and host liver of rats.

The turnover of messenger RNA (mRNA) in two intrahepatically transplantable hepatoma (5123 and 19) and host livers of Buffalo rats was evaluated with four different approaches. [14C]Orotic acid incorporation into the rapidly labeled peak between 18S and 4S of total polyribosomal RNA was measured. In vitro RNase assay of [14C]orotic acid-labeled mRNA of polyribosomes was utilized. The decay of mRNA as reflected by disaggregation of free and membrane-bound polyribosomes at intervals after actinomycin D treatment was determined. The incorporation of [14C]orotic acid into polyadenylic acid-mRNA of free and membrane-bound polyribosomes was assayed. The results revealed that the turnover of mRNA of total, free, and membrane-bound polyribosomes was greater in the host livers than it was in the two hepatomas. In host livers the turnover of mRNA of the free polyribosomes was greater than that of the membrane-bound polyribosomes. In the two hepatomas the turnover of mRNA of free polyribosomes was at a similar rate as that of membrane bound polyribosomes. Hepatoma 19, which grow more rapidly and is less differentiated morphologically than is hepatoma 5123, appeared to have a slower turnover of mRNA than did hepatoma 5123. Measurement of RNase activity revealed greater activity in host livers than in hepatomas.

Animals↗
PubMed1997

Molecular cloning of the human UMP synthase gene and characterization of point mutations in two hereditary orotic aciduria families.

Uridine monophosphate (UMP) synthase is a bifunctional enzyme catalyzing the last two steps of de novo pyrimidine biosynthesis, orotate phosphoribosyltransferase (OPRT) and orotidine-5'-monophosphate decarboxylase (ODC). Loss of either enzymatic activity results in hereditary orotic aciduria, a rare autosomal recessive disorder characterized by retarded growth, anemia, and excessive urinary excretion of orotic acid. We have isolated the UMP synthase chromosomal gene from a lambdaEMBL-3 human genomic library and report a single-copy gene spanning approximately 15 kb. The UMP synthase genomic structure encodes six exons ranging in size from 115 bp to 672 bp, and all splicing junctions adhere to the canonical GT/AG rule. Cognate promoter elements implicated in glucocorticoid- and cAMP-mediated regulation as well as in liver-, myeloid-, and lymphocyte-specific expression are located within the 5' flanking sequence. Molecular investigation of UMP synthase deficiency in a Japanese orotic aciduria patient revealed mutations R96G (A-to-G transition; nt 286) and G429R (G-to-C transversion; nt 1285) in one allele and V109G (T-to-G transversion; nt 326) in the other allele. Expression of human UMP synthase cDNAs containing these mutations in pyrimidine auxotrophic Escherichia coli and in recombinant baculovirus-infected Sf21 cells demonstrates impaired activity presumably associated with the urinary orotic acid substrate accumulations observed in vivo. We further establish the identity of two polymorphisms, G213A (v = .26) and 440Gpoly (v = .27) located in exons 3 and 6, respectively, which did not significantly compromise either OPRT or ODC function.

Adult↗
PubMed1990

Carrier detection in a partially dominant X-linked disease: ornithine transcarbamylase deficiency.

Ornithine transcarbamylase (OTC) deficiency is an X-linked disease responsible for lethal neonatal hyperammonemia in males. Partial OTC deficiency also occurs in females and can be responsible for life-threatening hyperammonemic comas in heterozygotes (15%). Increased orotic acid excretion occurs in both symptomatic and asymptomatic carriers, especially under protein loading tests. The disease is therefore partially dominant with neonatal lethality in the hemizygous male; the fraction of new mutations has previously been estimated to be low in males (point estimation = 0, upper bound of the confidence interval = 0.16) and 57% in females. Genetic counseling in this disease is difficult because it is not clear whether a negative protein loading test rules out carrier status. In an attempt to determine how reliable the test is for carrier detection, we investigated ten obligate carriers for orotic acid excretion; considering all data available, we concluded that the test is rarely negative in obligate carriers (8%). Consequently, a negative test in a mother decreases the minimum risk of being a carrier from 84% a priori to 30% if she had an affected son and from 43% a priori to 5% if she had a heterozygous daughter. Finally, the diagnosis of a new mutation in the germ cells of the maternal grandfather in one particular family could be ascertained by extensive DNA analysis.

Bayes Theorem↗
PubMed1993

Pyrimidine nucleotide metabolism in rat hepatocytes: evidence for compartmentation of nucleotide pools.

Pyrimidine nucleotide metabolism in rat hepatocytes was studied by measurement of the labelling kinetics of the various intermediates after double labelling with [14C]orotic acid and [3H]cytidine, the precursors for the de novo and the salvage pathways respectively. For the uridine nucleotides, differences were found for the 14C/3H ratios in the UDP-sugars, in UMP (of RNA) and in their precursor UTP, suggesting the existence of separated flows of the radioactive precursors through the de novo and the salvage pathways. Higher ratios in the UDP-sugars, which are synthesized in the cytoplasm, and a lower ratio in UMP (of RNA) relative to the 14C/3H ratio in UTP indicated that UTP derived from orotic acid is preferentially used for the cytoplasmic biosynthesis of the UDP-sugars. Uridine, derived from cytidine, is preferentially used for the nuclear-localized synthesis of RNA. In contrast to these findings, the 14C/3H ratios in the cytidine derivatives CMP-NeuAc and CMP (of RNA), and in the liponucleotides CDP-choline and CDP-ethanolamine, were all lower than that in the precursor CTP. This indicates a preferential utilization of the salvage-derived CTP for the synthesis of the liponucleotides as well as for RNA and CMP-NeuAc. Similar conclusions could be drawn from experiments in which the intracellular amounts of several uridine- and cytidine-nucleotide-containing derivatives were increased by preincubating the hepatocytes with unlabelled pyrimidine nucleotides or ethanolamine. Based on these data, we propose a refined model for the intracellular compartmentation of pyrimidine nucleotide biosynthesis in which three pools of UTP are distinguished: a pool of de novo-derived molecules and a pool of salvage-derived molecules, both of which are channelled to the site of utilization; in addition an 'overflow' pool exists, consisting of molecules having escaped from channelling. An overflow pool could also be distinguished for CTP, but no discrimination between de novo and salvage-derived molecules could be made.

Animals↗
PubMed1968

Isolation of two distinct classes of polysomes from a nuclear fraction of rat liver.

Isolated rat liver nuclei were washed with Triton-X-100 in the presence of liver cell sap. This treatment liberated a fraction of polysomes which were isolated by differential centrifugation and were designated "outer membrane polysomes." The outer membrane polysomes synthesized protein in vivo. Shortly after injection of orotic acid-(14)C, the RNA of outer membrane polysomes had a higher specific activity than that of cytoplasmic polysomes. It was postulated that outer membrane polysomes may be an intermediate in the transfer of newly synthesized RNA from the nucleus to the cytoplasm. In other experiments, Triton-washed rat liver nuclei were lysed in the presence of deoxycholate and deoxyribonuclease. A ribonucleoprotein fraction was isolated from the lysate by differential centrifugation. This fraction contained "intranuclear ribosomes," which sedimented like partially degraded polysomes in sucrose gradients. This degradation could be partially prevented if intranuclear ribosomes were purified by sedimentation through heavy sucrose. The resulting pellets were termed "intranuclear polysomes" because they contained some undergraded polysomes. Intranuclear polysomes were highly radioactive after a brief pulse with orotic acid-(14)C, but did not appear to synthesize protein rapidly in vivo. Intranuclear polysomes may represent the initial stage of assembly of polyribosomes in the nucleus.

Animals↗
PubMed1966

Light effects on the nucleic acids of excised cotton cotyledons.

The effects of light and glucose in the nutrient medium on the nucleic acid metabolism of excised 8-day cotton (Gossypium hirsutum var. Acala 44) cotyledons were determined. The rates of synthesis as affected by light and glucose were determined by brief exposures to C(14)-labeled orotic acid. The nucleic acids were fractionated by homogenizing in Tris-HCl buffer and centrifuging to obtain soluble and microsomal RNA (20,000 x g supernatant) and a particulate nucleic acid fraction (20,000 x g precipitate) or by extracting in phenol followed by 10% NaCl extraction at 100 degrees . The phenol extract was analyzed by density gradient centrifugation. Light and glucose caused parallel changes in nucleic acid levels of the various fractions, in orotic acid-6-C(14) absorption and in rates of synthesis of nucleic acids. Light and glucose appear to enhance binding of the ribosome nucleic acid so that it becomes less extractable in Tris-HCl buffer or phenol. The bound nucleic acids were labeled at a slightly higher rate than the total nucleic acids extracted by Tris-HCl or phenol. However, light treatment for 48 hours promoted a very high labeling rate in the soluble, low molecular weight nucleic acid as shown by density gradient centrifugation of the phenol extractable fraction.It was concluded that a part of the nucleic acid changes were brought about by light acting through the photosynthetic production of carbohydrate. This conclusion was strengthened by the observation that herbicide inhibitors of photosynthesis and limited atmospheric CO(2) concentrations partially inhibited the nucleic acid changes. However, glucose did not cause changes in nucleic acid levels as large as those caused by light and changes were observed to occur in light even though the endogenous sugar levels were maintained at a low level by the inhibition of photosynthesis with herbicides. The data indicated that light may produce changes in nucleic acid levels by other mechanisms additional to those regulating the sugar level in the tissue.

Gossypium↗
PubMed1968

Polyamines and nucleic acid metabolism in chick embryo. Incorporation of labelled precursors into nucleic acids of subcellular fractions and polyribosomal patterns.

1. An increase in polyamine concentration, caused by inhibiting the amine oxidase activities with iproniazid, increased the incorporation of [(3)H]orotic acid into chick-embryo RNA and DNA. On the other hand, a decrease in polyamine concentration, obtained by causing an increase in amine oxidase activities, decreased [(3)H]orotic acid incorporation into nucleic acids. This was particularly evident for nuclear DNA and ribosomal RNA. 2. Polyribosomal patterns obtained by sucrose-density-gradient centrifugation showed highest radioactivity in the regions of 259s and 280s aggregates in those embryos in which the polyamine contents were enhanced, whereas a decrease in the radioactivity was observed when the polyamine concentrations were decreased. 3. The activity of DNA-dependent RNA polymerase, assayed in the same experimental conditions, also varied in the same fashion with changes in polyamine concentration.

Journal Article↗
PubMedDate not supplied

[Effect of the antineoplastic substance brotheophine on the insertion of precursors into nucleic acids and proteins from Plisse lymphosarcoma].

The effect of antitumor medicine Brotheophine to insertion of the marked precursors into DNA (14[C]-timidine), RNA (3[H]-orotic acid) and proteins (14[C]-leucine) of tumor cells (Pliss lymphosarcoma) as the indices of biosynthetic processes was investigated. It has been shown that Brotheophine acts as an antimethabolite of purine exchange and inhibits the vertical genetic information transfer in tumor cell (DNA-RNA-protein). Namely, a significant inhibition of 14[C]-timide insertion to DNA has been observed. less distinct is the inhibition of 3[H]-orotic acid to RNA und 14[C]-leucine to proteins. The above revealed allows to assume that during Brotheophine treatment certain changes in DNA biosynthesis take place leading to synthesis of slightly transformed RNA with subsequent impairment of proteins synthesis.

Animals↗
PubMed1978

[Isolation and genetic study of Aspergillus nidulans mutants defective in pyrimidine biosynthesis].

8 uridine-requiring pyr mutants were isolated from Aspergillus nidulans under nitrosoguanidine treatment. All the mutants are capable to grow on the medium containing 20 mkg/ml of uridine or cytidine, or 100 mkg/ml of uracil, and they do not utilize thymidine, thymine, cytosine and deoxyuridine. Their ability to grow in the presence of orotic acid demonstrates that the pyrimidine synthesis in all the mutants is blocked at stages preceding the conversion of orotic acid into orotidine monophosphate. All the pyr mutants are of nuclear nature, they are recessive and represent three complementation groups located in the VIII chromosome. Unlike U. maydis mutant, the requirement in pyrimidines does not increase the sensitivity of A. nidulans pyr mutants to UV-irradiation.

Aspergillus nidulans↗
PubMed1971

Pyrimidine metabolism in cotyledons of germinating alaska peas.

Cotyledons from Pisum sativum L. cv. Alaska seeds were excised 12, 36, 108, 132, and 156 hours after imbibition in aerated distilled water. They were then incubated under aseptic conditions for 6 hours in solutions containing either uridine-2-(14)C or orotic acid-6-(14)C. Uridine was more extensively degraded to (14)CO(2) at all germination stages than was orotate, and these rates remained essentially constant at each stage. Incorporation of each compound into RNA increased about 2-fold from the 12th to the 156th hour, although the total RNA present decreased slightly over this interval. Paper chromatography of soluble labeled metabolites produced from orotate showed that the capacity to metabolize this pyrimidine increased markedly as germination progressed. Radioactivity in uridine-5'-P, uridine diphosphate-hexoses, and uridine diphosphate increased most, while smaller or less consistent increases in uridine, uracil, uridine triphosphate, and an unidentified UDPX compound were also observed. The data suggest that orotate metabolism was initially limited by orotidine-5'-phosphate pyrophosphorylase or by 5-phosphoribosyl-1-pyrophosphate. Incorporation of uridine into RNA appeared to be limited at the earliest germination periods by conversion of uridine-5'-P to uridine diphosphate. Thus, during the 1st week of germination the orotic acid pathway and a salvage pathway converting uridine into RNA become activated.

Journal Article↗
PubMed2003

Crystallization and preliminary X-ray crystallographic analysis of orotate phosphoribosyltransferase from Helicobacter pylori.

Orotic acid phosphoribosyltransferase (PyrE) (EC 2.4.2.10) is a key enzyme in de novo uridine monophosphate (UMP) biosynthesis. It catalyzes the reaction between orotic acid and 5-phosphoribosyl-1-pyrophosphate (PRPP) to yield orotidine monophosphate (OMP), which is transformed to uridine monophosphate by decarboxylation. H. pylori PyrE was crystallized at 294 +/- 1 K by the hanging drop vapor-diffusion method. The crystals belong to the space group P2(1)2(1)2(1) with unit-cell dimensions a = 95.8, b = 104.9, c = 281.1 A, alpha = beta = gamma = 90 degrees. A set of diffraction data was collected to 3.29 A resolution using synchrotron X-ray radiation.

Crystallization↗
PubMed1989

Alterations in polyadenylic acid-containing messenger RNA synthesis of brain polysomes after seizures of seizure-susceptible E1 mice.

The effect of seizures on synthesis of the polyadenylic acid (poly(A]-containing messenger RNA (mRNA) isolated from brain polysomes in a genetically seizure-susceptible E1 mouse was studied in vivo. The seizure in the E1 mice was induced by tossed-up stimulation. Immediately after the seizure ceased, the labeled orotic acid was injected into the brain. The incorporation rates of labeled orotic acid into poly(A)-containing mRNA isolated from polysomes are represented as the specific radioactivity (SR) (dpm/mg RNA) and the relative specific radioactivity (RSR) (dpm/mg RNA/dpm/mumoles of acid soluble uridine-5'-monophosphate (UMP]. Both the rates were reduced to 70% in SR and 65% in RSR at 1 h after the seizures. This reduction was gradually recovered to the level of interictal E1 mice at 6 h. The seizure-induced alterations are not attributable to the difference in the uridine nucleotide pool because the SR of UMP was not significantly affected by the seizure. The peak of labeled poly(A)-containing mRNA by analysis of gel electrophoresis displaced towards a lower molecular weight at 1 h after the seizures. The RNA showed a higher ratio of AMP and UMP per GMP and CMP in nucleotide composition, implying that this RNA is identical with DNA. These results suggest that the temporary decrease found in cytoplasmic mRNA synthesis induced by the seizures of E1 mice appears to be a result of impaired transcriptional processes in heterogeneous nuclear RNA synthesis and that the smaller mRNA coding for protein associated with seizures is newly synthesized during the postictal period.

Animals↗
PubMed1988

Excess crude protein for nongravid gilts.

Twenty-five nongravid crossbred gilts (avg initial wt, 126 kg) were placed on either a high (38%) or a low (13%) crude protein (CP) diet and fed either at the rate of 1.82 kg/d or had ad libitum access to feed. In addition, a fifth group was pair-fed the 13% CP diet to the average intake of the gilts fed high CP ad libitum. The experimental period lasted 30 d. Corn-soybean meal diets were used and CP levels were varied by altering the corn:soybean meal ratio. Gain and gain/feed were reduced (P less than .01) in gilts fed 1.82 kg/d compared with the gilts fed ad libitum or pair-fed gilts. Gain (P less than .03) and feed intake (P less than .01) of gilts with ad libitum access to the 13% CP diet were higher than those of gilts with ad libitum access to the 38% CP diet. Gain/feed was not different (P greater than .10) between the two groups, however. Rate of gain and feed efficiency of gilts pair-fed the 13% CP diet were similar (P greater than .10) to those of gilts with ad libitum access to the 38% CP diet. Plasma total free amino acids, NH3 and total protein were not (P greater than .10) affected by treatment. Plasma urea-N and urinary total N, urea-N and orotic acid were increased (P less than .01) in gilts fed the high CP diet regardless of feed intake level. However, urinary NH3 was higher (P less than .01) in gilts fed the low-protein diet. These results indicate that excess dietary CP for nongravid gilts decreases gain and feed intake and has no effect on efficiency of feed utilization, but it increases plasma urea-N and urinary total N, urea-N and orotic acid.

Amino Acids↗
PubMed1996

Inhibition of liver RNA breakdown during acute inflammation in the rat.

Liver RNA- and protein-degradation rates were measured after the induction of acute inflammation in the rat. A preliminary study determined changes in hepatic RNA and protein content 12, 18 and 24 h after a turpentine oil injection. The RNA content in turpentine-treated rats compared with pair-fed animals increased significantly and sharply from 12 h (+ 11%) to 18 h (+ 32%) and slightly thereafter (+ 37% at 24 h). The liver protein content was significantly enhanced only at 24 h (+ 11%) in response to inflammation. RNA-degradation rates were determined in livers perfused cyclically in situ for 15 min by measuring the accumulation of radioactive cytidine in the medium 60 h after in vivo labelling of RNA by [5-3H]cytidine instead of [6-14C]orotic acid, the most commonly used radioactive marker. Several validation procedures showed that the method employed was a valid alternative to the use of radioactive orotic acid. RNA-degradation rates, which mainly reflect rRNA breakdown, were significantly lower in the turpentine-treated rats than in respective pair-fed animals at 18 and 24 h (57 and 45% decrease respectively). Proteolysis rates measured at 24 h together with RNA breakdown by valine accumulation in the perfusion medium were not modified after turpentine treatment. The main factors known to regulate RNA degradation (amino acids, insulin/glucagon ratio) were measured in the portal blood 24 h after induction of acute inflammation. Of the known regulatory amino acids, only glutamine and to a lesser extent methionine were increased in the turpentine-treated rats as compared with their pair-fed counterparts. The insulin/glucagon molar ratio was similar in both groups. In conclusion, the reduced breakdown of RNA, especially rRNA, is largely responsible for the accumulation of hepatic RNA during acute inflammation. This inhibition of RNA degradation could possibly be related to the increase in glutamine.

Amino Acids↗