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D Richards

Publications and source records attributed to D Richards.

At least 145 records · Page 8Linked to original sources
PubMed1986

Conformational transitions in fluorescein-labeled (Na,K)ATPase reconstituted into phospholipid vesicles.

Fluorescein-labeled (Na,K)ATPase reconstituted into phospholipid vesicles has been used to study conformational transitions. Addition of K+ or Na+ to the vesicle medium induces fluorescence changes characteristic of the E2(K) or E1Na states of fluorescein-labeled (Na,K)ATPase (Karlish, S.J.D. (1980) J. Bioenerg. Biomembr. 12, 111-136). The cation effects are exerted from the cytoplasmic surface of inside-out-oriented pumps. Equilibrium cation titrations and measurements of rates of conformational transitions have led to the following observations. 1) The rate of E2(K)----E1Na or E2(T1)----E1Na is 4-6-fold faster and E1K----E2(K) is about 2-fold slower in vesicles compared to enzyme. In equilibrium titrations the K0.5 for K+ is higher and that for Na+ is lower for vesicles compared to enzyme. The conformational equilibrium E(1)2K----E2(2K) is apparently shifted toward E(1)2K in vesicles compared to enzyme. 2) Diffusion potentials, positive-outside, induced with valinomycin or Li+ ionophore AS701, do not affect the rates of E2(T1)----E1Na or E1K----E2(K), or equilibrium cation titrations. This demonstrates that the conformational transitions E(1)2K----E2(2K) are voltage-insensitive steps, confirming a prediction based on transport experiments. 3) In vesicles containing choline, K+, Na+, or Li+, the rate of E2(T1)----E1Na increases in the order given. Vesicles with reconstituted fluorescein-labeled (Na,K)ATPase provide a convenient system for correlating directly properties of conformational transitions with cation transport.

Animals↗
PubMed1986

Hypothermia among resort skiers: 19 cases from the Snowy Mountains.

Even in relatively temperate environments, accidental hypothermia is a potentially lethal complication of exposure. We have reviewed our experience of accidental hypothermia among recreational alpine skiers at an Australian resort during the 1983 and 1984 seasons. There were 19 cases of accidental hypothermia, which occurred in 10 men and nine women who were aged between six and 47 years (mean age, 15.9 years) and who had rectal temperatures that ranged from less than 35 degrees C to 36 degrees C. The temperature at presentation to the Ski Injury Clinic was less than 35 degrees C in seven cases. One patient presented to the Clinic with a gastrointestinal haemorrhage in addition to hypothermia, and one was initially thought to be suffering from alcohol intoxication. Two patients were lost in the snow overnight. All patients were removed from the snow, changed into warm dry clothes where necessary, and their body temperatures allowed to return to normal spontaneously (17 patients), or were exposed to heat actively by means of inhaled, heated, humidified air (two severely obtunded patients). All patients responded satisfactorily. There were no deaths and no sequelae. We conclude that all skiers should be advised to wear effective thermal insulation, and to ski with a partner to ensure that adequate care is taken to prevent accidental hypothermia. Inhalational "warming" is effective in the treatment of hypothermia in obtunded patients.

Accidents↗
PubMed1986

The use of monoclonal and polyspecific antibodies in the IgE sandwich ELISA.

The use of antibody to link antigen to microtitre plates in the enzyme-linked immunosorbent assay (ELISA) has been extended to include mouse monoclonal antibodies and polyspecific rabbit antibodies. Small amounts of both reagents could be used. The capacity of the microtitre plate for the antibody was found to be critical and irradiated plates generally gave better results. Both rabbit anti-IgE conjugated to horseradish peroxidase, and monoclonal mouse anti-IgE with alkaline phosphatase-conjugated rabbit anti-mouse IgG could be used as detection reagents. Comparison with the radioallergosorbent (RAST) test showed a good agreement with the sandwich ELISA. The sandwich ELISA using polyspecific rabbit antibody was substantially more sensitive than conventional ELISA and also marginally more sensitive than RAST.

Allergens↗
PubMed1986

The influence of dwarfing interstocks on the distribution and metabolism of xylem-applied [h]gibberellin a(4) in apple.

The influence of an interstock of the dwarfing cultivar M9 and the nondwarfing cultivar MM115 on the distribution and metabolism of labeled gibberellic acid A(4) ([(3)H]GA(4)) of high specific radioactivity (5.18 x 10(10) becquerel per millimole) applied to the xylem of the rootstock in grafted apple (Malus x domestica Borkh.) trees was compared. Free [(3)H] GA-like metabolites of [(3)H]GA(4), including putative GA(1), GA(2), GA(3), and GA(34), as well as various (3)H-putative GA glucosyl conjugates were detected in stem segments from both cultivars. M9 interstocks reduced the total uptake of [(3)H]GA(4) and decreased the proportion of (3)H metabolites transported to the shoots and leaves of scions. The M9 interstock tissue and adjacent rootstock and scion tissue retained a much greater amount and a higher proportion of the label than did comparable tissue of the nondwarfing MM115 interstock. In addition, the amount and proportion of free [(3)H]GAs was higher, and the proportion of putative [(3)H]GA glucosyl conjugates lower, in M9 interstocks compared to MM115. These effects of the dwarfing interstock on GA distribution and metabolism indicate a significant role for GAs in any satisfactory explanation of the dwarfing mechanism in apple.

Journal Article↗
PubMed1986

Sub-class of IgG anti-bee venom antibody produced during bee venom immunotherapy and its relationship to long-term protection from bee stings and following termination of venom immunotherapy.

The IgG sub-class antibody response to bee venom in the four sub-classes was investigated in ten patients during and after venom immunotherapy. All patients tolerated a bee sting challenge 1, 2 and 3 years after the start of treatment as well as 1 and 2 years after treatment was stopped. Anti-phospholipase A2 (PLA2) antibodies were of IgG1 and IgG4 sub-class and rose early in treatment, IgG1 anti-PLA2 fell to pre-treatment levels after 3 years in contrast to IgG4 anti-PLA2 levels, which remained high during maintenance therapy and declined relatively little in the 2 years after the termination of treatment. This data shows that IgG4 antibodies are maintained in the absence of monthly maintenance injections and suggests that they may provide long lasting clinical protection from insect stings.

Adolescent↗
PubMed1986

Sub-class of IgG in allergic disease. I. IgG sub-class antibodies in immediate and non-immediate food allergy.

Previous studies have suggested that, apart from IgE-mediated reactions, some of the symptoms of food allergy may be caused by IgG antibodies to food proteins. This study was carried out to see if there were any distinctive features of the IgG sub-class antibody response to dietary antigens which occurs in food allergic patients. IgG sub-class antibodies were measured using a quantitative enzyme-linked immuno-sorbent assay (ELISA) to wheat gliadin, ovalbumin and bovine casein in twenty patients who had coeliac disease and in twenty-eight egg allergic patients. These were compared with twenty-one atopic dermatitis patients who did not have food allergy and twenty-six healthy control subjects. Coeliac disease patients tended to have raised IgG antibody levels (especially IgG1) to all three antigens but these overlapped considerably with that seen in egg allergic and atopic dermatitis patients. Coeliacs who avoided gluten had anti-gliadin antibody levels which did not differ from those seen in healthy subjects but nevertheless had raised anti-ovalbumin and casein-specific antibodies. The IgG antibody was largely restricted to IgG1 and IgG4 sub-class although the relative amount of each varied with the antigen. Although gliadin-specific antibodies were mainly IgG1, ovalbumin-specific antibodies were mainly IgG4. The increased antibody levels to all three antigens in coeliacs were caused by a raised IgG1 response, IgG4 antibodies were usually normal. Egg allergic patients also had raised IgG1 but not IgG4 antibodies to ovalbumin. These data show that the response to different dietary antigens can vary with the antigen. The fact that IgG1 and not IgG4 antibodies were raised to all three antigens in patients with coeliac disease suggests that they are a secondary consequence of the disease, perhaps reflecting increased transport of antigens across a damaged gut mucosa rather than a specific immunopathological reaction. However, the observation that antibodies to gliadin, and not ovalbumin or casein, fell following gluten avoidance shows that the response to gliadin, at least, is dependent upon continued exposure to antigen.

Adult↗
PubMed1986

Gliadin IgG subclass antibodies in patients with coeliac disease.

We have measured IgG1, 2, 3 and 4 subclass antibodies to gliadin and casein in normal controls, treated and untreated coeliac patients. The amount of antibody in each subclass was determined separately as a titre and as a concentration, by reference to a standard curve. Both methods of quantitation, carried out in separate assays, yielded comparable results (IgG antibody r = 0.86, IgG1 antibody r = 0.89). In all the coeliac patients, the majority of antibody was IgG1, and little IgG2, 3, 4 antibody was detected. Untreated coeliac patients had higher titres of IgG and IgG1 antibodies to gliadin and casein compared with normal controls. In the treated coeliac patients compared with the normal controls, there was no significant difference between the titres of IgG and IgG1 antibodies to gliadin but there were higher titres to casein.

Antibodies↗
PubMed1986

Biotransformation of polycyclic aromatic compounds by fungi.

Incubations of several polycyclic aromatic hydrocarbons (PAHs) and heteroaromatic compounds with a series of common micro-organisms have been performed. The PAHs were not metabolized by any of the fungi studied. The sulphur-containing heterocyclic aromatic compounds dibenzothiophene, thioxanthone and thiochromanone were oxidized at sulphur by C. elegans. Other fungi are capable of oxidation at the sulphur atom of dibenzothiophene and thioxanthone. C-1 and C-3 methyl substituted thioxanthones are hydroxylated at the methyl group by C. elegans.

Biotransformation↗
PubMed1985

Increased sensitivity and specificity of a sandwich ELISA for measurement of IgE antibodies.

We have developed a sandwich enzyme-linked immunosorbent assay (ELISA) for measurement of IgE antibodies to the bee venom allergens phospholipase A2 (PLA2) and hyaluronidase (HYAL). The assay is 10-20 times more sensitive than conventional indirect ELISA or radioallergosorbent test (RAST). Furthermore, by using affinity purified rabbit antibodies to these allergens, the specificity of the test was increased compared to RAST. The use of antibodies to link the antigen to the solid phase removes the dependence on the individual protein's ability to bind to the microtitre plate. The increased sensitivity of the sandwich assay seems to be due to better presentation and retention of antigen on the solid phase.

Antibody Specificity↗
PubMed1985

A comparison of diuresis renography, the Whitaker test and renal pelvic morphology in idiopathic hydronephrosis.

Diuresis renography and the Whitaker test are established methods of diagnosing obstruction in dilated renal pelves. These techniques have been compared in 36 patients with radiologically demonstrated idiopathic hydronephrosis and evaluated, where possible, against renal pelvic morphological features. The agreement between the results of the tests was as follows: diuresis renography/Whitaker test 67%; diuresis renography/renal pelvic morphology 74%; Whitaker test/renal pelvic morphology 58%. Both diuresis renography and the Whitaker test are indicated in some cases of idiopathic hydronephrosis.

Diuresis↗
PubMed1985

Absorption of delayed-release prednisolone in ulcerative colitis and Crohn's disease.

The absorption and urinary excretion of [3H]prednisolone after oral ingestion was examined using hard gelatin capsules with and without a coating of Eudragit-S which delayed release of the contents. In 6 patients with ulcerative colitis absorption was delayed until the preparation reached the colon but the total absorption was unchanged. In 6 patients with Crohn's disease and ileal strictures the preparation broke proximal to the stricture and absorption was delayed until this occurred (within 4-12 h). In 5 patients with Crohn's disease and intestinal resections the capsules broke unreliably.

Adult↗
PubMed1985

Improved sensitivity and specificity of sandwich, competitive and capture enzyme-linked immunosorbent assays for allergen-specific antibodies.

Indirect ELISA is widely used to detect specific antibodies but can suffer from high non-specific binding-particularly of IgG. The use of affinity-purified rabbit antibody-coated microtitre plates to bind antigen greatly increases sensitivity without a significant increase in non-specific binding of IgG. Capture, competitive and sandwich assay procedures gave comparable results for IgG antibodies; but only the sandwich assay was suitable for detection of IgE antibodies.

Allergens↗