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Biomedical subjects

S E Clarke

Publications and source records attributed to S E Clarke.

At least 19 recordsLinked to original sources
PubMed2006

A critical evaluation of the experimental design of studies of mechanism based enzyme inhibition, with implications for in vitro-in vivo extrapolation.

The published literature on mechanism based inhibition (MBI) of CYPs was evaluated with respect to experimental design, methodology and data analysis. Significant variation was apparent in the dilution factor, ratio of preincubation to incubation times and probe substrate concentrations used, and there were some anomalies in the estimation of associated kinetic parameters (k(inact), K(I), r). The impact of the application of inaccurate values of k(inact) and K(I) when extrapolating to the extent of inhibition in vivo is likely to be greatest for those compounds of intermediate inhibitory potency, but this also depends on the fraction of the net clearance of substrate subject to MBI and the pre-systemic and systemic exposure to the inhibitor. For potent inhibitors, the experimental procedure is unlikely to have a material influence on the maximum inhibition. Nevertheless, the bias in the values of the kinetic parameters may influence the time for recovery of enzyme activity following re-synthesis of the enzyme. Careful attention to the design of in vitro experiments to obtain accurate kinetic parameters is necessary for a reliable prediction of different aspects of the in vivo consequences of MBI. The review calls for experimental studies to quantify the impact of study design in studies of MBI, with a view to better harmonisation of protocols.

Algorithms↗
PubMed2003

Endaural or postaural incision for myringoplasty: does it make a difference to the patient?

The aim of this study was to find out from patients who had undergone a myringoplasty via either an endaural or postaural approach whether or not they had experienced problems or symptoms relating to their scar, and if these differed depending on which incision had been used. A questionnaire was sent to 91 patients who had undergone myringoplasty between 18 and 62 months earlier. Thirty-four patients who had undergone previous or subsequent ear surgery were excluded. The only statistically significant difference found between groups having either a postauricular or endaural incision was in how likely others were to comment on their scar. There seem to be few long-term sequealae relating to the scar from myringoplasty. From a patient perspective, between 18 and 62 months postoperatively, there was very little difference in symptoms or problems whether or not an endaural or a postaural incision had been used for the surgery.

Adolescent↗
PubMed2003

Localized breeding of the Anopheles gambiae complex (Diptera: Culicidae) along the River Gambia, West Africa.

A study was undertaken to identify the major larval habitats of the Anopheles gambiae (Giles) complex in rural Gambia. Mosquito larvae and pupae were sampled along transects and in specific habitats in the central region of the country during the rainy seasons of 1996 and 1997. The sampling showed that the major breeding sites were located on the flooded alluvial soils bordering the river. The largest numbers of larvae were found during September, one month after the peak rains. Polymerase chain reaction analysis of specimens showed that Anopheles melas (Theobald) was the dominant species in the flooded areas (81.5%), followed by A. gambiae sensu stricto (Giles) (18.0%) and A. arabiensis (Patton) (0.5%). By sampling in specific habitats it was evident that A. arabiensis was mainly breeding in rain-fed rice fields along the edge of the alluvial soils. Anopheles melas and A. gambiae s.s. often coexisted but whereas A. melas were found in water with a salinity of up to 72% sea water (25.2 g NaCl l(-1)), A. gambiae s.s. only occurred in water with up to 30% sea water (10.5 g NaCl l(-1)). Anopheles melas larvae were found in association with plant communities dominated by sedges and grasses (Eleocharis sp., Paspalum sp., Sporobolus sp.) and sea-purslane Sesuvium portulacastrum (L.) and the presence of cattle hoof prints, whereas A. gambiae s.s. larvae mainly occurred in association with Paspalum sp. and Eleocharis sp. The study showed that even during the peak rainy season, breeding of the A. gambiae complex is almost entirely restricted to the extensive alluvial areas along the river.

Animals↗
PubMed2001

Expression, purification, and biochemical characterization of a human cytochrome P450 CYP2D6-NADPH cytochrome P450 reductase fusion protein.

Cytochrome P450 CYP2D6 metabolizes a wide range of pharmaceutical compounds. A CYP2D6 fusion enzyme (CYP2D6F), containing an amino-terminal human CYP2D6 sequence and a carboxyterminal human NADPH-cytochrome P450 oxidoreductase (CPR) moiety, was constructed. High levels of expression were achieved in Escherichia coli (60-100 nmol/liter) and the enzyme was catalytically active with optimal activities achieved in the presence of the antioxidant, GSH. Turnover values for bufuralol 1'-hydroxylation, metoprolol alpha-hydroxylation, O-desmethylation, and dextromethorphan O-demethylation, using membranes expressing the fusion enzyme, were 5.6, 0.4, 0.72, and 6.19 min(-1), respectively. These values were similar to E. coli membranes which coexpressed human CYP2D6 and CPR (CYP2D6/R). The K(m) and k(cat) values for bufuralol metabolism were estimated to be 10.2 microM and 4.1 min(-1), respectively. The enzyme was purified using ion-exchange chromatography, affinity chromatography (2'-5' ADP-Sepharose), and gel filtration. Estimated turnover rates for bufuralol 1'-hydroxylation, metoprolol alpha-hydroxylation, O-desmethylation, and dextromethorphan O-demethylation were 1.2, 0.52, 0.79, and 0.76 min(-1), respectively. Bufuralol 1'-hydroxylase activity by purified CYP2D6F was enhanced by phospholipids and added CPR. The CYP2D6F enzyme was able to stimulate CYP3A4 testosterone 6beta-hydroxylase activity in a reconstitution system indicating that electron transfer may be largely intermolecular. The catalytically self-sufficient CYP2D6F enzyme will facilitate investigations of P450-CPR interactions and the development of new biocatalysts.

Cytochrome P-450 CYP2D6↗
PubMed2001

Phenyl benzenesulfonamides are novel and selective 5-HT6 antagonists: identification of N-(2,5-dibromo-3-fluorophenyl)-4-methoxy-3-piperazin-1-ylbenzenesulfonamide (SB-357134).

Substituted N-phenyl-4-methoxy-3-piperazin-1-ylbenzenesulfonamides and conformationally restricted analogues have been identified as high affinity and selective 5-HT6 antagonists. Compounds from this series had a range of pharmacokinetic profiles in rat and in general there was a correlation between clearance and CNS penetration. Based on its overall biological profile 2 (SB-357134) was selected for further pre-clinical evaluation.

Animals↗
PubMed2001

Isotopic evaluation and therapy in patients with malignant endocrine disease.

The contribution of nuclear medicine to the diagnosis and treatment of endocrine malignancy is increasing. Advances in molecular biology offer new opportunities for tumour targeting via surface receptor recognition and tumour-specific metabolic markers. Imaging the biodistribution of these markers allows quantitative, in vivo characterization of tumour function. There is growing interest in the therapeutic potential of nuclear medicine targeting, substituting therapeutic beta-emitting radionuclides for the gamma-emitters used in diagnostic imaging. Limited clinical experience supports the rationale of this approach in patients with inoperable or disseminated disease and controlled trials are in progress. This chapter outlines the place of nuclear medicine techniques in the routine management of endocrine malignancy and explores areas for further development.

Endocrine Gland Neoplasms↗
PubMed2001

Cribriform variant papillary thyroid cancer: a characteristic of familial adenomatous polyposis.

Inherited cancer syndromes may predispose to more than one type of cancer, and these characteristically develop at an earlier age than their sporadic counterparts. The occurrence in a single individual of multiple, early onset primary cancers may indicate an inherited cancer susceptibility. Familial adenomatous polyposis (FAP), an autosomal, dominantly inherited susceptibility to colorectal adenomas and cancer also predisposes to childhood medulloblastomas and to a specific rare histologic type (cribriform variant) of papillary thyroid cancer. We describe a patient who developed a childhood medulloblastoma of the cerebellum, and subsequently a cribriform papillary thyroid cancer. These cancers predated the diagnosis of FAP in this patient, who was later found to have several relatives with FAP. The adenomatous polyposis coli (APC) mutation delineated in this family was in the region associated with those causing an increased risk of thyroid cancer. We submit that the diagnosis of the cribriform variant of papillary thyroid cancer in a young individual, especially after a previous cancer diagnosis, should alert the physician to the possibility of a diagnosis of FAP.

Adenocarcinoma, Papillary↗
PubMed2001

Carotid plaque characterization by magnetic resonance imaging: review of the literature.

Magnetic resonance imaging (MRI) of carotid plaque has undergone significant improvements in the last decade. Early studies utilizing ex vivo specimens and spin-echo or fast spin-echo imaging led to the conclusion that T2 weighting is the best single contrast to characterize carotid plaque morphology. On these images, the fibrous plaque appears bright and the lipid core is dark; thrombus can have variable intensity. There can be an overlap in T2-weighted signal intensities among the various plaque components, which can be partially offset by the use of multispectral analysis of multiple contrast images. With improvements in coil design, sequence design, and main field and gradient capabilities, accurate in vivo differentiation and measurement of these various carotid plaque components should be possible in 3 to 5 years. Ex vivo and in vivo studies have yielded high-resolution measurements of the complex three-dimensional lumen geometry, which are being used to predict hemodynamic forces acting on the lumenal surface. Carotid plaque burden can be accurately measured in vivo today; ongoing longitudinal studies should lead to a better understanding of the relationship between plaque burden and the risk of thromboembolic complications, as well as the effect of diet and drug therapy in hyperlipidemic patients. With these developments in place or soon to be available, MRI of the diseased carotid artery wall may prove to be even more important than magnetic resonance angiography.

Carotid Artery Diseases↗
PubMed2001

Role of radionuclide imaging in hyperthyroid patients with no clinical suspicion of nodules.

The purpose of the study was to assess the role of radionuclide imaging of hyperthyroid patients with no suspicion of nodules. (99)Tc(m) radionuclide scans performed on all 190 patients referred over a 2-year period with symptoms and thyroid function tests compatible with thyrotoxicosis were retrospectively reviewed. 73% of patients were referred by a hospital clinician and 27% were referred direct from the general practitioner. Referral letters and, where applicable, the clinical notes were reviewed and patients with suspected thyroid nodules were excluded. The results of (99)Tc(m) thyroid scans of the 190 patients (age range 23-93 years, mean 48 years) were reviewed. 152 (80%) patients had Graves' disease, 10 (5.3%) had Graves' disease with nodules, 6 (3.2%) had Graves' disease with non-functioning nodules, 5 (2.6%) had viral thyroiditis, 5 (2.6%) had autonomously functioning nodules and 12 (6.3%) were normal studies. Therefore, 20% of patients had a diagnosis other than solely Graves' disease. These diagnoses are important with respect to clinical management. In conclusion, routine radionuclide imaging is worthwhile, as a significant proportion of patients with clinical "Graves' disease" in this study would have received incorrect treatment without the result of this scan.

Adult↗
PubMed2001

Effect of passive zooprophylaxis on malaria transmission in The Gambia.

The effect of zooprophylaxis on malaria transmission has not been studied on the African continent despite that the World Health Organization has recommended this intervention method since 1982. The effect of passive zooprophylaxis on malaria vector abundance, mosquito feeding preferences, and infectivity was studied in an area of moderate seasonal transmission in The Gambia. A paired cohort of 204 children <7 yr of age was selected and matched in groups for presence or absence of cattle (Bos taurus) within 20 m of their bedroom. Comparisons were made between mosquitoes collected from the bedrooms of the two groups of children. Other ruminants and equines were present in both groups of compounds. Most of the anopheline mosquitoes (98.5%) collected were Anopheles gambiae sensu lato. There was no difference in the geometric mean number of An. gambiae s.l. mosquitoes caught in houses near or far from cattle. The species composition of the An. gambiae complex was similar in both groups. Blood meal analysis of specimens collected in houses without cattle showed a human blood index (HBI) of 82% for An. Arabiensis (Patton), 56% for An. gambiae sensu stricto (Giles), and 36% for Anopheles Melas (Theobald), indicating that each of these sibling species fed readily on animals. The presence of cattle reduced the HBI of An. arabiensis but did not significantly alter the HBI of An. gambiae s.s. or An. melas. There was no significant difference between the groups in the sporozoite rates of An. gambiae s.l. nor in the estimated malaria transmission risk. These findings suggest that passive zooprophylaxis using cattle does not alter the individual exposure to malaria parasites in The Gambia.

Animals↗
PubMed2001

Multisite kinetic models for CYP3A4: simultaneous activation and inhibition of diazepam and testosterone metabolism.

Some substrates of cytochrome P450 (CYP) 3A4, the most abundant CYP in the human liver responsible for the metabolism of many structurally diverse therapeutic agents, do not obey classical Michaelis-Menten kinetics and demonstrate homotropic and/or heterotropic cooperativity. The unusual kinetics and differential effects observed between substrates of this enzyme confound the prediction of drug clearance and drug-drug interactions from in vitro data. We have investigated the hypothesis that CYP3A4 may bind multiple molecules simultaneously using diazepam (DZ) and testosterone (TS). Both substrates showed sigmoidal kinetics in B-lymphoblastoid microsomes containing a recombinant human CYP3A4 and reductase. When analyzed in combination, TS activated the formation of 3-hydroxydiazepam (3HDZ) and N-desmethyldiazepam (NDZ) (maximal activation 374 and 205%, respectively). For 3HDZ, V(max) values remained constant with increasing TS, whereas the S(50) and Hill values decreased, tending to make the data less sigmoidal. Similar trends were observed for the NDZ pathway. DZ inhibited the formation 6beta-hydroxytestosterone (maximal inhibition, 45% of control), causing a decrease in V(max) but no significant change to the S(50) and Hill values, suggesting that DZ may inhibit via a separate effector site. Multisite rate equation models have been derived to explore the analysis of such complex kinetic data and to allow accurate determination of the kinetic parameters for activation and inhibition. The data and models presented are consistent with proposals that CYP3A4 can bind and metabolize multiple substrate molecules simultaneously; they also provide a generic solution for the interpretation of the complex kinetic data derived from CYP3A4 substrates.

Algorithms↗
PubMed2000

Biarylcarbamoylindolines are novel and selective 5-HT(2C) receptor inverse agonists: identification of 5-methyl-1-[[2-[(2-methyl-3-pyridyl)oxy]- 5-pyridyl]carbamoyl]-6-trifluoromethylindoline (SB-243213) as a potential antidepressant/anxiolytic agent.

The evolution, synthesis, and biological activity of a novel series of 5-HT(2C) receptor inverse agonists are reported. Biarylcarbamoylindolines have been identified with excellent 5-HT(2C) affinity and selectivity over 5-HT(2A) receptors. In addition, (pyridyloxypyridyl)carbamoylindolines have been discovered with additional selectivity over the closely related 5-HT(2B) receptor. Compounds from this series are inverse agonists at the human cloned 5-HT(2C) receptor, completely abolishing basal activity in a functional assay. The new series have reduced P450 inhibitory liability compared to a previously described series of 1-(3-pyridylcarbamoyl)indolines (Bromidge et al. J. Med. Chem. 1998, 41, 1598) from which they evolved. Compounds from this series showed excellent oral activity in a rat mCPP hypolocomotion model and in animal models of anxiety. On the basis of their favorable biological profile, 32 (SB-228357) and 40 (SB-243213) have been selected for further evaluation to determine their therapeutic potential for the treatment of CNS disorders such as depression and anxiety.

Administration, Oral↗
PubMed2000

Characterization of SB-271046: a potent, selective and orally active 5-HT(6) receptor antagonist.

SB-271046, potently displaced [(3)H]-LSD and [(125)I]-SB-258585 from human 5-HT(6) receptors recombinantly expressed in HeLa cells in vitro (pK(i) 8.92 and 9.09 respectively). SB-271046 also displaced [(125)I]-SB-258585 from human caudate putamen and rat and pig striatum membranes (pK(i) 8.81, 9.02 and 8.55 respectively). SB-271046 was over 200 fold selective for the 5-HT(6) receptor vs. 55 other receptors, binding sites and ion channels. In functional studies on human 5-HT(6) receptors SB-271046 competitively antagonized 5-HT-induced stimulation of adenylyl cyclase activity with a pA(2) of 8.71. SB-271046 produced an increase in seizure threshold over a wide-dose range in the rat maximal electroshock seizure threshold (MEST) test, with a minimum effective dose of < or =0.1 mg kg(-1) p.o. and maximum effect at 4 h post-dose. The level of anticonvulsant activity achieved correlated well with the blood concentrations of SB-271046 (EC(50) of 0.16 microM) and brain concentrations of 0.01-0.04 microM at C(max). These data, together with the observed anticonvulsant activity of other selective 5-HT(6) receptor antagonists, SB-258510 (10 mg kg(-1), 2-6 h pre-test) and Ro 04-6790 (1-30 mg kg(-1), 1 h pre-test), in the rat MEST test, suggest that the anticonvulsant properties of SB-271046 are likely to be mediated by 5-HT(6) receptors. Overall, these studies demonstrate that SB-271046 is a potent and selective 5-HT(6) receptor antagonist and is orally active in the rat MEST test. SB-271046 represents a valuable tool for evaluating the in vivo central function of 5-HT(6) receptors.

Adenylyl Cyclases↗
PubMed2000

Comparison of two blood pool contrast agents for 0.5-T MR angiography: experimental study in rabbits.

PURPOSE: To evaluate two experimental blood pool agents for potential use in equilibrium phase abdominal magnetic resonance (MR) angiography. MATERIALS AND METHODS: MR imaging at 0.5 T was performed in 37 rabbits before and after intravenous injection of a gadolinium-based blood pool contrast agent (SH L 643 A), superparamagnetic iron oxide blood pool agent (SH U 555 C), or gadopentetate dimeglumine. T1-weighted fast spoiled gradient-echo images from the renal arteries to below the iliac bifurcation were obtained. The aorta-to-tissue signal difference-to-noise ratio (SDNR) was measured over time. RESULTS: Both blood pool agents yielded excellent demonstration of the rabbit abdominal aorta. At a dose of 0.1 mmol/kg, both provided a statistically significant increase in aorta-to-tissue SDNR in comparison with that achieved with gadopentetate dimeglumine (200% increase for SH L 643 A, 95% increase for SH U 555 C; P < .05). A 0.1 mmol/kg dose of SH L 643 A provided a 24% increase in SDNR relative to the increase with a 0.37 mmol/kg dose of gadopentetate dimeglumine. Time-dependent enhancement properties of the blood pool agents differed due to differences in elimination method. CONCLUSION: Both blood pool agents were found to be promising contrast agents for 0.5-T MR angiography; however, their clinical applicability warrants further investigation. The gadolinium-based agent had several advantages over the iron oxide compound, including less T2* dephasing, lack of susceptibility artifacts, and fast renal elimination.

Abdomen↗
PubMed2000

The effectiveness of topical treatment in discharging ears with in-dwelling ventilation tubes.

The efficacy and tolerability of topical treatment (both drops and spray) in the treatment of otorrhoea in those ears with ventilation tubes in-situ was studied. Sixty patients were randomised into two treatment groups. One group used an antibacterial/anti-inflammatory ear drop preparation (Otosporin) and the other group a similar preparation administered as an ear spray (Otomize). A 'blind' clinical assessment was made one and three weeks after commencing treatment. Both treatments (drops and spray) significantly improved patients' symptoms and signs after one week (p < 0.001). The spray was significantly easier to administer (p < 0.01) and caused less discomfort on application (p < 0.02).

Administration, Topical↗