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H Mukasa

Publications and source records attributed to H Mukasa.

At least 19 recordsLinked to original sources
PubMed2003

Classification of oral streptococci by two-dimensional gel electrophoresis with direct activity stain for glycosyltransferases.

Thirty eight strains of oral streptococci were divided into six types by two-dimensional gel electrophoresis (2-DE) followed by glycosyltransferase (GLT) activity stain: Type 1, Streptococcus mutans; Type 2, Streptococcus rattus; Type 3, Streptococcus sobrinus and Streptococcus downei; Type 4, Streptococcus cricetus; Type 5, Streptococcus salivarius; and Type 6, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii. In Types 1, 2 and 5, two glucosyltransferases synthesizing water-insoluble (GTF-I) and water-soluble glucans (GTF-S) and a fructosyltransferase (FTF) were detected. In Types 3 and 4, GTF-I and two GTF-Ss were detected. Only one GTF-S was detected in Type 6. The 2-DE patterns for these six types were clearly distinguished from each other based on the kind, number and location of GLTs in gel.

Bacterial Typing Techniques↗
PubMed2001

Dextran acceptor reaction of Streptococcus sobrinus glucosyltransferase GTF-I as revealed by using uniformly 13C-labeled sucrose.

A sucrose glucosyltransferase GTF-I from cariogenic Streptococcus sobrinus transferred the uniformly 13C-labeled glucosyl residue ([U-(13)C]Glc) from [U-(13)C]sucrose to exogenous dextran T500 at the non-reducing-end, mostly by alpha-(1-->6) linkages and partially by alpha-(1-->3) linkages, as revealed by the 13C-(13)C NMR coupling pattern. With increasing amounts of [U-(13)C]sucrose, transfer of [U-(13)C]Glc to the alpha-(1-->3)-linked chain became predominant without increase in the number of chains. The transfer of [U-(13)C]Glc to an isomaltopentaose acceptor occurred similarly to its transfer to T500. alpha-(1-->3)-branches in the [U-(13)C]dextran, specifically synthesized from [U-(13)C]sucrose by a Streptococcus bovis dextransucrase, were not formed by GTF-I, as judged by the observation that a newly-formed alpha-1,3,6-branched [U-(13)C]Glc was not detected, which could have been formed by transferring the unlabeled Glc from sucrose to the internal alpha-(1-->6)-linked [U-(13)C]Glc at C-3. The 13C-(13)C one-bond coupling constants (1J) were also recorded for the C-1--C-6 bond of the internal alpha-(1-->6)-linked [U-(13)C]Glc and of the non-reducing-end [U-(13)C]Glc.

Bacterial Proteins↗
PubMed2000

Nigerooligosaccharide acceptor reaction of Streptococcus sobrinus glucosyltransferase GTF-I.

Nigerose and nigerooligosaccharides served as acceptors for a glucosyltransferase GTF-I from cariogenic Streptococcus sobrinus to give a series of homologous acceptor products. The soluble oligosaccharides (dp 5-9) strongly activated the acceptor reaction, resulting in the accumulation of water-insoluble (1-->3)-alpha-D-glucan. The enzyme transferred the labeled glucosyl residue from D-[U-13C]sucrose to the 3-hydroxyl group at the non-reducing end of the (1-->3)-alpha-D-oligosaccharides, as unequivocally shown by NMR 13C-13C coupling patterns. The values of the 13C-13C one-bond coupling constant (1J) are also presented for the C-1-C-6 of the 13C-labeled alpha-(1-->3)-linked glucosyl residue and of the non-reducing-end residue.

Bacterial Proteins↗
PubMed1996

Purification and properties of extracellular glucosyltransferase from Streptococcus bovis.

Eight Streptococcus bovis strains were classified into 3 types on the basis of isoelectric point (pI) and molecular mass (M(r)) of extracellular glucosyltransferase. Strains ATCC 9809, 35034 and 43143 produced glucosyltransferase of pI 3.7 and M(r) 165 kDa; strains ATCC 15351, 27960 and 33317 produced glucosyltransferase of pI 4.1 and M(r) 140 kDa; strains ATCC 43085 and 43144 did not produce any glucosyltransferase. The glucosyltransferase form S. bovis 9809 was purified by Bio-Gel hydroxyapatite chromatography and DEAE-Toyopearl chromatography. The S. bovis 9809 glucosyltransferase was immunologically identical with the other 5 S. bovis glucosyltransferases and not related to mutants streptococcal glucosyltransferases. The specific activity, the optimum pH and the Km value for sucrose were 17.9 U/mg protein, 6.0 and 5.0 mM, respectively. The first 11 N-terminal amino acid residues of the glucosyltransferases were DETSAVTLTRE, and the region was hydrophilic. The glucosyltransferases from S. bovis 9809 and 3317 synthesized from sucrose 1, 6-alpha-D-glucan with 9 and 2 mol%, 1, 3, 6-alpha-branched glucose, respectively.

Amino Acid Sequence↗
PubMed1994

Renaturation and activity staining of glycosidases and glycosyltransferases in gels after sodium dodecyl sulfate-electrophoresis.

Glycosidases and glycosyltransferases were electrophoresed in the presence of sodium dodecyl sulfate (SDS) in a thin-layer gel supported by a glass plate, treated with the nonionic detergent Triton X-100, and specifically stained for the sugar-releasing activity of these enzymes. Staining is based on conversion of monosugars or a sugar phosphate to glucose-6-phosphate by the appropriate intermediary enzymes, reduction of NADP+ to NADPH, and accumulation of reduced Nitroblue Tetrazolium in the gel. Among the enzymes tested, alpha-glucosidase, beta-glucosidase and beta-mannosidase could not be renatured, whereas beta-fructofuranosidase and alpha-mannosidase could be renatured unless heated before electrophoresis. Sucrose phosphorylase, glucosyltransferase and fructosyltransferase, which are single-peptide proteins with no cystine bond, could be renatured even after pretreatment with SDS and/or mercaptoethanol at 100 degrees C for 10 min. However, exclusive heating remarkably decreased the activities of these enzymes. Two-dimensional separation of the five renaturable enzymes was done in a single thin-layer gel, using SDS-electrophoresis in the first dimension and isoelectric focusing in the second dimension.

Electrophoresis, Polyacrylamide Gel↗
PubMed1994

Glucose, fructose, mannose and/or glucose-1-phosphate-releasing activity stains for glycosidases and glycosyltransferases in gels after isoelectric focusing.

beta-Fructofuranosidase, alpha-glucosidase, beta-glucosidase, alpha-mannosidase, beta-mannosidase, sucrose phosphorylase, glucosyltransferase and fructosyltransferase were separated by isoelectric focusing and sensitively detected to be slightly diffuse and insoluble spots in thin-layer gels, supported by a glass plate, by release of monosugars or a sugar phosphate, followed by conversion to glucose-6-phosphate (G6P) and then by reduction of NADP+ to NADPH, terminated by the formation of reduced Nitroblue Tetrazolium (NBT). Approximately 1-10 mU of enzyme was focused and the gel, after washing with a buffer, was partially dried and directly stained by uniformly spreading on the gel surface a staining medium containing sucrose or nitrophenyl glycosides as substrates, intermediary enzymes such as hexokinase, mutase and/or isomerase, NADP+, ATP, Mg+, phenazine methosulfate (PMS) and NBT. Specific staining procedures for each of these activities, on sucrose or on the glycosides as substrates, and staining procedures for multiple activities are described, with the conditions necessary for optimal development.

Fructose↗
PubMed1994

Identification of amino acid residues in Streptococcus mutans glucosyltransferases influencing the structure of the glucan product.

The glucosyltransferases (GTFs) of mutans streptococci are important virulence factors in the sucrose-dependent colonization of tooth surfaces by these organisms. To investigate the structure-function relationship of the GTFs, an approach was initiated to identify amino acid residues of the GTFs which affect the incorporation of glucose residues into the glucan polymer. Conserved amino acid residues were identified in the GTF-S and GTF-I enzymes of the mutans streptococci and were selected for site-directed mutagenesis in the corresponding enzymes from Streptococcus mutans GS5. Conversion of six amino acid residues of the GTF-I enzyme to those present at the corresponding positions in GTF-S, either singly or in multiple combinations, resulted in enzymes synthesizing increased levels of soluble glucans. The enzyme containing six alterations synthesized 73% water-soluble glucan in the absence of acceptor dextran T10, while parental enzyme GTF-I synthesized no such glucan product. Conversely, when residue 589 of the GTF-S enzyme was converted from Thr to either Asp or Glu, the resulting enzyme synthesized primarily water-insoluble glucan in the absence of the acceptor. Therefore, this approach has identified several amino acid positions which influence the nature of the glucan product synthesized by GTFs.

Amino Acid Sequence↗
PubMed1992

Vitamin B12 accelerates re-entrainment of activity rhythms in rats.

The effects of methyl vitamin B12 (5-6 mg/kg, p.o.) on the entrainment of circadian running wheel activity rhythm to a new lighting schedule were measured in rats. After the light-dark (LD) cycle was abruptly reversed, rats given vitamin B12 took less time to entrain their circadian locomotor activity rhythm to the new cycle than did controls. This result indicates that vitamin B12 accelerates the reentrainment of the mammalian circadian activity rhythm following an abrupt change in the environmental LD cycle.

Administration, Oral↗
PubMed1992

Effects of acute and repeated alcohol ingestion on hypothalamic-pituitary-gonadal and hypothalamic-pituitary-adrenal functioning in normal males.

We investigated the effects of acute and repeated alcohol ingestion on plasma levels of hormones associated with the functioning of the hypothalamic-pituitary-gonadal (HPG) and hypothalamic-pituitary-adrenal (HPA) systems in normal males. In the first experiment, 7 normal male subjects were given ethanol (1.3 g/kg) in the form of a 43% alcohol solution of whiskey and water over a 30-min period (from 19:00 h to 19:30 h); blood samples were collected 30 min and immediately before the beginning of alcohol ingestion and then at intervals of 30 min for 180 min. Blood ethanol levels rose sharply and reached their maximum at 60 min, remaining above 1.0 mg/ml until 180 min. Prolactin levels increased, reaching a peak at 60 min, gradually returning to the initial value at 180 min. Decreased testosterone levels were observed only at 30 min. Luteinizing hormone (LH), adrenocorticotrophic hormone (ACTH) and cortisol levels did not show any increases. In the second experiment, 9 normal males were given the same dose of alcohol, but this was given on 7 consecutive evenings and the hormonal changes were examined on the 1st and 7th days, only at 30 and 60 min after alcohol ingestion began (during the period that blood ethanol levels were ascending to their peak). The results on the 1st day reconfirmed the findings in the first experiment and on the 7th day, the last alcohol ingestion produced increases in prolactin levels and decreases in testosterone levels at 30 and 60 min, but did not change other hormone levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone↗
PubMed1992

Effects of thienodiazepine derivatives, etizolam and clotiazepam on the appearance of Fm theta.

The effects of new thienodiazepine anxiolytics, etizolam and clotiazepam, on the appearance of frontal midline theta activity (Fm theta) were studied by the double-blind crossover method. The results were as follows; 1) Both clotiazepam and placebo tended to increase the Fm theta appearance in all subjects, but etizolam showed no such tendency. 2) Clotiazepam significantly increased the Fm theta appearance as compared with placebo in subjects with a high neurotic tendency (N-scale of MPI above 19). 3) Clotiazepam tended to increase the Fm theta appearance as compared with placebo and etizolam in subjects with a high anxiety level (MAS score above 14). 4) Apparently more subjects complained of drowsiness after the administration of etizolam than after clotiazepam or placebo.

Adult↗
PubMed1991

Primary and secondary depression in alcoholism--clinical features and family history.

One hundred thirty-six alcoholic inpatients were studied with regard to the association between alcoholism and depression. They fell into three groups: primary depression, secondary depression and non-depressive alcoholism. The main results obtained are as follows: (1) Depression was found in 46 patients (33.8%) in the present or past history; 13 were regarded as primary depression and 33 as secondary depression. (2) The clinical symptoms and the background of the primary group resembled those of endogenous depression. (3) The clinical features, including the duration of depressive episodes in the secondary depression group differed from those in the primary group. (4) No statistically significant differences were noted as to the rates of family-history of alcoholism and depression among the three groups. Clinical and biological differences between the primary and secondary groups were discussed on the basis of these results.

Adult↗
PubMed1990

Platelet monoamine oxidase activity and personality traits in alcoholics and methamphetamine dependents.

Personality traits of alcoholics and methamphetamine dependents were examined by Karolinska scales of personality (KSP). The two groups differed with respect to Aggression, but, they were basically same in Impulsiveness. The biological marker platelet monoamine oxidase (MAO) activity showed a significant difference between the two groups. The low platelet MAO activity in alcoholics may suggest a certain biological basis to be involved in the etiology of dependence whereas the higher platelet MAO activity observed in methamphetamine dependents may reflect the prolonged use of methamphetamine and/or treatment with neuroleptics, or some biological basis.

Adult↗
PubMed1989

Purification and characterization of extracellular glucosyltransferase synthesizing water-insoluble glucan from Streptococcus rattus.

An extracellular glucosyltransferase synthesizing water-insoluble glucan (GTF-I) was purified from the culture supernatant of Streptococcus rattus strain BHT (mutans serotype b) by hydroxylapatite chromatography, DEAE-Toyopearl chromatography and preparative isoelectric focusing. The Mr of GTF-I was 155,000 by SDS-PAGE and the isoelectric point was pH 4.9. The specific activity, the optimum pH and the Km value for sucrose were 10.0 i.u. (mg protein)-1, 6.5 and 2.4 mM, respectively. The enzyme synthesized a water-insoluble glucan consisting of 69.4 mol% 1,3-alpha-linked glucose, 23.6 mol% 1,6-alpha-linked glucose, 2.6 mol% 1,3,6-alpha-branched glucose and 4.4 mol% non-reducing terminal glucose, and also a small amount (3% of the total glucan) of soluble glucan with 82.4 mol% 1,6-alpha-linked glucose. The Mr and pI values of purified GTF-I were identical with those of the enzyme in the culture supernatant.

Chromatography, Gel↗