Biomedical subjects

L Tao

Publications and source records attributed to L Tao.

At least 163 records · Page 9Linked to original sources

PubMed1993

Cloning and expression of the multiple sugar metabolism (msm) operon of Streptococcus mutans in heterologous streptococcal hosts.

The multiple sugar metabolism (msm) operon of Streptococcus mutans is responsible for the uptake and metabolism of a variety of sugars. In order to further characterize the substrate specificities of the transport system, a 12-kb region of DNA containing the entire msm operon was cloned, via a novel two-step integration strategy, into the chromosomes of two heterologous streptococcal strains, Streptococcus gordonii Challis and Streptococcus anginosus Is57, as well as the chromosome of a natural isolate of S. mutans with a deletion of the msm region. These strains are unable to transport or ferment melibiose, raffinose, or isomaltosaccharides, but the newly constructed recombinants gained the ability to ferment all of these sugars. The S. gordonii Challis construct containing msm was shown to transport radiolabelled melibiose, raffinose, isomaltotriose, and isomaltotetraose, and the transport function was also subjected to induction by raffinose, an inducer of the msm operon in S. mutans. The results confirm the role of the msm operon in the transport and metabolism of melibiose, raffinose, and isomaltosaccharides.

Biological Transport↗

PubMed1993

MsmE, a lipoprotein involved in sugar transport in Streptococcus mutans.

Metabolic labelling by [14C]palmitic acid showed that growth of Streptococcus mutans LT11 in raffinose, an inducer of the msm operon, resulted in increased production of a 45-kDa lipoprotein corresponding to MsmE, which is believed to be a sugar-binding protein. MsmE was also labelled when an msmE clone was expressed in Escherichia coli. The presence of a lipid anchor on MsmE provides a likely explanation of how the sugar-binding protein component of the msm binding protein-dependent multiple sugar transport system is retained at the cell surface.

Amino Acid Sequence↗

PubMed1993

Long-term effects on pancreatic function of feeding a HC formula to rats during the preweaning period.

We have investigated pancreatic changes associated with hyperinsulinemia and an insulin secretory response to an oral glucose load in a new rat model for obesity. Male Sprague-Dawley pups were reared on a high-carbohydrate (HC) or high-fat (HF) formula by gastrostomy during the suckling period and were weaned onto a stock diet. These animals remained either nutritionally unchallenged or challenged with a high-sucrose diet during the postweaning period. The HC formula-fed animals showed increased insulin concentrations in the plasma and pancreas and also showed impaired insulin secretory response compared with mother-fed control or HF animals in adult life. Immunocytochemical and morphometric studies revealed that hyperinsulinemia in the HC animals during the preweaning period and also in adult life was associated with hypertrophy of beta-cells in the pancreas. The results show that consumption of a HC formula during the suckling period influences pancreatic islet morphology resulting in hyperinsulinemia which eventually leads to the development of obesity later in adult life.

Animals↗

PubMed1993

Transformation efficiency of EMS-induced mutants of Streptococcus mutans of altered cell shape.

Some Streptococcus mutans strains change shape from bacillary to coccal or ellipsoid form in response to the ratio of bicarbonate to potassium or of borate to potassium in growth media. So that insight into determinants of shape of these streptococci could be gained, and future genetic studies facilitated, the shapes of a series of transformable and nontransformable strains of S. mutans were studied and attempts made to isolate a mutant of augmented transformability. Several strains were mutagenized by ethylmethane sulfonate and mutants with altered colonial and cellular morphologies isolated. Cell shapes were studied by Gram stain and Nomarski interference microscopy, and by scanning and transmission electron microscopy. Diverse shape-altered mutants were isolated from seven transformable and two nontransformable strains of S. mutans. Among these, length-to-width ratios ranged from > 10 to about 0.25. Regulation of timing of cell division, septum formation, or septum completion events may have been altered in these mutants. While most mutants substantially or completely lost transformability, mutant LT11 had transformation efficiency of 1.3 x 10(-4) to 2.3 x 10(-3), more than two to three orders of magnitude greater than its parental UA159 and the well-known transformable strain GS5(HK), respectively. There was no evidence of production of competence factor by LT11. Competence of LT11 was maintained for at least six months upon storage at -70 degrees C, facilitating its use for genetic studies. While the morphologies of several shape-altered mutants were no longer responsive to changes of the bicarbonate/potassium, unlike those of their parentals, the morphology of LT11 persisted in its response to this condition.(ABSTRACT TRUNCATED AT 250 WORDS)

Ethyl Methanesulfonate↗

PubMed1993

Transport of sugars, including sucrose, by the msm transport system of Streptococcus mutans.

The range of substrates transported by the sugar-binding protein-dependent msm (multiple sugar metabolism) system of S. mutans was investigated. By determining the ability of unlabeled sugar to compete with radiolabeled melibiose transport, we have demonstrated that the transported sugars included a number of carbohydrates structurally related to raffinose. A model accommodating these results has been devised which accounts for the sugars transported by the msm transport system. Competition with radiolabeled melibiose transport indicated sucrose to be an msm substrate. This was confirmed by examination of uptake of radiolabeled sucrose in scrAB mutants lacking the sucrose-specific phosphotransferase system.

Biological Transport↗

PubMed1993

Sterilization of human teeth: its effect on permeability and bond strength.

Extracted human teeth were randomly divided into three groups. Group 1 teeth were untreated and served as controls. Group 2 teeth were sterilized in a steam autoclave. Group 3 teeth were sterilized by exposure to ethylene oxide gas. Crown segments were then prepared from these teeth by resection of the roots at the CEJ and the occlusal enamel to expose a flat occlusal plane of dentin. The permeability of the dentin was determined to be similar in all three groups. The shear bond strength of Prisma Universal Bond 3 and C&B Metabond was also similar in all groups indicating that sterilization of teeth does not alter dentin permeability or bond strength.

Boron Compounds↗

PubMed1993

[Long-term follow up of dacryorhinostomy in 14 cases with fibroendoscopy].

The orifice of anastomosis and the lower aperture of the nasolacrimal duct in 14 cases (16 eyes) after dacryorhinostomy were followed up with the fibroendoscope. It was found that the size of the office and positioning of the bony window were key factors for a successful operation. Postsurgical management was advised to continue for at least 3 months or preferably 6 months. Deviation of the nasal septum toward the affected side was not regarded as contraindication for the operation, and the authors recommended fibroendoscopy in the diagnosis and postsurgical examination of lacrimal passage diseases.

Adolescent↗

PubMed1993

Effects of oxalate on dentin bonding.

The purpose of this study was to determine how much topical treatment of dentin with potassium oxalate would interfere with subsequently placed dentin bonding resins. The results indicated that oxalate lowered the bond strengths of Superbond, All Bond and Scotchbond 2, but that normal bond strengths could be obtained for Superbond and All Bond after pumicing the oxalate-treated dentin.

Acid Etching, Dental↗

PubMed1992

Induction of mucosal and systemic immunity to a recombinant simian immunodeficiency viral protein.

Heterosexual transmission through the cervico-vaginal mucosa is the principal route of human immunodeficiency virus (HIV) infection in Africa and is increasing in the United States and Europe. Vaginal immunization with simian immunodeficiency virus (SIV) had not yet been studied in nonhuman primates. Immune responses in macaques were investigated by stimulation of the genital and gut-associated lymphoid tissue with a recombinant, particulate SIV antigen. Vaginal, followed by oral, administration of the vaccine elicited three types of immunity: (i) gag protein p27-specific, secretory immunoglobulin A (IgA) and immunoglobulin G (IgG) in the vaginal fluid, (ii) specific CD4+ T cell proliferation and helper function in B cell p27-specific IgA synthesis in the genital lymph nodes, and (iii) specific serum IgA and IgG, with CD4+ T cell proliferative and helper functions in the circulating blood.

Administration, Oral↗

PubMed1992

Novel streptococcal-integration shuttle vectors for gene cloning and inactivation.

Seven new streptococcal integration shuttle vectors have been constructed which contain different antibiotic-resistance-encoding genes capable of expression in both Streptococcus sp. and Escherichia coli. These plasmids can replicate in E. coli, but not in streptococci because of the absence of a streptococcal origin of replication. The size, antibiotic resistance, and number of unique restriction sites available for cloning for each plasmid are as follows: pSF141 (7.6 kb, CmR and KmR, 7 sites), pSF143 (5.7 kb, TcR, 6 sites), pSF148 (7.3 kb, CmR and SpR, 7 sites), pDL285 (3.4 kb, KmR, 3 sites), pDL286 (3.1 kb, SpR, 4 sites), pSF151 (3.5 kb, KmR, 10 sites), pSF152 (3.2 kb, SpR, 9 sites). If these plasmids carry a fragment of streptococcal DNA they can specifically integrate into the chromosome via Campbell-like, homologous recombination. Therefore, they should be useful for gene inactivation, cloning, chromosomal walking, or linkage analysis in streptococci. The availability of these integration plasmids resistant to different antibiotics, along with the previously described plasmid, pVA891 (ErR), should also allow the construction of mutants possessing multiple insertionally inactivated genes useful for a variety of genetic studies.

Cloning, Molecular↗

PubMed1992

Rapid and sensitive anion-exchange high-performance liquid chromatographic determination of radiolabeled inositol phosphates and inositol trisphosphate isomers in cellular systems.

A rapid and sensitive high-performance liquid chromatographic method for the determination of multiple inositol phosphates and inositol trisphosphate isomers was developed. The separation of inositol phosphates was optimized by controlling the ionic strength with stepped gradient programs and the pH of mobile phase. Six inositol phosphates were determined within 22 min or the six compounds plus an inositol trisphosphate isomer within 24 min using a single anion-exchange column containing the quaternary ammonium functional group. This technique was successfully applied to the determination of inositol phosphatide turnover by AlF4-stimulation in a small amount (5.10(5)-1.10(6) cells) of cultured retinal capillary pericytes. Because of its efficiency, accuracy and applicability to the separation of inositol phosphates from biological samples, this method may be useful in signal transduction studies in cellular systems.

Animals↗

PubMed1992

A binding protein-dependent transport system in Streptococcus mutans responsible for multiple sugar metabolism.

An 11-kilobase gene region of Streptococcus mutans has been identified which contains eight contiguous genes involved with the uptake and metabolism of multiple sugars (the msm system). Sequence analysis of this region indicates that several of these genes specify proteins with strong homology to components of periplasmic binding protein-dependent transport systems of Gram-negative bacteria. Additionally, this operon is controlled by a regulatory gene (msmR) that acts as a positive effector. The proteins specified by the structural genes of the msm operon include alpha-galactosidase (aga), a "periplasmic-like" sugar-binding protein (msmE), two membrane proteins (msmF, msmG), sucrose phosphorylase (gtfA), an ATP-binding protein (msmK), and dextran glucosidase (dexB). Insertional inactivation of each of these genes along with uptake data indicate that this system is responsible for the uptake of melibiose, raffinose, and isomaltotriose and the metabolism of melibiose, sucrose, and isomaltosaccharides.

Amino Acid Sequence↗

PubMed1992

Relationship between bond strength and microleakage measured in the same Class I restorations.

Microleakage measurements were made by use of a pressurized fluid method in Class I restorations prepared in extracted human teeth just prior to measuring the tensile bond strengths of the same restorations. The restorative materials included dentin bonding systems that are applied to smear layers as well as those which remove the smear layer. A light-cured glass-ionomer cement was also included. The results demonstrated that there was an inverse relationship between dentin bond strength and microleakage in some materials and that the bond strengths made to three-dimensional Class I cavities were much lower than those made to flat dentin surfaces. Measurement of microleakage by fluid filtration had no apparent effect on bond strength.

Adhesives↗

PubMed1992

Comparison and characterization of retinal pericytes and retinal pigment epithelial cells on subcellular IP3-sensitive Ca2+ pools.

A comparative study of inositol 1,4,5-trisphosphate (IP3)-induced Ca2+ mobilization in bovine retinal capillary pericytes (BRCP) and bovine retinal pigment epithelial cells (BRPE) was carried out. Both cells were permeabilized with saponin. The two cell types had similar basal levels of [Ca2+]i (130 nM for BRCP, 132 nM for BRPE) and responded to IP3 in a dose-dependent manner. However, when stimulated by various concentrations of IP3 (1-10 microM), the increase in [Ca2+]i of BRCP was always two- to threefold higher than that in BRPE. Subcellular-fractionation studies showed that a single population of IP3 binding site with a high affinity and high specificity of IP3 mainly localized to plasma membrane in these two cell types. Although the dissociation constant of specific [32P]-IP3 binding sites (Kd 1.9-2.8 nM) was similar, the profile of maximal binding capacity (Bmax) of each fraction was markedly different. In comparison, plasma membrane fractions of BRCP were with Bmax of 165 fmol/mg protein versus 90 fmol/mg protein for BRPE membranes. The ATP-dependent Ca2+ uptake and IP3-dependent Ca2+ release were observed in the both plasma membrane fractions. With quantitative correlation, the membrane fraction (2 mg) of BRCP released 0.2 nmol Ca2+ whereas BRPE only released 0.07 nmol Ca2+ with the same dose of IP3 (5 microM). The selectively higher density of IP3 binding sites in coupling to the larger Ca(2+)-release in the membrane of BRCP suggests that the quantity of Ca2+ mobilized is determined by the spatially preferential distribution of membrane-associated IP3 binding sites. These findings may provide an explanation for the differences observed between BRCP and BRPE in IP3-induced DNA replication.

Binding Sites↗

PubMed1992

Differential binding of (+) and (-) gossypol to plasma protein and their entry into rat testis.

Concentrations of (+) and (-) gossypol were measured by high performance liquid chromatography after they were incubated with plasma proteins in vitro. The concentration of (-) gossypol decreased more than the concentration of (+) gossypol. A similar decrease in free gossypol concentrations in the blood plasma of rats was observed after intravenous infusion of gossypol enantiomers. The concentration of (-) gossypol was also found to be lower than the concentration of (+) gossypol at the blood-testis barrier. The biological effect of (-) gossypol probably results from its stereospecific binding to extra- and intracellular proteins in vivo and inhibition of the biological activity of some proteins.

Animals↗

PubMed1991

Fluid and protein flux across the pulpodentine complex of the dog in vivo.

Dentinal fluid volume and protein concentrations were measured from cavities prepared in the dentine of dog molars. Fluid was collected under spontaneous conditions, during the application of negative pressures, and during a recovery period. The data allowed the calculation of dentine permeability as hydraulic conductances, fluid flux, protein flux, reflection coefficients and pulpal tissue pressures. In general, the protein concentration of dentinal fluid was about one-fifth that of plasma during spontaneous conditions and it fell when fluid flux was increased by the application of external negative pressures. The collection and analysis of dentinal fluid looks promising as a non-invasive method of assessing the state of the underlying pulp.

Animals↗

PubMed1991

Reduced absolute rate of myo-inositol biosynthesis of cultured bovine retinal capillary pericytes in high glucose.

De novo biosynthesis of myo-inositol (MI) by permeabilized cultured bovine retinal capillary pericytes (BRCP) and feline retinal pigment epithelial cells (FRPE), grown in different concentrations of glucose, were studied. After incubation with a physiological concentration of [14C]glucose 6-phosphate (G6P), the radioactive G6P derivatives were quantitated by a single HPLC column. Based on the determined specific activity of [14C]G6P, activities of inositol 1-phosphate synthase (MI synthase) were calculated. The activity of MI synthase was reduced 48% by growing BRCP in a high-glucose medium (20 mM) in comparison with that in the normal medium (glucose 5 mM). In contrast, the de novo MI biosynthesis by FRPE was not changed with increasing concentrations of glucose in the medium. As compared with MI uptake previously studied, the synthesized MI contributes a substantial proportion of cellular MI pool in BRCP. Therefore, in BRCP growing in high glucose the reduced MI biosynthesis aggravates the low MI content resulting from the inhibited MI uptake, and thus leads to altered inositol phospholipid metabolism.

Animals↗

PubMed1991

Shear bond strength of Scotchbond in vivo.

The shear bond strengths of Scotchbond, HEMA/Scotchbond, and Scotchbond 2 were measured in vivo in dog canine and molar enamel and dentin. Dentin bond strengths were compared in superficial, middle, and deep dentin. The acid-etched enamel bond strengths of the three bonding systems ranged from 10 to 11 MPa and were not statistically different. Scotchbond/Silux bonds to superficial and middle cuspid and molar dentin were 3 MPa and were not statistically different. HEMA-treated dentin did not consistently improve Scotchbond strengths to either tooth type at any dentin depth. Deep dentin from either tooth type yielded significantly lower bond strengths. Scotchbond 2/Silux shear bond strengths were significantly higher (6-8 MPa) in superficial and middle cuspid dentin but were not different from Scotchbond bonds made to deep cuspid dentin or to any depth of molar dentin. The observation that molar bond strengths are lower than those made to cuspid dentin indicates that there are important substrate differences between teeth as well as within dentin as a function of depth. The dog model may be useful for the screening of new dentin bonding systems prior to clinical trials.

Animals↗