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Biomedical subjects

B A Stone

Publications and source records attributed to B A Stone.

At least 73 records · Page 4Linked to original sources
PubMed1986

Production of steroids and release of prostaglandins by spherical pig blastocysts in vitro.

Levels of pregnenolone and progesterone in spherical pig blastocysts (near 4 and 15 microM respectively) exceeded respective levels in histotroph by about 400-fold. When blastocysts were cultured for 5 days in a synthetic medium containing pregnenolone sulfate (1 microM), daily rates of release of pregnenolone, progesterone, androstenedione, testosterone, oestrone and oestradiol were determined to be near 320, 45, 26, 27, 0.8 and 9.2 fmol per blastocyst respectively. Daily outputs of progesterone and testosterone (fmol per blastocyst) diminished (P less than 0.05) to 1.3 and undetectable levels (less than 2) respectively in the presence of Trilostane (94 microM). Increasing the content of pregnenolone sulfate in the culture medium (to 4.5 microM) resulted in higher daily rates of release of pregnenolone and progesterone (to near 1740 and 380 fmol per blastocyst respectively), verifying activity of 3 beta-hydroxy-delta 5-steroid dehydrogenase, and of arylsulfatase, in tissues of intact spherical pig blastocysts. Prostaglandin E2 was the predominant prostaglandin (PG) released by cultured blastocysts (about 1 fmol per blastocyst per hour), hourly rates of release of PGH2 (derived) and PGF2 alpha being near 0.1 and less than 0.06 fmol per blastocyst respectively. The data establish a capacity for spherical pig blastocysts to release a range of steroids and PGs of possible significance to embryonic growth and development in vivo.

Androstenedione↗
PubMed1986

Detection of Corynebacterium kutscheri in animal tissues by DNA-DNA hybridization.

Corynebacterium kutscheri, a pathogen of experimental rats and mice, is difficult to detect in the early stages of infection. A diagnostic assay was developed to reveal the organism in tissues of asymptomatic, infected animals. Three fragments of DNA specific to C. kutscheri were isolated, radiolabeled with 32P, and used in DNA-DNA hybridization assays with blots of impression smears of animal tissues on membrane filters. The C. kutscheri probes did not cross-hybridize with 15 DNA isolates from nonpathogenic corynebacteria. The method described is more rapid and more specific than conventional immunological and culture procedures used to detect C. kutscheri; the organism was detected in tissues of animals infected for only 1 week.

Animals↗
PubMed1986

Concentrations of steroid hormones, and of prolactin, in washings of the human uterus during the menstrual cycle.

Levels of steroid hormones, prolactin and protein were determined in trans-cervical flushings of uteri of 73 consenting women presenting for reversal of sterilization. Median total levels of steroids (pmol), prolactin (mu i.u.) and protein (mg) in the washings were: pregnenolone, 4.22; pregnenolone sulphate, 15.1; progesterone, 1.01; dehydroepiandrosterone (DHEA), 8.92; DHEA sulphate, 368; androstenedione, 2.23; testosterone, 1.04; oestrone, less than 0.7; oestrone sulphate, 0.49; oestradiol, 0.08; prolactin, 23.8; and protein, 5.75. Levels of these components of uterine flushings did not vary significantly between Days 6-10, 11-14, 15-20 and 21-28 after the onset of the previous menstrual period (P greater than 0.05). Uniform levels of free steroids in uterine washings throughout the menstrual cycle, and low free steroid/total protein ratios (all less than 3 pmol/mg), support other evidence for a paucity of steroid-binding proteins in human histotroph. The predominance of DHEA sulphate and of pregnenolone sulphate in human uterine washings is in accord with their abundance in plasma, and may provide an important precursor pool for de-novo steroidogenesis by human embryos before implantation. Our results support the view that human histotroph is a filtrate of plasma.

Dehydroepiandrosterone↗
PubMed1985

Activity and action pattern of Bacillus licheniformis alpha-amylase in aqueous ethanol.

A purified B. licheniformis alpha-amylase in a mixture of ethanol-aqueous buffer (1:1, v/v) retains half the activity shown in water alone. In ethanol-aqueous buffer (7:3, v/v) about 20% of the activity is retained. The pattern of oligosaccharides produced from amylose changed with ethanol concentration; in aqueous buffer the products are: DP 1 and 2, 33.7%; DP 3, 28.5%; DP 4, 4.4% and DP 5, 33.4%. Whereas in ethanol-aqueous buffer (7:3, v/v) the products are DP 1 and 2, 66.8%; DP 3, 17.3%; DP 4, 4.1% and DP 5, 11.8%. These results suggest that a change in substrate affinity at the active centre subsites is induced in the ethanol-aqueous buffer medium.

Bacillus↗
PubMed1985

Steroid hormones in uterine washings and in plasma of gilts between days 9 and 15 after oestrus and between days 9 and 15 after coitus.

Between Days 9 and 15 after oestrus, concentrations of pregnenolone, pregnenolone sulphate, dehydroepiandrosterone (DHEA), DHEA sulphate, androstenedione, oestrone and oestrone sulphate in free uterine fluid collected from non-pregnant gilts were greater than respective values in plasma (P less than 0.05). The total contents of pregnenolone, progesterone, DHEA, testosterone, oestrone and oestradiol in washings from pregnant uteri exceeded (P less than 0.05) respective non-pregnancy levels during this same period. Concentrations of pregnenolone, pregnenolone sulphate, DHEA, DHEA sulphate, androstenedione, oestrone, oestrone sulphate and oestradiol in free uterine fluid recovered from gravid uteri were also higher (P less than 0.05) than respective plasma values. By contrast, the progesterone concentration in uterine fluid from pregnant animals was lower (P less than 0.001) than the plasma value. Concentrations of DHEA, DHEA sulphate, androstenedione and oestrone sulphate in plasma of pregnant gilts between Days 9 and 15 after mating exceeded (P less than 0.05) the respective concentrations in unmated gilts between Days 9 and 15 after oestrus. Plasma levels of pregnenolone sulphate were lower (P less than 0.05) in the pregnant animals. We therefore suggest that the endometrium of the pig can concentrate steroid hormones in uterine fluid and that increases in steroid levels in this milieu between Days 9 and 15 after coitus reflect steroidogenesis by embryonic tissues and modification of enzyme activities within uterine tissues under the influence of progestagens. The pool of steroid sulphoconjugates present in uterine fluid between Days 9 and 15 post coitum could serve as an important precursor source for progestagen, androgen and oestrogen synthesis by tissues of pig embryos before implantation.

Androstenedione↗
PubMed1984

An improved procedure for the methylation analysis of oligosaccharides and polysaccharides.

An improved procedure for the methylation analysis of oligosaccharides and polysaccharides is described. Steps in the procedure were examined and optimised for quantitative recovery and speed. Methylation was shown to be complete by using [14C]methyl iodide. All operations were performed in the same tube and the need to concentrate solutions containing acetylated alditols of methylated sugars was eliminated, thus minimising losses due to volatilization. The method is convenient, gives high recoveries of acetylated alditols of methylated sugars, and allows analysis of the glycosyl linkages of oligo- or poly-saccharides to be completed within a working day. A wide range of oligo- and poly-saccharides were methylated by this procedure.

Chromatography, Gas↗
PubMed1983

Localization of beta-glucan synthases on the membranes of cultured Lolium multiflorum (ryegrass) endosperm cells.

The distribution of beta-glucan synthases between plasma membranes and intracellular membranes of suspension-cultured Italian-ryegrass (Lolium multiflorum Lam.) endosperm cells was examined. Highly purified plasma membranes prepared from protoplasts were only slightly enriched in beta-glucan synthases assayed at 10 microM- and 1 mM-UDP-glucose. Most beta-glucan synthase was associated with intracellular membranes. These membranes were fractionated on a linear sucrose density gradient and were resolved into different membrane fractions containing beta-glucan synthases. Beta-Glucan synthases assayed at 10 microM-UDP-glucose were found in a fraction banding at a density of 1.11 g . cm-3, but most of the beta-glucan synthase assayed at 1 mM-DDP-glucose was at a density of 1.04 g . cm-3.

Cell Membrane↗
PubMed1983

Detection of neutral and aminosugars from glycoproteins and polysaccharides as their alditol acetates.

A new method for the preparation and separation of alditol acetates from neutral sugars has been applied to aminosugars. Reduced aminosugars were rapidly acetylated using 1-methylimidazole as the catalyst without removal of borate formed during reduction. The alditol acetates were separated by glass capillary gas chromatography on Silar 10C. The alditol acetates of aminosugars had retention times much longer than those of neutral sugars. However, the alditol acetates of the deamination products of aminosugars had shorter retention times and were resolved from those of neutral sugars. This method was used for the simultaneous detection of neutral and aminosugars in acid hydrolysates of chitin and the glycoproteins, ovalbumin and peroxidase.

Acetates↗
PubMed1982

Isolation of plasma membrane from protoplasts of Lolium multiflorum (ryegrass) endosperm cells.

Plasma membranes have been isolated from protoplasts of suspension-cultured ryegrass (Lolium multiflorum) endosperm cells. The protoplast membrane is coated before cell disruption with murine myeloma protein J539, a galactose-binding immunoglobulin A. The plasma membrane is labelled with 125I by using chemically or enzymically catalysed iodination techniques, or, more conveniently, by using 125I-labelled myeloma protein J539, which enables the membrane to be simultaneously coated and labelled. Protoplast lysis is effected by gentle mechanical means after swelling in hypo-osmotic medium. The plasma-membrane fraction is recovered at low centrifugal forces by fractionation of cell lysates on a discontinuous sucrose/sorbitol gradient. The plasma-membrane fraction is enriched 96-fold on a protein basis with respect to the specific radioactivity of 125I-labeled myeloma protein J539 in the homogenate. Electron microscopy showed long membrane profiles often associated with one another.

Carrier Proteins↗
PubMed1982

Solubilization of beta-glucan synthases from the membranes of cultured ryegrass endosperm cells.

beta-Glucan synthases were solubilized by treating membrane preparations from suspension-cultured ryegrass (lolium multiflorum) endosperm cells with detergents. Of the seven detergents tested only digitonin and octyl glucoside dissociated active synthases from the membranes. The digitonin-solubilized enzymes produced 1,4-beta-glucans and 1,3:1,4-beta-glucans, whereas the digitonin-insoluble enzymes produced, in addition, 1,3-beta-glucans. Chromatography of the digitonin-solubilized beta-glucan synthases on DEAE-Sepharose resulted in their partial purification. The octyl glucoside-solubilized enzymes produced more 1,3-beta-glucans than did the membrane-bound preparations. These results suggest that the 1,3-beta-glucan synthase is a separate enzyme and is not involved in 1,3:1,4-beta-glucan synthesis. Digitonin not only dissociated synthases from the membranes, but also stimulated synthase activity. This effect may be related to the inhibition by digitonin of glucosyl transfer from UDP-glucose to form steryl glucosides.

Cell Membrane↗
PubMed1982

Factors Influencing beta-Glucan Synthesis by Particulate Enzymes from Suspension-Cultured Lolium multiflorum Endosperm Cells.

Particulate enzymes from suspension-cultured ryegrass (Lolium multiflorum Lam.) endosperm cells incorporated glucosyl residues from UDP-glucose and GDP-glucose into beta-glucans. Three types of beta-glucans were produced from UDP-glucose: 1,3-beta-glucan; 1,4-beta-glucan; and mixed-linkage 1,3;1,4-beta-glucan. As in other systems, relatively more 1,4-beta-glucan was produced from a low (10 micromolar) UDP-glucose concentration, and relatively more 1,3-beta-glucan was produced from a high (1 millimolar) UDP-glucose concentration. However, in ryegrass, 1,3;1,4-beta-glucan represented a major proportion of the products at both low and high UDP-glucose concentrations. The arrangement of linkages in the 1,3;1,4-beta-glucan was different at the two concentrations; at the low UDP-glucose concentration, more sequences of three consecutive 1,4-linkages were produced.The effects of pH, temperature, and metal ion concentrations on incorporation were dependent on the UDP-glucose concentration. At the low UDP-glucose concentration, incorporation into all three types of beta-glucan increased with increasing pH. At the high UDP-glucose concentration, 1,3-beta-glucan was the major product at pH 7 and below; 1,4-beta-glucan synthesis was optimal at pH 8; and synthesis of 1,3;1,4-beta-glucan was greatest above pH 8.With 10 micromolar GDP-glucose as substrate, 1,4-beta-glucan, but no 1,3;1,4-beta-glucan, was produced. Incorporation from either UDP-glucose or GDP-glucose was not influenced by the presence of the other.

Journal Article↗
PubMed1981

Thermal characteristics of the testis and epididymis of the boar.

Intra-testicular temperatures of conscious boars were in the range 35.0-36.5 degrees C; values for the caput epididymidis were 35.5-37.0 degrees C and for the cauda epididymidis 35.3-37.0 degrees C. The modal temperatures of these tissues were approximately 2.5, 1.5 and 1.9 degrees C respectively below rectal temperature (38.2 degrees C) and each tissue temperature varied diurnally. With increasing environmental temperature (from 23 to 34 degrees c), testicular and epididymal temperatures increased, and the differential between tissue and rectal temperature levels diminished. The temperature of the caput epididymidis changed most with rising environmental temperature. It is possible to predict testis or epididymal temperature from environmental or rectal temperature.

Animals↗
PubMed1974

Chemical and physical properties of an arabinogalactan-peptide from wheat endosperm.

1. An arabinogalactan-peptide from wheat endosperm was studied by using physicochemical techniques and some aspects of its chemical structure were determined. 2. The arabinogalactan-peptide is a non-associating, polydisperse macromolecule ([unk]=22000) which exhibits only minor non-ideal effects in aqueous solution. 3. Examination of the products of partial acid hydrolysis of the polysaccharide component showed that arabinose is present in the alpha-l-arabinofuranosyl configuration, and i.r.-absorption spectroscopy and optical-rotation studies suggest that the d-galactopyranose residues are linked by glycosidic linkages in the beta-anomeric configuration. 4. The arabinogalactan is linked to a peptide which represents 8% (w/w) of the arabinogalactan-peptide and which may be present as a molecular core. Partial degradation of the polymer by successive treatment with oxalic acid and NaOH showed that the linkage between polysaccharide and peptide involves galactose and hydroxyproline residues and is glycosidic in nature. A tentative model is proposed for the structure of the wheat endosperm arabinogalactan-peptide. 5. The subcellular location and function of the arabinogalactan-peptide is discussed in relation to previous work with related molecules.

Amino Acids↗