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Biomedical subjects

R Kelly

Publications and source records attributed to R Kelly.

At least 145 records · Page 8Linked to original sources

Organ blood flow haemodynamics and metabolism of the albacore tuna Thunnus alalunga (Bonnaterre).

Metabolic haemodynamic, and organ blood flow measurements were made in tabled, partially anaesthetized albacore Thunnus alalunga. Heart rates were 115 +/- 9 beats/min: blood pressure 98/75 mm Hg (systolic/diastolic): cardiac output 36.1 +/- 5 (ml/min/kg): oxygen consumption 3.4 +/- 0.7 (ml O2/min/kg) and cardiac contractility (dP/dt) 6342 +/- 822 mm Hg/s. Organ blood flows were measured with radiolabelled microspheres. The red muscle, kidney, and spleen received the highest flows and the white-muscle the least. There was a flow gradient in the white-muscle with the inner portion near the red-muscle receiving the highest flows. Arterial and venous blood gas measurements showed a reverse temperature effect on arterial PO2 and a P50 of 15.9 Torr corrected to 37 degrees C. Red-muscle temperature was 7 degrees C higher than ambient water temperature. These measurements record the albacore's markedly high cardiovascular capability.

Animals↗

Mouse DNA 'fingerprints': analysis of chromosome localization and germ-line stability of hypervariable loci in recombinant inbred strains.

Human minisatellite probes cross-hybridize to mouse DNA and detect multiple variable loci. The resulting DNA "fingerprints" vary substantially between inbred strains but relatively little within an inbred strain. By studying the segregation of variable DNA fragments in BXD recombinant inbred strains of mice, at least 13 hypervariable loci were defined, 8 of which could be regionally assigned to mouse chromosomes. The assigned loci are autosomal, dispersed and not preferentially associated with centromeres or telomeres. One of these minisatellites is complex, with alleles 90 kb or more long and with internal restriction endonuclease cleavage sites which produce a minisatellite "haplotype" of multiple cosegregating fragments. In addition, one locus shows extreme germ-line instability and should provide a useful system for studying more directly the rates and processes of allelic variation of minisatellites.

Animals↗

Chemotherapy influencing the course of nephrotic syndrome in colonic carcinoma.

A 50-year-old man presented with the Nephrotic Syndrome (NS). Subsequent investigations showed this to be due to a membranous glomerulonephritis. He was found to have a primary colonic carcinoma with extensive intra-abdominal spread. He was treated with 5-fluorouracil (5FU) with excellent resolution of the NS for a period of 6 months. The disease eventually progressed and the NS recurred on cessation of chemotherapy. This is the first report of such a response to 5FU of which we are aware and emphasizes the usefulness of palliative chemotherapy even in 'resistant' solid tumors when they are complicated by the NS.

Adenocarcinoma, Mucinous↗

Directed mutagenesis in Candida albicans: one-step gene disruption to isolate ura3 mutants.

A method for introducing specific mutations into the diploid Candida albicans by one-step gene disruption and subsequent UV-induced recombination was developed. The cloned C. albicans URA3 gene was disrupted with the C. albicans ADE2 gene, and the linearized DNA was used for transformation of two ade2 mutants, SGY-129 and A81-Pu. Both an insertional inactivation of the URA3 gene and a disruption which results in a 4.0-kilobase deletion were made. Southern hybridization analyses demonstrated that the URA3 gene was disrupted on one of the chromosomal homologs in 15 of the 18 transformants analyzed. These analyses also revealed restriction site dimorphism of EcoRI at the URA3 locus which provides a unique marker to distinguish between chromosomal homologs. This enabled us to show that either homolog could be disrupted and that disrupted transformants of SGY-129 contained more than two copies of the URA3 locus. The A81-Pu transformants heterozygous for the ura3 mutations were rendered homozygous and Ura- by UV-induced recombination. The homozygosity of a deletion mutant and an insertion mutant was confirmed by Southern hybridization. Both mutants were transformed to Ura+ with plasmids containing the URA3 gene and in addition, were resistant to 5-fluoro-orotic acid, a characteristic of Saccharomyces cerevisiae ura3 mutants as well as of orotidine-5'-phosphate decarboxylase mutants of other organisms.

Candida albicans↗

Role of protein synthesis in decay and accumulation of mRNA during spore germination in the cellular slime mold Dictyostelium discoideum.

Spore germination in Dictyostelium discoideum is a particularly suitable model for studying the regulation of gene expression, since developmentally regulated changes in both protein and mRNA synthesis occur during the transition from dormant spore to amoeba. The previous isolation of three cDNA clones specific for mRNA developmentally regulated during spore germination allowed for the quantitation of the specific mRNAs during this process. The three mRNAs specific to clones pLK109, pLK229, and pRK270 have half-lives much shorter (minutes) than those of constitutive mRNAs (hours). Using spore germination as a model, we studied the roles of ribosome-mRNA interactions and protein synthesis in mRNA degradation by using antibiotics that inhibit specific reactions in protein biosynthesis. Cycloheximide inhibits the elongation step of protein synthesis. Polysomes accumulate in inhibited cells because ribosomes do not terminate normally and new ribosomes enter the polysome, eventually saturating the mRNA. Pactamycin inhibits initiation, and consequently polysomes break down in the presence of this drug. Under this condition, the mRNA is essentially free of ribosomes. pLK109, pLK229, and pRK270 mRNAs were stabilized in the presence of cycloheximide, but pactamycin had no effect on their normal decay. Since it seems likely that stability of mRNA reflects the availability of sites for inactivation by nucleases, it follows that in the presence of cycloheximide, these sites are protected, presumably by occupancy by ribosomes. No ribosomes are bound to mRNA in the presence of pactamycin, and therefore mRNA degrades at about the normal rate. The data further indicate that a labile protein is probably not involved in mRNA decay or stabilization, since protein synthesis is inhibited equally by both antibiotics. We conclude that it may be important to use more than one type of protein synthesis inhibitor to evaluate whether protein synthesis is required for mRNA decay. The effect of protein synthesis inhibition on mRNA synthesis and accumulation was also studied. mRNA synthesis continues in the presence of inhibitors, albeit at a diminished rate relative to that of the uninhibited control.

Cycloheximide↗

Encapsidation of 18 S rRNA by tobacco mosaic virus coat protein.

It has been reported that tobacco mosaic virus capsid protein encapsidates discretesized truncated portions of host 18 S rRNA in vitro. This paper presents further information concerning the nature and specificity of this reaction. We have found that it is only the 5' portions of 18 S rRNA that are encapsidated. The structure recognized by capsid protein is highly conserved; bovine as well as plant 18 S rRNA becomes encapsidated. It is further demonstrated that assembly of 18 S rRNA is slow in comparison to assembly of TMV RNA and that this is due to a slow rate of initiation. Synthetic 18 S rRNA, prepared by in vitro transcription of an 18 S rRNA coding sequence, differs from native 18 S rRNA in that full length, rather than a truncated portion, is encapsidated. The possible reasons for this are discussed.

Base Sequence↗

Dictyostelium discoideum mRNAs developmentally regulated during spore germination have short half-lives.

mRNA decay was studied during spore germination in Dictyoselium discoideum by the use of three previously isolated cDNA clones, pLK109, pLK229, and pRK270, which are specific for mRNAs developmentally regulated during spore germination. The half-life of a constitutive mRNA, pLK125, which is present throughout germination, growth, and development, as also determined. Nogalamycin, a DNA-intercalating compound, was used to inhibit RNA synthesis. Total RNA was isolated at intervals after addition of the drug, and the decay of mRNAs specific for the cDNA clones was determined by both Northern blot and RNA dot hybridization. If nogalamycin was added immediately after activation of dormant spores, neither pLK229 nor pLK109 mRNA decayed, but pLK125 mRNA did decay. Although pLK109 mRNA did not decay under these conditions, the RNA was smaller 1 h after activation than in dormant spores, indicating that it was processed normally. At 1 h after activation, pLK229-, pLK125-specific mRNAs decayed exponentially, with half-lives of 24, 39, and 165 min, respectively. Under the same conditions, decay of pLK109-specific mRNA was biphasic. Thirty-eight percent of the mRNA decayed with a half-life of 5.5 min, and the remainder decayed with a half-life of 115 min. It seems likely that nogalamycin inhibits the synthesis of an unstable component of the mRNA degradative pathway which is needed continuously for the decay of pLK109 mRNA. By extrapolating the curve representing the rapidly decaying component, a half-life of 18 min was calculated for pLK109-specific mRNA. The mRNAs developmentally regulated during spore germination have half-lives shorter than that of the constitutive messenger and shorter than the average half-life of 3 to 4 h previously determined for total Dicyostelium polyadenylated mRNA.

Dictyostelium↗

The Neuman Systems Model in practice. Planning phase.

How do you choose and plan for the implementation of a conceptual model for professional nursing practice? This article discusses one institution's experience with the Neuman Systems Model. The process discussed in this article will serve as a guide for other nursing administrators considering this significant undertaking.

Curriculum↗

Development of a gas chromatographic-mass spectrometric method using a stable isotope internal standard for quantitation of thromboxane B2.

Tetradeuterated 19, 19',20,20'-2H4-thromboxane B2 was synthesized and used as a stable isotope internal standard for the development of a gas chromatographic/mass spectrometric (GC/MS) method to quantitate thromboxane B2. Quantitation was based on the detection of fragment ions at m/z 301 for thromboxane B2 and m/z 305 for the corresponding tetradeuterated internal standard. At m/z 301/305 a response equivalent to a protium/deuterium ratio of 0.2% thromboxane B2 could be measured with a standard deviation of 15% when 100 nanograms of the tetradeuterated internal standard was analyzed. For measurement of cellular biosynthesis of thromboxane B2 in whole blood or platelet rich plasma, the internal standard and naturally occurring thromboxane B2 were i) isolated by solvent extraction, or sequestration to XAD-2 columns; ii) purified by reversed phase high performance liquid chromatography; and iii) converted to methoxime methyl ester trimethylsilyl ethers prior to analysis. A limited comparison was made using both radioimmunological and mass spectrometric quantitation of thromboxane B2.

Animals↗

Multiple sclerosis in the Republic of San Marino.

Previous studies on the prevalence of multiple sclerosis in Italy have grossly underestimated the prevalence of the disease. The prevalence in the Republic of San Marino (near Rimini), in Sicily, and no doubt in the rest of Italy, is of the same order of magnitude as in Europe--that is, 40-60/100 000. The contrast of this with the very low prevalence in Malta (only 60 miles (96 km) away from Sicily) of 4/100 000 should provide a clue to the genetic and environmental factors responsible for multiple sclerosis.

Adult↗

Inhibition of gastric acid secretion in the gastric brooding frog, Rheobatrachus silus.

The female gastric brooding frog Rheobatrachus silus broods its young in its stomach. A substance that inhibits gastric acid secretion in a toad stomach preparation in vitro appears to be secreted by the developing young. This substance has been identified as prostaglandin E2. Rheobatrachus silus may thus have developed a mechanism whereby prostaglandin secreted by the larvae inhibits acid secretion in the stomach of the female until the larvae have completed development and emerged as juvenile frogs by way of the female's mouth.

Animals↗

Characterization of cDNA clones specific for sequences developmentally regulated during Dictyostelium discoideum spore germination.

Spore germination in the slime mold Dictyostelium discoideum was used as a model to study the developmental regulation of protein and mRNA synthesis. Changes in the synthesis of these macromolecules occur during the transition from dormant spore to amoebae. The study of the mechanisms which regulate the quantity and quality of protein synthesis can best be accomplished with cloned genes. cDNA clones which hybridized primarily with mRNAs from only spores or germinating spores and not with growing amoebae were collected. Three such clones, denoted pLK109, pLK229, and pRK270, were isolated and had inserts of approximately 500, 1,200, and 690 base pairs, respectively. Southern blot hybridization experiments suggested that each of the genes is present in multiple copies in the D. discoideum genome. RNA blot hybridizations were performed to determine the sizes of the respective mRNAs and their developmental regulation. The mRNA that hybridized to pLK109 DNA was present predominantly in spores and at 1 h after germination but was absent in growing amoebae. Its concentration dramatically dropped at 3 h. The mRNA present in spores is apparently larger (approximately 0.5 kilobase) than in the later stages of germination (0.4 kilobase), indicating processing of the RNA during germination. The mRNA that hybridized to pLK229 DNA was approximately 1.0 kilobase and was present in very low amounts during growth. Its concentration rose until 1 h after spore germination and decreased thereafter. pRK270-specific RNA was approximately 2.7 kilobases and was found predominantly at 1 h after germination. It was present in lower concentrations at 2 and 3 h after germination and was absent in spores and amoebae. In vitro translation of mRNA selected from 1-h polyadenylated RNA which was hybridized to pLK109 or pLK229 DNA gave proteins of molecular weights consistent with the sizes of the mRNAs as determined by the RNA blot analysis.

Cloning, Molecular↗