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Biomedical subjects

V Doyle

Publications and source records attributed to V Doyle.

13 recordsLinked to original sources

Comparative efficacy of the combination fipronil-(S)-methoprene and the combination permethrin-imidacloprid against Dermacentor reticulatus, the European dog tick, applied topically to dogs.

This study compared the efficacy of two of the most widely veterinary-dispensed topical products for control of ticks on dogs: fipronil-(S)-methoprene and imidacloprid-permethrin. Eighteen healthy beagle dogs of both sexes were divided into three groups of six dogs. Group 1 served as the untreated control. Dogs in group 2 were treated with fipronil 10% + (S)-methoprene 9% w/v applied once on day 0, and those in group 3 were treated with imidacloprid 8.8% and permethrin 44% w/v, applied once on day 0. All dogs were infested with approximately 50 unfed Dermacentor reticulatus ticks on days 1, 7, 14, 21, 28, 35, and 42, and ticks remaining were counted and removed 48 hours after each infestation. The 48-hour efficacy of the fipronil-(S)-methoprene combination remained at 100% at all assessment points through and including day 37, declining to 95.30% on day 44. The 48-hour efficacy of the imidacloprid-permethrin combination peaked at 86.46% on day 9, decreased to 73.37% by the third week, and fell to 63.53% by the end of the study (day 44). Significantly (P < .05) fewer ticks were recovered from dogs treated with fipronil-(S)-methoprene or imidacloprid-permethrin than from untreated control dogs at each infestation. Notably, the dogs treated with fipronil-(S)-methoprene had significantly (P < .05) lower tick counts at every assessment than the dogs treated with imidacloprid-permethrin.

Administration, Topical↗

Evidence that cyclophilin-A protects cells against oxidative stress.

Cyclophilin-A is the cytosolic isoform of a family of peptidylproline cis-trans-isomerases that bind cyclosporin A. This study investigates the role of cyclophilin-A in necrotic cell death, induced by 'chemical ischaemia' and by t-butylhydroperoxide. An 18-mer antisense phosphorothioate oligodeoxynucleotide was used to target a translated region of cyclophilin-A mRNA in rat neonatal cardiomyocytes. After a 24 h exposure to the oligonucleotide, the amount of cyclophilin-A in the cells was decreased by at least 93% as judged by immunological and enzymic criteria. For the enzyme assays, peptidyl proline cis-trans-isomerase activity was measured fluorimetrically in small (10 microl) volumes of cell extract. Immunoblots were developed with a polyclonal anti-cyclophilin-A antibody after sample isoelectric focusing and SDS/PAGE. Cyclophilin-A suppression had no effect on cyanide-plus-2-deoxyglucose-induced cell death. However, cyclophilin-A-suppressed cells were markedly more sensitive to t-butylhydroperoxide. Cyclosporin A conferred some resistance to the peroxide in both types of cell, but protection was greater in cyclophilin-A-suppressed cells, where cyclosporin A increased the survival time 2-fold. It is concluded that two cyclophilin isoforms are involved, in quite different ways, in peroxide-induced cell death. Cyclophilin-A has a protective role. Another isoform, possibly mitochondrial cyclophilin-D, has a deleterious role, such that blockade by cyclosporin A leads to protection.

Animals↗

The mitochondrial permeability transition pore.

This chapter reviews recent advances in the identification of the structural elements of the permeability transition pore. The discovery that cyclosporin A (CsA) inhibits the pore proved instrumental. Various approaches indicate that CsA blocks the pore by binding to cyclophilin (CyP)-D. In particular, covalent labelling of CyP-D in situ by a photoactive CsA derivative has shown that pore ligands have the same effects on the degree to which CsA both blocks the pore and binds to CyP-D. The recognition that CyP-D is a key component has enabled the other constituents to be resolved. Use of a CyP-D fusion protein as affinity matrix has revealed that CyP-D binds very strongly to 1:1 complexes of the voltage-dependent anion channel (from the outer membrane) and adenine nucleotide translocase (inner membrane). Our current model envisages that the pore arises as a complex between these three components at contact sites between the mitochondrial inner and outer membranes. This is in line with recent reconstitutions of pore activity from protein fractions containing these proteins. The strength of interaction between these proteins suggests that it may be a permanent feature rather than assembled only under pathological conditions. Calcium, the key activator of the pore, does not appear to affect pore assembly; rather, an allosteric action allowing pore flicker into an open state is indicated. CsA inhibits pore flicker and lowers the binding affinity for calcium. Whether adenine nucleotide translocase or the voltage-dependent anion channel (via inner membrane insertion) provides the inner membrane pore has not been settled, and data relevant to this issue are also documented.

Animals↗

A specific quantitative assay for collagen synthesis by cells seeded in collagen-based biomaterials using sirius red F3B precipitation.

The measurement of collagen synthesis by seeded cells in vitro is a prerequisite for the assessment of biocompatibility of many biomaterials. Existing methods are either complicated or not applicable to systems utilizing collagen-based materials, and the development of a rapid and simple technique would be an advantage. In the current paper, a method is described which relies on the radiolabelling of newly synthesized protein with [3H]-proline followed by specific precipitation of collagen using 1% sirius red dissolved in water. The results indicate that collagen binding to sirius red is unaffected by using water rather than picric acid as a solvent and the dye binds in a similar fashion to collagen type I, II and III. Cycloheximide treatment of the gels indicated that precipitated [3H]-proline was restricted to macromolecular protein. Collagenase treatment eradicated labelled precipitation formation when using 1% sirius red in water, indicating a high degree of specificity for collagen whilst specificity was poor when using 1% sirius red in picric acid. The method described is both simple and rapid and shows a high degree of specificity and sensitivity. For these reasons it is highly suited for the assessment of collagen synthesis by cells in collagen-based materials.

Journal Article↗

Introducing quality assurance to health service delivery--some approaches from South Africa, Ghana and Kenya.

The following report combines the contributions of four health care professionals engaged in quality improvement activities in three different areas of Africa--Ghana in West Africa, Kenya in the East, and the Republic of South Africa. These reports provide current and vivid accounts of the impact of quality assurance and quality improvement activities on areas of change in each country's rapidly expanding health care system.

Delivery of Health Care↗

A direct two-site peroxidase-based enzyme-linked immunosorbent assay for proinsulin.

A non-competitive sandwich assay using an anti-C-peptide IgG and an anti-insulin IgG was developed to measure fasting levels of proinsulin in human serum. The former antibody provided the lower layer in a sandwich immunoassay, the upper layer being composed of an anti-insulin IgG-horse radish peroxidase conjugate. The assay showed negligible cross reactivity at supraphysiological levels of insulin and C-peptide. The method enabled the estimation of proinsulin in fasting non-diabetic control subjects [13.7 +/- 1.6(4) pM] and in type 2 non-insulin-dependent diabetic patients [23.2 +/- 1.1(8) pM].

C-Peptide↗

Cardiac sensitivity to isoprenaline, lymphocyte beta-adrenoceptors and age.

The heart rate response to isoprenaline in 11 subjects aged 19-46 years was compared with lymphocyte beta-adrenoceptor numbers and lymphocyte cyclic AMP responsiveness. The dose of isoprenaline required to increase heart rate by 25 beats/min ( CD25 ) increased as a function of age (r 0.79; P less than 0.01). Lymphocyte receptor numbers also correlated directly with age (r 0.61; P less than 0.05), but there was no true correlation between CD25 and lymphocyte receptor numbers. Baseline and maximum lymphocyte cyclic AMP concentrations in response to isoprenaline stimulation in vitro were unrelated to age, CD25 or receptor numbers. This study confirms previous findings of a reduced responsiveness with age and a rise in receptor numbers in young and middle-aged adults. However, the decline in the heart response to isoprenaline was unrelated to any measurable change in beta-adrenoceptor concentration or responses, at least in the isolated lymphocyte.

Adult↗

Human lymphocyte beta-adrenoceptor density in relation to age and hypertension.

beta-Adrenoceptor density was measured in lymphocytes from neonates and from normotensive and hypertensive adults aged from 18 to 85 years. Receptor density did not differ in young and elderly people. In normotensive but not in hypertensive people there was a peak in beta-adrenoceptor density in middle life; otherwise, receptor densities were similar in normotensive and hypertensives. Neonates had a lower lymphocyte receptor density than adults. Decreases in beta-adrenoceptor-mediated responses in old age may be due to alterations in postreceptor events.

Adolescent↗