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Biomedical subjects

T T Wheeler

Publications and source records attributed to T T Wheeler.

54 records · Page 3Linked to original sources

Modeling the relationship between clinical, microbiologic, and immunologic parameters and alveolar bone levels in an elderly population.

A cross-sectional periodontal study of 74 subjects aged 65 to 75 years was performed. Clinical data were collected and related to microbiological and immunological data. A statistical model (step-wise multiple regression) of factors related to bone loss was created initially using clinical data only; then by adding either the microbiologic or immunologic data; and then by using clinical, microbiologic, and immunologic data together. When only clinical data were considered, three factors were found to have significant positive correlations with bone loss. Tooth mobility accounted for 17% of the variability in the alveolar bone level measurements, probing depth for 12%(r2), and plaque index for 3%, for a total of 32% of the variability explained by these clinical factors. Tooth mobility and probing depth were clinical factors which remained significant in the model when the microbiological data were also considered. As percentages of the total cultivable microbiota, E. corrodens (r2 = 14%) and black-pigmenting Prevotella intermedia (r2 = 4%) correlated positively with alveolar bone loss. The addition of the microbiologic data only increased the r2 to 33%. When immunological data were considered with the clinical data, pocket depth and tooth mobility were the clinical parameters which remained in the model. IgG antibody levels to P. gingivalis W83 and/or 381 (r2 = 24%) A. actinomycetemcomitans 627 (r2 = 2%) were the significant immunologic measures having a positive correlation with bone loss. Anti-F. nucleatum levels had a significant negative correlation. A total of 50% of the variability in alveolar bone level was accounted for in the model by the addition of specific serum antibody levels to subgingival plaque microorganisms. When clinical, microbiological, and immunological measurements were all considered together, antibody to P. gingivalis W83 and/or 381 (r2 = 42%), percentage of B-lymphocytes (r2 = 3%), probing depth (r2 = 4%), anti-E. corrodens levels (r2 = 2%), and anti-P. gingivalis 33277 levels (r2 = 4%) all had significant positive correlation with loss of alveolar bone. The number of enteric bacteria, anti-F. nucleatum levels, and anti-P. intermedia levels each had a significant negative correlation with alveolar bone heights. The r2 for this model was 75%. These results indicated that antibody levels to subgingival plaque microorganisms and tooth mobility were the best predictors of bone loss in the elderly patients tested in this study.

Age Factors↗

Traumatic root resorption in dentine-immunized mice.

Traumatic root resorption in a mouse model has been shown to coincide with a decline in naturally occurring serum antibody levels to dentin. It has been proposed that root resorption may be dentin antibody mediated. The purpose of this study was to examine the traumatic root resorption response in mice after hyperimmunization with a crude tooth extract (dentin). The hypothesis of this study was that elevated dentin antibody titers would positively correlate with root resorption. Mice were immunized with mouse dentin and controls were sham immunized. All mice were boosted 4 weeks later with or without mouse dentin as appropriate. All mice were then boosted two more times at weekly intervals with mouse dentin and then twice at weekly intervals with rat dentin. The change to rat dentin was made to increase mouse serum antibody titers to dentin. Serum samples were obtained before the initial immunization and weekly after each boost and were examined for antibody-to-dentin antigen by the enzyme-linked immune sorbent assay (ELISA). One week after the second boost with rat dentin, all animals were exposed to the cryoprobe procedure. Mice were killed 10 days later, and serum tested for antibody to dentin antigen. The incisors were examined by scanning electron microscopy, and root resorption quantified. Root resorption was observed on the incisors in the sham-immunized mice but not in the dentin-immunized mice. A trend toward increased serum antibody titers to dentin in immunized mice was observed over time.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Gene expression during 3T3-L1 adipocyte differentiation. Characterization of initial responses to the inducing agents and changes during commitment to differentiation.

The mouse 3T3-L1 fibroblastic cell line rapidly differentiates to an adipocyte phenotype when post-confluent cells are treated for 48 h in fetal calf serum-containing medium supplemented with 1 microM dexamethasone (D), 0.5 mM methylisobutylxanthine (M) and 10 micrograms/ml insulin (I). D and I act synergistically to commit the cells to differentiate 24-48 h after initiating treatment, and this is blocked by the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate. In order to identify cellular proteins involved in the differentiation process we analyzed differentiating 3T3-L1 cells using two-dimensional electrophoresis on large format gels. We observed changes in over 300 proteins during differentiation (over 100 within 5 h of initiating differentiation) and many of these are also changed at the level of mRNA (by analysis of in vitro translation products). About 75% of the initial changes were maximally induced by treatment with a combination of M and I, while no more than 10 proteins and their corresponding mRNAs were maximally induced by D within 3.5 h. Another 10 proteins were synergistically regulated by the combination of all three agents (DMI) within 3.5 h. Additional species were induced at later times. Five of these were synergistically induced by treatments that lead to differentiation, were first expressed at elevated levels during commitment and remained elevated in fully differentiated adipocytes. One or more of these proteins could well have a functional role in the commitment to and/or expression of the adipocyte differentiation program.

3T3 Cells↗

Suppression of tumorigenicity of a Wilms' tumor cell line is associated with a decrease in synthesis of two proteins.

Wilms' tumor has been associated with deletions in two loci on chromosome 11, and the introduction of a translocated human chromosome [t(X;11)] into a Wilms' tumor cell line (G401.6TG.6) by microcell hybridization suppresses tumor formation in nude mice. The tumorigenic phenotype is restored in segregants of these microcell hybrids, in which the introduced chromosome is lost. We have used ultrahigh-resolution 'giant' two-dimensional gel electrophoresis of metabolically labeled cellular proteins and in vitro translation products of isolated mRNA to identify changes in cellular gene expression that occur in these cell lines. The changes in gene expression associated with these chromosomal manipulations per se are quite minimal. However, we have identified two proteins (p16 and p28) whose synthesis is consistently decreased in three non-tumorigenic (suppressed) microcell hybrid clones relative to parental and segregant tumorigenic lines. They are also decreased at the level of mRNA in at least two of the non-tumorigenic clones. The decrease of these proteins represents markers of the suppressed phenotype, and their down-regulation may conceivably mediate the suppression of tumorigenicity.

Child, Preschool↗

The timing of treatment for Class II malocclusions in children: a literature review.

Two basic strategies for the timing of treatment for Class II malocclusions in children are common: (1) correction achieved in two phases, one during pre-adolescence (early treatment) and the other during the teen years; and (2) correction accomplished in one phase of active treatment during the adolescent years. The issues of efficacy and cost(risk)-benefit of these strategies have not been well delineated. Most clinical studies examining these issues have suffered serious methodological deficiencies, such as being retrospective, lacking adequate controls, and evaluating only successfully treated cases. However, despite a lack of objective data, clinicians have shown considerable interest in recent years in two-phase treatment. This paper reviews major issues of two-phase Class II treatment and concludes by delineating several important clinical questions which could be resolved by a carefully controlled prospective study.

Adolescent↗

A double-label two-dimensional procedure for the analysis of membrane proteins.

A previously described double-label two-dimensional electrophoresis procedure (Wheeler et al., Anal. Biochem. 1986 159, 1-7) for the analysis of differences between two complex mixtures of soluble proteins has been modified to allow analysis of proteins requiring detergent for aqueous solubility. The samples are first disrupted by sonication and the insoluble proteins concentrated by high-speed centrifugation. The proteins are then solubilized with sodium dodecyl sulfate and further concentrated in a centrifugal concentrator to achieve protein mixtures suitable for labeling with 14C and 3H by reductive methylation and subsequent two-dimensional electrophoresis. The sample concentration step is quick, minimizes the concentration of sodium dodecyl sulfate in the final sample, and avoids the potential difficulties associated with lyophilization or precipitation. The modified procedure was applied to the analysis of erythrocyte membranes, platelets and isolated placental microvilli. The high resolving power of two-dimensional gel electrophoresis is retained and the procedure is sensitive because the conditions of labeling allow substantial incorporation of radioactivity into protein despite the presence of detergent.

Blood Platelets↗

Search for abnormal proteins in erythrocytes and plasma from patients with a major depressive episode.

A sensitive double-label two dimensional gel electrophoresis procedure has been used to search for abnormal proteins in plasma and erythrocyte plasma membranes from patients with major depressive illness. For both plasma and erythrocytes, minor qualitative and quantitative differences between the proteins in pooled samples from depressed and normal subjects were observed; however, these were shown to be artifacts and no consistent differences were found.

Blood Protein Electrophoresis↗

A search for protein abnormalities in erythrocyte membranes and platelets from patients with multiple sclerosis using double-label two-dimensional electrophoresis.

Proteins from erythrocyte membranes and whole platelets from patients with definite multiple sclerosis (MS) and normal subjects were compared using a double-label two-dimensional procedure. No consistent differences were found in the proteins adherent or intrinsic to erythrocyte membranes. The results of initial studies with platelets showed the absence of a group of three protein spots in the electrophoretic gels of the MS specimens. Further experiments suggested that these results were probably due to minor differences in the handling of the MS and normal specimens although the possibility of a difference in protease activity was not completely eliminated.

Blood Platelets↗

Double-label two-dimensional electrophoresis of serum and plasma proteins from patients with multiple sclerosis.

A double-label two-dimensional electrophoretic procedure has been used to search for abnormal proteins in the serum and plasma of patients with definite multiple sclerosis. The procedure possesses high resolving power and is particularly valuable in comparative studies of complex mixtures of proteins since differences due to gel-to-gel variations in protein mobilities are eliminated. Proteins present in serum and plasma at a concentration of 5-10 micrograms/ml are detected routinely. No consistent differences were observed between patients with multiple sclerosis and normal subjects in comparisons using pooled or individual specimens of serum or plasma. The absence of consistent differences in serum and plasma proteins between normal subjects and MS patients applied even when the sensitivity of the procedure was increased several-fold and when the possibly obscuring effect of albumin in the electrophoretic gels was eliminated. To our knowledge, this study combines the most extensive and sensitive search for consistent abnormalities in serum and plasma proteins in multiple sclerosis sera thus far reported.

Autoradiography↗

A double-label two-dimensional gel electrophoresis procedure specifically designed for serum or plasma protein analysis.

A double-label two-dimensional electrophoresis procedure has been developed which is specifically designed for the comparison of serum or plasma proteins in two different samples. Proteins are labeled by reductive methylation with [14C]- or [3H] formaldehyde. The procedure is economical because small quantities of relatively inexpensive isotopes are used and it is at least as sensitive as silver staining in detecting proteins. A fourfold increase in the sensitivity of autoradiography over existing methods was obtained by performing autoradiography before processing the gel for fluorography. A spot in the electrophoretic gel that contains 17-28 ng of labeled protein is detectable. This corresponds to proteins present in serum at a concentration of 5-10 micrograms/ml. Even greater sensitivity can be achieved, at greater expense, by increasing the quantities of the radioisotopes in the labeling reaction. The particular value of the double label approach is that complex mixtures from two different sources are resolved together thus eliminating the possibility of differences arising from the resolving procedure itself. The procedure was applied to a mixture of serum and plasma from a single subject and a number of qualitative and quantitative differences were observed.

Autoradiography↗

Actinomyces adsorption mediated by type-1 fimbriae.

Monospecific antibody against the type-1 fimbriae of Actinomyces viscosus T14V reacted with 12 strains of A. viscosus (serotype 2), 12 strains of A. naeslundii (serotype 2 or serotype 3), but not with 11 serotype 1 strains of A. naeslundii. All strains positive for the type-1 fimbrial antigen adsorbed strongly to saliva-treated hydroxyapatite, and in many instances (18 of 24 strains) this interaction was inhibited by the monospecific antibody. These findings strongly suggest that only those strains of actinomyces which possess type-1 fimbriae adhere to the tooth surfaces, and that these fimbriae constitute the principal adhesin involved in this interaction.

Actinomyces↗

Relative hydrophobicities of Actinomyces viscosus and Actinomyces naeslundii strains and their adsorption to saliva-treated hydroxyapatite.

The present study examined 42 strains of Actinomyces spp. to determine whether adsorption to saliva-treated hydroxyapatite (SHA) of the selected strains of this prominent group of dental-plaque bacteria correlated with hydrophobicity. The relative hydrophobicity of the strains was determined by their adsorption to hydrophobic gels (i.e., phenyl-Sepharose) and their aggregation in ammonium sulfate. Within serogroups the relative hydrophobicity for the strains was similar. The relative adsorption of strains to SHA was also similar within the respective serogroups. Strains which were relatively hydrophobic, as judged by their binding to the hydrophobic gel and aggregation in low concentrations of ammonium sulfate, adsorbed well to SHA. Strains which adsorbed poorly to SHA were relatively hydrophilic since they did not bind well to the hydrophobic gel and were only aggregated in relatively high concentrations of ammonium sulfate. Tween 80, a nonionic detergent known to inhibit hydrophobic interactions, blocked binding of cells to the hydrophobic gel, suggesting that hydrophobic interactions had been inhibited. However, Tween 80 exhibited no influence on the adsorption of cells to SHA. Thus, although there was a strong statistical correlation between the relative hydrophobicity of a strain and its adsorption to SHA, the data were consistent with the view that other interactions, such as ionic bonds and interactions between complimentary macromolecules, are involved in adsorption of the Actinomyces strains to SHA.

Actinomyces↗

Specific inhibition of adsorption of Actinomyces viscosus T14V to saliva-treated hydroxyapatite by antibody against type 1 fimbriae.

Specific antibodies and their Fab fragments were used to study adsorption of Antinomyces viscosus T14V to saliva-treated hydroxyapatite. Antibody against A. viscosus T14V blocked adsorption of the strain to saliva-treated hydroxyapatite, and this activity was removed after preincubation of the immune immunoglobulin G (IgG) with purified type 1 fimbriae in amounts near equivalence but not after preincubation with type 2 fimbriae in antigen excess. Specific IgG or Fab fragments against type 1 fimbriae inhibited adsorption of strain T14V to saliva-treated hydroxyapatite and promoted desorption of cells already attached to the apatite surface. Anti-type 2 IgG or Fab fragments did not possess these activities. These data demonstrate that type 1 but not type 2 fimbriae mediate adsorption of strain T14V to the saliva-treated hydroxyapatite surface.

Actinomyces↗

Influence of physicochemical parameters on adsorption of Actinomyces viscosus to hydroxyapatite surfaces.

Adsorption of Actinomyces viscosus strains T14V and T14AV to saliva-treated and untreated hydroxyapatite (HA) under various environmental conditions was studied. Strain T14V adsorption to saliva-treated HA was not influenced by pH, whereas strain T14AV adsorbed in higher numbers under acidic conditions. The addition of cations inhibited adsorption of strains T14V and T14AV to saliva-treated HA. Strain T14V possessed a greater affinity for hydrophobic gels than did strain T14AV, but incorporation of non-ionic detergents, which inhibit hydrophobic interactions, did not influence adsorption of strain T14V to saliva-treated HA. Adsorption of strain T14V to saliva-treated HA increased in the presence of an anionic detergent; however, strain T14AV adsorption was not affected. Strain T14V adsorption decreased in the presence of a cationic detergent, whereas strain T14AV adsorption increased. Collectively, these data suggest that electrostatic interactions, but not hydrophobic interactions, are of major importance in the adsorption of strain T14V to saliva-treated HA.

Actinomyces↗

Role of surface fimbriae (fibrils) in the adsorption of Actinomyces species to saliva-treated hydroxyapatite surfaces.

We studied the adsorption, morphological, and serological characteristics of selected Actinomyces and related species. Evaluation of uranyl acetate-stained cells by electron microscopy revealed wide variations among strains in the frequency of surface fimbriae. These variations did not always correlate with the percent adsorption to saliva-treated hydroxyapatite of the various Actinomyces strains. However, two strains of Rothia dentocariosa possessing no surface fimbriae and five strains of A. israelii possessing very few surface fimbriae exhibited feeble adsorption to saliva-treated hydroxyapatite. Although the calculated number of adsorption sites on saliva-treated hydroxypatite did not vary widely among the strains tested, significant differences were observed in the affinities calculated for some species or serotypes. The mean affinities for strains of A. viscosus serotype 2 and A. naeslundii serotype 3 were similar, and these strains adsorbed well to saliva-treated hydroxyapatite. The mean adsorption and affinity for the A. naeslundii strain serotype 1 and all strains of A. israelii tested were significantly less than those determined for the A. viscosus serotype 2 or A. naeslundii serotype 3 strains. Adsorption inhibition activity of antiserum to strain T14V, previously shown to be solely related to antibodies in immune serum directed against the VA1 fimbria (fibril) antigen, was removed by preadsorption of the antiserum with most A. viscosus and A. naelundii strains, but not with A. israelii strains. This suggests some cross-reactivity among strains of A. viscosus and A. naeslundii but not A. israelii. Adsorption to saliva-treated hydroxyapatite of all A. viscosus and A. naeslundii strains tested was strongly inhibited by fimbriae isolated from A. viscosus strain T14V. Collectively, these data suggest that the adsorption of certain A. viscosus and A. naeslundii strains is mediated by surface fimbriae, many of which appear serologically cross-reactive with strain T14V fimbriae.

Actinomyces↗

Fibril-mediated adherence of Actinomyces viscosus to saliva-treated hydroxyapatite.

Fibril-mediated adherence of Actinomyces viscosus strain T14V cells to saliva-treated hydroxyapatite was studied. Fibrils were purified by ammonium sulfate precipitation and differential centrifugation from the crude supernatant of whole cells that were sheared by one passage through a French pressure cell. Purified fibrils and crude supernatant inhibited strain T14V adherence to saliva-treated hydroxyapatite to similar extents. However, anti-strain T14V serum and antifibril specific antibody completely abolished strain T14V adherence. The blocking immunoglobulin could be adsorbed from anti-T14V serum by strain T14V whole cells, by purified fibrils, and, to a lesser extent, by cell walls. It was concluded that fibrils mediate adherence of strain T14V cells to saliva-treated hydroxyapatite. In addition, fibril preparations were shown to contain more than 95% protein and to be antigenically homogeneous by immunodiffusion and Laurell rocket immunoelectrophoresis. Purified fibril preparations showed serological identity with the virulence-associated 1 antigen of Lancefield-extracted T14V cells, whereas crude supernatants contained both virulence-associated 1 and virulence-associated 2 antigens, as shown by rocket immunoelectrophoresis.

Actinomyces↗

Adherence of Actinomyces viscosus T14V and T14AV to hydroxyapatite surfaces in vitro and human teeth in vivo.

Adsorption of Actinomyces viscosus strains T14V and T14AV to hydroxyapatite (HA) surfaces was studied, using an adsorption model based on the Langmuir adsorption isotherm. Data generally followed the adsorption model as judged by high correlation coefficients obtained for both strains to most of the treated surfaces studied. The number of binding sites for strains T14V and T14AV cells to human saliva-treated HA was similar to that for untreated HA. The affinity of strain T14V for saliva-treated HA was tenfold greater than the affinity of strain T14AV for that surface. To approximate the pellicle of the gingival crevice and margin and to determine whether adherence by strain T14V was to specific saliva or serum receptors, experimental pellicles were formed on HA by saliva/serum mixtures. The number of binding sites on the saliva/serum-treated HA remained the same as for the saliva-treated surface. Although the affinity of strain T14V cells for the saliva/serum HA surface remained generally the same as the affinity for the HA treated with saliva alone, the affinity of strain T14AV cells decreased further as the serum content increased. Strain T14V cell numbers adsorbed to serum-treated HA, and albumin-treated HA were less than those adsorbed to saliva-treated HA, indicating that the adherence by strain T14V was to specific saliva receptors. In vivo results from streptomycin-resistant mutants of both strains T14V and T14AV confirmed in vitro results using saliva-serum pellicles. Pretreatment of strain T14V with proteolytic enzymes and heat inhibited adherence to saliva-treated HA, suggesting that the adherence receptor(s) on the cell surface of strain T14V is protein in nature.

Actinomyces↗