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Biomedical subjects

E C Reynolds

Publications and source records attributed to E C Reynolds.

At least 73 records · Page 4Linked to original sources
PubMed1992

The effect of acetylsalicylic acid on orthodontic tooth movement in the guinea pig.

This study examined the influence of acetylsalicylic acid an inhibitor of prostaglandin synthesis, on orthodontic tooth movement induced with light spring forces in the guinea pig. The animal model was shown to permit reliable and accurate recording of tooth movement up to 28 days. Tooth movement was found to be highly correlated with spring forces, indicating that the model provided a sensitive test of the effect of aspirin on tooth movement. Aspirin was administered orally at the rate of 65 mg/kg per day in three divided doses and was found to effectively inhibit prostaglandin synthesis at the level of the bronchioles. However, aspirin did not appear to significantly affect tooth movement. Thus prostaglandins may not be the only mediators of the bone resorption associated with tooth movement induced by light orthodontic forces under these experimental conditions.

Analysis of Variance↗
PubMed1992

Efficient solution-phase synthesis of multiple O-phosphoseryl-containing peptides related to casein and statherin.

The multiple Ser(P)-containing peptides, H-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe.TFA, H-Asp-Ser(P)-Ser(P)-Glu-Glu-NHMe.TFA and H-Glu-Ser(P)-Ser(P)-Glu-Glu-NHMe.TFA were prepared by the use of Boc-Ser(PO3Ph2)-OH in the Boc mode of solution phase peptide synthesis followed by platinum-mediated hydrogenolytic de-protection of the Ser(PO3Ph2)-containing peptides. The protected peptides were assembled using the mixed anhydride coupling methods with 40% TFA/CH2Cl2 used for removal of the Boc group from intermediate Boc-protected peptides.

Amino Acid Sequence↗
PubMed1992

pH regulation by Streptococcus mutans.

The intracellular pH (pHi) optimum for glycolysis in Streptococcus mutans Ingbritt was determined to be 7.0 by use of the ionophore gramicidin for manipulation of pHi. Glycolytic activity decreased to zero as the pHi was lowered from 7.0 to 5.0. In contrast, glycolysis had an extracellular pH (pHo) optimum of 6.0 with a much broader profile. The relative insensitivity of glycolysis to the lowering of pHo was attributed to the ability of S. mutans to maintain a transmembrane pH gradient (delta pH, inside more alkaline) at low pHo. At a pHo of 5.0, glycolyzing cells of S. mutans maintained a delta pH of 1.37 +/- 0.09 units. The maintenance of this delta pH was dependent on the concentration of potassium ions in the extracellular medium. Potassium was rapidly taken up by glycolyzing cells of S. mutans at a rate of 70 nmol/mg dry weight/min. This uptake was dependent on the presence of both ATP and a proton motive-force (delta p). The addition of N-N'-dicyclohexylcarbodiimide (DCCD) to glycolyzing cells of S. mutans caused a partial collapse of the delta pH. Growth of S. mutants at pHo 5.5 in continuous culture resulted in the maintenance of a delta pH larger than that produced by cells grown at pH 7.0. These results suggest the presence of a proton-translocating F1Fo-ATPase in S. mutans whose activity is regulated by the intracellular pH and transmembrane electrical potential (delta psi). The production of an artificial delta p of 124 mV across the cell membrane of S. mutans did not result in proton movement through the F1Fo-ATPase coupled to ATP synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Acids↗
PubMed1991

A synthetic beta-casein phosphopeptide and analogues as model substrates for casein kinase-1, a ubiquitous, phosphate directed protein kinase.

The phosphopeptide Ser (P)-Ser(P)-Ser-(P)-Glu-Glu-Ser22-Ile-Thr, reproducing the 17-24 segment of beta-casein A2 including the seryl residue (Ser-22) which is targeted by casein kinase-1 was synthesized and used as model substrate for this enzyme. Its phosphorylation efficiency is actually higher than that of intact beta-casein (similar Vmax and 14 microM Km). Conversely the fully dephosphorylated peptide SSSEESIT is not affected by CK-1 to any detectable extent and its glutamyl derivative EEEEESIT displays a more than 50-fold higher Km and a 5-fold lower Vmax as compared to the parent phosphopeptide. The relevance of the individual phosphoseryl residues has been assessed by comparing the phosphorylation efficiencies of the phosphopeptides EESpEESIT, ESpEEESIT and SpEEEESIT: while the first is a substrate almost as good as the tris Ser (P)-peptide (Km = 62 microM), and the third one is almost as poor as EEEEESIT (Km = 1.55 mM), ESpEEESIT displays a intermediate efficiency (Km = 277 microM). These data in conjunction with the finding that the phosphopentapeptide Ser(P)-Ser(P)-Ser(P)-Ser-Ser(P), but neither Ser(P)-Ser(P)-Ser-Ser(P) nor Ser-Ser(P)-Ser(P)-Glu-Glu and Ser-Ala-Ala-Ser(P)-Ser(P), is readily phosphorylated by CK-1, support the concept that CK-1 is a phosphate directed protein kinase recognizing the Ser(P)-X-X-Ser-X and, less efficiently, the Ser(P)-X-X-X-Ser-X motifs.

Adenosine Triphosphate↗
PubMed1991

Phosphotyrosine as a specificity determinant for casein kinase-2, a growth related Ser/Thr-specific protein kinase.

The motif Ser-Ser-Ser-Glu-Glu is readily phosphorylated by casein kinase-2 (CK-2), a growth-related protein kinase whose consensus sequence is Ser(Thr)-Xaa-Xaa-Glu(Asp) [(1990) Biochim. Biophys. Acta 1054, 267-283]. Here we show that phosphotyrosine can replace carboxylic acids as specificity determinant for CK-2 phosphorylation, the phosphotyrosyl peptide Ser-Ser-Ser-TyrP-TyrP actually being a substrate more efficient than Ser-Ser-Ser-Glu-Glu itself both in terms of Km (0.69 vs 2.43 mM) and Vmax. Prior dephosphorylation of phosphotyrosine entirely prevents the subsequent phosphorylation of serine by CK-2. While Ser-Ser-Ser-TyrP-TyrP is a better substrate than Ser-Ser-Ser-SerP-SerP, which in turn is better than Ser-Ser-Ser-Glu-Glu, Ser-Ser-Ser-ThrP-ThrP is a less efficient substrate than Ser-Ser-Ser-Glu-Glu. Thus the order of efficiency of phosphoamino acids as specificity determinants for CK-2 appears to be TyrP greater than SerP much greater than ThrP.

Amino Acid Sequence↗
PubMed1991

Fmoc/solid-phase synthesis of Tyr(P)-containing peptides through t-butyl phosphate protection.

The synthesis of Tyr(P)-containing peptides by the use of Fmoc-Tyr(PO3Me2)-OH in Fmoc/solid phase synthesis is complicated since, firstly, piperidine causes cleavage of the methyl group from the -Tyr(PO3Me2)-residue during peptide synthesis and, secondly, harsh conditions are needed for its final cleavage. A very simple method for the synthesis of Tyr(P)-containing peptides using t-butyl phosphate protection is described. The protected phosphotyrosine derivative, Fmoc-Tyr(PO3tBu2)-OH was prepared in high yield from Fmoc-Tyr-OH by a one-step procedure which employed di-t-butyl N,N-diethyl-phosphoramidite as the phosphorylation reagent. The use of this derivative in Fmoc/solid phase peptide synthesis is demonstrated by the preparation of the Tyr(P)-containing peptides, Ala-Glu-Tyr(P)-Ser-Ala and Ser-Ser-Ser-Tyr(P)-Tyr(P).

Amino Acid Sequence↗
PubMed1990

The utilization of casein and amino acids by Streptococcus sanguis P4A7 in continuous culture.

Streptococcus sanguis P4A7 was grown in glucose limited conditions in continuous culture at pH 7.0 in a chemically defined medium containing either free amino acids or casein as the organic nitrogen source. Apart from aspartate and threonine, which were poorly utilized at the higher dilution rates, all amino acids in the free-amino-acid medium were utilized to various extents. At the higher dilution rates, aspartate actually increased in concentration, probably due to deamidation of asparagine. The amino acid most utilized at all dilution rates was arginine, with up to 99% of the amino acid being consumed. Both casein and its alpha s1-casein fraction supported growth at a level only slightly lower than that obtained with the free-amino-acid medium, provided that either cysteine or thioglycollate was present. With the exception of tyrosine, nearly all of the amino acyl residues of alpha s1-casein were utilized to some degree. In general, the higher the concentration of each amino acid in the medium (whether free or as part of alpha s1-casein) the higher the extent of utilization by S. sanguis P4A7. Only 50% of the arginyl residues (0.16 mM) of alpha s1-casein were utilized compared with 99% of free arginine (1.5 mM) under similar conditions, suggesting that only 50% of the alpha s1-casein arginine was accessible to the organism.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1990

Characterization of transmembrane movement of glucose and glucose analogs in Streptococcus mutants Ingbritt.

The transmembrane movement of radiolabeled, nonmetabolizable glucose analogs in Streptococcus mutants Ingbritt was studied under conditions of differing transmembrane electrochemical potentials (delta psi) and pH gradients (delta pH). The delta pH and delta psi were determined from the transmembrane equilibration of radiolabeled benzoate and tetraphenylphosphonium ions, respectively. Growth conditions of S. mutants Ingbritt were chosen so that the cells had a low apparent phosphoenolpyruvate (PEP)-dependent glucose:phosphotransferase activity. Cells energized under different conditions produced transmembrane proton potentials ranging from -49 to -103 mV but did not accumulate 6-deoxyglucose intracellularly. An artificial transmembrane proton potential was generated in deenergized cells by creating a delta psi with a valinomycin-induced K+ diffusion potential and a delta pH by rapid acidification of the medium. Artificial transmembrane proton potentials up to -83 mV, although producing proton influx, could not accumulate 6-deoxyglucose in deenergized cells or 2-deoxyglucose or thiomethylgalactoside in deenergized, PEP-depleted cells. The transmembrane diffusion of glucose in PEP-depleted, KF-treated cells did not exhibit saturation kinetics or competitive inhibition by 6-deoxyglucose or 2-deoxyglucose, indicating that diffusion was not facilitated by a membrane carrier. As proton-linked membrane carriers have been shown to facilitate diffusion in the absence of a transmembrane proton potential, the results therefore are not consistent with a proton-linked glucose carrier in S. mutans Ingbritt. This together with the lack of proton-linked transport of the glucose analogs suggests that glucose transmembrane movement in S. mutans Ingbritt is not linked to the transmembrane proton potential.

Acetone↗
PubMed1989

Protein dissimilation by human salivary-sediment bacteria.

Proteins of known composition and structural characteristics were incubated (1.0 mg/mL) with re-suspended salivary sediment (2.5% v/v) in a lactate-salt medium with an initial pH of 5.2 for two hr at 37 degrees C. Hydrolysis of the proteins was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Hydrogen ion, amines, and ammonia were measured by use of a combined pH electrode, high-performance liquid chromatography, and glutamate dehydrogenase, respectively. Of the proteins studied, the caseins alpha s1, beta, and kappa and the histones H1 and H3 were extensively hydrolyzed by the salivary-sediment bacteria. The hydrolysis of these proteins was attributed to their relative lack of tertiary (folded) structure. The only amine detected was the polyamine putrescine arising from the catabolism of arginine following the hydrolysis of the arginine-rich histone H3. None of the other proteins extensively hydrolyzed by salivary sediment, although containing arginyl and lysyl residues, served as substrates for putrescine or cadaverine production. Pre-hydrolysis of the arginine-rich histone H3 and poly-L-arginine with trypsin resulted in a marked increase in putrescine produced, suggesting that the salivary-sediment proteolytic activity was not "trypsin-like". Incubation of salivary-sediment bacteria with the caseins and the histone H3 resulted in an increase in ammonium ion concentration and an associated decrease in hydrogen ion concentration. The increase in ammonium ion concentration not attributed to arginine hydrolysis was correlated with the content of glutaminyl plus asparaginyl residues of the proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Colonization of rat molar teeth by mutans streptococci with different salivary agglutination characteristics.

The oral implantation of salivary agglutination-positive and -negative mutans streptococci was studied using streptomycin resistant (StrR) organisms. StrR Streptococcus mutans strains Ingbritt and NCTC 10449 are agglutinated by rat saliva and the StrR strains Streptococcus sobrinus 6715-13 and Strep. mutans GS5 are not. Four groups of Sprague-Dawley rats were inoculated orally with each organism (one per group) and fed a sucrose diet. A further two groups of animals were similarly inoculated with either the agglutination-positive Strep. mutans Ingbritt or the agglutination-negative Strep. sobrinus 6715-13 and fed a glucose diet. StrR streptococci were recovered from smooth-surface dental plaque of all animals on the sucrose diet with no significant difference in the recovery of agglutination-positive Strep. mutans strains Ingbritt and NCTC 10449 and agglutination-negative Strep. mutans GS5. However, the recovery of agglutination-negative Strep. sobrinus 6715-13 from smooth-surface plaque of animals on either the sucrose or the glucose diets was significantly lower than that of the other strains. Agglutination-positive Strep. mutans Ingbritt colonized smooth enamel surfaces of animals on the sucrose and the glucose diets in numbers that were not significantly different. However, the colonization of such surfaces by agglutination-negative Strep. sobrinus 6715-13 was significantly enhanced by the sucrose diet. Agglutination-positive and -negative StrR mutans streptococci were recovered from fissure plaque of all inoculated sucrose-fed animals in numbers that were not significantly different. Successful colonization of smooth enamel surfaces by the StrR streptococci resulted in increased smooth-surface caries.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination↗
PubMed1987

Characterization of a rat salivary sialoglycoprotein complex which agglutinates Streptococcus mutans.

Rat saliva agglutinated Streptococcus mutans Ingbritt and NCTC 10449 and Streptococcus sanguis NCTC 7864 but not S. mutans NCTC 10921, GS 5, or LM 7, Streptococcus sobrinus 6715-13 or OMZ 65, or Streptococcus cricetus HS 6, as measured turbidometrically. The specificity of agglutination by rat saliva was the same as that by human saliva. Agglutination was associated with a mucin complex (rat salivary agglutinin complex [rS-A]) of sulfated sialoglycoproteins, with a trace of associated lipid and an apparent Mr of 1.6 X 10(6), isolated by gel-filtration Fast Protein Liquid Chromatography. The complex was dissociated in a high-ionic-strength buffer containing 6 M urea and then fractionated by gel filtration and anion-exchange Fast Protein Liquid Chromatography into four sulfated sialoglycoprotein components, designated rS-A-1Q1, rS-A-1Q2, rS-A-1Q3, and rS-A-2, with rS-A-1Q2 being polydisperse through differential glycosylation of the polypeptide backbone. The dissociation destroyed agglutination activity. The polypeptide backbones contained up to 42% serine plus threonine and up to 40% glycine plus alanine plus proline plus valine. The carbohydrate moiety of the rS-A sialoglycoproteins consisted of N-acetylgalactosamine, sialate, galactose, fucose, N-acetylglucosamine, and small amounts of mannose, with the predominant sugar being N-acetylgalactosamine. Agglutination was inhibited by 1 mM EDTA but was restored by 1.5 mM CaCl2. Agglutination was also inhibited by 5 mM CaCl2; nonimmune sera; cationic polymers; and wheat germ, lentil, soybean, and peanut lectins. However, agglutination was not affected by lipoteichoic acid, various anionic proteins, or various sugars. Neuraminidase treatment of rS-A did not affect activity, but tryptic digestion of S. mutans did prevent agglutination. The results are consistent with calcium bridging the negative groups within the rS-A complex and allowing the approach of rS-A to the bacterial cell surface to effect a specific conformational attachment.

Agglutinins↗
PubMed1987

The prevention of sub-surface demineralization of bovine enamel and change in plaque composition by casein in an intra-oral model.

The ability of bovine milk phosphoprotein (casein) to be incorporated into plaque, prevent enamel sub-surface demineralization, and affect bacterial composition was determined using a modified intra-oral caries model. The intra-oral model consisted of a removable appliance containing a left and right pair of bovine enamel slabs placed to simulate an approximal area. Supragingival plaque was collected and impacted into the left and right inter-enamel spaces. The left side of the appliance was exposed to various sugar and salt solutions, while the right side was exposed to sugar and casein solutions. Sodium caseinate, the major fraction alpha s1-casein, and a tryptic digest of alpha s1-casein (TD-casein) were studied. Sodium caseinate at a level of 2% w/v in a 3% sucrose-3% glucose-salt solution (pH 7.0) prevented sub-surface enamel demineralization over a ten-day period as shown by microradiography and microhardness. Two exposures of a 2% w/v sodium caseinate, alpha s1-casein, or TD-casein solution (pH 7.0) per day prevented sub-surface enamel demineralization caused by six exposures of a 3% sucrose-3% glucose-salt solution per day over a ten-day period. Intact alpha s1-casein and tryptic peptides were shown immunochemically to be incorporated into the inter-enamel plaque. The incorporation of casein and its breakdown in plaque did not produce a significant change in the amount or composition of plaque bacteria. The ability of casein and tryptic peptides to prevent enamel demineralization was related to their incorporation into plaque, thereby increasing plaque calcium phosphate and acid-buffering capacity by the phosphoseryl, histidyl, glutamyl, and aspartyl residues and indirectly through catabolism by plaque bacteria.

Adolescent↗
PubMed1984

Effect of casein and whey-protein solutions on caries experience and feeding patterns of the rat.

Casein (bovine milk phosphoprotein) at 2 per cent (w/v) in drinking water reduced the extent of fissure and smooth-surface caries of male Sprague-Dawley rats consuming a solid cariogenic diet. Whey protein (the non-phosphorylated protein group of bovine milk) also at 2 per cent (w/v) in the drinking water produced a smaller reduction and only of fissure caries. There was no significant difference in salivary-gland function (as determined by protein concentration), or in the amount or frequency of cariogenic diet consumed. The finding that a 2 per cent solution of whey protein reduced the extent of fissure caries in animals consuming a solid diet containing 26 per cent whey protein suggests that the anticariogenic action is mediated by the protein being in solution. These results suggest a topical anticariogenic action for dietary protein.

Animals↗
PubMed1983

Effect of adsorbed protein on hydroxyapatite zeta potential and Streptococcus mutans adherence.

The adherence of Streptococcus mutans PK1 to hydroxyapatite disks pretreated with various acidic and basic proteins in imidazole buffer was studied. Adsorption of a basic protein onto an hydroxyapatite disk enhanced or had no effect on bacterial adherence, whereas adsorption of an acidic protein reduced adherence. The effect of adsorbed protein on bacterial adherence was of both short and long range. The long-range effect of the acidic proteins in reducing the number of bacteria adhering to hydroxyapatite was related to protein adsorption causing an increase in surface net negative charge, as shown by zeta potential measurement. Basic protein produced a net positive surface charge which facilitated adherence. Within the acidic protein group, the acidic residue percentage of the adsorbed protein was negatively correlated with the number of bacteria adhering, whereas the nonpolar residue percentage was positively correlated with bacterial adherence. Within the basic protein group, the basic residue percentage was correlated with the number of cells adhering. These results indicate the involvement of short-range hydrophobic and ionic interactions in bacterial adherence to protein-coated hydroxyapatite.

Adhesiveness↗