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Biomedical subjects

S Petry

Publications and source records attributed to S Petry.

At least 19 recordsLinked to original sources

Identification of Taylorella equigenitalis responsible for contagious equine metritis in equine genital swabs by direct polymerase chain reaction.

A direct-PCR assay was developed for the rapid detection of Taylorella equigenitalis, a Gram-negative bacterium responsible for contagious equine metritis (CEM) in Equidae. The bacteria may be detected in equine genital swabs without need for a preliminary step of DNA extraction or bacterial isolation. Specificity was determined with 125 isolates of T. equigenitalis, 24 isolates of Taylorella asinigenitalis, five commensal bacteria of the genital tract and a facultative intracellular pathogen of foals found in large concentration in soil. Our PCR is specific and amplified a 413-bp 16S ribosomal DNA product only in all T. equigenitalis.

Animals↗

Identification of binding proteins for cholesterol absorption inhibitors as components of the intestinal cholesterol transporter.

To identify protein components of the intestinal cholesterol transporter, rabbit small intestinal brush border membrane vesicles were submitted to photoaffinity labeling using photoreactive derivatives of 2-azetidinone cholesterol absorption inhibitors. An integral membrane protein of M(r) 145.3+/-7.5 kDa was specifically labeled in brush border membrane vesicles from rabbit jejunum and ileum. Its labeling was concentration-dependently inhibited by the presence of cholesterol absorption inhibitors whereas bile acids, D-glucose, fatty acids or cephalexin had no effect. The inhibitory potency of 2-azetidinones to inhibit photolabeling of the 145 kDa protein correlated with their in vivo activity to inhibit intestinal cholesterol absorption. These results suggest that an integral membrane protein of M(r) 145 kDa is (a component of) the cholesterol absorption system in the brush border membrane of small intestinal enterocytes.

Animals↗

Factors affecting exocellular polysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus grown in a chemically defined medium.

We developed a chemically defined medium (CDM) containing lactose or glucose as the carbon source that supports growth and exopolysaccharide (EPS) production of two strains of Lactobacillus delbrueckii subsp. bulgaricus. The factors found to affect EPS production in this medium were oxygen, pH, temperature, and medium constituents, such as orotic acid and the carbon source. EPS production was greatest during the stationary phase. Composition analysis of EPS isolated at different growth phases and produced under different fermentation conditions (varying carbon source or pH) revealed that the component sugars were the same. The EPS from strain L. delbrueckii subsp. bulgaricus CNRZ 1187 contained galactose and glucose, and that of strain L. delbrueckii subsp. bulgaricus CNRZ 416 contained galactose, glucose, and rhamnose. However, the relative proportions of the individual monosaccharides differed, suggesting that repeating unit structures can vary according to specific medium alterations. Under pH-controlled fermentation conditions, L. delbrueckii subsp. bulgaricus strains produced as much EPS in the CDM as in milk. Furthermore, the relative proportions of individual monosaccharides of EPS produced in pH-controlled CDM or in milk were very similar. The CDM we developed may be a useful model and an alternative to milk in studies of EPS production.

Culture Media↗

Identification of protein components of the microsomal glucose 6-phosphate transporter by photoaffinity labelling.

The glucose-6-phosphatase system catalyses the terminal step of hepatic glucose production from both gluconeogenesis and glycogenolysis and is thus a key regulatory factor of blood glucose homoeostasis. To identify the glucose 6-phosphate transporter T1, we have performed photoaffinity labelling of human and rat liver microsomes by using the specific photoreactive glucose-6-phosphate translocase inhibitors S 0957 and S 1743. Membrane proteins of molecular mass 70, 55, 33 and 31 kDa were labelled in human microsomes by [3H]S 0957, whereas in rat liver microsomes bands at 95, 70, 57, 54, 50, 41, 33 and 31 kDa were detectable. The photoprobe [3H]S 1743 led to the predominant labelling of a 57 kDa and a 50 kDa protein in the rat. Stripping of microsomes with 0.3% CHAPS retains the specific binding of T1 inhibitors; photoaffinity labelling of such CHAPS-treated microsomes resulted in the labelling of membrane proteins of molecular mass 55, 33 and 31 kDa in human liver and 50, 33 and 31 kDa in rat liver. Photoaffinity labelling of human liver tissue samples from a healthy individual and from liver samples of patients with a diagnosed glycogen-storage disease type 1b (GSD type 1b; von Gierke's disease) revealed the absence of the 55 kDa protein from one of the patients with GSD type 1. These findings support the identity of the glucose 6-phosphate transporter T1, with endoplasmic reticulum protein of molecular mass 50 kDa in rat liver and 55 kDa in human liver.

Adult↗

Analysis of lipid metabolism in adipocytes using a fluorescent fatty acid derivative. I. Insulin stimulation of lipogenesis.

Stimulation of lipid synthesis (lipogenesis) is one of the most pronounced metabolic actions of insulin. Here we demonstrate insulin-stimulated lipogenesis in isolated rat adipocytes using a fatty acid derivative which carries a fluorophore. Three major fluorescent lipid products (lipids 1, 2, 3) are generated as revealed by TLC analysis and subsequent fluorescent scanning or imaging. Lipolytic digestion and labeling studies suggest monoacylglycerol-3-phosphate and diacylglycerol (-3-phosphate) structures harboring a single fluorescent fatty acyl residue each for lipids 1 and 3 (2), respectively. Fluorescent triglycerides are not generated. Assaying acylation with isolated microsomes using the purified lipids 1 and 3 indicates that incorporation of one fluorescent fatty acyl residue into glycerol(-3-phosphate) interferes with subsequent esterification. Pretreatment of the adipocytes with insulin significantly stimulates synthesis of lipids 1 and 2, only. The insulin concentration-response relationship (EC50 = 0.5 nM) and the maximal insulin response for synthesis of lipid 1 (stimulation factor = 14- to 20-fold at low glucose and 3- to 7-fold at high glucose) are comparable with those for incorporation of [3-3H]glucose into total adipocyte lipids. Thus this fluorescence-based assay may be useful for studying insulin action and lipogenesis.

Adipocytes↗

Selective defucosylation by mercaptolysis. A potential step in analyzing branched oligosaccharides.

A method of stepwise chemical degradation was elaborated on a microgram quantity of 3-O-alpha-L-fucosyllactose. The key step, TiCl4-catalysed dithioacetal formation from the permethylated N-4-nitrophenyl triosylamine (4) was accompanied by quantitative defucosylation. [14C]Acetylation of the dried mercaptalation mixture gave radiolabelled 3,5-di-O-[14C]acetyl-4-O-(2,3,4,6-tetra-O-methyl-beta-D-galactopyranosyl )-2 , 6-di-O-methyl-D-glucose diethyl dithioacetal (7) and 5-O-[14C]acetyl-2,3,4-tri-O-methyl-L-fucose diethyl dithioacetal (8). The former was further degraded via the bis(sulfone), and thereby 2,3,4,6-tetra-O-methyl-D-galactose (13) was expelled. The monosaccharide branches, fucose and galactose, were identified as derivatives 8 and 13, respectively, by comparison with authentic samples. Isolation of microquantities of products was carried out by preparative TLC.

Acetylation↗

A spacer-modified disaccharide as a photoaffinity reagent for the acceptor-binding area of bovine (1----4)-beta-D-galactosyltransferase: comparison of its acceptor properties with those of other 2-acetamido-2-deoxy-beta-D-glucopyranosides.

The spacer-modified disaccharide 1,10-di-O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-2-azi-1,10- decanediol (10) that mimics the biantennary core heptasaccharide of N-glycoproteins has been synthesised. Compound 10 is an excellent acceptor in galactosyltransferase-catalysed galactosylation by UDP-galactose, is superior (7-8-fold) to analogues that have only one GlcNAc unit, and is an efficient photoaffinity reagent for galactosyltransferase. In the presence of UDP-Gal, no photoaffinity labelling by 10 takes place, which agrees with the mechanism of galactosyltransferase action.

Animals↗

Spacer-modified trisaccharide glycosides that mimic the biantennary Asn-linked oligosaccharide acceptor of (1----4)-beta-D-galactosyltransferase and can be used as competitive inhibitors and for irreversible deactivation.

The biantennary spacer-modified trisaccharide glycoside methyl 3,6-di-O-(2-acetamido-2-deoxy-beta-D-glucopyranosyloxyethyl)-alpha -D- mannopyranoside (5) was synthesised and used together with several 2-acylamino-2-deoxy-D-glucose derivatives in competition experiments with beta-D-galactosyltransferase. Compound 5 was an acceptor substrate (KM 0.18 mM) comparable to the biantennary core heptasaccharide of glycoproteins (KM 0.13 mM). Replacing the N-acetyl group by other N-acyl groups did not alter the kinetic parameters significantly. When the N-acyl group was iodoacetyl, the compound was an irreversible inhibitor.

Asparagine↗

Evidence for independent processing of subjective contour brightness and sharpness.

Subjective contours have been of considerable interest because of their importance to theories and physiological models of form perception. In particular, they have recently been characterized as the result of magnocellular cortical processing. There is, however, a paucity of parametric data relating to basic psychophysical parameters in this field. Two experiments are reported in which the roles of subjective contour size, retinal eccentricity, and flicker rate in subjective contour salience were investigated. Eleven observers estimated subjective contour magnitude using an Ehrenstein configuration. Configurations ranging in size from 0.25 to 3 deg were presented to three retinal loci (fovea, 2 deg, and 4 deg) at flicker rates ranging from 5 to 15 Hz. Subjective contour brightness and distinctness were measured separately. Brightness was greatest at a subjective contour size of about 1.25 deg, at flicker rates of 5-7 Hz, and at 3 deg peripheral for all flicker rates and all but the smallest stimulus sizes. Distinctness decreased with eccentricity and flicker, but remained high at small diameters (thus implicating spatially sensitive mechanisms). Taken together, the results support a magnocellular processing of subjective contours with respect to brightness, but also suggest that there is a parvocellular contribution to subjective contour sharpness.

Adult↗

Comparison of four methods for testing high-level aminoglycoside resistance in enterococci.

In a prospective study the prevalence of high-level aminoglycoside resistance (MIC greater than or equal to 2,000 micrograms/ml) among 62 clinically significant enterococci was investigated. A total of 10(5) organisms were inoculated a) onto a plate containing 2,000 micrograms/ml of gentamicin or streptomycin; b) into a microtube for dilution MIC determinations for gentamicin, amikacin, tobramycin and streptomycin; and c) into a single tube containing 500 micrograms/ml of gentamicin, amikacin, tobramycin or streptomycin in supplemented Mueller-Hinton broth. In addition, tubes containing 500 micrograms/ml of gentamicin, amikacin, tobramycin or streptomycin were inoculated with five enterococcal colonies ("crude" method). For 45 of the 62 isolates, MICs of gentamicin, amikacin and tobramycin were less than or equal to 500 micrograms/ml, while 17 (27%) showed high-level resistance. The MICs of streptomycin were less than or equal to 500 micrograms/ml for 42 of 62 isolates, and greater than or equal to 2,000 micrograms/ml for 20 (32.3%). For 8 of the 17 (47%) isolates showing high-level gentamicin resistance, MICs of streptomycin were less than or equal to 500 micrograms/ml. There was complete agreement between the results of the plate method, the microtube dilution MIC and the tube inoculated with 10(5) CFU, but the crude method gave discordant results for two isolates. It is concluded that a tube containing 500 micrograms/ml of aminoglycoside is a simple, accurate and inexpensive method for determining high-level aminoglycoside resistance.

Amikacin↗

Personality alterations in multi-infarct dementia.

Multi-infarct dementia (MID) produces changes in intellectual function and in overt behavior, but alterations in demeanor have received little systematic investigation. In this study, an assessment of personality change occurring after the onset of the dementia was performed using an inventory completed by the subjects' spouses. Personality alterations in MID were compared to those occurring in a group of healthy elderly individuals. Twelve of the 18 inventory items changed more in the MID patients than in normal control subjects. Personality changes are a consistent part of the clinical syndrome of MID and occur early in the course of the disease.

Aged↗

Organic personality disorder in dementia syndromes: an inventory approach.

Personality alterations were investigated in patients with dementia of the Alzheimer type (DAT), multi-infarct dementia (MID), and normal control subjects, using a standardized informant-rated personality inventory. Personality alterations were observed in all patients, and the two diagnostic groups exhibited many similar behavioral alterations. DAT patients had significantly greater alterations in maturity and exhibited less personal control compared with the MID patients; the latter manifested more apathy and remained more affectionate and easy-going. Few correlations were found between the severity of dementia and the magnitude of behavioral changes. Personality alterations are an important and consistent aspect of the phenomenology of dementing disorders.

Aged↗

Personality alterations in dementia of the Alzheimer type.

Personality alterations in dementia of the Alzheimer type (DAT) are common but have received little systematic or quantitative investigation. In this study, changes in personality in patients with DAT were compared with those of nondemented retirees using a personality inventory. The inventory used information obtained from each subject's spouse. No personality changes were identified in the control subjects when pre- and post-retirement inventory scores were compared, whereas patients with DAT had highly significant alterations on 12 of the 18 inventory items. The results demonstrate that patients with DAT become more passive, more coarse, and less spontaneous as a result of the disease. Personality changes are a consistent part of the clinical syndrome of DAT and occur early in the course of the disease.

Aged↗

Rocket-ship psychophysics. Assessing visual functioning in young children.

A set of psychophysical procedures is described for rapidly measuring visual functions such as spatial contrast sensitivity, flicker fusion, and rod and cone two-color increment thresholds in young children. The psychophysical procedures, disguised as electronic space games, use a forced-choice paradigm combined with the method of descending limits. Because children enjoy the tasks and are highly motivated, the tests yield good sensory thresholds in a short period of time. However, when thresholds from 6-8-year-old children are compared with those of adults, the children have slightly higher thresholds for all of the functions tested. The differences are ascribed to the tendency of children to adopt a guessing strategy when stimuli are close to threshold.

Adolescent↗