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Biomedical subjects

S A Thompson

Publications and source records attributed to S A Thompson.

At least 73 records · Page 4Linked to original sources

Shaping ability of Profile rotary nickel-titanium instruments with ISO sized tips in simulated root canals: Part 1.

The aim of this study was to determine the shaping ability of ProFile 0.04 taper rotary nickel-titanium instruments with ISO sized tips in simulated canals. A total of 40 simulated root canal made up of four different shapes in terms of angle and position of curvature were prepared by ProFile instruments using the 'crown down' approach recommended by the manufacturer. Part 1 of this two-part report describes the efficacy of the instruments in terms of preparation time, instrument failure, canal blockages, loss of canal length and three-dimensional canal form. The time necessary for canal preparation was on average 5.2 min and was not influenced significantly by canal shape. Three instrument fractures occurred and a further 3 instruments deformed; size 35 instruments failed the most (4) followed by size 30 (2), four of the failures occurred in canals with 40 degrees curves. None of the canals became blocked with debris and change in working distance was minimal. Intra-canal impressions of canal form demonstrated that most canals had apical stops and smooth canal walls whereas all canals had good flow and taper. Under the conditions of this study, ProFile 0.04 taper rotary nickel-titanium instruments with ISO sized tips prepared simulated canals rapidly and created good three-dimensional form.

Dental Instruments↗

Acid-induced expression of an LPS-associated gene in Helicobacter pylori.

To investigate urease-independent mechanisms by which Helicobacter pylori resists acid stress, subtractive RNA hybridization was used to identify H. pylori genes whose expression is induced after exposure to acid pH. This approach led to the isolation of a gene that encoded a predicted 34.8kDa protein (WbcJ), which was homologous to known bacterial O-antigen biosynthesis proteins involved in the conversion of GDP-mannose to GDP-fucose. An isogenic wbcJ null mutant strain failed to express O-antigen and Lewis X or Lewis Y determinants and was more sensitive to acid stress than was the wild-type strain. Qualitative differences in LPS profiles were observed in H. pylori cells grown at pH 5 compared with pH 7, which suggests that H. pylori may alter its LPS structure in response to acidic pH. This may be an important adaptation facilitating H. pylori colonization of the acidic gastric environment.

Amino Acid Sequence↗

Campylobacter fetus surface layer proteins are transported by a type I secretion system.

The virulence of Campylobacter fetus, a bacterial pathogen of ungulates and humans, is mediated in part by the presence of a paracrystalline surface layer (S-layer) that confers serum resistance. The subunits of the S-layer are S-layer proteins (SLPs) that are secreted in the absence of an N-terminal signal sequence and attach to either type A or B C. fetus lipopolysaccharide in a serospecific manner. Antigenic variation of multiple SLPs (encoded by sapA homologs) of type A strain 23D occurs by inversion of a promoter-containing DNA element flanked by two sapA homologs. Cloning and sequencing of the entire 6.2-kb invertible region from C. fetus 23D revealed a probable 5.6-kb operon of four overlapping genes (sapCDEF, with sizes of 1,035, 1,752, 1,284, and 1,302 bp, respectively) transcribed in the opposite direction from sapA. The four genes also were present in the invertible region of type B strain 84-107 and were virtually identical to their counterparts in the type A strain. Although SapC had no database homologies, SapD, SapE, and SapF had predicted amino acid homologies with type I protein secretion systems (typified by Escherichia coli HlyBD/TolC or Erwinia chrysanthemi PrtDEF) that utilize C-terminal secretion signals to mediate the secretion of hemolysins, leukotoxins, or proteases from other bacterial species. Analysis of the C termini of four C. fetus SLPs revealed conserved structures that are potential secretion signals. A C. fetus sapD mutant neither produced nor secreted SLPs. E. coli expressing C. fetus sapA and sapCDEF secreted SapA, indicating that the sapCDEF genes are sufficient for SLP secretion. C. fetus SLPs therefore are transported to the cell surface by a type I secretion system.

Amino Acid Sequence↗

Muscle fiber type distribution in the normal human levator veli palatini muscle.

OBJECTIVE: This study examined the muscle fiber type distribution within the normal adult levator veli palatini muscle. METHODS: Levator veli palatini muscle tissue was harvested from the palates of 12 (seven female, five male) adult noncleft cadavers. Adjacent sections were stained for adenosine triphosphatase at pH 10.4 or 4.2. After mounting, magnifying, and photographing, Type I versus Type II fiber types were differentiated by the intensity of, or by the inhibition of, staining of matched fibers at each pH level. Type I fibers stained light at pH 10.4 and dark at pH 4.2, while Type II fibers stained light at pH 4.2 and dark at pH 10.4. MAIN OUTCOME MEASURES: The number of fibers counted for each specimen ranged from 60 to 616. The numbers of Type I and Type II stained fibers appearing in each muscle tissue sample were determined and expressed as a percentage of the total number of fibers identified. A few identified fibers could not be labelled as either Type I or Type II. RESULTS: The overall proportion of Type I fibers, averaged across all specimens, was 59.8%. Male specimens had 67.4% Type I fibers and 31.8% Type II fibers, while female specimens had 54.4% Type I fibers and 44.4% Type II fibers. CONCLUSIONS: Observed fiber type distributions were similar to those reported for other articulatory muscles, but differed slightly from previously reported distributions for normal levator veli palatini. The distributions observed in this study provide a baseline against which to relate fiber type data from the levator veli palatini of cleft palates to the functional status of the velopharyngeal mechanism.

Adenosine Triphosphatases↗

Alterations in immune parameters associated with low level methylmercury exposure in mice.

Methylmercury (MeHg) is a known toxicant and continues to be a significant environmental contaminant. While the neurotoxicity and developmental toxicity of MeHg are well established, the immunotoxic effects of MeHg are just now being studied and described. This study evaluated strain and gender specific effects of low level, prolonged MeHg exposure in mice. Mice were exposed to MeHg in the drinking water (0, 3 or 10 ppm) for 4 weeks. Splenocytes and thymocytes were evaluated for alterations in immunophenotype, GSH levels, and intracellular Ca2+ flux after mitogen stimulation. MeHg exposure resulted in alterations in splenocyte and thymocyte subsets and a dose dependent decrease in GSH levels (as measured by monochlorobimane fluorescence and flow cytometry) of all splenocyte subsets. This decrease in GSH was further confirmed by biochemical assay in splenocytes. In addition, there was a dose response related decrease in mitogen stimulated Ca2+ flux and in the percentages of CD4+ splenocytes and CD8+ splenocytes from mice exposed to 10 ppm of MeHg. These results suggest that low-level chronic MeHg exposure may cause immune disfunction by disturbing thiol redox balance, transmembrane signaling and splenic cellularity.

Analysis of Variance↗

Regulation of IL-18 (IFN-gamma-inducing factor) gene expression.

IL-18 (also known as IFN-gamma-inducing factor), although structurally unrelated to IL-12, shares with it the role of activating NK cells and polarizing T cells toward Th1 cell function. To understand how the IL-18 gene (and consequently Th1 function) is regulated, we have determined the gene structure and investigated the mechanisms of transcriptional control and cell type expression. The mouse IL-18 gene comprises seven exons distributed over 26 kb. Exons 1 and 2 of this gene are 5'-noncoding exons. Promoter activity was detected upstream of these noncoding exons in two distinct regions. Both promoters are TATA-less and not G+C rich. The promoter activity located upstream of exon 2 was shown to act constitutively, while the activity located upstream of exon 1 was up-regulated in activated macrophage and T cell lines. IL-18 gene expression may be regulated in a wide range of cell types by the activities of these two distinct promoters. IL-18 is known to be synthesized as a precursor, pro-IL-18, and its maturation is controlled by IL-1beta-converting enzyme (ICE). We observed concordant expression of IL-18 and ICE mRNAs in a wide range of cell types, unlike the more restricted expression of IL-12 p40 mRNA. The widespread IL-18 mRNA distribution and the special relationship with ICE lead us to the hypothesis that IL-18 expression may be coupled with apoptotic processes involving activation of ICE or ICE-like proteinase.

Animals↗

Amphiregulin antisense oligonucleotide inhibits the growth of T3M4 human pancreatic cancer cells and sensitizes the cells to EGF receptor-targeted therapy.

Human pancreatic cancers overexpress the epidermal growth factor (EGF) receptor (EGFR) and all 5 ligands that bind to this receptor, including amphiregulin. It is not known, however, whether amphiregulin contributes in an autocrine manner to enhance pancreatic cancer cell growth. Therefore, we used an amphiregulin antisense oligonucleotide (AR-AS) to suppress amphiregulin expression in T3M4 human pancreatic cancer cells. These cells express high levels of EGFR and amphiregulin. AR-AS abolished amphiregulin immunoreactivity in T3M4 cells, decreased amphiregulin release into the medium and inhibited cell growth in a dose-dependent manner. Exogenous amphiregulin reversed AR-AS-mediated growth inhibition. A random oligonucleotide (AR-R) did not alter either cell growth or cellular amphiregulin immunoreactivity. AR-AS also increased cellular EGFR protein levels and enhanced the growth-inhibitory actions of TP40, a chimeric protein consisting of transforming growth factor-alpha coupled to Pseudomonas exotoxin that internalizes into cells via EGFR. These findings indicate that there is an important EGFR/ amphiregulin autocrine loop in T3M4 cells and raise the possibility that modalities aimed at abrogating amphiregulin action may prove useful in pancreatic cancer, especially when used in conjunction with EGFR-targeted therapy.

Amino Acid Sequence↗

Neuronally restricted RNA splicing regulates the expression of a novel GABAA receptor subunit conferring atypical functional properties [corrected; erratum to be published].

We report the isolation and characterization of a cDNA encoding a novel member of the GABA receptor gene family, epsilon. This polypeptide is 506 amino acids in length and exhibits its greatest amino acid sequence identity with the GABAA receptor gamma3 subunit (47%), although this degree of homology is not sufficient for it to be classified as a fourth gamma subunit. The epsilon subunit coassembles with GABAA receptor alpha and beta subunits in Xenopus laevis oocytes and transfected mammalian cells to form functional GABA-gated channels. alpha1beta1epsilon GABAA receptors, like alpha1beta1gamma2s receptors, are modulated by pentobarbital and the steroid 5alpha-pregnan-3alpha-ol-20-one but, unlike alpha1beta1gamma2s receptors, are insensitive to flunitrazepam. Additionally, alpha1beta1epsilon receptors exhibit rapid desensitization kinetics, as compared with alpha1beta1 or alpha1beta1gamma2s. Northern analysis demonstrates widespread expression of a large epsilon subunit transcript in a variety of non-neuronal tissues and expression of a smaller transcript in brain and spinal cord. Sequence analysis demonstrated that the large transcript contained an unspliced intron, whereas the small transcript represents the mature mRNA, suggesting regulation of expression of the epsilon subunit via neuronally restricted RNA splicing. In situ hybridization and immunocytochemistry reveal a pattern of expression in the brain restricted primarily to the hypothalamus, suggesting a role in neuroendocrine regulation, and also to subfields of the hippocampus, suggesting a role in the modulation of long term potentiation and memory.

Amino Acid Sequence↗

Induction of growth arrest and DNA damage-inducible genes Gadd45 and Gadd153 in primary rodent embryonic cells following exposure to methylmercury.

Methylmercury (MeHg) is recognized as a significant environmental hazard, particularly to the development of the nervous system. Studies on the mechanism of MeHg-induced toxicity reveal that inhibition of cell cycle progression may be one way by which MeHg interferes with normal development. In this study, we utilized primary rodent embryonic neuronal cell (CNS) and limb bud (LB) cultures to determine the mRNA expression level of two genes involved in cell cycle arrest, Gadd45 and Gadd153, both during cellular differentiation and in response to MeHg exposure. A differential expression pattern of Gadd45 and Gadd153 was observed during CNS and LB differentiation in culture. However, both CNS and LB cells responded to MeHg exposure with a concentration-dependent increase in Gadd45 and Gadd153 mRNA. Previous studies have shown that MeHg exposure (2 microm) of CNS cells for 24 hr causes a fourfold decrease in the number of cells passing through the cell cycle. The present study shows that at the same exposure concentration, a five- to eightfold increase in Gadd45 mRNA levels and a two- to fourfold increase of Gadd153 was observed. Induction of Gadd45 was also noted in adult female mice chronically exposed to 10 ppm MeHg, a dose that caused developmental toxicity in vivo. Based on the known involvement of the Gadd genes in cell cycle arrest, activation of these genes could be one mechanism by which MeHg interferes with the cell cycle in adult and developing organisms.

Animals↗

Shaping ability of Lightspeed rotary nickel-titanium instruments in simulated root canals. Part 2.

The aim of this laboratory based study was to determine the shaping ability of Lightspeed nickel-titanium rotary instruments in simulated root canals. A total of 40 canals with four different shapes in terms of angle and position of curve were prepared with Lightspeed instruments using the stepback technique recommended by the manufacturer. This report describes the efficacy of the instruments in terms of prevalence of canal aberrations, the amount and direction of canal transportation, and overall postoperative shape. Preoperative and postoperative images of the canals were taken using a video camera attached to a computer with image analysis software. The preoperative and postoperative views were superimposed to highlight the amount and position of material removed during preparation. Only one (2.5%) zip and one elbow were created, with no ledges, perforations, danger zones, or blockages being produced. At specific points along the canal length there were highly significant differences (p < 0.001) between the canal shapes in total canal width and in the amount of resin removed from the inner and outer aspects of the curve. The direction of canal transportation at the end point of preparation was most frequently toward the outer aspect of the curve; although in half of the canals, transportation was either directed toward the inner aspect or not observed. At the apex of the curve, the beginning of the curve, and halfway to the orifice, transportation was reversed with the majority of canals being transported toward the inner aspect of the curve. Mean absolute transportation was small and was below 0.06 mm at every position except the orifice. Overall, Lightspeed rotary instruments prepared canals well and would appear to be a valuable addition to the endodontic armamentarium.

Dental Alloys↗

Shaping ability of Lightspeed rotary nickel-titanium instruments in simulated root canals. Part 1.

The aim of this study was to determine the shaping ability of Lightspeed nickel-titanium rotary instruments in simulated canals. Forty canals consisting of four different shapes in terms of angle and position of curvature were prepared by Lightspeed instruments using the technique recommended by the manufacturer. This report describes the efficacy of the instruments in terms of preparation time, instrument failure, canal blockages, loss of canal length, and three-dimensional canal form. Overall, the mean preparation time for all canals was 8.12 min with canal shape having no significant effect on the speed of preparation. No instruments deformed or separated during the study, and none of the canals became blocked with debris. Seventeen canals retained their original working length, but 16 gained in length and 7 lost length. There was no significant difference between the canal shapes in terms of the mean loss of distance or category of distance change. Apical stops as judged from intracanal impressions were present in 23 of the canals but they were all judged to be of poor quality. The canals were found to be smooth in the apical half of the canal in 36 specimens and in the coronal half of 24 specimens. All the canals had poor taper characteristics, and only 16 specimens showed good flow characteristics. Under the conditions of this study, Lightspeed instruments prepared canals rapidly, with no fractures, canal blockages, and with minimal change in working length. The three dimensional form of the canals was compromised as flow and taper were less than ideal, presumably as a result of an ineffective stepback procedure. The results imply that either the stepback sequence should be modified or another instrument with increased taper should be used to refine the canal walls before obturation.

Dental Alloys↗

Shaping ability of NT Engine and McXim rotary nickel-titanium instruments in simulated root canals. Part 1.

The aim of this study was to determine the shaping ability of NT Engine and McXim nickel-titanium rotary instruments in simulated root canals. In all, 40 canals consisting of four different shapes in terms of angle and position of curvature were prepared by a combination of NT Engine and McXim instruments using the technique recommended by the manufacturer. Part 1 of this two-part report describes the efficacy of the instruments in terms of preparation time, instrument failure, canal blockages, loss of canal length and three-dimensional canal form. Overall, the mean preparation time for all canals was 6.01 min, with canal shape having a significant effect (P < 0.01) on the speed of preparation. One instrument fractured and only four instruments deformed, with most of the failures occurring in canals with curves which began 12 mm from the orifice, that is, in short acute curves. None of the canals became blocked with debris. Following preparation, 20 canals retained their original working length but 19 lost length and one gained in length; there were significant differences (P < 0.05) between the canal shapes in terms of mean loss of distance and in the category of distance change. Apical stops as determined from intracanal impressions were present in 37 of the canals; 16 were judged to be of good quality and 21 of poor quality. The canals were found to be smooth in the apical half of the canal in 33 specimens and in the coronal half of 39 specimens. All canals had good taper characteristics and 35 had good flow characteristics. Under the conditions of this study, NT Engine and McXim instruments prepared canals rapidly, with few deformations, no canal blockages and with minimal change in working length. The three-dimensional form of the canals demonstrated good flow and taper characteristics.

Dental Alloys↗

Shaping ability of NT Engine and McXim rotary nickel-titanium instruments in simulated root canals. Part 2.

The aim of this laboratory-based study was to determine the shaping ability of NT Engine and McXim nickel-titanium rotary instruments in simulated root canals. A total of 40 canals with four different shapes in terms of angle and position of curve were prepared with NT Engine and McXim instruments, using the technique recommended by the manufacturer. Part 2 of this report describes the efficacy of the instruments in terms of prevalence of canal aberrations, the amount and direction of canal transportation and overall postoperative shape. Pre- and postoperative images of the canals were taken using a video camera attached to a computer with image analysis software. The pre- and postoperative views were superimposed to highlight the amount and position of material removed during preparation. No zips, elbows, perforations or danger zones were created during preparation. Forty-two per cent of canals had ledges on the outer aspect of the curve, the majority of which (16 out of 17) occurred in canals with short acute curves. There were significant differences (P < 0.001) between canal shapes in terms of the incidence of ledges. There were highly significant differences (P < 0.001) between the canal shapes in total canal width at specific points along the canal length and in the amount or resin removed from the inner and outer aspects of the curve. The direction of canal transportation at the end-point of preparation was most frequently towards the outer aspect of the curve, especially in canals with 40 degrees curves. At the beginning of the curve, transportation in the majority of canals was towards the inner aspect of the curve. Mean absolute transportation was less than 0.03 mm throughout the curve and towards the end-point, with significant differences between canal shapes occurring at the apex (P < 0.05) and at the beginning of the curve (P < 0.001). Under the conditions of this study, NT Engine and McXim rotary nickel-titanium instruments created no aberrations other than ledges and produced only minimal transportation. The overall shape of canals was good.

Dental Alloys↗

Shaping ability of ProFile.04 Taper Series 29 rotary nickel-titanium instruments in simulated root canals. Part 2.

The aim of this study was to determine the shaping ability of ProFile.04 Taper Series 29 rotary nickel-titanium instruments in simulated canals. A total of 40 simulated root canals made up of four different shapes in terms of angle and position of curvature were prepared by ProFile instruments using a step-down approach. Part 2 of this two-part report describes the efficacy of the instruments in terms of prevalence of canal aberrations, the amount and direction of canal transportation and the overall postoperative shape. No zips, perforations or danger zones were created although 24 specimens (60%) had ledges on the outer wall of the canal. The incidence of ledges differed significantly (P < 0.001) between the canal shapes. At specific points along the canal length there were highly significant differences (P < 0.001) in total canal width and in the amount of material removed from the inner and outer aspects of the curve between the various canal shapes. Overall, 40 degrees canals were wider than 20 degrees canals and canals with curves which began 8 mm from the orifice were wider than 12 mm canals. The direction of canal transportation at the end-point of preparation was balanced between inner and outer in the 8 mm canals, but more often towards the outer in the 12 mm canals. At the apex of the curve, transportation was invariably towards the outer aspect of the curvature. At the beginning of the curve, transportation was more balanced between inner and outer. Mean absolute transportation, ignoring direction, was generally greater in 40 degrees canals and in those with the curve beginning 8 mm from the orifice. Of particular importance was the finding that excessive resin was removed from the outer aspect of the canal at the apex of the curve which was often associated with irregular widened areas or ledges. This is in contrast to the pattern of tissue removal found with stainless steel hand instruments where more resin is removed from the inner aspect of the canal at the apex, to create a danger zone.

Dental Alloys↗

Shaping ability of ProFile.04 Taper Series 29 rotary nickel-titanium instruments in simulated root canals. Part 1.

The aim of this study was to determine the shaping ability of ProFile.04 Taper Series 29 nickel-titanium instruments in simulated canals. A total of 40 simulated root canals made up of four different shapes in terms of angle and position of curvature were prepared by ProFile instruments using a step-down approach. Part 1 of this two-part report describes the efficacy of the instruments in terms of preparation time, instrument failure, canal blockages, loss of canal length and three-dimensional canal form. The time necessary for canal preparation was not influenced significantly by canal shape. No instrument fractures occurred but a total of 52 instruments deformed. Size 6 instruments deformed the most followed by sizes 5, 3 and 4. Canal shape did not influence significantly instrument deformation. None of the canals became blocked with debris and loss of working distance was on average 0.5 mm or less. Intracanal impressions of canal form demonstrated that most canals had definite apical stops, smooth canal walls and good flow and taper. Under the conditions of this study, ProFile.04 Taper Series 29 rotary nickel-titanium instruments prepared simulated canals rapidly and created good three-dimensional form. A substantial number of instruments deformed but it was not possible to determine whether this phenomenon occurred because of the nature of the experimental model or through an inherent design weakness in the instruments.

Dental Alloys↗

Key amino acids in the gamma subunit of the gamma-aminobutyric acidA receptor that determine ligand binding and modulation at the benzodiazepine site.

Pharmacological analyses of gamma-aminobutyric acidA (GABAA) receptor subtypes have suggested that both the alpha and gamma subunits, but not the beta subunit, contribute to the benzodiazepine binding site. We took advantage of the different pharmacological properties conferred by the inclusion of different gamma subunits in the receptor macromolecule to identify amino acids gamma2Phe77 and gamma2Met130 as key determinants of the benzodiazepine binding site. gamma2Phe77 was required for high affinity binding of the benzodiazepine site ligands flumazenil, CL218,872, and methyl-beta-carboline-3-carboxylate but not flunitrazepam. This amino acid was, however, required for allosteric modulation by flunitrazepam, as well as other benzodiazepine site ligands. In contrast, gamma2Met130 was required for high affinity binding of flunitrazepam, clonazepam, and triazolam but not flumazenil, CL218, 872, or methyl-beta-carboline-3-carboxylate and did not affect benzodiazepine efficacy. Introduction of the phenylalanine and methionine into the appropriate positions of gamma1 was not sufficient to confer high affinity for the benzodiazepine site ligand zolpidem. These data show that gamma2Phe77 and gamma2Met130 are necessary for high affinity binding of a number of benzodiazepine site ligands. Although most previous studies have focused on the contribution of the alpha subunit, we demonstrated a critical role for the gamma subunit at the benzodiazepine binding site, indicating that this modulatory site is located at the interface of these two subunits. Furthermore, gamma2Phe77 is homologous to alpha1Phe64, which has been previously shown to be a key determinant of the GABA binding site, suggesting a conservation of motifs between different ligand binding sites on the GABAA receptor.

Amino Acid Sequence↗