Biomedical subjects

R L Wilson

Publications and source records attributed to R L Wilson.

At least 19 recordsLinked to original sources

PubMed2004

Isomeric interconversion in the linear Cl(-)-HD anion complex.

The rotationally resolved infrared photodissociation spectrum of Cl(-)-HD is measured in the HD stretch region. Two Sigma-Sigma bands are observed, corresponding to transitions from the ground state [the (nuHD = 0, n = 0) level] and first excited intermolecular bend state [the (nuHD = 0, n = 1) level]. The (nuHD = 0, n = 0) and (nuHD = 0, n = 1) states are predominantly associated with the linear Cl-...DH and Cl-...HD geometries, respectively. The spectrum is complicated by perturbative interactions between levels of the (nuHD = 0, n = 0) and (nuHD = 0, n = 1) rotational manifolds and between levels of the (nuHD = 1, n = 0) and (nuHD = 1, n = 1) rotational manifolds. A global fit to the transition frequencies, taking the lower and upper state perturbations into account, yields zero-order rotational and centrifugal distortion constants and allows us to establish that the (nuHD = 0, n = 1, J" = 0) level lies 13.7 cm(-1) above the (nuHD = 0, n = 0, J" = 0) level. Rovibrational energy level calculations performed using a recent ab initio potential energy surface confirm the picture emerging from the experimental data and provide good agreement with measured molecular parameters. The results emphasize the importance of quantum mechanical interconversion between two isomeric structures of a simple anion complex.

Journal Article↗

PubMed2001

Fis, a DNA nucleoid-associated protein, is involved in Salmonella typhimurium SPI-1 invasion gene expression.

The ability of Salmonella enterica serovar Typhimurium to cause disease depends upon the co-ordinated expression of many genes located around the Salmonella chromosome. Specific pathogenicity loci, termed Salmonella pathogenicity islands, have been shown to be crucial for the invasion and survival of Salmonella within host cells. Salmonella pathogenicity island 1 (SPI-1) harbours the genes required for the stimulation of Salmonella uptake across the intestinal epithelia of the infected host. Regulation of SPI-1 genes is complex, as invasion gene expression responds to a number of different signals, presumably signals similar to those found within the environment of the intestinal tract. As a result of our continued studies of SPI-1 gene regulation, we have discovered that the nucleoid-binding protein Fis plays a pivotal role in the expression of HilA and InvF, two activators of SPI-1 genes. A S. typhimurium fis mutant demonstrates a two- to threefold reduction in hilA:Tn5lacZY and a 10-fold reduction in invF:Tn5lacZY expression, as well as a 50-fold decreased ability to invade HEp-2 tissue culture cells. This decreased expression of hilA and invF resulted in an altered secreted invasion protein profile in the fis mutant. Furthermore, the virulence of a S. typhimurium fis mutant is attenuated 100-fold when administered orally, but has wild-type virulence when administered intraperitoneally. Expression of hilA:Tn5lacZY and invF:Tn5lacZY in the fis mutant could be restored by introducing a plasmid containing the S. typhimurium fis gene or a plasmid containing hilD, a gene encoding an AraC-like regulator of Salmonella invasion genes.

Animals↗

PubMed2001

Identification of Listeria monocytogenes in vivo-induced genes by fluorescence-activated cell sorting.

Listeria monocytogenes is a gram-positive, intracellular, food-borne pathogen capable of causing severe infections in immunocompromised or pregnant individuals, as well as numerous animal species. Genetic analysis of Listeria pathogenesis has identified several genes which are crucial for virulence. The transcription of most of these genes has been shown to be induced upon entry of Listeria into the host cell. To identify additional genes that are induced in vivo and may be required for L. monocytogenes pathogenesis, a fluorescence-activated cell-sorting technique was initiated. Random fragments of the L. monocytogenes chromosome were cloned into a plasmid carrying a promoterless green fluorescent protein (GFP) gene, and the plasmids were transformed into the L. monocytogenes actA mutant DP-L1942. Fluorescence-activated cell sorting (FACS) was used to isolate L. monocytogenes clones that exhibited increased GFP expression within macrophage-like J774 cells but had relatively low levels of GFP expression when the bacteria were extracellular. Using this strategy, several genes were identified, including actA, that exhibited such an expression profile. In-frame deletions of two of these genes, one encoding the putative L. monocytogenes uracil DNA glycosylase (ung) and one encoding a protein with homology to the Bacillus subtilis YhdP hemolysin-like protein, were constructed and introduced into the chromosome of wild-type L. monocytogenes 10403s. The L. monocytogenes 10403s ung deletion mutant was not attenuated for virulence in mice, while the yhdP mutant exhibited a three- to sevenfold reduction in virulence.

Amino Acid Sequence↗

PubMed2001

Hha is a negative modulator of transcription of hilA, the Salmonella enterica serovar Typhimurium invasion gene transcriptional activator.

An early step in the establishment of Salmonella enterica serovar Typhimurium murine infection is the penetration of the intestinal mucosa of the small intestine. The majority of the genes responsible for the Salmonella invasive phenotype are encoded on Salmonella pathogenicity island 1, and their transcription is controlled by the hilA transcriptional activator. The expression of hilA is regulated by environmental signals including oxygen, osmolarity, pH, and growth phase such that the presence of any one suboptimal condition results in repression of hilA expression and the invasive phenotype. We have conducted a search for negative regulators of hilA by introduction of a Salmonella enterica serovar Typhimurium chromosomal DNA gene bank into a Salmonella enterica serovar Typhimurium hilA::Tn5lacZY reporter strain. This screen has identified the hha gene as a regulator that exerts a negative influence on hilA expression. Plasmid-encoded hha significantly reduces hilA::Tn5lacZY chromosomal expression, as well as expression of the invasion genes invF, prgH, and sipC. An hha null mutation results in substantial derepression of both chromosomally encoded and plasmid-encoded hilA::Tn5lacZY expression. Introduction of plasmid-encoded hha into strain SL1344 results in attenuation of invasion using in vitro and in vivo assays. Importantly, purified Hha protein was found to bind to a hilA DNA promoter fragment, suggesting that the regulatory activity of the Hha protein occurs at the hilA promoter. These data add detail to the developing model of the regulation of Salmonella invasion genes.

Animals↗

PubMed2000

Transient expression of bacterial gene fragments in eukaryotic cells: implications for CD8(+) T cell epitope analysis.

CD8(+) T cells are potent effectors of acquired immunity against some viruses and intracellular bacterial pathogens. Antigens recognized by CD8(+) T cells are small, 8-9 amino acid peptides derived from proteins produced by the pathogen. These peptides are presented by MHC class I molecules on the surface of the infected cell. When characterizing the CD8(+) T cell response to a bacterial or viral pathogen, it is often necessary to express an antigenic protein in a eukaryotic host cell that is capable of processing and presenting peptide epitopes to antigen-specific CD8(+) T cells. We describe a system designed to transiently express bacterial polypeptides and MHC class I molecules in eukaryotic cells. Recognition of these peptide-MHC complexes stimulates TNF production by antigen-specific CD8(+) T cell lines. This system should be useful for analysis of CD8(+) T cell epitope-containing bacterial gene fragments when expression of the entire bacterial protein is detrimental to the eukaryotic cell, or when overexpression of the bacterial gene is detrimental to the bacterial cloning strain. Furthermore, this system can be used for the rapid mapping of CD8(+) T cell epitopes within a protein.

Animals↗

PubMed2000

Salmonella enterica serovars gallinarum and pullorum expressing Salmonella enterica serovar typhimurium type 1 fimbriae exhibit increased invasiveness for mammalian cells.

Salmonella enterica serovars Gallinarum and Pullorum are S. enterica biotypes that exhibit host specificity for poultry and aquatic birds and are not normally capable of causing disease in mammalian hosts. During their evolution toward host restriction serovars Gallinarum and Pullorum lost their ability to mediate mannose-sensitive hemagglutination (MSHA), a phenotype correlated with adherence to certain cell types. Because adherence is an essential requirement for invasion of cells by bacterial pathogens, we examined whether MHSA type 1 fimbriae would increase the ability of serovars Pullorum and Gallinarum to invade normally restrictive cells. Serovars Gallinarum and Pullorum expressing S. enterica serovar Typhimurium strain LT2 type 1 fimbriae exhibited a 10- to 20-fold increased ability to adhere to and a 20- to 60-fold increased invasion efficiency of the human epithelial HEp-2 cell line. Invasion was accompanied by extensive ruffling of the membranes of the HEp-2 cells. In a murine ligated ileal loop model, a 32% increase in the number of M-cell ruffles was seen when serovar Gallinarum expressed serovar Typhimurium type 1 fimbriae.

Animals↗

PubMed2000

Conventional resistance of experimental maize lines to corn earworm (Lepidoptera: Noctuidae), fall armyworm (Lepidoptera: Noctuidae), southwestern corn borer (Lepidoptera: Crambidae), and sugarcane borer (Lepidoptera: Crambidae).

Plant resistance is a useful component of integrated pest management for several insects that are economically damaging to maize, Zea mays L. In this study, 15 experimental lines of maize derived from a backcross breeding program were evaluated for resistance to corn earworm, Helicoverpa zea (Boddie); fall armyworm, Spodoptera frugiperda (J. E. Smith); southwestern corn borer, Diatraea grandiosella Dyar; and sugarcane borer, Diatraea saccharalis (F.). Experimental line 100-R-3 was resistant in the field to leaf feeding by fall armyworm and line 116-B-10 was resistant in the field to leaf feeding by fall armyworm and leaf and stalk feeding by southwestern corn borer. When corn earworm larvae were fed field harvested silks from experimental line 81-9-B in the laboratory, their pupal weights were significantly lower than the pupal weights of larvae that were fed silks from the resistant control, Zapalote Chico. Maysin levels lower than those commonly associated with corn earworm resistance were present in the resistant experimental line, 107-8-7, indicating a new basis confers resistance to corn earworm in this line. These resistant experimental lines will provide plant breeders with new sources of resistance to lepidopterous insects for the development of improved maize breeding populations.

Animals↗

PubMed2000

Evaluation of conventional resistance to European corn borer (Lepidoptera: Crambidae) and western corn rootworm (Coleoptera: Chrysomelidae) in experimental maize lines developed from a backcross breeding program.

Plant resistance is a promising control method for the two most damaging insect pests of maize, Zea mays L.: the European corn borer, Ostrinia nubilalis (Hübner), and the western corn rootworm Diabrotica virgifera virgifera LeConte. Fifteen experimental lines of maize, derived from a backcross breeding program designed to introgress resistance to European corn borer from Peruvian maize into two U.S. Corn Belt adapted inbred lines, were evaluated for resistance to European corn borer and western corn rootwonrm. The experimental lines were in the second generation of backcrossing. All experimental lines were resistant to leaf blade feeding by European corn borer. These lines had low levels of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one, a chemical commonly associated with leaf blade feeding resistance, indicating that this was not the mechanism of resistance to leaf blade feeding in these lines. Eleven experimental lines were resistant to leaf sheath and collar feeding by European corn borer. Useful sources of European corn borer ovipositional nonpreference and root feeding resistance to western corn rootworm were not identified. Some of the lines evaluated in this study may provide useful sources of resistance to both leaf blade and leaf sheath and collar feeding by European corn borer.

Animals↗

PubMed1996

Extensor tendon problems in rheumatoid arthritis.

Extensor tenosynovitis is a common presenting upper extremity problem and, unless it resolves with medical management, preventative tenosynovectomy is indicated to prevent tendon rupture. When a rupture has occurred, tendon reconstruction with either a transfer or a graft has a reasonable chance of restoring function as long as the number of tendons involved is limited. Rupture of a single extensor tendon requires surgical treatment to eliminate the cause and prevent further damage, as well as repairing the injured tendon. It is important to give consideration to associated wrist and MP joint problems before, during, or after treating tendon abnormalities.

Arthritis, Rheumatoid↗

PubMed1995

DNA binding sites of the LysR-type regulator GcvA in the gcv and gcvA control regions of Escherichia coli.

The GcvA protein is a LysR family regulatory protein necessary for both activation and repression of the Escherichia coli glycine cleavage enzyme operon (gcv) and negative regulation of gcvA. Gel shift assays indicated that overexpressed GcvA in crude extracts is capable of binding specifically to DNA containing the gcv and gcvA control regions. DNase I footprint analysis of the gcvA control region revealed one region of GcvA-mediated protection overlapping the transcription initiation site and extending from -28 to +20. Three separate GcvA binding sites in gcv were identified by DNase I footprint analysis: a 29-bp region extending from positions -271 to -242, a 28-bp region extending from -242 to -214, and a 35-bp region covering positions -69 to -34 relative to the transcription initiation site. PCR-generated mutations in any of the three GcvA binding sites in gcv decreased GcvA-mediated activation and repression of gcv.

Amino Acid Oxidoreductases↗

PubMed1994

Intelligibility and nonspeech orofacial strength and force control following traumatic brain injury.

Objective measures of nonspeech orofacial strength and force control were obtained for 20 individuals with traumatic brain injury. The dynamic and static force generating abilities of the upper lip, lower lip, tongue, and jaw were assessed. Based on sentence intelligibility scores, the subjects were divided into two groups, more and less intelligible. Force measures included reaction time, slope, derivative, peak overshoot, and first- and second-half mean hold and standard deviation. Groups differed only in the ability to sustain the 2 N force level with the tongue. Other potential contributors to the differences in intelligibility are discussed.

Adolescent↗

PubMed1994

DNA sequence and characterization of GcvA, a LysR family regulatory protein for the Escherichia coli glycine cleavage enzyme system.

The gene encoding GcvA, the trans-acting regulatory protein for the Escherichia coli glycine cleavage enzyme system, has been sequenced. The gcvA locus contains an open reading frame of 930 nucleotides that could encode a protein with a molecular mass of 34.4 kDa, consistent with the results of minicell analysis indicating that GcvA is a polypeptide of approximately 33 kDa. The deduced amino acid sequence of GcvA revealed that this protein shares similarity with the LysR family of activator proteins. The transcription start site was found to be 72 bp upstream of the presumed translation start site. A chromosomal deletion of gcvA resulted in the inability of cells to activate the expression of a gcvT-lacZ gene fusion when grown in the presence of glycine and an inability to repress gcvT-lacZ expression when grown in the presence of inosine. The regulation of gcvA was examined by constructing a gcvA-lacZ gene fusion in which beta-galactosidase synthesis is under the control of the gcvA regulatory region. Although gcvA expression appears to be autogenously regulated over a two- to threefold range, it is neither induced by glycine nor repressed by inosine.

Amino Acid Sequence↗

PubMed1993

Roles of the GcvA and PurR proteins in negative regulation of the Escherichia coli glycine cleavage enzyme system.

When Escherichia coli was grown in medium containing both inosine and glycine, the PurR repressor protein was shown to be responsible for a twofold reduction from the fully induced glycine cleavage enzyme levels. This twofold repression was also seen by measuring beta-galactosidase levels in cells carrying a lambda gcvT-lacZ gene fusion. In this fusion, the synthesis of beta-galactosidase is under the control of the gcv regulatory region. A DNA fragment carrying the gcv control region was shown by gel mobility shift assay and DNase I footprinting to bind purified PurR protein, suggesting a direct involvement of the repressor in gcv regulation. A separate mechanism of purine-mediated regulation of gcv was shown to be independent of the purR gene product and resulted in an approximately 10-fold reduction of beta-galactosidase levels when cells were grown in medium containing inosine but lacking the inducer glycine. This additional repression was dependent upon a functional gcvA gene, a positive activator for the glycine cleavage enzyme system. A dual role for the GcvA protein as both an activator in the presence of glycine and a repressor in the presence of inosine is suggested.

Amino Acid Oxidoreductases↗

PubMed1993

Positive regulation of the Escherichia coli glycine cleavage enzyme system.

A new mutation in Escherichia coli, designated gcvA1, that results in noninducible expression of both gcv and a gcvT-lacZ gene fusion was isolated. A plasmid carrying the wild-type gcvA gene complemented the mutation and restored glycine-inducible gcv and gcvT-lacZ gene expression. These results suggest that gcvA encodes a positive-acting regulatory protein that acts in trans to increase expression of gcv.

Amino Acid Oxidoreductases↗

PubMed1993

Common hand and wrist injuries in basketball players.

Multiple injuries to the hand are relatively common in basketball. This article explains the importance of an early and thorough examination, preferably by a hand specialist. It is important to obtain a history of prior injury during the assessment because treatment and prognosis may be very different. Delayed treatment often yields an inferior result and lost time. If an injury is not treated initially, it may require operative treatment or even be untreatable.

Basketball↗

PubMed1991

Effect of level of endophyte infection, nitrogen fertilization rate, grazing period, and paddock exchange on some chemical properties of four bovine tissues.

Three grazing experiments were conducted to determine the effect of level of endophyte infection, rate of N fertilization of tall fescue grass, grazing period, and paddock exchange on selected chemical properties of four bovine carcass tissues. Samples of semitendinosus and longissimus muscle and of subcutaneous and perinephric adipose tissue were excised from the left side of each carcass. In Exp. 1, percentage of stearic acid was higher (P less than .05) and percentages of palmitoleic and oleic acid were lower (P less than .05) in all tissues from steers grazed on LELN Au-Triumph fescue than in tissues from steers grazed on LELN KY-31 fescue. Grazing periods of 175 or 245 d or paddock exchange (Exp. 2) had no significant effect on proximate composition of semitendinosus and longissimus muscles; however, moisture content was higher (P less than .05) in the semitendinosus muscle from steers grazed on 100% (100E) endophyte-infected KY-31 fescue. Forage treatment, grazing period, and paddock exchange (Exp. 2 and 3) had no significant effect on percentage of saturated fatty acids in the subcutaneous, semitendinosus, and longissimus tissue samples. In Exp. 2 and 3, percentages of saturated fatty acids were higher (P less than .05) in subcutaneous and perinephric adipose tissue samples from steers grazed on 100E than in samples from steers grazed on zero endophyte (OE) fescue.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

PubMed1990

Modulation of hippocampal primed burst potentiation by anesthesia.

This study demonstrates that the anesthetics urethane and pentobarbital differentially affect a low threshold form of long-lasting synaptic plasticity, termed primed burst (PB) potentiation, in the CA1 area of rat hippocampus. PB potentiation was generated by the delivery of a 5-pulse patterned stimulus train, consisting of one priming pulse followed 170 ms later by a burst of 4 pulses at 200 Hz. PB potentiation could not be reliably generated in urethane-anesthetized rats unless stimulus currents were raised to 150% of baseline levels during the stimulus train. In pentobarbital-anesthetized rats, PB potentiation could always be evoked at baseline stimulus intensities. Differences between the anesthetics which could contribute to their varying effects upon PB potentiation are discussed.

Action Potentials↗

PubMed1990

Interference by DOPAC and ascorbate during attempts to measure drug-induced changes in neostriatal dopamine with Nafion-coated, carbon-fiber electrodes.

The selectivity of Nafion-coated, carbon-fiber electrodes was evaluated for the voltammetric detection of dopamine in the presence of 3,4-dihydroxyphenylacetic acid (DOPAC) and ascorbate, which are interferant anions in the brain. Nafion coating was applied to both polished carbon-disk electrodes and electrochemically modified electrodes. During in vitro testing, polished disk electrodes showed the greatest selectivity for dopamine at the fastest scan rate tested (300 V s-1), but the drift in signal made slow changes in dopamine difficult to determine. Electrochemically modified electrodes already provide an ascorbate wave distinct from that for dopamine, but Nafion coating actually decreased dopamine selectivity with respect to DOPAC. In vivo testing was carried out in the neostriatum of urethane-anesthetized rats in response to drug- or stimulation-induced increases in dopamine transmission. Administration of haloperidol (0.5 mg kg-1) followed by GBR 12909 (20 mg kg-1), which is known to cause a 10-fold increase in extracellular dopamine, failed to produce a selective signal for dopamine when measured voltammetrically. Comparable results were obtained following administration of amphetamine (2.5 mg kg-1), which also increases dopamine overflow. Voltammetric detection of dopamine was possible, however, during electrical stimulation of dopaminergic afferents in the medial forebrain bundle. Thus, voltammetry with Nafion-coated electrodes is best suited to the measurement of transient changes in extracellular dopamine rather than the relatively prolonged changes in dopamine overflow produced by various drugs.

3,4-Dihydroxyphenylacetic Acid↗