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Biomedical subjects

R Edwards

Publications and source records attributed to R Edwards.

At least 217 records · Page 12Linked to original sources

Identification, purification, and characterization of S-adenosyl-L-methionine: isoliquiritigenin 2'-O-methyltransferase from alfalfa (Medicago sativa L.).

An O-methyltransferase (OMT) which methylates the 2'-hydroxyl of isoliquiritigenin (2',4,4'-trihydroxychalcone) was identified in alfalfa (Medicago sativa L.) seedlings and cell cultures. The OMT activity increased during early stages of seedling development and was predominantly located in roots. Treatment of alfalfa cell cultures with an elicitor from yeast resulted in a fivefold increase in chalcone OMT activity, whereas treatment of seedlings with CuCl2 caused a reduction in activity. The chalcone OMT was purified to near homogeneity from elicited alfalfa cell cultures. Only one form of the enzyme was found. It consisted of an active monomer of subunit Mr 43,000 which could be photoaffinity labeled with S-adenosyl-L-[methyl-3H]methionine. The purified OMT had a pH optimum of 9.0, pI of 4.7, and was highly specific for the 2'-hydroxyl of 2',4,4'-trihydroxychalcone, with essentially no activity toward narigenin chalcone, caffeic acid, or daidzein. Kinetic analysis indicated a sequential bi bi mechanism with Km values of 2.2 and 17.7 microM for 2',4,4'-trihydroxychalcone and S-adenosyl-L-methionine, respectively. S-Adenosyl-L-homocysteine was a potent inhibitor. The chalcone OMT represents the third distinct OMT isolated from alfalfa cell cultures.

Affinity Labels↗

The comparative activity of fosfomycin trometamol against organisms isolated from infected urines.

Five hundred urinary pathogens, collected from patients of general practitioners and hospital in-patients, were identified and tested for susceptibility to fosfomycin, ampicillin, cephalexin, nalidixic acid, nitrofurantoin, trimethoprim and sulfamethoxazole. Overall, 83% of the isolates were sensitive to fosfomycin, comprising 89% of the out-patient strains and 77% of the in-patient isolates. This degree of sensitivity was similar to that of cephalexin, nalidixic acid and trimethoprim, but higher than that observed with ampicillin, nitrofurantoin and sulfamethoxazole. Fosfomycin generally showed a broad spectrum of activity, but was less active than some other compounds against Klebsiella spp. and streptococci. More than 70% of strains resistant to ampicillin, sulfamethoxazole or trimethoprim were sensitive to fosfomycin indicating that cross resistance is not presently a problem.

Anti-Bacterial Agents↗

The influence of anaerobiosis on the activity of fosfomycin trometamol.

MICs of fosfomycin trometamol were estimated for 40 strains of bacteria (20 gram-positive cocci, 20 gram-negative bacilli) by the agar incorporation method (Iso-Sensitest agar) in the presence of the potentiating agent, glucose-6-phosphate (25 mg/l). Titrations were carried out in duplicate under aerobic and anaerobic conditions. For 22 strains (12 gram-negative bacilli), a fourfold or greater reduction in MIC was observed in tests conducted under anaerobic conditions. The effect was particularly marked with Klebsiella spp., four of five strains of which showed a 16- to 32-fold reduction in MIC in anaerobic conditions. To investigate the reasons for the effect of anaerobiosis, selected strains were examined in an opacity monitoring device in which cultures can be grown in aerobic or anaerobic atmosphere. Surprisingly, the effect of anaerobiosis observed by continuous turbidimetric monitoring was much less than that seen in agar incorporation MIC titrations: under anaerobic conditions, there was little or no reduction in the concentration of fosfomycin trometamol required to cause a lytic effect on dense bacterial cultures, and a small, but variable effect on the emergence of resistant variants.

Anaerobiosis↗

Risk factors for melanoma: site variation in minimal erythema dose.

Skin types 1 and 2, increased numbers of moles, and excessive intermittent sun exposure are known risk factors for cutaneous melanoma, but the inter-relationship between UV radiation exposure, moles and melanoma remains unclear. There is a noteworthy site variation in melanoma, it being more common on the lower leg in women and on the back in men. In order to determine whether this site variation could provide further clues to the pathogenesis of melanoma, we examined site variation in photosensitivity and its relationship to other known melanoma risk factors (number of moles, skin type and skin colour) in 25 healthy volunteers. A marked site variation in photosensitivity was found. The pale skin of the volar aspect of the forearm was markedly less photosensitive than the darker skin of the back. Females were more photoresistant than males on the lower legs even though this is the more common site for melanoma in women. There was some correlation between the number of moles and photosensitivity at the two sites.

Adult↗

An investigation of beta-lactamases from clinical isolates of Bacteroides species.

Among a group of 116 clinically significant isolates of Bacteroides spp., 24 exhibited beta-lactamase activity greater than the basal level characteristic of most Bacteroides strains. Investigation of specific enzyme activity, iso-electric point and enzyme inhibition profiles revealed that the beta-lactamases involved could be divided into four groups, some showing similarity to those described in previous studies. Seven of the enzymes were able to hydrolyse cefoxitin, latamoxef or imipenem, and eight enzymes degraded penicillin in the presence of clavulanic acid. Five strains showed reduced susceptibility to cefoxitin, latamoxef or imipenem which was not associated with beta-lactamase activity.

Anti-Bacterial Agents↗

The role of complement, platelet-activating factor and leukotriene B4 in a reversed passive Arthus reaction.

1. The mechanisms underlying oedema formation induced in a reversed passive Arthus (RPA) reaction and, for comparison, in response to zymosan in rabbit skin were investigated. 2. Oedema formation at skin sites was quantified by the accumulation of intravenously-injected 125I-labelled human serum albumin. 3. Recombinant soluble complement receptor type 1 (sCR1), administered locally in rabbit skin, suppressed oedema formation induced in the RPA reaction and by zymosan. 4. The platelet-activating factor (PAF) antagonists, WEB 2086 and PF10040 administered locally, inhibited oedema formation induced in the RPA reaction and by PAF but not by zymosan. 5. A locally administered leukotriene B4 (LTB4) antagonist, LY-255283, inhibited oedema formation induced by LTB4 but did not inhibit oedema responses to PAF, zymosan or the RPA reaction. 6. The results demonstrate a role for complement in oedema formation in both the RPA reaction and in response to zymosan. An important contribution by PAF is indicated in the RPA reaction but not in response to zymosan whereas no evidence was obtained to suggest a role for LTB4 in either inflammatory response.

Animals↗

Effect of inoculum size on bacteriolytic activity of cefminox and four other beta-lactam antibiotics against Escherichia coli.

MICs and turbidimetric experiments revealed a negligible inoculum effect with two Escherichia coli strains exposed to cefminox and cefoxitin, whereas a marked inoculum effect was revealed after exposure to cefotaxime, ceftizoxime, and imipenem. The activities of the cephamycins were associated with spheroplast formation and bacteriolysis at concentrations close to the MIC, whereas the other agents induced the formation of filaments or, in the case of imipenem, rounded cells.

Anti-Bacterial Agents↗

Immunoreactive beta-core-like material in normal postmenopausal urine: human chorionic gonadotrophin or LH origin? Evidence for the existence of LH core.

Material with the immunochemical properties of the beta-core of human chorionic gonadotrophin (hCG) can be found in the urine of normal postmenopausal women. However, we have been unable to detect intact hCG (using an assay which is specific for the alpha-beta heterodimer of intact hCG) in serum of such subjects. The levels of serum LH and urinary beta-core were compared in matched samples from 28 women (serum LH: median 27 U/l, range 4-70 U/l, urinary beta-core: median 0.27 microgram/l, range less than 0.05-0.645 microgram/l). Urine (4 litres) from three postmenopausal women was concentrated, dialysed and subjected to gel exclusion chromatography on Sephadex G-100. Fractions were analysed by specific assays for LH, intact hCG, total beta-hCG (free beta-subunit and intact hCG), free alpha-subunit and beta-core. Material eluting at the expected position of the beta-core fragment of hCG was detected in all three samples by the beta-core, beta-hCG and LH assays, despite the fact that the LH antibody does not recognize the authentic beta-core of pregnancy. Electrophoresis and Western blotting of the concentrated urines revealed that material of the same molecular size as beta-core was recognized by the antibody to LH but not by a monoclonal antibody raised to free beta-hCG which also recognizes the beta-core molecule of hCG. We conclude that the predominant core-like material identified in postmenopausal urine is probably derived from the beta-subunit of LH.

Aged↗

Health visiting on a playbus: a community approach.

Inappropriate and overcrowded housing, poverty and lack of social support can affect the health and development of young children and their mothers. Such factors may prevent those families who most need health resources from using them. Rosalind Edwards and Stephanie Ramsey describe a project that attempts to encourage the use of health-related information by families with young children through taking an inter-agency and community-based approach.

Community Health Nursing↗

A chromatographic and spectroscopic study of photodegraded metronidazole in aqueous solution.

Preparative reversed-phase HPLC has been used to isolate a yellow photodegradation (2) product from aqueous metronidazole (1) solutions buffered with citrate:phosphate. Compound 2 can be further degraded into colorless materials by light, heat, or the addition of the nonaqueous solvents. These products have been characterized by UV, IR, proton NMR, mass spectroscopy, and melting point determination. It is proposed that the initial yellow degradation product (2) is an "excimer ion-pair" formed by the stabilization of 1 in its first electronic excited state by the citrate molecule.

Chromatography, High Pressure Liquid↗

Stress responses in alfalfa (Medicago sativa L.) 11. Molecular cloning and expression of alfalfa isoflavone reductase, a key enzyme of isoflavonoid phytoalexin biosynthesis.

The major phytoalexin in alfalfa is the isoflavonoid (-)-medicarpin (or 6aR, 11aR)-medicarpin. Isoflavone reductase (IFR), the penultimate enzyme in medicarpin biosynthesis, is responsible for introducing one of two chiral centers in (-)-medicarpin. We have isolated a 1.18 kb alfalfa cDNA (pIFRalf1) which, when expressed in Escherichia coli, converts 2'-hydroxyformononetin stereospecifically to (3R)-vestitone, as would be predicted for IFR from alfalfa. The calculated molecular weight of the polypeptide (35,400) derived from the 954 bp open reading frame compares favorably to estimated Mrs determined for IFR proteins purified from other legumes. The transcript (1.4 kb) is highly induced in elicited alfalfa cell cultures. The kinetics of induction are consistent with the appearance of IFR activity, the accumulation of medicarpin, and the observed induction of other enzymes in the pathway. Low levels of IFR transcripts were found in healthy plant parts (roots and nodules) which accumulate low levels of a medicarpin glucoside. IFR appears to be encoded by a single gene in alfalfa. The cloning of IFR opens up the possibility of genetic manipulation of phytoalexin biosynthesis in alfalfa by altering isoflavonoid stereochemistry.

Amino Acid Sequence↗

Purification and characterization of S-adenosyl-L-methionine: caffeic acid 3-O-methyltransferase from suspension cultures of alfalfa (Medicago sativa L.).

Caffeic acid O-methyltransferase (COMT) is one of a group of proteins present in alfalfa cell cultures which can be photoaffinity labeled with S-adenosyl-L-[methyl-3H]methionine. The enzyme was purified to homogeneity from elicitor-treated suspension cultures and shown to exist as an active monomer of subunit Mr 41,000. COMT could be separated into two forms on the basis of their isoelectric points and relative affinities for S-adenosyl-methionine and S-adenosylhomocysteine. Both forms had equal affinities for caffeic acid, were highly specific for the 3-hydroxyl group of substituted cinnamic acids, and exhibited negligible activity toward flavonoid substrates. An antiserum raised against COMT from aspen immunoprecipitated alfalfa COMT activity. Peptide mapping studies indicated that the two forms of COMT and an isoflavone O-methyltransferase from alfalfa are closely related proteins. The extractable activity of COMT doubled over a 48-h period following exposure of alfalfa cell suspensions to a yeast elicitor preparation, and this was associated with a small change in the relative proportions of the two forms of the enzyme.

Affinity Labels↗