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Biomedical subjects

R Shaw

Publications and source records attributed to R Shaw.

At least 19 recordsLinked to original sources

Characterisation of the Historic Demographic Decline of the British European Polecat Population.

The European polecat (Mustela putorius) has a widespread distribution across many countries of mainland Europe but is documented to be declining within these ranges. In Britain, direct persecution led to a severe decline of the polecat population during the 19th century. Unlike European mainland populations, it is now recovering across much of its former British range. The genomic and conservation implications of such a severe demographic decline, followed by the current recovery, have still to be characterised in the European polecat in Britain. Here we carry out population-level whole-genome analyses of 65 polecats from Britain (Wales and England) and the European mainland. Our analyses reveal that Welsh polecats show genetic variability from both English and European polecats, while British polecats as a whole exhibit signs of genetic isolation from mainland European populations. We also reconstructed the demographic history of the Welsh polecat to quantify the magnitude of the bottleneck. Our analyses confirmed the drastic decline of the Welsh polecat's effective population size, with a severe genetic bottleneck around 30-40 generations ago (1854-894). We investigated whether whole-genome diversity reflected this demographic event and found that Welsh polecats had significantly less genetic diversity than English polecats, but not European polecats. Runs of homozygosity and genetic load present in Welsh and English polecat genomes also indicated recent historic inbreeding. Our findings suggest that the increase in the British polecat population size may be attributed to admixture events. Additionally, we demonstrate that the Welsh polecat constitutes a genetically distinct population, which could be crucial for the overall conservation of European polecats by preserving unique genetic diversity.

Genetics, Population

Fluorocytosine causes uncoupled dissipation of the proton gradient and behaves as an imperfect substrate of the yeast cytosine permease.

At pH 5-6 ATP-depleted washed cell preparations of strain NC233-10b[pII4-9], in which the cytosine permease was overexpressed, absorbed cytosine, hypoxanthine or fluorocytosine stoichiometrically with, respectively, about 1, 1.4 and 5 proton equivalents. The cellular pH fell proportionately. The membrane depolarization caused by each compound was assayed in the presence of glucose with a voltage-sensitive dye and increased in the same order. Fluorocytosine significantly lowered the growth yield that a 'petite' strain of the yeast formed at limiting glucose concentrations. At pH 5.6 with extracellular [K+] below 1 mM, each of the three substrates was accumulated about 200-fold from a dilute solution at the expense of the proton gradient. This concentration ratio corresponds to a solute gradient (delta mu(s)) of 13 kJ mol-1. Raising [K+]o systematically lowered the substrate accumulation ratio and delta muH. The mean ratio delta mu(s)/delta muH was 0.82 for all three substrates. It was concluded that whereas the behaviour of cytosine approximated to that expected for a symport of unit proton stoichiometry, the absorption of protons with fluorocytosine and, to a lesser extent, hypoxanthine, was only partly conserved as useful work. A possible mechanism of this novel phenomenon is outlined.

Adenosine Triphosphate

Bile acids. LIX. Purification of 5 alpha-anhydrocyprinol by preparative high-performance liquid chromatography.

26,27-Oxido-5 alpha-cholestane-3 alpha,7 alpha,12 alpha-triol can be obtained in a homogeneous state in gram quantities by passing it through one PrepPak-500/Silica cartridge mounted in a Waters Assoc. preparative liquid chromatograph. The elution solvent was methanol-chloroform (1:14). The isolated material was analyzed for purity by several chromatographic means and by elemental analysis, and was finally characterized by the usual spectroscopic means. Gas-liquid chromatography of its trimethylsilyl ether indicated the formation of a tetrakis-trimethylsilyl-26-chloro derivative, in addition to the expected tris-trimethylsilylated substance. The structure of the former compound is deduced from the fragmentation and isotope abundance in its mass spectrum and from chemical principles.

Animals

Competitive inhibitors of rabbit hepatic microsomal 12 alpha-steroid hydroxylase.

Rabbit hepatic microsomal 12 alpha-steroid hydroxylase which is stable to storage at -70 degrees C in the pellet form was assayed for activity with [5 alpha,6 alpha-3H2]cholestane-3 alpha,7 alpha-diol solubilized with Tween 80 since methanol was incapable of maintaining the sterol in aqueous solution. Under optimized conditions in phosphate buffer, pH 7.4, containing nicotinamide, magnesium chloride, and NADPH, the enzyme conversion appeared linear for the initial 10 min. The rate of hydroxylation was proportional to protein concentration up to 4 mg/ml. Apparent Km and Vmax were 71 microM and 323 pmol of product/mg of protein/min. Based on the known structural requirements of the enzyme system, competitive inhibitors were prepared with the C-12 position derivatized as an alkene, hydroxyl, or oxo functional group. A Dixon plot revealed that 5 alpha-cholest-11-ene-3 alpha,7 alpha,26-triol was the best inhibitor with an apparent Ki of 26 microM.

Animals

Effects of cyclic AMP altering drugs on endotoxin-induced termination of pregnancy.

The effects of dibutyryl-cyclic AMP (DBcAMP), propranolol, and theophylline on endotoxin (LPS) challenged pregnant mice was determined by comparing the patterns of termination and times of fetal expulsion. Endotoxin isolated from wild type (WT) and Re chemotype mutant cells of Salmonella typhimurium was used. Wild type LPS caused the expulsion of the fetuses, while Re LPS primarily caused the resorption of the fetuses. The major change in pattern occurred in the propranolol treated Re LPS challenged group with a trend toward more expulsions and a decrease in uterine cyclic AMP (cAMP) levels to that approaching the group challenged with wild type LPS. The time of expulsion was also affected in the wild type LPS treated group, with propranolol hastening and DBcAMP and theophylline delaying the expulsion. These results support the hypothesis that uterine cAMP is involved in the expulsion of the fetuses.

Abortion, Spontaneous

Chemical synthesis of three 14 alpha-hydroxymethyl cholestenols.

Reported herein are chemical syntheses of 14 alpha-hydroxymethyl-5 alpha-cholest-8-en-3 beta-ol, 14 alpha-hydroxymethyl-5 alph-cholest-7-en-3 beta-ol, and 14 alpha-hydroxymethyl-5 alpha-cholest-6-en 3 beta-ol. These compounds were obtained in pure form after repeated medium-pressure column chromatography of the mixture obtained by treatment of 3 beta-acetoxy-7 alpha,32-epoxy-14 alpha-methyl-5 alpha-cholestane with pyridine hydrochloride in refluxing acetic anhydride followed by reduction with lithium aluminum hydride. The compounds were characterized by their chromatographic properties and by the results of infrared, optical rotation, nuclear magnetic resonance, and low and high resolution mass spectral studies.

Cholestenes

Gel permeation chromatography as a stability-indicating assay for human serum albumin.

It has been known for a considerable period of time that preparations of normal human serum albumin contain a amount of polymers and their formation appears to be related to the stability of human albumin solutions. This report describes a gel permeation chromatography technique using Sephadex G-150 to separate albumin molecules into monomers, imers, and polymers. In order to determine if this system could detect small changes in albumin solutions, 5% solutions of albumin were heat denatured and assayed. The changes which resulted from the heat denaturation were easily measurable. Additional experiments using 5% and 25% albumin solutions showed that this system could also detect changes in the amount of aggregation which occurred in these solutions with time. It therefore appears that this technique may be of value in assessing the stability of albumin solutions as measured by the formation of protein aggregates.

Chromatography, Gel

Bile acids. LIV--mass spectra of conjugated bile acids.

The electron impact mass spectra of conjugated bile acids, their 5alpha-analogs and methyl esters of glyco conjugates were determined by direct insertion into the ion source and their fragmentation patterns were found to be basically similar to those of methyl esters of the free bile acids. The conjugates are additionally characterized by a significant loss of [NH2CH2CO2H] from the glycine moiety and [CH2 = CHSO3 H] from the taurine group. Several 5 alpha- and 5 beta-isomers can be differentiated, but no general pattern of recognition is discernable. Field desorption mass spectra contain [M + Na]+ and [m + Na]2+ ions.

Bile Acids and Salts

Bile acids LVII. Analysis of bile acids by high pressure liquid chromatography and mass spectrometry.

Several high pressure liquid chromatographic methods for the separation of conjugated and free bile acids are presented. A mixture of synthetic conjugated bile acids has been separated by reverse-phase systems consisting of either a Waters Associated' "fatty-acid analysis" or a muBondapak/C18 column eluted with a mixture of 2-propanol/potassium phosphage buffer (pH 2.5 or 7.0). The major conjugated bile acids present in the gallbladder bile of obese subjects have been analyzed each in less than 30 min and quantitated with a U.V. detector set at 193 nm. Some of the 5 alpha- and 5 beta-isomers of conjugated bile salts could be resolved in straight-phase systems on Corasil II or muPorasil columns. Mass spectra of the conjugated bile acids obtained by electron impact were characteristics of the type of amino acid attached to the side chain, and the number of hydroxyl substituents on the nucleus. Most of the isomers could readily be differentiated by the relative intensities of the fragment ions.

Amino Acids

Nitrous oxide contamination in dental surgeries using relative analgesia.

The concentrations of nitrous oxide were determined at various positions in four dental surgeries in which nitrous oxide sedation was used without scavenging. Contamination was present in all the surgeries, the greatest concentration being recorded at the dentist's nose and the least in the peripheral air. The reception area adjoining the surgeries was found also to be contaminated by nitrous oxide.

Air Pollutants

Comparison of sampling techniques for studies of nitrous oxide pollution.

The retention of nitrous oxide in samples of nitrous oxide in air held in disposable plastic syringes, glass syringes and custom-made nylon film bags was studied by means of gas chromatography at intervals of 90 min, 24, 48 and 120 h after filling the containers. After 24 h the nylon bags retained a mean concentration of 94.5% of the original concentration of nitrous oxide. The disposable plastic syringes and glass syringes were unsatisfactory and retained only 80% of the original concentration of nitrous oxide after 90 min. These values decreased to 51% and 31% of the original concentration of nitrous oxide respectively at 24 and 48 h.

Air Pollution

Calcific constrictive pericarditis in Degos' disease.

A 31-year-old woman presented with symptoms and signs of constrictive pericarditis. She had a history of Degos' disease, a rare disorder characterized by skin and bowel lesions thought to be secondary to vasculitis. A chest roentgenogram showed extensive calcification of the pericardium. Although pleural and pericardial involvement has been reported in this disease, constrictive pericarditis is most unusual, and radiographically demonstrable calcification of the pericardium has not been reported previously.

Adult

Effect of endotoxin on uterine cycle AMP in pregnant mice.

The effect of endotoxin (LPS) on uterine cAMP was determined by measuring the levels of cAMP and cAMP phosphodiesterase after challenge. Two LPS preparations, isolated from wild type (WT) and Re chemotype mutant cells of Salmonella typhimurium were used. The pattern of termination differed with WT LPS resulting in expulsion of the fetuses, and Re LPS primarily causing the resorption of the fetuses. The animals challenged with WT LPS showed a decrease in uterine cAMP when the mice were starting to expel the fetuses while the Re LPS treated group maintained control levels of cAMP. Cyclic AMP phosphodiesterase also decreased in WT LPS and not the Re LPS group. These results suggest the possibility that uterine cAMP is involved in the expulsion of the fetuses.

3',5'-Cyclic-AMP Phosphodiesterases