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Biomedical subjects

C E Shaw

Publications and source records attributed to C E Shaw.

At least 55 records · Page 3Linked to original sources
PubMed1996

Association of apolipoprotein E epsilon 4 allele with bulbar-onset motor neuron disease.

BACKGROUND: Variants of the apolipoprotein E (APOE) gene influence the age of onset of Alzheimer's disease. APOE may influence the presentation of other neurological diseases. We investigated the relationship between the allelic variants of apolipoprotein E and clinical presentation in motor neuron disease. METHODS: 123 patients with motor neuron disease and 121 controls were studied. Diagnosis, location of onset and date of onset were recorded prospectively. Genotyping was performed blind to clinical information. FINDINGS: Possession of at least one epsilon 4 allele was significantly more common in patients with bulbar onset motor neuron disease (14/33, 42%) than in limb onset patients (20/90, 22%) and controls (26/121, 21%) (chi 2 = 4.93, p = 0.026 and chi 2 = 5.91, p = 0.015, respectively). INTERPRETATION: These results suggest that the apolipoprotein E epsilon 4 allele may influence the pattern of motor neuron loss in motor neuron disease and that it may affect neuronal function in ways unrelated to the deposition of beta-amyloid or accumulation of neurofibrillary tangles.

Age of Onset↗
PubMed1995

Measurement of immune markers in the serum and cerebrospinal fluid of multiple sclerosis patients during clinical remission.

Magnetic resonance imaging of multiple sclerosis (MS) patients often shows active inflammatory lesions despite clinical remission. No immunological marker of disease activity has been identified in these patients. Concentrations of neopterin, interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R) and tumour necrosis factor-alpha (TNF-alpha) were measured in the serum and cerebrospinal fluid of 19 clinically-inactive MS patients and compared with those of 19 non-inflammatory controls. Cerebrospinal fluid (CSF) neopterin concentrations were significantly higher in the MS group than in controls (mean 9.1 mM vs 3.4 nM, P < 0.01) and 10 of 19 MS patients had levels above the control range. This finding provides evidence of ongoing T-cell-directed and interferon-gamma-mediated macrophage activation in the central nervous system. Analysis of IL-2, sIL-2R and TNF-alpha concentrations revealed no significant differences between MS patients and controls. We conclude that CSF neopterin concentration may correlate with disease activity in asymptomatic patients.

Adult↗
PubMed1995

Physical determinants of independence in mature women.

The purpose of this study was to determine the relationship in mature women between muscle strength and whole body oxidative capacity and the ability to perform activities of daily living (ADL). Sixty-one women (mean age 69 years) without major disease or disability were recruited from either a community exercise center or a personal care facility. Physiological measurements consisted of peak oxygen consumption on a cycle ergometer (VO2peak) and one repetition maximum strength of nine muscle groups (1-RM). Ability to perform ADL was measured with a balance and gait test, "Bag Carrying Test", and ADL questionnaires. Significant correlations were found with VO2peak and calf muscle strength and ability to perform ADL, with weaker or no correlations for other muscle groups. For some relationships, it was possible to identify the minimum level of physiological functioning associated with successful performance of independence tasks. In summary, physiological capacities, particularly VO2peak and strength of the calf muscles, predicted ability to perform activities needed for functional independence in healthy mature women.

Activities of Daily Living↗
PubMed1993

Biogenesis and regulation of the Vibrio cholerae toxin-coregulated pilus: analogies to other virulence factor secretory systems.

Biogenesis of the toxin-coregulated pilus (TCP) of Vibrio cholerae 01 is essential for successful bacterial colonization of the small intestine. Pilus assembly requires the products of at least seven genes located on the chromosome adjacent to the pilin-encoding gene, tcpA. Previously reported TnphoA insertions in the TCP-assembly-deficient V. cholerae strains, KP2.21 and KP4.2, were isolated from the chromosome for further analysis. Nucleotide sequencing of the tcpE::phoA and tcpF::phoA fusions and corresponding clones of the region containing the intact genes revealed the presence of two open reading frames (ORFs) of 340 and 338 amino acids, designated TcpE and TcpF, respectively. The partial sequence of an ORF downstream from the TcpF coding sequence was determined to correspond to the global virulence regulator, ToxT. Proteins corresponding to the observed ORFs were visualized with the T7 promoter/RNA polymerase expression system. Computer-generated alignment algorithms predict that a homology exists between TcpE and the Klebsiella pneumoniae pullulanase secretion proteins PulD and PulF, the Xanthomonas campestris extracellular enzyme secretion factor XpsF, the Bacillus subtilis DNA competence protein ComG-ORF2, and the Yersinia enterocolitica Yop secretion determinant YscC. These observations provide a model to investigate further the relationship between the secretion mechanisms utilized by these seemingly diverse virulence determinants. Additionally, an extreme C-terminal segment of TcpE shows striking homology to the transmembrane segment of the eukaryotic integrin beta-1 chain, which could imply a role for TcpE in not only TCP secretion, but also host cell interaction.

Amino Acid Sequence↗
PubMed1993

3 beta-hydroxysteroid dehydrogenase activity in tissues of the human fetus determined with 5 alpha-androstane-3 beta,17 beta-diol and dehydroepiandrosterone as substrates.

3 beta-Hydroxysteroid dehydrogenase (3 beta-HSD)/delta 5-->4-isomerase activity in steroidogenic tissues is required for the synthesis of biologically active steroids. Previously, by use of dehydroepiandrosterone (3 beta-hydroxy-5-androsten-17-one, DHEA) as substrate, it was established that in addition to steroidogenic tissues 3 beta-HSD/delta 5-->4-isomerase activity also is expressed in extraglandular tissues of the human fetus. In the present study, we attempted to determine whether the C-5,C-6-double bond of DHEA serves to influence 3 beta-HSD activity. For this purpose, we compared the efficiencies of a 3 beta-hydroxy-5-ene steroid (DHEA) and a 3 beta-hydroxy-5 alpha-reduced steroid (5 alpha-androstane-3 beta,17 beta-diol, 5 alpha-A-diol) as substrates for the enzyme. The apparent Michaelis constant (Km) for 5 alpha-A-diol in midtrimester placenta, fetal liver, and fetal skin tissues was at least one order of magnitude higher than that for DHEA, viz the apparent Km of placental 3 beta-HSD for 5 alpha-A-diol was in the range of 18 to 40 mumol/l (n = 3) vs 0.45 to 4 mumol/l for DHEA (n = 3); for the liver enzyme, 17 mumol/l for 5 alpha-A-diol and 0.60 mumol/l for DHEA, and for the skin enzyme 14 and 0.18 mumol/l, respectively. Moreover, in 13 human fetal tissues evaluated the maximal velocities obtained with 5 alpha-A-diol as substrate were higher than those obtained with DHEA. A similar finding in regard to Kms and rates of product formation was obtained by use of purified placental 3 beta-HSD with DHEA, pregnenolone, and 3 beta-hydroxy-5 alpha-androstan-17-one (epiandrosterone) as substrates: the Km of 3 beta-HSD for DHEA was 2.8 mumol/l, for pregnenolone 1.9 mumol/l, and for epiandrosterone 25 mumol/l. The specific activity of the purified enzyme with pregnenolone as substrate was 27 nmol/mg protein.min and, with epiandrosterone, 127 nmol/mg protein.min. With placental homogenate as the source of 3 beta-HSD, DHEA at a constant level of 5 mumol/l behaved as a competitive inhibitor when the radiolabeled substrate, [3H]5 alpha-A-diol, was present in concentrations of 20 to 60 mumol/l, but at lower substrate concentrations the inhibition was of the mixed type; similar results were obtained with [3H]DHEA as the substrate at variable concentrations in the presence of a fixed concentration of 5 alpha-A-diol (40 mumol/l).(ABSTRACT TRUNCATED AT 400 WORDS)

3-Hydroxysteroid Dehydrogenases↗
PubMed1991

3 beta-Hydroxysteroid dehydrogenase activity in glandular and extraglandular human fetal tissues.

The expression of 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) in steroidogenic tissues is an absolute requirement for mammalian reproduction, fetal growth, and life maintenance. We sought to identify extraglandular tissue sites in the human fetus where 3 beta HSD is expressed. To this effect, we conducted in vitro studies by use of homogenates prepared from second trimester fetal tissues. To facilitate the determination of 3 beta HSD activity, an abbreviated technique was developed that consisted in the use of [3 alpha-3H]dehydroepiandrosterone [( 3 alpha-3H]DHEA) as the substrate and NAD+ as the cofactor. With these reagents, the enzymatic reaction leads to the production of both nonradiolabeled androstenedione and NAD3H in equimolar amounts, and the radioactivity associated with NAD3H is used for quantification of 3 beta HSD activity. The kinetic isotope effect introduced by substitution of tritium for hydrogen at the C-3 alpha position of DHEA, determined with six different tissues, was 2.5 +/- 0.7 (mean +/- SD). The specific activities of the enzyme in peripheral tissues and ovary were relatively low, in the range of 0.03 nmol/mg protein.h for stomach (n = 2) to 0.18 +/- 0.14 nmol/mg protein.h for liver (mean +/- SD; n = 13), while in fetal testis and placenta the specific activities were relatively high, viz. 3.4 +/- 0.7 nmol/mg protein.h (mean +/- SD; n = 4) and 2.8 +/- 1.8 nmol/mg protein.h (mean +/- SD; n = 13), respectively. The findings of this study serve to demonstrate that 3 beta HSD is distributed widely among tissues of the human fetus. Although the enzymatic activity was easily demonstrated in peripheral tissues by the use of radiolabeled DHEA as the substrate, 3 beta HSD protein was not readily detected by Western analysis.

3-Hydroxysteroid Dehydrogenases↗
PubMed1990

Vibrio cholerae O395 tcpA pilin gene sequence and comparison of predicted protein structural features to those of type 4 pilins.

Vibrio cholerae O1 expresses a pilus that is coordinately regulated with cholera toxin production and hence termed TCP, for toxin-coregulated pilus. Insertion of Tn5 IS50L::phoA (TnphoA) into the major pilin subunit gene, tcpA, has previously been shown to render the strain avirulent as a result of its inability to colonize. One such insertion was isolated and used as a probe to screen for clones containing the intact tcpA gene. The DNA sequence of tcpA was determined by using the intact gene and several tcpA-phoA gene fusions. The deduced protein sequence agreed completely with that previously determined for the TcpA N terminus and with the size of the mature pilin protein. The reported homology with N-methylphenylalanine (type 4) pilins near the N terminus was extended and shown to include components of the atypical leader peptide as well as overall predicted structural similarities in other regions of the pilins. In contrast to the modified N-terminal phenylalanine residue found in all characterized type 4 pilins, the corresponding position in tcpA contains a Met codon, thus implying that the previously uncharacterized amino acid corresponding to the N-terminal position of the mature TcpA pilin is a modified form of methionine. Except for this difference, mature TcpA has the overall predicted structural motifs shared among type 4 pilins.

Amino Acid Sequence↗
PubMed1987

Transfusion related HIV infection: the Wellington experience.

A case of transfusion-related AIDS is described which is believed to be the first published case to occur in New Zealand in a nonhaemophiliac patient. The human immunodeficiency virus (HIV) positive donor was shown to be the source of infection in five further HIV positive recipients.

Acquired Immunodeficiency Syndrome↗
PubMed1987

In vitro activity of Ro 15-8074, Ro 19-5247, A-56268, and roxithromycin (RU 28965) against Neisseria gonorrhoeae and Chlamydia trachomatis.

In vitro Ro 15-8074 and Ro 19-5247 (T2525), two new oral cephalosporins, were active against 410 penicillin-susceptible and -resistant isolates of Neisseria gonorrhoeae. Two new macrolides, A-56268 and to a lesser extent roxithromycin (RU 28965), were active against Chlamydia trachomatis. A-56268 had activity against N. gonorrhoeae similar to that of erythromycin.

Anti-Bacterial Agents↗
PubMed1987

Cell culture compared with broth for detection of Trichomonas vaginalis.

Trichomonas vaginalis can be grown in cell culture. We studied the growth kinetics of T. vaginalis in McCoy cell culture compared with that in a conventional broth medium (Diamond TYI-S-33 medium supplemented with 10% heat-inactivated bovine serum [TYI]). In the presence of McCoy cells and two parts cell culture medium to one part TYI, a peak concentration of 2 X 10(6) to 6 X 10(6) T. vaginalis per ml was consistently achieved with inocula as low as three T. vaginalis cells per ml. Without cells, this medium did not support growth of T. vaginalis. T. vaginalis in TYI in 1-ml vials with or without McCoy cells demonstrated poor growth. In tubes containing 10 ml of TYI, inocula grew to 2 X 10(6) to 6 X 10(6) T. vaginalis per ml, but at least 3 X 10(3) T. vaginalis per tube was required to initiate growth. Thus, in vitro, cell culture was more sensitive than TYI broth in detecting low numbers of T. vaginalis. In a subsequent clinical comparison of broth and cell culture for isolation of T. vaginalis from 188 vaginal specimens and 21 urethral specimens from men, the results were in agreement for 206 specimens (98.6%). There were no situations in which culture was negative and a saline preparation showed motile trichomonads. For women, using a positive culture as the indicator of true positivity, the sensitivity of detection of T. vaginalis was 83% with the Pappenheim stain and 77% with saline preparations. These studies show that cell culture can be used for isolation of T. vaginalis from clinical specimens; it gave results comparable to those of broth culture for the group of mainly symptomatic women. Further studies should be performed to determine its utility in clinical populations such as asymptomatic women and men with and without symptoms, in which T. vaginalis is more likely to be present in low numbers.

Animals↗
PubMed1986

In-vitro susceptibility of 400 isolates of Neisseria gonorrhoeae in Vancouver, 1982-84.

Consecutive isolates of Neisseria gonorrhoeae obtained at a sexually transmitted disease clinic in Vancouver between June 1982 and June 1984 were tested for in-vitro susceptibility to eight antimicrobial agents. Of the 400 isolates 6 (1.5%) were penicillinase-producing N. gonorrhoeae, and for 25 (6.2%) the minimum inhibitory concentrations (MICs) of penicillin were 1.0 to 4.0 micrograms/ml. Ceftriaxone sodium was the most active agent. The MICs were higher than those reported in a Canadian study in 1973-74, except for tetracycline hydrochloride. The patterns of susceptibility of the isolates to one antimicrobial agent correlated significantly with those to each other agent, although the relation was weakest for trimethoprim-sulfamethoxazole and spectinomycin. The results reinforce the need to evaluate local in-vitro susceptibility patterns, especially since the proportion of isolates with relative and absolute resistance to penicillin is increasing.

Anti-Bacterial Agents↗
PubMed1986

In vitro activity of difloxacin hydrochloride (A-56619), A-56620, and cefixime (CL 284,635; FK 027) against selected genital pathogens.

Management of sexually transmitted diseases is facilitated by having antimicrobial agents with activity against all of the major genital pathogens. Newer quinolones show promise of being active against Neisseria gonorrhoeae and Chlamydia trachomatis. Two quinolones, difloxacin (A-56619) and A-56620, and an oral cephalosporin, cefixime (CL 284,635; FK 027), were evaluated in vitro. All three were highly active against 400 isolates of N. gonorrhoeae, including penicillinase-producing N. gonorrhoeae, N. gonorrhoeae with chromosomally mediated resistance, and isolates with penicillin MICs of less than 1 microgram/ml. Susceptibilities to one antimicrobial agent were usually strongly correlated with susceptibilities to the other antimicrobial agents evaluated, but isolates with increasing resistance to beta-lactams were least likely to show increasing resistance to quinolones. Difloxacin and, to a lesser extent, A-56620 were active against all 10 strains of C. trachomatis, and both had moderate activity against over 200 strains of Gardnerella vaginalis. Based on in vitro activity, difloxacin and A-56620 merit in vivo assessment for management of both C. trachomatis and N. gonorrhoeae infections, and cefixime shows considerable promise for treatment of N. gonorrhoeae infections.

Anti-Bacterial Agents↗
PubMed1976

Study of bound phospholipase activities of fungal mycelia using an organic solvent system.

It has been possible to demonstrate and characterize high phospholipase activities in mycelia of Rhizopus arrhizus and Mucor javanicus by use of a system in which substrates were dissolved in diisopropyl ether. Such activities were associated with bound enzymes and would have been difficult to detect using aqueous assay systems. In both cases, phosphatidylcholine hydrolysis was by phospholipase A1 (EC 3.1.1.32) activity followed by the action of lysophospholipase (EC 3.1.1.5). Phospholipase D (EC 3.1.4.4) activity was also detected. The methods used appear to be of general applicability for the detection and study of insoluble phospholipases.

Animals↗
PubMed1972

Production of diagnostic pigment by phenoloxidase activity of cryptococcus neoformans.

Cryptococcus neoformans produces brown pigmented colonies when grown on agar media made from an extract of potatoes and carrots, broad beans (Vicia faba), or Guizotia abyssinica seeds. Since other yeasts do not produce the pigment, these media are useful as differential isolation media for C. neoformans. Similar specific pigment was produced by C. neoformans on chemically defined agar media which contained six different substrates of phenoloxidase (o-diphenol: oxygen oxidoreductase EC 1.10.3.1) an enzyme which catalyses the oxidation of o-diphenols to melanin. Substrates were incorporated singly into the media and included L-3, 4-dihydroxyphenylalanine (L-DOPA), chlorogenic acid, protocatechuic acid, catechol, norepinephrine, and 3-hydroxytyramine hydrochloride (dopamine). No pigment was produced on media without substrate. Phenoloxidase activity in (NH(4))(2)SO(4) precipitates of C. neoformans cell-free extract was assayed by measuring increases in absorbance at 480 nm produced in solutions of L-DOPA. This reaction showed oxygen uptake and was effectively inhibited by copper chelators, but not by catalase. The enzyme also oxidized the five other substrates which induced pigment formation. Electron micrographs of cells incubated in L-DOPA showed deposition of the pigment in the cell wall.

Agar↗
PubMed1969

Improvement in laboratory diagnosis of pulmonary cryptococcosis.

A new differential colour medium prepared with an extract of potatoes and carrots produced ochre-brown pigmentation in Cryptococcus neoformans colonies, identical to that obtained with Guizotia abyssinica seed extract. The pigment formation was observed to be specific for this pathogenic yeast in a survey of 534 yeast-positive specimens during routine diagnostic work; among 47 C. neoformans-positive specimens were 27 sputa and bronchial aspirations, many overgrown by common yeasts.Detection of C. neoformans in the contaminated specimens was credited largely to the differential medium used. Twelve cases of cryptococcosis were investigated between 1962 and 1968, five of which presented involvement of the respiratory tract only.

Adult↗