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Elevation of the salvage synthesis of ribonucleic acid in the rat liver during the induction of hepatoma with 3'-methyl-4-(dimethylamino)azobenzene.

Incorporation of uracil and orotic acid into the ribonucleic acid (RNA) fraction of rat liver during carcinogenesis induced with 3'-methyl-4-(dimethylamino)azobenzene was investigated. Uracil incorporation was found to be gradually elevated during the early stage (about 2 weeks) of the carcinogenesis, although not in the normal rat liver homogenates contacted with the carcinogen for a short hours, and the elevated uptake was maintained until tumor induction. On the other hand, orotic acid incorporation reverted to the original level after a temporary increase during the early stage. In a good agreement with the increased uracil incorporation, activities of both uridine phosphorylase and uridine kinase involved in the salvage pathway of RNA synthesis also increased during the early stage, and their activities in the liver were maintained at elevated levels after discontinuance of the carcinogen feeding. The activity of uridine monophosphate (UMP) pyrophosphorylase, converting uracil to UMP, was not detected during the early stage. Significance of the activation of the salvage pathway of RNA synthesis during the early stage of an axo dye-induced carcinogenesis were discussed.

Animals↗

Advanced renal cell carcinoma: treatment with xenogeneic immune ribonucleic acid and appropriate surgical resection.

Herein we describe the first clinical treatment of renal cell carcinoma in humans with xenogeneic immune ribonucleic acid. Twelve patients with advanced renal cell carcinoma have been treated by appropriate operations to remove tumor bulk followed by specific passive immunotherapy. Xenogeneic specific immune ribonucleic acid was prepared from the spleen of normal sheep that had received 4 weekly injections of a homogenate of renal cell carcinoma. Results indicate that 1) xenogeneic specific immune ribonucleic acid can safely be given to humans without local or systemic toxicity, 2) there is a suggestion of clinical benefit, since only 2 patients have had progression of known metastases during treatment with immune ribonucleic acid and 3) xenogeneic immune ribonucleic acid can enhance the immune response to renal cell carcinoma, as demonstrated by in vitro lymphocytoxicity tests.

Adenocarcinoma↗

The quantitative histochemistry of ribonucleic acid using gallocyanin.

A method for the cytophotometric estimation of ribonucleic acid in tissue sections using gallocyanin-chrome alum is described. The dye obeys Beer's law in gelatin sections. The effect of deoxyribonuclease on the staining of ribonucleic acid is also investigated. The results indicate that this method is of value in the quantitation of ribonucleic acid.

Alum Compounds↗

Enhanced transformability with heterospecific deoxyribonucleic acid in a Streptococcus sanguis mutant impaired in ribonucleic acid polymerase activity.

We have induced with nitrosoguanidine in Streptococcus sanguis a mutation conferring inability to grow and synthesize ribonucleic acid (RNA) at 42 C, the optimal temperature for growth and RNA synthesis in the parental strain. The mutation (ts) is transferable via transforming deoxyribonucleic acid (DNA) and is replaceable by its wild-type allele with fairly high efficiency in transformation reactions. The ts mutation is unlinked to the sites of mutation conferring resistance of rifampin (rifr) and streptolydigin (stgr), known to affect the beta subunit of DNA-dependent RNA polymerase. Extracts from strains carrying the ts mutation are more sensitive to elevated temperatures than are parental extracts when assayed for DNA-dependent RNA polymerase. The conclusion that the mutation causes a temperature-sensitive defect in some component of this enzyme (other than beta) is supported by the finding that the polymerase activity of a heat-inactivated ts stgr extract cannot be increased by addition of an unheated ts stgs extract, which is itself inactivated by streptolydigin. S. sanguis recipients carrying the ts mutation are highly transformable with heterospecific DNA, especially at the restrictive temperature.

Aminoglycosides↗

Prostaglandin endoperoxide H synthase type 1 and type 2 messenger ribonucleic acid in human fetal tissues throughout gestation and in the newborn infant.

OBJECTIVE: We determined the relative abundance of prostaglandin endoperoxide H synthase type 1 and type 2 messenger ribonucleic acid levels in the human fetus and newborn infant. STUDY DESIGN: We used ribonuclease protection assays and normalized values to messenger ribonucleic acid of cyclophilin. Tissues were obtained from all 3 trimesters and in the first 9 days of the newborn period. RESULTS: Prostaglandin endoperoxide H synthase type 1 and type 2 messenger ribonucleic acid is present in every fetal tissue examined (lung, kidney, intestine, heart, brain, and stomach). Kidney and lung demonstrated no changes in the expression of prostaglandin endoperoxide H synthase type 1 messenger ribonucleic acid with gestational age, whereas postnatal levels in lung were one third those in the first trimester (P <.05). A large increase (5-fold to 30-fold; P <.05) occurred throughout gestation for the expression of prostaglandin endoperoxide H synthase type 2 messenger ribonucleic acid in intestine, lung, and kidney, which extended into the newborn period for lung and kidney. CONCLUSIONS: These data imply that the expression of prostaglandin endoperoxide H synthase type 2 messenger ribonucleic acid may be responsible for prostaglandin-related effects and is coordinated in several human fetal tissues in late gestation.

Brain↗

Ribonucleic acid synthesis during morphogenesis in Myxococcus xanthus.

Ribonucleic acid synthesis was measured during the morphogenesis of Myxococcus xanthus. After induction of microcyst formation by the addition of glycerol to an exponential culture, net ribonucleic acid (RNA) synthesis was immediately terminated (measured either chemically or by the accumulation of acid-insoluble radioactivity). Extensive RNA turnover did take place, however, including RNA made both before and after induction. Sucrose gradient centrifugation revealed that ribosomes and ribosomal RNA were synthesized during microcyst formation even though there was no net RNA synthesis. Base analyses of the total RNA of vegetative cells and 120-min microcysts were indistinguishable.

Adenine↗

Unusual deficiency of the modified purine base queuine in transfer ribonucleic acid from the human placenta as tested by enzymatic assay.

Transfer ribonucleic acid from rapidly growing tissues, particularly from neoplasia, is partially deficient in queuine, a highly modified transfer ribonucleic acid constituent. By means of an enzymatic assay we also found a queuine deficiency (14%) in human placenta transfer ribonucleic acid despite its high concentrations in the amniotic fluid. Proposed cause and significance of the results are discussed.

Female↗

Control of dihydrofolate reductase messenger ribonucleic acid production.

We used methotrexate-resistant mouse cells in which dihydrofolate reductase levels are approximately 500 times normal to study the effect of growth stimulation on dihydrofolate reductase gene expression. As a result of growth stimulation, the relative rate of dihydrofolate reductase protein synthesis increased threefold, reaching a maximum between 25 and 30 h after stimulation. The relative rate of dihydrofolate reductase messenger ribonucleic acid production (i.e., the appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm) increased threefold after growth stimulation and was accompanied by a corresponding increase in the relative steady-state level of dihydrofolate reductase ribonucleic acid in the nucleus. However, the increase in the nuclear level of dihydrofolate reductase ribonucleic acid was not accompanied by a significant increase in the relative rate of transcription of the dihydrofolate reductase genes. These data indicated that the relative rate of appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm depends on the relative stability of the dihydrofolate reductase ribonucleic acid sequences in the nucleus and is not dependent on the relative rate of transcription of the dihydrofolate reductase genes.

Animals↗

Double-stranded ribonucleic acid in Agaricus bisporus.

Double-stranded ribonucleic acid present in virus-infected mushrooms of Agaricus bisporus has been resolved through polyacrylamide gel electrophoresis into six molecular-weight forms. Identification of these six double-stranded ribonucleic acids in mushrooms by this procedure has proven to be a useful and diagnostic method for viral infection in the cultivated mushroom.

Agaricales↗

Nature of ribonucleic acid synthesis during early sporulation in Saccharomyces cerevisiae.

Phosphate uptake in sporulating cultures of Saccharomyces cerevisiae has been found to occur approximately 2 h after the transfer to sporulation medium. Early ribonucleic acid synthesis begins at approximately 4 h and continues to 8 h. Incorporation of phosphate into acid-extractable precursor pools parallels phosphate uptake. In triple-labeling experiments it was observed that the breakdown of vegetatively synthesized ribonucleic acid is not a significant source of precursors for ribonucleic acid synthesis during sporulation. The majority of the ribonucleic acid made in a 10-min period during sporulation does not migrate on gels with precursor or mature ribosomal ribonucleic acid.

Adenine↗

Immunoprotective activity of capsular polysaccharide in Klebsiella pneumoniae ribosomal preparations does not involve ribonucleic acid.

Two peaks were obtained by cesium chloride density gradient ultracentrifugation of Klebsiella pneumoniae ribosomal preparations. Peak I contained capsular polysaccharide, lipopolysaccharide, protein, and less than 0.5% ribonucleic acid. Peak II consisted mainly of ribonucleic acid, with low amounts of protein and capsular polysaccharide. Expressed as capsular polysaccharide content, the 50% protective dose of peak I and of nonfractionated ribosomal preparations was nearly constant (2.6 and 1.2 ng, respectively). Since peak I contained less than 0.5% ribonucleic acid, these results provide evidence that ribosomal ribonucleic acid is not required for protection of mice by K. pneumoniae capsular polysaccharide which contaminates ribosomal preparations.

Animals↗

Expression of transforming growth factor-beta 1 messenger ribonucleic acid and the modulation of deoxyribonucleic acid synthesis by transforming growth factor-beta 1 in human endometrial cells.

OBJECTIVE: The purpose of this study was (1) to evaluate the potential sites of transforming growth factor-beta 1 synthesis in human endometrium by analyzing separated endometrial glands and stromal cells for transforming growth factor-beta 1 messenger ribonucleic acid by Northern analysis of total ribonucleic acid and (2) to investigate the effects of transforming growth factor-beta 1 on deoxyribonucleic acid synthesis in endometrial epithelial and stromal cells in culture. STUDY DESIGN: Endometrial glands and stroma from proliferative and secretory endometrium were isolated after collagenase treatment of endometrial tissue minces and were analyzed for transforming growth factor-beta 1 messenger ribonucleic acid by Northern analysis. We studied the effects of estradiol-17 beta and transforming growth factor-beta 1 on deoxyribonucleic acid synthesis in endometrial epithelium and transforming growth factor-beta 1 on stromal cells in culture by evaluating tritiated thymidine incorporation into trichloroacetic acid-precipitable material. RESULTS: Transforming growth factor-beta 1 messenger ribonucleic acid was detected for Northern analysis in separated endometrial stromal cells in levels that were greatest during the secretory phase and in greater levels than in epithelial cells from that same tissue. Transforming growth factor-beta 1 messenger ribonucleic acid in glandular epithelium in culture was not increased to detectable levels by treatment with transforming growth factor-beta 1. Deoxyribonucleic acid synthesis in endometrial glandular epithelium was inhibited by transforming growth factor-beta 1, but transforming growth factor-beta 1 stimulated deoxyribonucleic acid synthesis in endometrial stromal cells in culture. After treatment for 5 days with estradiol-17 beta (10(-8) mol/L), deoxyribonucleic acid synthesis in endometrial glands in culture was decreased by 40%. Transforming growth factor-beta 1 (1 ng/ml) did not alter this effect of estradiol-17 beta on deoxyribonucleic acid synthesis. CONCLUSIONS: Transforming growth factor-beta 1 acts to decrease deoxyribonucleic acid synthesis in epithelial cells and to increase it in stromal cells isolated from human endometrium and maintained in monolayer culture. Transforming growth factor-beta 1, potentially of stromal cell origin, could participate in the regulation of endometrial cell proliferation and differentiation in vivo.

Cells, Cultured↗

Developmental changes in atrial natriuretic factor content and localization of its messenger ribonucleic acid in ovine fetal heart.

OBJECTIVES: The current study investigated the developmental changes in atrial natriuretic factor peptide content and messenger ribonucleic acid localization in the atria and ventricles of the ovine fetus throughout the second half of gestation. STUDY DESIGN: Ovine fetuses from 67 to 146 days' gestation (term 147 days) and newborn lambs were used for the study. Tissue atrial natriuretic factor contents were determined by radioimmunoassay, and atrial natriuretic factor messenger ribonucleic acid distribution was determined by in situ hybridization. RESULTS: In fetal atria, atrial natriuretic factor peptide levels were much greater than those in the ventricles. The levels in the atria increased with advancing gestation from 70 to 140 days, reflecting an increase in weight of the atrial chambers. A similar trend was not observed in the ventricles. In the atria, atrial natriuretic factor peptide content (per unit protein) reached high levels at 100 to 110 days' gestation; this was associated with an increase in level of atrial natriuretic factor gene expression. In the ventricles, atrial natriuretic factor peptide content and gene expression were very low throughout the second half of gestation, except for a peak in content that occurred at 100 days. Atrial natriuretic factor messenger ribonucleic acid abundance was much greater in the atria than in the ventricles in fetuses from 90 to 130 days' gestation. The distribution of atrial natriuretic factor messenger ribonucleic acid was homogeneous throughout the thickness of the atria and ventricles of the fetal heart. CONCLUSION: During the second half of gestation in the ovine fetus, the expression of atrial natriuretic factor messenger ribonucleic acid in the atria and ventricles paralleled the appearance of the peptide.

Animals↗

Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.

Intact ribonucleic acid (RNA) has been prepared from tissues rich in ribonuclease such as the rat pancreas by efficient homogenization in a 4 M solution of the potent protein denaturant guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol to break protein disulfide bonds. The RNA was isolated free of protein by ethanol precipitation or by sedimentation through cesium chloride. Rat pancreas RNA obtained by these means has been used as a source for the purification of alpha-amylase messenger ribonucleic acid.

Animals↗

Mutant of vesicular stomatitis virus which allows deoxyribonucleic acid synthesis and division in cells synthesizing viral ribonucleic acid.

Some temperature-sensitive mutants of vesicular stomatitis virus were tested for their ability to block the initiation of deoxyribonucleic acid (DNA) synthesis and division in serum-stimulated hamster embryo fibroblasts at the nonpermissive temperature. Although the parental strain blocked these processes, one particular mutant allowed essentially normal DNA synthesis and division. By autoradiography, it was shown that individual cells infected with this mutant could synthesize viral ribonucleic acid and at the same time initiate DNA synthesis and divide. Cells infected with such conditional defective mutants appear to be suitable for studies on the effects of persistent viral infections on molecular and cellular functions in proliferating cell populations.

Animals↗

Transforming growth factor-alpha and epidermal growth factor messenger ribonucleic acid and protein levels in human placentas from early, mid, and late gestation.

OBJECTIVE: Human placenta expresses receptors for transforming growth factor-alpha and epidermal growth factor throughout pregnancy. Experiments were performed to determine whether epidermal growth factor or transforming growth factor-alpha might be synthesized by placental cells and act through an autocrine mechanism to influence functioning of placental cells in vivo. STUDY DESIGN: Human placentas from early, mid, and late gestations were analyzed for transforming growth factor-alpha and epidermal growth factor messenger ribonucleic acid and proteins. Polyadenylic acid-positive ribonucleic acid was isolated from placentas from 10, 11, 13, 21, 32, 38, 39, and 40 weeks of gestation and analyzed by Northern analysis for hybridization with complementary deoxyribonucleic acid probes specific for epidermal growth factor or transforming growth factor-alpha. Levels of immunoreactive epidermal growth factor and transforming growth factor-alpha were measured by specific radioimmunoassays in pools of placentas from early, mid, and late gestations, and levels of epidermal growth factor and transforming growth factor-alpha receptor-active protein were measured by radioreceptor assay. RESULTS: All placentas had a strong transforming growth factor-alpha hybridization band at 4.5 kb and a weak epidermal growth factor hybridization band at 5.2 kb. High levels of transforming growth factor-alpha immunoreactive protein (90 to 180 ng/mg protein) and low levels of immunoreactive epidermal growth factor (3 to 9 pg/mg protein) were detected in pools of placentas from early, mid, and late gestations. High levels of epidermal growth factor and transforming growth factor-alpha receptor-active protein (250 ng/mg protein) were also detected. CONCLUSION: Human placentas contain relatively high levels of immunoreactive and receptor-active transforming growth factor-alpha, as well as transforming growth factor-alpha messenger ribonucleic acid, throughout gestation. This finding suggests that transforming growth factor-alpha may act by an autocrine system to influence human placental cell function in vivo.

Blotting, Northern↗

Cytochrome p450 3A4 messenger ribonucleic acid induction by rifampin in human peripheral blood mononuclear cells: correlation with alprazolam pharmacokinetics.

OBJECTIVE: There is significant interest in the assessment of the individual cytochrome p450 (CYP) 3A4 activity. We analyzed whether CYP3A4 messenger ribonucleic acid (mRNA) concentrations in leukocytes reflect CYP3A activity in the liver measured by alprazolam as an in vivo probe drug. We also wanted to identify whether genetically determined high CYP3A5 expression is associated with increased alprazolam clearance. METHODS: Alprazolam plasma concentrations were measured 10 hours after intake of 1 mg alprazolam. CYP3A4 mRNA concentrations in peripheral blood mononuclear cells were quantified in 96 healthy volunteers before and after 5-day treatment with 450 mg rifampin (INN, rifampicin) daily. Genetic polymorphisms in CYP2C19, CYP3A4, and CYP3A5 were analyzed by polymerase chain reaction, restriction fragment length polymorphism, and sequencing. RESULTS: The median alprazolam concentration measured 10 hours after dosage was 8.1 mug/L (range, 4.5-14.6 mug/L) before and 1.7 mug/L (range, 0.3-4.1 mug/L) after rifampin treatment. Leukocyte CYP3A4 mRNA was detectable in all samples with a median of 28 molecules per 1 ng total ribonucleic acid before (range, 10-128 molecules per 1 ng total ribonucleic acid) and 50 molecules per 1 ng total ribonucleic acid after (range, 9-484 molecules per 1 ng total ribonucleic acid) rifampin treatment (P <.001). However, mRNA concentrations before and during rifampin induction were largely overlapping, and there was a poor correlation between mRNA concentrations and alprazolam 10-hour trough concentrations reflecting CYP3A4 activity (r = -0.4, P <.001). Alprazolam kinetics did not differ between genetically determined expressers of CYP3A5 (genotype CYP3A5*1/*3) compared with homozygous carriers of the splice site variant. A marginally significant dependence of alprazolam concentrations from the CYP2C19 allele *2 was found (P =.04). CONCLUSIONS: CYP3A4 mRNA concentrations in blood cells were very low and did not reflect systemic drug clearance mediated by CYP3A enzymes. The CYP3A5 genetic polymorphism does not appear relevant for alprazolam kinetics.

Adult↗