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At least 163 records · Page 9Linked to original sources
PubMed1971

Changes in the transcription pattern of flax cotyledons after inoculation with flax rust.

The rate of (32)P incorporation into RNA fractions of flax cotyledons (Linum usitatissimum L. var. Bison) was found to increase two- to three-fold by 48h after inoculation with flax rust [Melampsora lini (Pers.) Lev., race no. 3]. This was accompanied by a change in the nucleotide composition of the newly transcribed sodium chloride-soluble RNA fraction. A comparison of the nucleotide composition of the RNA synthesized in the host-parasite complex at different stages of development indicated the preferential synthesis of one or more molecular species of RNA with a high A+U/G+C ratio at a relatively early stage of infection. Treatment of healthy plants with indol-3-ylacetic acid also resulted in a substantial stimulation in the rate of (32)P incorporation into RNA but this was not accompanied by a detectable change in the nucleotide ratios of the newly synthesized RNA. These results suggest that the synthesis of one of more additional RNA species or the augmented synthesis of certain species of RNA may be a specific phenomenon elicited by host-pathogen interaction.

Adenosine Monophosphate↗
PubMed1996

Transcriptionally active chromosomes (TACs) of barley chloroplasts contain the alpha-subunit of plastome-encoded RNA polymerase.

Transcriptionally active chromosomes (TACs) were isolated from mature chloroplasts of barley, from proplastids enriched in basal segments of barley primary foliage leaves, and from ribosome-deficient plastids of heat-bleached barley leaves. Immunological analysis with a specific antibody raised against the plastid rpoA gene product revealed that chloroplasts contain an immunoreactive protein of 38 kDa in the TAC fraction which appears to be identical to the alpha-subunit contained in the soluble RNA polymerase (sRNAP) fraction of the same chloroplasts. However, only traces of immunoreactive protein were detected in a TAC preparation derived from "proplastids". A positive correlation could be demonstrated between transcriptional activity and the amount of immunoreactive 38-kDa protein by analyzing different TAC fractions eluting at different times during gel filtration of a standard TAC preparation as well as in TAC preparations obtained under various detergent conditions.

Chloroplasts↗
PubMed1995

Deprivation of dietary nucleotides results in a transient decrease in acid-soluble nucleotides and RNA concentration in rat liver.

This study examines the contribution of dietary nucleotides to liver nucleotide pools in rats. Liver acid-soluble nucleotides, DNA and RNA concentrations were monitored in two groups of rats fed either a diet supplemented with nucleotides or a diet free of nucleotides for 3 wk. Significantly lower concentrations of ATP, ADP, GTP and CDP as well as of RNA were found after 1 wk in the rats fed a nucleotide-free diet compared with those fed the nucleotide-supplemented diet; concentrations remained lower after 2 wk except for ATP and ADP. No changes over time were observed in the rats fed the nucleotide-supplemented diet. Between wk 2 and 3 an increase in both acid-soluble nucleotides and RNA was observed in the rats fed the nucleotide-free diet, reaching the values found in the rats fed the nucleotide-supplemented diet. These findings, which indicate that dietary nucleotides are utilized at least in part by the liver to maintain the cell nucleotide pools and that diets devoid of nucleotides affect hepatic nucleotide metabolism and RNA, support the hypothesis that liver nucleotide metabolism is modulated by the availability of dietary nucleotides.

Animals↗
PubMed2000

Increasing cerebrospinal fluid chemokine concentrations despite undetectable cerebrospinal fluid HIV RNA in HIV-1-infected patients receiving antiretroviral therapy.

Only limited data on cerebrospinal fluid (CSF) HIV-1 RNA responses and markers of local inflammation in CSF during antiretroviral therapy are available. HIV-RNA, soluble tumor necrosis factor (TNF)-receptor (sTNFr)-II, monocyte chemoattractant protein (MCP)-1, and interferon-gamma-inducible protein (IP)-10 were measured in the peripheral blood and CSF of 26 antiretroviral-naive HIV-1-positive patients, who were treated with ritonavir (RTV)/saquinavir (SQV) (n = 5), RTV/SQV/stavudine (d4T; n = 8) or zidovudine (AZT)/lamivudine (3TC)/abacavir/nevirapine/indinavir (n = 13). After 8 to 12 weeks of treatment, CSF HIV-RNA dropped to <400 copies/ml in 1 of 5 patients in the RTV/SQV group, 8 of 8 patients in the RTV/SQV/d4T group, and 9 of 10 patients in the five-drug group. CSF sTNFr-II and IP-10 levels increased in patients with detectable CSF HIV-RNA. However, increases in CSF chemokine and sTNFr-II concentrations were also observed in some patients with good CSF HIV-RNA responses. Moreover, CSF MCP-1 concentrations increased in the whole population after 2 months of treatment. Ongoing residual HIV replication in the central nervous system, which cannot be detected with CSF HIV-RNA measurements, may account for this phenomenon.

Anti-HIV Agents↗
PubMed1978

[Protein metabolism in vitamin A deficient rats. I. Protein and DNA turn-over in rat liver].

The effect of a vitamin A deficient diet on liver proteosynthesis was studied with in vivo and in vitro methods. Weanling rats obtained from vitamin A rstricted female were fed "separate diet". Control animals received vitamin A in protein diet. Deficient rats were used at the weight plateau stage. The RNA, DNA and protein liver content was unaffected by vitamin A deficiency and the same result was found for free amino acid pool. The turn-over rate of protein and RNA of cellular fractions and total liver was studied. (14C) leucine and (3H) uridine was injected intraperitoneally. Animals were killed at 20-40 minutes and 2-4-6-8-10 days after injection. Synthesis rate of liver proteins was the same in both groups. Our results indicate no difference in the average rate of (14C) protein degradation between control and deficient animals. Vitamin A deficiency decrease the turn-over rate of total ribosomal and soluble RNA. Polyribosome and postribosomal supernatant from deficient rats liver were less active for protein synthesis in vitro than corresponding fractions from well nourished rats. The lesion was located in the post ribosomal supernatant fraction by "crossing over" experiments. Polyribosome desimentation analysis on sucrose gradients revealed no difference between the two groupe. The results indicate that vitamin A deficiency decrease cell proliferation without degeneracy. The limitative agent of proteosynthesis is situated at RNA level. However protein synthesis is unaffected in our experimental procedure.

Amino Acids↗
PubMed1987

Nucleic acid extraction and virus detection in grapevine.

Three extraction media for the isolation of nucleic acids from grapevines, a tissue high in polyphenols and other materials that interfere with nucleic acid extraction, were compared. When phenol was present in the initial extraction media only a small yield of soluble RNA and no high molecular weight rRNA was obtained. In the absence of phenol in conventional salt and detergent-based extraction media, rRNAs were extracted, but a major proportion of the RNAs were broken down. Using Na-perchlorate, a chaotropic salt, in a rapid procedure, it was possible to extract both high and low molecular weight RNA efficiently. This procedure enabled the detection of viral RNA, which could not be detected following phenol extraction, at the picogram levels, by dot-blot hybridization.

Nucleic Acid Hybridization↗
PubMed1965

The sedimentation behaviour of ribonuclease-active and -inactive ribosomes from bacteria.

1. The ;30s' and ;50s' ribosomes from ribonuclease-active (Escherichia coli B) and -inactive (Pseudomonas fluorescens and Escherichia coli MRE600) bacteria have been studied in the ultracentrifuge. Charge anomalies were largely overcome by using sodium chloride-magnesium chloride solution, I 0.16, made 0-50mm with respect to Mg(2+). 2. Differentiation of enzymic and physical breakdown at Mg(2+) concentrations less than 5mm was made by comparing the properties of E. coli B and P. fluorescens ribosomes. 3. Ribonuclease-active ribosomes alone showed a transformation of ;50s' into 40-43s components. This was combined with the release of a small amount of ;5s' material which may be covalently bound soluble RNA. Other transformations of the ;50s' into 34-37s components were observed in both ribonuclease-active and -inactive ribosomes at 1.0-2.5mm-Mg(2+), and also with E. coli MRE600 when EDTA (0.2mm) was added to a solution in 0.16m-sodium chloride. 4. Degradation of ribonuclease-active E. coli B ribosomes at Mg(2+) concentration 0.25mm or less was coincident with the formation of 16s and 21s ribonucleoprotein in P. fluorescens, and this suggested that complete dissociation of RNA from protein was not an essential prelude to breakdown of the RNA by the enzyme. 5. As high Cs(+)/Mg(2+) ratios cause ribosomal degradation great care is necessary in the interpretation of equilibrium-density-gradient experiments in which high concentrations of caesium chloride or similar salts are used. 6. The importance of the RNA moiety in understanding the response of ribosomes to their ionic environment is discussed.

Escherichia coli↗
PubMed1966

Inhibition by soluble ribonucleic acid of stimulatory effect of liver template ribonucleic acid.

1. Liver soluble RNA (s-RNA) and, to a smaller extent, Escherichia coli s-RNA inhibited the stimulation of [(14)C]leucine incorporation into protein in an E. coli S-30 system brought about by liver microsomal RNA or polyuridylic acid as template. 2. The inhibitory activity was associated with the fraction of the s-RNA possessing transfer-RNA activity. 3. The inhibition was exercised at a stage after charging of the s-RNA with amino acid. 4. Neither the method of preparation of the s-RNA nor its state of amino acid acylation affected its inhibitory action. 5. Stimulation of [(14)C]phenylalanine incorporation by polyuridylic acid or by liver microsomal RNA was not inhibited by addition of s-RNA. 6. It appears that excess of s-RNA inhibits the ambiguous incorporation of leucine with polyuridylic acid and also that something similar occurs with a natural template. 7. Estimation of messenger activity of samples of RNA should be carried out only after removal of s-RNA.

Animals↗
PubMed1968

Occurrence of 7-methylguanine in nucleic acids of rat liver.

1. Microsomal and soluble RNA of rat liver have been studied by column and paper chromatography after administration of [Me-(14)C]methionine; evidence was obtained for the occurrence of 7-methylguanine, the methyl group being derived from methionine. 2. No evidence was obtained for the occurrence of 7-methylguanine in DNA.

Animals↗
PubMed1961

Chromatographic separation, and characteristics of nucleic acids from HeLa cells.

The application of the phenol-duponol method to extraction of nucleic acids from HeLa cells is described. Chromatography of the phenol extract on an esterified bovine serum albumin column with a salt gradient of sodium chloride gives separation of soluble RNA, DNA, and two different high molecular RNA fractions. Ultracentrifugation of the DNA eluted from the column gives a sedimentation coefficient (s(20) (o),w) of 38, which agrees with ultracentrifugation data on the phenol extract. The eluted RNA appears polydisperse at low ionic strength, but at high ionic strength and after alcohol precipitation two fractions with the sedimentation coefficients of 16 and 25 to 29, respectively, were obtained.

Chromatography↗
PubMed1967

Effects of Phosfon-S on Nucleic Acid Metabolism in Pisum sativum Alaska.

Phosfon-S, a substance which inhibits stem elongation, alters nucleic acid metabolism in Pisum sativum Alaska. Methylated albumin kieselguhr (MAK) columns were used to fractionate (32)P-labeled nucleic acids. Phosfon-S treatment of the plants resulted in a decrease in soluble RNA and an increase in ribosomal RNA. Specific activities of the various nucleic acid fractions were lower as a result of treatment. The nucleic acids from treated tissues were more resistant to RNase degradation, and endogenous RNase activity was lower in treated tissues. When RNase treated nucleic acids were fractionated on MAK columns, the DNA-RNA fractions from treated plants had a higher specific activity than that of the control, which was not true before nuclease treatment. Spectrophotometric examination of this fraction revealed a difference in absorption spectra, possibly indicating a Phosfon-S nucleic acid complex. It is suggested that these alterations in nucleic acid metabolism could in turn alter a wide variety of metabolic processes, resulting in retarded growth.

Journal Article↗
PubMed1969

Changes in Nucleic Acid Fractions of Seed Components of Red Pine (Pinus resinosa Ait.) during Germination.

Changes in nucleic acid fractions of Pinus resinosa during seed germination were studied. At various stages of seed germination, embryos and megagametophytes were surgically separated and nucleic acids were extracted separately by a phenol-method. Total nucleic acids were fractionated on single-layer methylated albumin kieselguhr (MAK) columns. Total nucleic acids in embryos increased significantly 2 days after seeds were moistened, whereas, in megagametophytes, total nucleic acids stayed almost at a constant level until they degenerated at the time of shedding. In embryos, ribosomal RNAs (rRNA) increased 2 days after seeds were sown, whereas soluble RNA (sRNA) increased at 3 days. By comparison, nucleic acid fractions of megagametophytes did not show any quantitative changes during germination, except that rRNA fractions decreased shortly before shedding of seed coats. In dormant embryos the proportion of DNA was high and the proportions of sRNA and rRNA were low, whereas in megagametophytes at dormancy the proportions were completely reversed. As seed germination progressed, proportions of nucleic acid fractions in embryos changed significantly. In megagametophytes, although proportions of individual fractions remained almost constant throughout the experimental period, incorporation of (32)P into sRNA and rRNA of megagametophytes indicated turnover of these fractions.

Journal Article↗
PubMed1974

Nucleic Acid and protein metabolism of senescing and regenerating soybean cotyledons.

An alternative to the leaf disk system for studies of the metabolism of senescence is described. The progress of senescence of soybean (Glycine max L.) cotyledons is arrested when the epicotyl is removed. Epicotyl removal at 16 or 17 days reversed the decline in nucleic acid, protein, and chlorophyll content in the cotyledon. Epicotyl removal at 18 days did not reverse the decline in the above components, and therefore the progress of cotyledon has passed the point of no return. Cotyledons lost 90% of their nucleic acid and 80% of their protein before senescence became irreversible. The rate of recovery in various macromolecular components after epicotyl removal did not occur in an equal manner. Nucleic acid was regenerated at a faster rate than chlorophyll, which was regenerated at a faster rate than soluble protein. The heavy nucleic acid components (ribosomal and heavy ribosomal messenger fractions) regenerated at greater rates than did the soluble RNA or DNA. No label from (14)CO(2) was incorporated into DNA of the cotyledons when the epicotyl was present but label was incorporated into DNA after epicotyl removal.The parallels between the mechanisms of cotyledon senescence and apical dominance are discussed.

Journal Article↗
PubMed1982

Studies on the regeneration of the skin barrier and the changes in 32P incorporation into the epidermis after stripping.

The epidermis of the rabbit ear was stripped and the effects on percutaneous absorption and 32P incorporation into lipid, TCA-soluble, RNA and DNA fractions in the epidermis was studied for 21 days. Skin penetration by 14C-propylene glycol increased 300-fold immediately after stripping twenty times. Three to five days after stripping, penetration had returned almost to normal levels, but it required several additional days for the barrier function to become normal. The process of regeneration of the skin barrier paralleled the morphological changes in the epidermis but was inconsistent with the biochemical data. A significant increase in 32P incorporation was observed at 10 days, when the hyperplasia had disappeared and percutaneous absorption had returned to normal. The discrepancy between 32P incorporation and morphology or percutaneous absorption after stripping is discussed.

Animals↗
PubMed2006

Cytokine accumulation in photochemically treated and gamma-irradiated platelet concentrates during storage.

BACKGROUND: Photochemical treatment (PCT) for pathogen reduction of platelet concentrates (PCs) affects all cells containing DNA and/or RNA. Soluble mediators, which may cause transfusion reactions, are determined by the balance between secretion and/or cell destruction and binding and/or degradation. STUDY DESIGN AND METHODS: Ten double-dose single-donor leukoreduced PCs were split in two identical units. Two study arms were created: Study Arm A consisting of five PCT PCs with corresponding untreated control PCs and Study Arm B consisting of five PCT PCs with corresponding gamma-irradiated control PCs. PCs that had added PAS-III (Intersol) were treated with amotosalen and ultraviolet A light. Corresponding controls PCs, to which PAS-II (T-sol) were added, received no treatment or were gamma-irradiated before storage. Platelet (PLT)-derived (CCL5/RANTES, CXCL4/PF4, CCL3/MIP-1alpha, transforming growth factor [TGF]-beta, CXCL8/interleukin [IL]-8, IL-1beta) as well as white blood cell (WBC)-associated (IL-6, IL-10, IL-11, IL-12, tumor necrosis factor, interferon-gamma) cytokines were investigated by enzyme-linked immunosorbent assay and cytometric bead array during storage for up to 12 days. RESULTS: Independent of previous treatment we observed that all concentrates showed low levels of WBC-associated cytokines. PLT-derived cytokines were detected at higher levels and showed significant increase during storage. Statistical analysis showed lower PLT content per unit in PCT PCs, higher levels of activation variables in PCT PCs, and higher levels and accumulation rate of CCL5, CXCL4, TGF-beta, and CXCL8 in PCT PCs. CONCLUSION: PLTs are the main source of released cytokines during storage of untreated, gamma-irradiated, and PCT PCs. PCT may affect the level of PLT-derived cytokines in PCs. No additional reduction of WBC-associated cytokines were observed after PCT in prestorage leukoreduced PCs.

Blood Platelets↗
PubMed1965

Sulfur: incorporation into the transfer fraction of soluble ribonucleic acid.

When S(35)-labeled soluble RNA from Escherichia coli K 38 is subjected to gel filtration, four fractions of RNA are obtained by elution. Only one RNA fraction, the transfer RNA, contains sulfur, presumably as thionucleotides. Treatment with ribonuclease suggests that the incorporated sulfur is an integral part of the polynucleotide chain; digestion with alkali yields a mixture of products containing sulfur, the major one being eluted in a position similar to uridine diphosphate upon Dowex-l chromatography. Analysis by countercurrent distribution of S(35)-labeledtransfer RNA from E. coli B reveals that the incorporated sulfur is found in many RNA's that accept amino acids, but the possibility remains that not all acceptor RNA's contain sulfur.

Chromatography↗