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Biomedical subjects

P Rowell

Publications and source records attributed to P Rowell.

At least 19 recordsLinked to original sources

Pituitary suppression in ultrasound-monitored frozen embryo replacement cycles. A randomised study.

BACKGROUND: This study was designed to assess the value of using a gonadotrophin-releasing hormone (GnRH) agonist prior to exogenous steroid supplementation for endometrial preparation in frozen-thawed embryo replacement (FER) cycles. METHODS: A prospective randomized trial of 234 patients undergoing FER cycles was conducted. The study population was randomly divided into two groups according to a computer-generated list. In group A (n = 117), a daily dose of 6 mg of oral estradiol valerate was initiated on menstrual day 1 following pituitary suppression using 400 mcg buserelin acetate daily. In group B (n = 117), the same dose of estradiol valerate was initiated on day 1 of bleeding without prior GnRH agonist therapy. In both groups, ovulation monitoring was not undertaken and progesterone pessaries (800 mg daily) were administrated when the endometrial thickness had reached 8 mm or more with embryo transfer taking place 2 days later. RESULTS: The two groups were comparable with respect to cause of infertility, age at stimulation (32.8 +/- 4 vs 33.2 +/- 3.9 years, P = 0.4), basal FSH level (6.3 +/- 1.7 vs 6.4 +/- 2 IU/l, P = 0.5), number of oocytes collected (16.9 +/- 7.3 vs 16.5 +/- 7.4, P = 0.7) and fertilized normally in the retrieval cycle (11.5 +/- 4.9 vs 11 +/- 4.9, P = 0.4) and number of embryos cryopreserved (6.6 +/- 3.6 vs 6.2 +/- 3.6, P = 0.3). There was no significant difference between the two groups in age at frozen replacement (33.6 +/- 4.2 vs 34 +/- 3.9 years, P = 0.4), duration of the proliferative phase (20.7 +/- 8.6 vs 21 +/- 9.2 days, P = 0.7) and number of thawed embryos replaced (2.3 +/- 0.6 vs 2.2 +/- 0.6, P = 0.2). However, compared with group B, group A achieved significantly higher pregnancy (37.6% vs 24%, OR 1.8, 95%CI 1.1-3.4), clinical pregnancy (24% vs 11.3%, OR 2.5, 95%CI 1.2-5.5) and live birth rates (20% vs 8.5%, OR 2.9, 95%CI 1.2-8). CONCLUSION: Medicated frozen embryo replacement cycles timed by endometrial thickness measurement alone without monitoring or suppression of ovarian activity are associated with reduced outcome.

Adult↗

A pregnancy following PGD for X-linked dominant [correction of X-linked autosomal dominant] incontinentia pigmenti (Bloch-Sulzberger syndrome): case report.

Incontinentia Pigmenti (Bloch-Sulzberger syndrome) is a rare multisystem, ectodermal disorder associated with dermatological, dental and ocular features, and in <10% of cases, severe neurological deficit. Pedigree review suggests X-linked dominance with lethality in affected males. Presentation in female carriers is variable. Following genetic counselling, a mildly affected female carrier diagnosed in infancy with a de novo mutation was referred for preimplantation sexing, unusually selecting for male gender, with an acceptance of either normality or early miscarriage in an affected male. Following standard in-vitro fertilization and embryo biopsy, fluorescence in situ hybridization (FISH) unambiguously identified two male and two female embryos. A single 8-cell, grade 4 male embryo was replaced. A positive pregnancy test was reported 2 weeks after embryo transfer, although ultrasonography failed to demonstrate a viable pregnancy. Post abortive fetal tissue karyotyping diagnosed a male fetus with trisomy 16. This is an unusual report of preimplantation genetic diagnosis (PGD) being used for selection of males in an X-linked autosomal dominant disorder and demonstrates the value of PGD where amniocentesis or chorion villus sampling followed by abortion is not acceptable to the patient. This case also demonstrates the importance of follow-up prenatal diagnosis.

Adult↗

Assisted conception in HIV discordant couples: evaluation of semen processing techniques in reducing HIV viral load.

Artificial insemination with motile spermatozoa prepared from HIV-infected men using standard procedures has been employed with many HIV-discordant couples. We have demonstrated that processing semen from HIV positive men can reduce HIV levels, measured as HIV1 RNA copies/ml using nucleic acid based sequence amplification (NASBA), to undetectable levels (less than 400 copies/ml) but not in all samples. We believe that all processed samples should be tested prior to insemination.

Evaluation Studies as Topic↗

Continence nurses: a survey of who they are and what they do.

This survey is the first report on the characteristics, functions, and barriers to practice reported by continence care nurses in the United States. The data obtained provide valuable information regarding a new subspecialization in nursing.

Adolescent↗

Presynaptic nicotinic receptors and the modulation of transmitter release.

Nicotine is increasingly recognized to promote transmitter release in the brain by a direct action on presynaptic terminals. Pharmacological evidence indicates that this action is mediated by nicotinic receptors. From their sensitivity to mecamylamine, neosurugatoxin and neuronal bungarotoxin these presynaptic receptors can be distinguished from alpha-bungarotoxin-sensitive muscle-type nicotinic receptors, and can be correlated with [3H] nicotine binding sites in the brain. The release of many transmitters in different brain regions is susceptible to stimulation by nicotine, but this effect is not ubiquitous. However, lesioning and subcellular fractionation studies suggest that the majority of brain nicotine receptors are located presynaptically, so that a direct influence of nicotine on transmitter release assumes considerable importance. Although the sensitivity of presynaptic receptors is such that they are likely to be partially activated by doses of nicotine obtained by smoking, the desensitization-induced up-regulation of nicotinic binding sites that follows chronic nicotine treatment raises questions about their functional status during tobacco usage. Chronic administration of the agonist (+)anatoxin-a also up-regulated [3H] nicotine binding sites, and led to increased nicotine-evoked transmitter release in vitro. This could have implications for the involvement of these receptors during withdrawal.

Animals↗

Structure-activity relationships of enkephalins containing serially replaced thiomethylene amide bond surrogates.

An isomeric series of four leucine-enkephalin analogs containing the thiomethylene ether unit as an amide bond replacement in all positions have been prepared by solid phase methods. The resulting pseudopeptides divulged widely differing retentive behaviors on reversed phase high performance liquid chromatography (HPLC). An analog containing the Phe psi[CH2S]Leu dipeptide replacement at the 4-5 position exhibited binding close to the parent, leucine enkephalin; its guinea pig ileum (GPI) activity was the highest of the analogs tested. Another compound, Tyr psi[CH2S]Gly1-2]-Leu-enkephalin, also displaced 3H-etorphine well in the binding assay, but caused increased contractions in the GPI assay at low concentrations. The Phe psi[CH2S]Leu results are not compatible with the necessity of a beta-turn structure for agonist activity in the GPI assay.

Amides↗

Purification and characterization of thioredoxin from the N2-fixing cyanobacterium Anabaena cylindrica.

Thioredoxin has been purified to homogeneity from the cyanobacterium Anabaena cylindrica. The protein consists of a single polypeptide chain with a relative molecular mass of about 11 680 which has two cysteine residues (residues 31 and 34) in the sequence-Cys-Gly-Pro-Cys- and an isoelectric point at pH 4.55. The N-terminal amino acid sequence of 39 residues shows distinct homologies with the sequences of Escherichia coli and Corynebacterium nephridii thioredoxins. Anti-(A. cylindrica thioredoxin) antiserum was used to quantify the thioredoxin which constituted about 0.22% of the soluble protein in cell-free extracts of N2-fixing, NO3- -grown or NH4+-grown A. cylindrica. Activation of fructose-1,6-bisphosphatase of A. cylindrica, activation of glutamine synthetase and NADP+-dependent malate dehydrogenase of the green alga Scenedesmus obliquus but not of A. cylindrica, and deactivation of glucose-6-P dehydrogenase of the cyanobacterium Anabaena variabilis were all achieved using the same thioredoxin species. No other thioredoxin species were detected in extracts of A. cylindrica when examined for the activation of these enzymes.

Amino Acid Sequence↗

The role of specific cations in regulation of cyanobacterial glutamine synthetase.

Purified glutamine synthetase from the cyanobacterium Anabaena cylindrica required a divalent cation for activity. Maximum biosynthetic activity required Mg2+ (25 mM when supplied alone). Co2+ and Mn2+ each supported up to 20% of this activity; 12 other cations tested were ineffective. At 2.5 - 10 mM Mg2+, 0.1 mM Co2+ or ethylene glycol-bis-(beta-aminoethyl ether) N,N'-tetraacetic acid (EGTA) stimulated GS activity to maximum rates; other divalent cations (particularly Mn2+) inhibited Mg2+-dependent activity. At 5 mM Mg2+ the Kappm for NH+4 (0.05 mM) was 20-fold lower than at 25 mM Mg2+; added Co2+ did not markedly alter this low Km for NH+4; this could be physiologically important.

Ammonia↗

Nitrogenase activity and membrane electrogenesis in the cyanobacterium Plectonema boryanum.

The relationships between nitrogenase activity (acetylene reduction) and the transplasmalemma proton electrochemical potential gradient (delta muH+) have been investigated using the non-heterocystous filamentous cyanobacterium Plectonema boryanum. By selectively modifying the chemical (delta pH) and electrical (delta psi) components of delta muH+ under conditions in which the size of the ATP pool remained unaffected, nitrogenase activity was found to be dependent on, or regulated by delta psi. When the ATP pool decreased, concomitant with a decreased internal pH (pHi) the requirement for delta psi was modified. The observed reduction in the intracellular ATP pool and the decreased ATP:ADP ratio also correlated with an inhibition of nitrogenase activity. The data are consistent with a model in vivo in which reductant supply to nitrogenase is regulated by, or dependent on an energised plasmalemma and where there is a fine balance between the supply of reductant and ATP for nitrogenase activity. The correlation observed between delta psi and nitrogenase activity extends our previous observations using the heterocystous cyanobacterium Anabaena variabilis.

Adenosine Diphosphate↗

Characterization of the transport of potassium ions in the cyanobacterium Anabaena variabilis Kütz.

Interrelationships between potassium-ion transport and transplasmalemma electrical-potential difference (delta psi m) have been investigated in Anabaena variabilis (ATCC 29413) by measuring K+ translocation and membrane potential in parallel. At pH 7.0, 5 mmol . dm-3 external K+, there was a thirtyfold accumulation of K+. The K+ equilibrium potential was lower (more negative) than the measured membrane potential by up to 20 mV, (delta psi K+ = -90 mV; delta psi m = -70 mV to -75 mV, respectively). Dark pretreatment and low temperature (4 degrees C) reduced internal K+ and depolarized delta psi m. External pH affected K+ translocation and membrane potential; delta psi m was hyperpolarized at high external pH; transplasmalemma K+ fluxes and internal K+ concentration were also increased at high pH. The effects of pH upon delta psi m, coupled with the finding that the membrane potential was relatively insensitive to external K+, suggest that delta psi m is unlikely to be due primarily to a diffusion potential of K+, but that the membrane potential is maintained by an electrogenic proton-extrusion mechanism. There was no close (obligate) link between K+ transport and changes in delta psi m. Carbonylcyanide m-chlorophenylhydrazone decreased K+ fluxes, internal K+ and delta psi m when added in amounts up to 100 mumol . dm-3. However, delta psi K+ was always more negative than delta psi m. Valinomycin up to concentrations of 50 mumol . dm-3 increased transplasmalemma K+ fluxes by up to 300%, while changes in delta psi m were negligible. Internal K+ was unaffected by valinomycin. N,N'-Dicyclohexylcarbodiimide at concentrations up to 100 mumol . dm-3, reduced K+ flux rates and caused a hyperpolarization of delta psi m. These observations suggest that delta psi m is primarily due to electron transport reactions at the plasmalemma and that K+ transport is energy-dependent. In the presence of dicyclohexylcarbodiimide, internal K+ redistributed in accordance with the membrane potential, suggesting that passive uniport in response to delta psi m (i.e. secondary active transport) is not usually important but may operate when primary active mechanisms are blocked.

Biological Transport, Active↗

Molecular heterogeneity of ferredoxin:NADP+ oxidoreductase from the cyanobacterium Anabaena cylindrica.

The enzyme ferredoxin:NADP+ oxidoreductase (EC 1.18.1.2) from whole filaments of Anabaena cylindrica can be separated into four major fractions by chromatography on phosphocellulose; chromatography using ferredoxin-Sepharose 4B proved to be less satisfactory in separating the fractions. The purified fractions, designated 1, 2, 3 and 4, all showed diaphorase and ferredoxin-dependent cytochrome c reductase activity. The major fractions present were 2 and 3 which were each obtained in an electrophoretically homogeneous state (forms 2 and 3) and represented 30-37% and 30-42%, respectively, of the total enzyme activity. Each was a monomeric species with a molecular weight of approx. 33 000 as determined by gel filtration and sodium dodecyl (SDS)-polyacrylamide gel electrophoresis. Evidence for the presence of a 70 000 molecular weight dimer was also obtained. Forms 2 and 3 had isoelectric points of 5.75 and 6.0, respectively, had similar kinetic properties and were flavoproteins. Extracts of isolated heterocysts showed no form 2 or 3 activity but contained a single form which closely resembled one of the species present in fraction 4; fraction 1 may have been a purification artifact because it was not detected in crude extracts of the cyanobacterium.

Cyanobacteria↗

Nitrogenase activity and membrane electrogenesis in the cyanobacterium Anabaena variabilis Kütz.

Relationships between nitrogenase activity and individual components of the proton electrochemical potential gradient (delta microH+) in Anabaena variabilis have been investigated. The ionophore nigericin was found to collapse delta pH in favour of the membrane potential (delta psi); hyperpolarization of delta psi was correlated with an increase in nitrogenase activity. A positive relationship between nitrogenase activity and membrane potential was also observed using the ionophore valinomycin and the uncoupler carbonylcyanide m-chlorophenylhydrazone. Furthermore, using the energy transfer inhibitor N,N'-dicyclohexylcarbodiimide, nitrogenase activity appeared to be limited by the supply of reductant rather than ATP. The data suggest, for intact filaments and isolated heterocysts, that delta psi may be important in regulating nitrogenase activity in vivo.

Cell Membrane↗