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PubMed1968

Hybridizable ribonucleic acid of rat brain.

1. Cerebral RNA of adult and newborn rats was labelled in vivo by intracervical injection of [5-(3)H]uridine or [(32)P]phosphate. Hepatic RNA of similar animals was labelled by intraperitoneal administration of [6-(14)C]orotic acid. Nuclear and cytoplasmic fractions were isolated and purified by procedures involving extraction with phenol and repeated precipitation with ethanol. 2. The fraction of pulse-labelled RNA from cerebral nuclei that hybridized to homologous DNA exhibited a wide range of turnover values and was heterogeneous in sucrose density gradients. 3. Base composition of the hybridizable RNA was similar to that of the total pulse-labelled material; both were DNA-like. 4. Pulse-labelled cerebral nuclear RNA hybridized to a greater extent than cytoplasmic RNA for at least a week after administration of labelled precursor. This finding suggested that cerebral nuclei contained a hybridizable component that was not transferred to cytoplasm. 5. The rates of decay of the hybridizable fractions of cerebral nuclei and cytoplasm were faster in the newborn animal than in the adult. Presumably a larger proportion of labile messenger RNA molecules was present in the immature brain. 6. Cerebral nuclear and cytoplasmic RNA fractions from newborn or adult rats, labelled either in vivo for periods varying from 4min. to 7 days or in vitro by exposure to [(3)H]-dimethyl sulphate, uniformly hybridized more effectively than the corresponding hepatic preparation. These data suggested that a larger proportion of RNA synthesis was oriented towards messenger RNA formation in brain than in liver.

Animals↗
PubMed1975

Age-dependent changes of uridine nucleotide and RNA metabolism in the brain of normal and hypothyroid rats.

The effects of neonatal thyroidectomy on the incorporation in vivo of labelled orotic acid into acid-soluble uridine nucleotides, nuclear RNA and microsomal RNA of the rat brain were studied at 10 and 30 days of age. It was found that the conversion of orotate into uridine nucleotides is high at the earliest stage and does not change upon maturation. At any stage, the neonatal lack of thyroid function does not effect the conversion of orotate into uridine nucleotides. Neonatal thyroidectomy led to a significant decrease in the synthesis of cerebral RNA only at 10 days after birth.

Age Factors↗
PubMed1973

The effects of diethylnitrosamine on ribonucleic acid and protein synthesis in the liver and lung of the Syrian golden hamster.

1. Syrian golden hamsters were treated with a single subcutaneous dose of 200mg of diethylnitrosamine/kg. In the liver the treatment produced a significant and early inhibition of the incorporation of orotic acid into RNA and of leucine into protein. Diethylnitrosamine also lowered basal and 20-methylcholanthrene-stimulated activities of hepatic aryl hydrocarbon hydroxylase. 2. RNA synthesis, protein synthesis and aryl hydrocarbon hydroxylase activity were also evaluated in the lungs of the same animals. In this organ only protein synthesis was affected by diethylnitrosamine, but not RNA synthesis or aryl hydrocarbon hydroxylase activity. 3. The incorporation of thymidine into DNA was inhibited in both organs early after diethylnitrosamine treatment and increased 2-3 days later. 4. Although diethylnitrosamine, injected subcutaneously, accumulates in liver and lung in toxicologically active amounts, the acute biochemical responses of the two organs are not identical.

Animals↗
PubMed1979

Selective lesions of acinar pancreatic cells in rats poisoned with abrin. A morphological and biochemical study.

Rats poisoned with abrin (2.5 micrograms/100 g body weight) died within 36 h with severe necrosis of acinar pancreatic cells. Incorporation in vivo of labelled amino acids into pancreatic protein was greatly impaired 6 h after poisoning. Microsomes isolated from the pancreas of poisoned rats at 6 h had a reduced capacity for protein synthesis in vitro. Incorporation in vivo of orotic acid into pancreatic RNA was decreased 12 h after poisoning.

Abrin↗
PubMed1975

Effect of narcotic drugs on ribonucleic acid and nucleotide metabolism in mouse brain.

Mice either were administered 10, 30 or 100 mg/kg of morphine sulfate acutely or were chronically implanted with pellets containing morphine, naloxone or pentazocine. They were then injected intraperitoneally or intracerebroventricularly with [5-3H] uridine or [5-3H] orotic acid either 30 minutes, 24 hours of 48 hours before sacrifice. The incorporation of the 3H into brain total homogenate, ribonucleic acid (RNA) and uridine nucleotides was measured. The RNA content of brain and liver was also assayed. When [3H] uridine was injected. i.p. 30 minutes before sacrifice, acute injection of 30 or 100 mg/kg of morphine sulfate or chronic implantation of morphine pellets decreased the incorporation of the [3H] uridine into brain RNA. However, neither the acute administration of 10 mg/kg of morphine sulfate, nor the chronic administration of naloxone or pentazocine, altered the amount of radioactivity incorporated into RNA. Chronic morphine treatment decreased the incorporation of 3H into uridine nucleotides and nucleotide sugars due partially to increased catabolism of the [3H] uridine. The brain and liver RNA concentration was unchanged by chronic morphine administration. Thus, chronic morphine treatment alters the metabolism of uridine nucleotides but does not appear to alter the net synthesis of the total brain RNA.

Animals↗
PubMed1984

Failure of protein loading tests to identify heterozygosity for ornithine carbamoyltransferase deficiency.

Protein loading tests for the diagnosis of heterozygous ornithine carbamoyltransferase deficiency were performed on two occasions on an asymptomatic woman whose daughter and two infant sons died of the disease. Neither loading test produced the expected increases in urinary orotic acid excretion and studies of other pyrimidine and purine metabolites in urine and plasma did not suggest that these would provide better discrimination from non-carriers. The results probably reflect an extensive inactivation of the mutant X chromosome in liver cells and reinforce the need for caution in interpreting negative test results.

Adult↗
PubMed1992

Normal N-acetylglutamate concentration measured in liver from a new patient with N-acetylglutamate synthetase deficiency: physiologic and biochemical implications.

N-Acetyl-L-glutamate synthetase (NAG synthetase) is a mitochondrial matrix enzyme which catalyzes the synthesis of N-acetyl-Lglutamate (NAG), a physiologic activator of the urea cycle enzyme carbamylphosphate synthetase I. Deficiency of NAG synthetase in humans has been reported only three times previously. Two cases presented with uncontrolable neonatal hyperammonemia leading to death, while a third child presented with hyperammonemia and a neurodegenerative picture at 15 months of age after previously being healthy. We report here a new case of NAG synthetase deficiency who presented at 4 years, 10 months of age with an episode of hyperammonemia. Diagnosis was made at age 5 years, 6 months when a liver biopsy showed 9.7% of normal activity. Urine orotic acid was low, and total NAG content in liver was normal. Liver pathology revealed micro- and macrovesicular fat and mitochondria of irregular size and shape with intracristae crystallizations. NAG content in liver in patients with NAG synthetase deficiency has not previously been reported. Its normal value in the face of NAG synthetase deficiency suggests an abnormal localization of NAG to the cytoplasm and the likelihood of aberrant cytoplasmic synthesis of this compound. Additional physiologic implications of this speculative abnormal compartmentalization are discussed.

Acetyltransferases↗
PubMed1982

[Hyperammonemia: a suggestion for diagnostic and therapeutic procedures].

Hyperammonemia in pediatrics leads to emergency situations. Adequate decisions for treatment have to be taken rapidly. These depend on the underlying disorder. While the patient's history and symptoms should lead to a search for hyperammonemia, the further steps will depend on biochemical results of aminoacid determinations in plasma and urine, and orotic acid excretion. In some cases where this latter metabolite is not increased, the determination of organic acids in urine and enzyme assays in liver are needed. A scheme for the diagnostic and therapeutic measures is proposed.

Amino Acids↗
PubMed1975

Effects of gamma-irradiation on biosynthesis of different types of ribonucleic acids in normal and regenerating rat liver.

1. The effect of gamma-irradiation (4000rd) on the synthesis of ribosomal (pre-rRNA) and heterogeneous nuclear RNA (pre-mRNA) in normal and in regenerating rat liver was studied by using 40 min labelling with [6(-14)C]orotic acid. 2. Partial hepatectomy caused a sharp transient increase in the specific radioactivity of the endogenous low-molecular-weight RNA precursors in the livers of both normal and irradiated rats. Irradiation of intact animals did not affect the pool. 3. Irradiation enhanced the synthesis of pre-rRNA for at least 12h. The synthesis of pre-mRNA was also enhanced, but only in the first 3h after irradiation. 4. Partial hepatectomy strongly stimulated the synthesis of both pre-rRNA and pre-mRNA. 5. The synthesis of pre-rRNA was enhanced also in regenerating liver of animals irradiated before or after the operation. The conclusion can be drawn that the early increase in the synthesis of ribosomal RNA is a non-specific cellular response to different injuring factors. 6. The only case where irradiation caused an early inhibition of RNA synthesis was that of pre-mRNA in regenerating liver. This supports the hypothesis that ionizing radiation does not suppress the transcription per se but affects the mechanisms of activation of new genes (cellular programming).

Animals↗
PubMed1997

Nutritional and metabolic effects and significance of mild orotic aciduria during dietary supplementation with arginine or its organic salts after trauma injury in rats.

The effects of acute food deprivation and subsequent refeeding with isonitrogenous oral liquid diets supplemented with arginine (ARG), ARG alpha-ketoglutarate (AKG), or ARG alpha-ketoisocaproate (AKIC) were examined in a Sprague-Dawley rat trauma model (bilateral femur fracture). Both control and trauma rats were starved for 2 days and then pair-fed for 4 days with one of four liquid isonitrogenous diets: diet 1 was a basal casein-based diet, and diets 2, 3, and 4 were the basal diet in which 10% of the nitrogen was replaced by ARG, AKG, or AKIC nitrogen. Two days of starvation resulted in a 13% loss of body weight and also a 27% decrease in the excretion of orotic acid (OA) in control and trauma rats. Although the ARG content of diets 2, 3, and 4 was the same, ARG- and AKIC-supplemented rats excreted significantly (P < .05) more OA than AKG-fed rats. The low level of OA excretion in AKG-fed rats indicates greater use of ARG for metabolic purposes, including efficient urea cycle operation. The metabolic adaptation and nutritional efficacy, i.e., Increased nitrogen retention, larger weight gain, and altered amino acid (AA) metabolism, of AKIC rats seem to be better than in ARG- or AKG-fed rats.

Amino Acids↗
PubMed1976

Studies on ribonucleic acid metabolism using nuclear columns. Release of rapidly labeled RNA from rat liver nuclei.

A method is described to study the effect of successively changing incubation conditions on the release of rapidly labeled RNA from isolated nuclei. Nuclear columns containing immobilized rat liver nuclei isolated after in vivo application of labeled orotic acid are perfused with different non-radioactive media. Within the course of one perfusion, the rate of RNA release can be repeatedly altered by variation of temperature, acidity and concentrations of nucleoside triphosphates, complexing agents, sodium chloride and manganese chloride. RNA release can be started and stopped, indicating that the reaction does not result from damage to nuclei. During 60 min perfusion the same product, labeled ribonucleoprotein (sigma = 1.43 g/cm3 in CsCl), is released. High release rates depend on the ratio of nucleoside triphosphate to divalent cation concentration, not on the concentration of the agents per se. Ribonucleoside and deoxyribonucleoside triphosphates exert the same effect as ATP. The SH reagents iodoacetamide and iodoacetate only slightly affect the ATP-induced reaction. In contrast, p-chloromercuribenzoate, after an initial stimulation, causes inhibition of RNA release.

Adenosine Triphosphate↗
PubMed1974

Studies on the form and synthesis of messenger ribonucleic acid in the rat ventral prostate gland, including its tissue-specific stimulation by androgens.

1. When prostate polyribosomes are labelled with radioactive precursors in vivo and subsequently dissociated with sodium dodecyl sulphate, a heterogeneous 6-15S RNA species may be identified that possesses all of the distinctive properties of mRNA. 2. Apart from the selective incorporation of 5'-fluoro-orotic acid into this 6-15S RNA component, it is bound by nitrocellulose filters under experimental conditions where only poly(A)-rich species of RNA are specifically retained. Most importantly, however, only the 6-15S RNA fraction is capable of promoting the incorporation of amino acids into peptide linkage in an mRNA-depleted cell-free system derived from ascites-tumour cells. 3. With the development of a simpler method for labelling the total RNA fraction of the prostate gland in vitro, the poly(A)-enriched RNA fraction may be readily isolated by adsorption and elution from oligo(dT)-cellulose. The synthesis of the poly(A)-enriched 6-15S RNA fraction is stringently controlled by androgens in a highly tissue- and steroid-specific manner. 4. From an analysis of the proteins synthesized in the ascites cell-free system in the presence of the poly(A)-rich RNA fraction, it appears that protein synthesis in the prostate gland is stimulated in a rather general way, even during the earliest phases of the androgenic response. This conclusion may require modification when more specific means of analysis are available than those used in the present investigation. 5. The implications of these findings to the mechanism of action of androgens are discussed.

Amino Acids↗
PubMed1976

DNA of Myxococcus phage MX-1. Pyrimidine isostichs and the recognition of a minor pyrimidine.

DNA was isolated from bacteriophage MX-1 and was hydrolysed with acids. Hydrolysis with 90% formic acid produced several interconvertable fluorescent artefacts. These substances were produced in greater quantity following hydrolysis with H2SO4. In the course of the course of the study it was found that DNA extracted from phage by 4-aminosalicylate and phenol contained 4-aminosalicylate and breakdown products derived from 4-aminosalicylate following acid hydrolysis. Pyrimidine oligonucleotides released by formic acid-diphenylamine hydrolysis were fractionated by two-dimensional paper chromatography and by ion-exchange chromatography. In this way, the pyrimidine isostichs were obtained and further fractionated. The distribution and with isostich patterns from a variety of DNA sources. MX-1 DNA was found to contain an unusually high frequency of cytosine-rich isostichs 5 and 6 and very low occurrence of the sequence R-Y-R. Based on its absorption spectrum and the fact that it can be labelled in vivo with [14C] orotic acid, we suggest that H-base is a pyrimidine.

Aminosalicylic Acids↗
PubMed2002

Fluorescence sensing of ionic analytes in water: from transition metal ions to vitamin B13.

The fluorescence chemosensor ATMCA has been realised by appending an anthrylmethyl group to an amino nitrogen of TMCA (2,4,6-triamino-1,3,5-trimethoxycyclohexane), a tripodal ligand selective for divalent first-row transition metal ions in water. The ATMCA ligand can act as a versatile sensor for ZnII and CuII ions. Its sensing ability can be switched by simply tuning the operating conditions. At pH 5, ATMCA detects copper(II) ions in aqueous solutions by the complexation-induced quenching of the anthracene emission. Metal ion concentrations < 1 microM can be readily detected and very little interference is exerted by other metal ions. At pH 7, ATMCA signals the presence of ZnII ions at concentrations < 1 microM by a complexation-induced enhancement of the fluorescence. Again the sensor is selective for ZnII over several divalent metal ions, with the exception of CuII, CoII and HgII. Most interestingly, the [ZnII(atmca)]2+ complex can act as a fluorescence sensor for specific organic species, notably selected dicarboxylic acids and nucleotides, by the formation of ternary ligand/zinc/substrate complexes. The oxalate anion is detected in concentrations <0.1 mM; however, no effects on the system's fluorescence is observed in the presence of monocarboxylic acids and long-chain dicarboxylic acids. Among the nucleotides, those containing an imide or amide function are readily detected and an unprecedented high sensitivity for guanine derivatives allows the determination of this nucleotide for 0.05-0.5 mM solutions. Moreover, [ZnII(atmca)]2+ is a very effective and selective sensor in the case of vitamin B13 (orotic acid) in sub-micromolar concentrations. The operative features of the systems investigated are also clearly suitable for intracellular analyses. The factors at the source of organic substrate recognition, here briefly discussed, are of paramount importance for further developments in the applicability of these sensing systems.

Adenosine Monophosphate↗
PubMed1976

Tape test as a simple new method for the study of compounds increasing the problem-solving ability of the rat.

A simple new method, the "tape test" has been developed for studying the enhancement of learning by drugs in "learning-dull" rats. A piece of adhesive tape is pressed on the left front pad of the rats. The time of tape removal by the animal, i.e., the problem-solving time is measured. In our experiments the selected learning-dull rats were used which were unable to remove the tape within 60 s observed on 3 consecutive days. The problem-solving ability of the rats was studied on 4 consecutive days, by posttrial administration. The problem-solving ability was found to be increased after treatment with different drugs such as para-chlorphenylalanine (PCPA), pemolin, orotic acid, vitamin B12. The stimulatory effect of vitamin B12 could be inhibited by vincristine.

Animals↗
PubMed1981

Arginine: an indispensable amino acid for mature dogs.

There studies examined the effect of dietary arginine deficiency in the mature dog. Deletion of arginine from the diet resulted in a slight but significant loss of body weight. Severe episodes of emesis were observed in all experiments. Muscle tremors and frothing around the mouth were also observed in the experiments where the arginine-free diet was force fed. Increasing the amount of diet force-fed to mature dogs accentuated the symptoms of emesis, muscle tremors and frothing. Elevated plasma ammonia and orotate were detected in dogs fed an arginine-deficient diet. Urinary citric and orotic acid was also increased in mature dogs fed a diet devoid of arginine. Nitrogen balance was not significantly altered by deletion of arginine from the diet. Based on the occurrence of emesis, loss of body weight and alterations in intermediary metabolism, we concluded that the mature dog does require a dietary source of arginine. Dietary inclusions of 0.28% arginine prevented the symptoms of arginine deficiency.

Animals↗
PubMed1991

The effect of lentinan on proliferative processes in parenchymal organs of rats--I. The effect on pyrimidine and nucleic acid syntheses.

The present study investigated the effect of Lentinan on the biochemical events associated with the pyrimidine and nucleic acid syntheses in the liver, kidney, thymus and spleen of rats. Lentinan was used at a dose of 4 mg/kg/day (twice) and in a single dose of 20 mg/kg. The following changes were observed. (1) The utilization of (14C)orotic acid for the synthesis of uridine components of liver acid-soluble extract and RNA uracil was activated after the administration of both doses of the drug. The specific activity of cytidine components of the acid-soluble extract and RNA were, on the other hand, not affected. The same holds true for the kidney. The ratio of the specific activity of cytidine:uridine components of the acid soluble extract as well as RNA decreased after the administration of both doses of the drug. The specific activity of DNA cytosine and thymine are slightly suppressed in the liver after the administration of a high dose of Lentinan; no effect was observed in the kidney. (2) The uptake of (14C)cytidine by the liver was not affected; the specific activity of DNA cytosine and thymine were increased after the administration of a high dose of Lentinan. (3) The uptake of (14C)thymidine by the liver was not affected; the specific activity of liver DNA thymine was increased after the administration of both doses of the drug. In the thymus an increase of specific activity of DNA thymine has also been observed. (4) Repeated doses of the drug (4 mg/kg for 6 consecutive days) increased the weight of the spleen. The specific activity of DNA thymine of the liver and spleen were significantly increased.

Animals↗
PubMed1984

Mechanism of arginine protection against ammonia intoxication in the rat.

To examine the beneficial effect of arginine on ammonia intoxication, rats were injected intraperitoneally with a single dose of NH4Cl (6.75 mmol/kg) with and without arginine (5.0 mmol/kg) or ornithine (5.0 mmol/kg). Arginine or ornithine reduced the blood ammonia nitrogen at 30 min after NH4Cl injection from 3,288 +/- 800 micrograms/dl (mean +/- SE) to 538 +/- 90 and 575 +/- 34 micrograms/dl, respectively. In rats administered this dose of NH4Cl, arginine or ornithine did not increase further the hepatic carbamoyl-phosphate synthetase (EC 6.3.4.16) activation by N-acetylglutamate beyond the effect of NH4Cl. However, arginine or ornithine did increase the hepatic citrulline and urea content as well as the plasma urea concentration in these NH4Cl-injected rats. In rats injected with four doses of NH4Cl (2.5 mmol/kg), arginine or ornithine pretreatment increased the urea excretion and normalized the orotic acid excretion. These results indicate that arginine mitigates ammonia intoxication in the rat by increasing ornithine carbamoyltransferase activity through increased ornithine availability and not via activation of N-acetylglutamate synthetase. By increasing ornithine carbamoyltransferase activity, ornithine enhances the conversion of ammonia to citrulline and urea.

Acetyltransferases↗