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Biomedical subjects

R Powell

Publications and source records attributed to R Powell.

At least 19 recordsLinked to original sources

Sequence of the gene encoding the 16S rRNA of the beer spoilage organism Megasphaera cerevisiae.

The 16S ribosomal RNA gene from the beer-spoilage organism, Megasphaera cerevisiae was polymerase chain reaction (PCR)-amplified and sequenced. Analysis confirmed the phylogenetic position of M. cerevisiae as a sister taxon of Megasphaera elsdenii, within the obligately anaerobic, Gram-negative cocci. The sequence obtained should facilitate the development of DNA probes for early detection of this spoilage organism.

Base Sequence

Recovery and phylogenetic analysis of novel archaeal rRNA sequences from a deep-sea deposit feeder.

In 1992, two independent reports based on small-subunit rRNA gene (SSU rDNA) cloning revealed the presence of novel Archaea among marine bacterioplankton. Here, we report the presence of further novel Archaea SSU rDNA sequences recovered from the midgut contents of a deep-sea marine holothurian. Phylogenetic analyses show that these abyssal Archaea are a paraphyletic component of a highly divergent clade that also includes some planktonic sequences. Our data confirm that this clade is a deep-branching lineage in the tree of life.

Animals

Sequence analysis and evolutionary aspects of piscine alpha-1-microglobulin/bikunin mRNA transcripts.

Studies on mammalian alpha-1-microglobulin/bikunin (AMBP) protein sequences suggest the ancestral AMBP gene arose between 270 and 80 million years ago. Here we report a similar study including recently reported piscine AMBP messenger RNA sequences. The analysis implies a far earlier data for the assembly of the ancestral AMBP gene, more than 450 million years ago. Similarly, a revised date for the origin of the ancestral Kunitz-type protein is estimated at approximately 875 million years ago. Evolutionary conservation is apparent among all the AMBP amino acid sequences. This implies a common functional significance in both teleosts and mammals. However, an unusually rapid rate of evolution is observed for domain II of the piscine bikunin sequences, suggesting that this domain is no longer truly orthologous in fish.

Alpha-Globulins

A novel line of transgenic mice (RSV/LTR-bGH) expressing growth hormone in cardiac and striated muscle.

In order to further investigate the deleterious effects of GH overexpression, we generated a novel line of transgenic mice featuring stable and specific expression of bovine GH in the heart and striated muscle. A DNA construct, containing a region with promoter activity from the Long Terminal Repeat of Rous Sarcoma Virus (RSV-LTR) and the entire structural gene of bovine GH (bGH), was microinjected by standard techniques in male pronuclei of fertilized mice eggs. Transgenic mice expressed bGH mRNA in the heart and striated muscle starting at 5-6 weeks of age. They featured circulating levels of a 22 kDa form of bGH up to 700 ng/ml and enhanced growth starting at 6 weeks of age. No pathologic changes of the myocardium and striated muscle fibers, other than hypertrophy, were noticed, although severe glomerulosclerosis and liver alteration occurred in older mice. Future studies on this new line of transgenic GH mice and integration with the existing data might improve our understanding of the molecular mechanism underlying the detrimental effects of elevated GH levels on various organs and functions.

Animals

Four new coccidia (Apicomplexia: Eimeriidae) from anoles (Lacertilia: Polychrotidae) in the Dominican Republic.

Fecal samples from 25 Anolis armouri, 2 Anolis bahorucoensis, 48 Anolis cybotes, and 21 Anolis olssoni (Lacertilia: Polychrotidae) from southern Hispaniola were examined for coccidian oocysts. Two eimerians and 2 isosporans are herein described as new species. Sporulated oocysts of Eimeria schwartzi n. sp. from A. armouri are ellipsoidal, 22.7 (20.8-25.0) x 15.7 (14.6-17.7) microns, with spherical to subspherical sporocysts, 7.9 (6.2-9.4) x 7.4 (6.2-8.3) microns. Sporulated oocysts of Isospora reui n. sp. from A. bahorucoensis are spherical to subspherical, 18.2 (15.6-20.0) x 17.8 (15.6-19.8) microns, with ovoid sporocysts, 11.9 (10.4-12.7) x 8.5 (7.5-9.4) microns. Sporulated oocysts of Isospora hendersoni n. sp. from A. armouri and A. cybotes are spherical to subspherical, 23.2 (20.8-26.0) x 21.1 (18.4-23.9) microns, with ellipsoidal sporocysts, 14.7 (12.5-15.6) x 10.0 (9.2-11.4) microns. Sporulated oocysts of Eimeria avilae n. sp. from A. olssoni are cylindrical, 29.3 (26.0-33.3) x 15.9 (13.5-18.9) microns, with ellipsoidal sporocysts 10.2 (9.4-11.4) x 6.8 (5.2-8.0) microns.

Animals

Sequence of a cDNA clone encoding the Atlantic salmon alpha 1-microglobulin/bikunin protein.

We report here the nucleotide sequence of a cDNA clone encoding the salmon (Salmo salar) alpha 1-microglobulin/bikunin precursor protein (sAMBP). The encoded precursor shows 36 and 42% amino acid (aa) similarity to the AMBP of pig and human, respectively. Signature aa motifs are conserved. The data infer that the ancestral AMBP gene arose more than 450 million years ago, before the tetrapod-fish divergence.

Alpha-Globulins

Detection of Aeromonas salmonicida, causal agent of furunculosis in salmonid fish, from the tank effluent of hatchery-reared Atlantic salmon smolts.

The fish pathogen, Aeromonas salmonicida, could be detected only by bacteriological culture from the kidney of dead or moribund fish in one tank in a hatchery rearing Atlantic salmon (Salmo salar L.) smolts. However, by using a DNA probe specific for this species, allied to a PCR assay, the pathogen could be detected in water, feces and effluent samples taken from this fish tank. Also, the presence of the pathogen was found in effluent samples from two fish tanks containing apparently healthy fish. Subsequently, the presence of pathogen in these tanks was confirmed by an increase in the daily mortality rate and by a plate culture from moribund fish.

Aeromonas

Dihydrolipoic acid--a universal antioxidant both in the membrane and in the aqueous phase. Reduction of peroxyl, ascorbyl and chromanoxyl radicals.

Thioctic (lipoic) acid is used as a therapeutic agent in a variety of diseases in which enhanced free radical peroxidation of membrane phospholipids has been shown to be a characteristic feature. It was suggested that the antioxidant properties of thioctic acid and its reduced form, dihydrolipoic acid, are at least in part responsible for the therapeutic potential. The reported results on the antioxidant efficiency of thioctic and dihydrolipoic acids obtained in oxidation models with complex multicomponent initiation systems are controversial. In the present work we used relatively simple oxidation systems to study the antioxidant effects of dihydrolipoic and thioctic acids based on their interactions with: (1) peroxyl radicals which are essential for the initiation of lipid peroxidation, (2) chromanoxyl radicals of vitamin E, and (3) ascorbyl radicals of vitamin C, the two major lipid- and water-soluble antioxidants, respectively. We demonstrated that: (1) dihydrolipoic acid (but not thioctic acid) was an efficient direct scavenger of peroxyl radicals generated in the aqueous phase by the water-soluble azoinitiator 2,2'-azobis(2-amidinopropane)-dihydrochloride, and in liposomes or in microsomal membranes by the lipid-soluble azoinitiator 2,2'-azobis(2,4-dimethylvaleronitrile); (2) both dihydrolipoic acid and thioctic acid did not interact directly with chromanoxyl radicals of vitamin E (or its synthetic homologues) generated in liposomes or in the membranes by three different ways: UV-irradiation, peroxyl radicals of 2,2'-azobis(2,4-dimethylvaleronitrile), or peroxyl radicals of linolenic acid formed by the lipoxygenase-catalyzed oxidation; and (3) dihydrolipoic acid (but not thioctic acid) reduced ascorbyl radicals (and dehydroascorbate) generated in the course of ascorbate oxidation by chromanoxyl radicals. This interaction resulted in ascorbate-mediated dihydrolipoic acid-dependent reduction of the vitamin E chromanoxyl radicals, i.e. vitamin E recycling. We conclude that dihydrolipoic acid may act as a strong direct chain-breaking antioxidant and may enhance the antioxidant potency of other antioxidants (ascorbate and vitamin E) in both the aqueous and the hydrophobic membraneous phases.

Animals

The kinetics of oocyte activation and polar body formation in bovine embryo clones.

The kinetics of polar body formation were examined in parthenogenetically activated, in vitro matured and aged bovine oocytes. Subsequently, the presence or absence of polar body formation was determined in bovine embryo clones. Polar body formation, defined as telophase II, occurred by 1 (13/40, 43%) and 2 h (15/21, 71%) postparthenogenetic activation of metaphase II stage oocytes. Parthenogenetically activated oocytes readily formed pronuclei by 4 h. Some oocytes had chromatin in a highly condensed state at 1, 2, and 4 h postactivation (13/72, 18%). These oocytes often (10/13, 77%) appeared to be "self-enucleated," as the condensed chromatin was found in a membrane-bound extrusion. The phenomenon was most prevalent when oocytes were handled at room temperature (25-27 degrees C). Nuclear transfer procedures were established to bring about synchronous blastomere fusion and oocyte activation conditions. Synchronous conditions were achieved only when oocytes were handled and manipulated at 37-39 degrees C. Embryo clones examined 2 h postfusion did not form a polar body. Conversely, nucleate demi-oocyte controls were at the late telophase II stage of meiosis. The results are discussed in relation to cell cycle effects on bovine nuclear transfer.

Animals

Gene expression following transfection of fish cells.

Various genes containing different transcriptional regulatory elements (TRE) and the bacterial marker gene coding for chloramphenicol acetyl transferase were transfected into several fish cell lines to evaluate the efficiency of expression in comparison with mammalian cells. The CMV and RSV TRE were the most efficient non-inducible promoters in directing reporter gene expression. RSV and CMV appeared of similar potency in a stable fish cell line. The human HSP-70 promoter showed high potency in a carp and in a trout cell line after thermal induction. This promoter also induced the synthesis of human growth hormone directed by the corresponding cDNA, but not by the gene. RSV TRE was also able to drive the synthesis of bovine growth hormone when attached directly to the cDNA but not to the gene. These data suggest that non-fish gene TRE can be used to express foreign genes in fish cells or transgenic fish; however, in most cases they are relatively inefficient. The data also suggest that the translation and secretion machinery of fish cells can express efficiently foreign genes but that mammalian introns might be not processed properly in some cases.

Animals

DNA probe for Aeromonas salmonicida.

A DNA fragment that is specific to Aeromonas salmonicida has been isolated from a genomic DNA library by differential hybridization. The specificity of this fragment as a DNA probe for A. salmonicida was shown by hybridization against reference strains and clinical isolates of A. salmonicida, related aeromonads, and species from several other bacterial genera. The sensitivity of detection by a polymerase chain reaction test, based on this fragment, was approximately two A. salmonicida cells.

Aeromonas

An adenosine at position 27 in the human immunodeficiency virus type 1 trans-activation response element is not critical for transcriptional or translational activation by Tat.

Tat protein binds to the trans-activation response (TAR) element of human immunodeficiency virus type 1 RNAs and activates gene expression at the level of transcription in mammalian cell lines and translation in Xenopus oocytes. Certain residues within TAR are important for Tat binding in vitro, including residue A-27, which appears to be able to be modified in a Tat-dependent manner in Xenopus oocytes (L. Sharmeen, B. Bass, N. Sonenberg, H. Weintraub, and M. Groudine, Proc. Natl. Acad. Sci. USA 88:8096-8100, 1991). Activation by Tat in oocytes occurs via a covalent modification of TAR-containing RNA. We have found that in both mammalian cells and Xenopus oocytes, conversion of A-27.U-38 or C-27.G-38 or C-27.G-38 reduces activation. However, conversion to G-27.U-38 or G-27.C-38 had little or no effect on activation, and in oocytes, these mutant RNAs were still covalently modified. These data exclude a specific role for the adenosine at residue 27 for Tat activation but suggest a requirement for a purine at this position.

Adenosine

The dieffenbachia plant. Case history.

When ingested, the common house plant dieffenbachia can lead to significant toxicity and possibly death if timely medical attention is not forthcoming following initial exposure. The deleterious effects should not be overlooked especially when children are involved who, naturally curious, may bite into this innocuous looking plant. Although the patient in this case was a type II diabetic on insulin, neither hypo- nor hyperglycemia developed. There was no other significant electrolyte imbalance despite the acute stressor placed on him.

Adult

The salmon gene encoding apolipoprotein A-I: cDNA sequence, tissue expression and evolution.

A cDNA encoding an apolipoprotein (Apo) has been isolated from the Atlantic salmon (Salmo salar) and sequenced. It encodes a peptide of 258 amino acids (aa), including a signal peptide of 18 aa, with 5'- and 3'-untranslated regions of the mRNA of 12 and 329 nucleotides, respectively. The protein has structural features in common with other Apo's of human and avian origin, including conserved sequences in the signal peptide and a series of internal repeats of 22 aa. The sequence has been identified as salmon Apo A-I (sApoA-I), and has 23% aa identity with human ApoA-I. Northern-blot analysis using the sApoA-I cDNA probe against total RNA prepared from several salmon tissues detects the expression of this gene in liver, intestine and muscle. A phylogenetic analysis reveals that the mammalian ApoA-I, ApoA-IV and Apo-E aa sequences are more closely related to each other than any of them are to sApoA-I. This suggests that the duplication events, from which A-I, A-IV and E arose, occurred after the divergence of the tetrapod and teleost ancestors.

Amino Acid Sequence

Use of the Branemark implant in the cleft palate patient.

This paper describes the clinical and surgical management of a patient presenting with a unilateral alveolar cleft and associated oronasal fistula. After orthodontic expansion of the maxilla, a secondary osseous graft was placed. A single Branemark implant was subsequently utilized to allow for prosthetic restoration of the dental arch. Through case presentation, a detailed course of treatment is outlined that effectively restores the anatomy, integrity, and function of both the alveolar and dental arches.

Adolescent