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Biomedical subjects

S A Thompson

Publications and source records attributed to S A Thompson.

At least 19 recordsLinked to original sources
PubMed2001

Bioisosteric determinants for subtype selectivity of ligands for heteromeric GABA(A) receptors.

The potency and efficacy of a series of bioisosterically modified GABA analogues were determined electrophysiologically using heteromeric GABA(A) receptors expressed in Xenopus oocytes. These agonist parameters were shown to be strongly dependent on the receptor subunit combination. On the other hand, the antagonist potencies of the classical GABA(A) antagonists SR 95531 (7) and BMC (8) and also of 5g and the phosphinic acid bioisosteres of 5a, compounds 5f and 6, were essentially independent of the receptor subunit combinations.

Animals↗
PubMed2001

Integrating composting and vermicomposting in the treatment and bioconversion of biosolids.

Traditional thermophilic composting is commonly adopted for treatment of organic wastes or for production of organic/natural fertilizers. A related technique, called vermicomposting (using earthworms to breakdown the organic wastes) is also becoming popular. These two techniques have their inherent advantages and disadvantages. The integrated approach suggested in this study borrows pertinent attributes from each of these two processes and combines them to enhance the overall process and improve the products qualities. Two approaches investigated in this study are: (1) pre-composting followed by vermicomposting, and (2) pre-vermicomposting followed by composting. The substrate was biosolids (activated sewage sludge) with mixed paper-mulch as the carbon base. Eisenia fetida (red wigglers) was the species of earthworms used in the vermicomposting processes. The results indicate that, a system that combines the two processes not only shortens stabilization time, but also improves the products quality. Combining the two systems resulted in a product that was more stable and consistent (homogenous), had less potential impact on the environment and for compost-vermicomposting (CV) system, the product met the pathogen reduction requirements.

Animals↗
PubMed2001

Campylobacter fetus uses multiple loci for DNA inversion within the 5' conserved regions of sap homologs.

Campylobacter fetus cells possess multiple promoterless sap homologs, each capable of expressing a surface layer protein (SLP) by utilizing a unique promoter present on a 6.2-kb invertible element. Each sap homolog includes a 626-bp 5' conserved region (FCR) with 74 bp upstream and 552 bp within the open reading frame. After DNA inversion, the splice is seamless because the FCRs are identical. In mutant strain 23D:ACA2K101, in which sapA and sapA2 flanking the invertible element in opposite orientations were disrupted by promoterless chloramphenicol resistance (Cm(r)) and kanamycin resistance (Km(r)) cassettes, respectively, the frequency of DNA inversion is 100-fold lower than that of wild-type strain 23D. To define the roles of a 15-bp inverted repeat (IR) and a Chi-like site (CLS) in the FCR, we mutagenized each upstream of sapA2 in 23D:ACA2K101 by introducing NotI and KpnI sites to create strains 23D:ACA2K101N and 23D:ACA2K101K, respectively. Alternatively selecting colonies for Cm(r) or Km(r) showed that mutagenizing the IR or CLS had no apparent effect on the frequency of the DNA inversion. However, mapping the unique NotI or KpnI site in relation to the Cm(r) or Km(r) cassette in the cells that changed phenotype showed that splices occurred both upstream and downstream of the mutated sites. PCR and sequence analyses also showed that the splice could occur in the 425-bp portion of the FCR downstream of the cassettes. In total, these data indicate that C. fetus can use multiple sites within the FCR for its sap-related DNA inversion.

5' Untranslated Regions↗
PubMed2000

IL-10 gene expression is controlled by the transcription factors Sp1 and Sp3.

IL-10 is an 18-kDa cytokine with a key role in homeostatic control of inflammatory and immune responses. We have investigated how transcription of the IL-10 gene is regulated, so as to be able to understand the circumstances of IL-10 expression in both health and disease. In the mouse, IL-10 gene expression is regulated by a TATA-type promoter with a critical cis-acting element containing GGA repeats located at -89 to -77. Its complementary sequence is similar to the cis-acting elements (TCC repeats) in the promoters of genes encoding epidermal growth factor receptor and CD58. All these elements comprise a common CCTCCT sequence with less conserved C + T-rich sequences. Eliminating this CCTCCT sequence results in a marked reduction in promoter activity, suggesting a necessary role in IL-10 gene expression. Despite its dissimilarity to the G + C-rich Sp1 consensus sequence (GC box), Sp1 and Sp3 transcription factors could be shown to bind to this motif. The requirement for Sp1 and Sp3 in transcription of IL-10 was confirmed using Drosophila SL2 cells, which lack endogenous Sp factors. These results suggest that the transcription of IL-10 is positively regulated by both Sp1 and Sp3.

5' Untranslated Regions↗
PubMed2000

Posttranscriptional regulation of IL-10 gene expression through sequences in the 3'-untranslated region.

IL-10 is an 18-kDa immunoregulatory cytokine the transcription of which is controlled by the ubiquitously expressed transcription factors Sp1 and Sp3. Although many cell types express IL-10 mRNA, not all make detectable amounts of protein, and levels of protein expression vary enormously. We show here that much of this variation can be accounted for by posttranscriptional mechanisms. Multiple copies of potential mRNA destabilizing motifs AUUUA and related sequences can be found to the 3'-untranslated region (UTR) of IL-10 mRNA distributed through three potential regulatory regions. Evidence of RNA-destabilizing activities in all three regions was deduced from luciferase reporter assays. The half-life of RNA containing the 3'-UTR of IL-10 mRNA was quite short in both nonstimulated (t1/2 = 1 h), and PMA-stimulated EL-4 cell (t1/2 = 3 h). In contrast, the half-life of RNA lacking the 3'-UTR was much longer (t1/2 = >12 h) whether cells were stimulated or not. This suggests that many cells are poised to secrete IL-10 and will do so if they receive appropriate posttranscriptional signals.

3' Untranslated Regions↗
PubMed2000

N-(indol-3-ylglyoxylyl)piperidines: high affinity agonists of human GABA-A receptors containing the alpha1 subunit.

A new class of N-(indol-3-ylglyoxylyl)piperidines are high affinity agonists at the benzodiazepine binding site of human GABA-A receptor ion-channels, with modest selectivity for receptors containing the alpha1 subunit over alpha2 and alpha3. All three receptor subtypes discriminate substantially between the two enantiomers of the chiral ligand 10.

GABA-A Receptor Agonists↗
PubMed2000

Allosteric modulators affect the efficacy of partial agonists for recombinant GABA(A) receptors.

Different alpha subunits of human gamma-aminobutyric acid type A (GABA(A)) receptors were transiently expressed together with beta(3) and gamma(2) subunits in Xenopus oocytes to examine the interactions of various GABA(A) agonists and representative allosteric modulators. Chloride currents elicited by agonists were measured using two electrode voltage clamp electrophysiology. Where compounds behaved as full agonists, i.e. GABA on all subtypes and 4,5,6, 7-tetrahydroisoxazolo [5,4-c]pyridin-3-ol (THIP) on alpha2beta(3)gamma(2) GABA(A) receptors, agonist concentration-response curves were shifted to the left by the benzodiazepine full agonist chlordiazepoxide and the anticonvulsant loreclezole, or to the right by the inverse agonist 6, 7-dimethoxy-4-ethyl-beta-carboline-3-carboxylic acid methyl ester (DMCM), with no effect on the maximal currents (I(max)). In contrast, maximal responses for different partial GABA(A) agonists on all benzodiazepine-sensitive alpha(x)beta(3)gamma(2) GABA(A) receptors were enhanced by chlordiazepoxide. I(max) values for piperidine-4-sulphonic acid (P4S) on alpha(1)beta(3)gamma(2), THIP on alpha(3)beta(3)gamma(2), and 5-(4-piperidyl)isothiazol-3-ol (thio-4-PIOL) on alpha(2)beta(3)gamma(2) and alpha(5)beta(3)gamma(2) GABA(A) receptors were increased by chlordiazepoxide, while that for P4S on alpha(1)beta(3)gamma(2) receptors was decreased by DMCM. The I(max) values for partial agonists were also enhanced by pentobarbitone, the neurosteroid allopregnanolone and loreclezole irrespective of receptor subtype or the nature of the partial agonist. In the light of models of ligand-gated ion channel receptor activation we suggest two possible mechanisms of action for the effects of allosteric modulators on partial agonist receptor activation: either selective modulation of agonist affinity for the open/closed state, or direct modulation of the gating process itself.

Allosteric Regulation↗
PubMed2000

Shaping ability of Hero 642 rotary nickel-titanium instruments in simulated root canals: Part 1.

AIM: To determine the general efficacy and shaping ability of Hero 642 nickel-titanium rotary instruments during the preparation of simulated canals. Part 1 of this two-part report describes the efficacy of the instruments in terms of three-dimensional canal form. METHODOLOGY: A total of 40 simulated root canals made up of four different shapes in terms of angle and position of curvature were prepared by Hero 642 instruments using a crown-down preparation sequence. The efficacy of the instruments was assessed in terms of preparation time, instrument failure, canal blockages and loss of canal length. Intra-canal impressions were taken of the prepared canals in order to assess three-dimensional canal form. RESULTS: The mean time for canal preparation was 8.6 min and was influenced significantly (P < 0.05) by canal shape. Two instruments fractured and eight instruments deformed; significant differences were observed between canal shapes (P < 0.05). All of the canals remained patent. Fifteen canals (39.5%) maintained the correct working distance, 15 lost distance and eight canals gained length. Examination of intracanal impressions demonstrated that the majority of canals (79%) had apical stops; canal shape had a significant influence (P < 0.001) on the quality of apical stops. With one exception all canals had smooth canal walls and all of the canals showed good flow characteristics. Taper was poor in 30 canals (79%) and good in eight canals (21%). CONCLUSIONS: Hero 642 rotary nickel-titanium instruments prepared simulated canals rapidly but with a three-dimensional form that lacked adequate taper.

Dental High-Speed Equipment↗
PubMed2000

Shaping ability of Hero 642 rotary nickel-titanium instruments in simulated root canals: Part 2.

AIM: To determine the shaping ability of Hero 642 nickel-titanium rotary instruments during the preparation of simulated canals. METHODOLOGY: A total of 40 simulated root canals made up of four different shapes, in terms of angle and position of curvature, were prepared by Hero 642 instruments using a crown-down preparation sequence. 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. 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. RESULTS: Four zips and four elbows were created during preparation, all in canals with 40 degrees, 12 mm curves. No perforations or danger zones were created. Highly significant differences (P < 0.001) were apparent between the canal shapes in total canal width at the apex and beginning of the curve, and in the amount of resin removed from the inner and outer aspects of the curve at the orifice. Canal transportation was most frequently directed toward the outer aspect of the curve at specific points along the canal, except at the orifice, where it was apparent that canals with 20 degrees curves transported toward the inner. Overall, mean absolute transportation was always less than 0.15 mm; however, significant differences occurred between canal shapes at the end-point (P < 0.01), apex of the curve (P < 0.01) and at the orifice (P < 0.01). CONCLUSIONS: Under the conditions of this study, Hero 642 rotary nickel-titanium instruments created canals with few aberrations and no perforations. The relatively high proportion of aberrations in canals with short, acute curves may indicate that instruments with increased taper should be used with caution at or near the full working distance. Further research in real teeth is necessary to elucidate the full potential of these new rotary instruments for use in root canal preparation.

Dental High-Speed Equipment↗
PubMed2000

An overview of nickel-titanium alloys used in dentistry.

The nickel-titanium alloy Nitinol has been used in the manufacture of endodontic instruments in recent years. Nitinol alloys have greater strength and a lower modulus of elasticity compared with stainless steel alloys. The super-elastic behaviour of Nitinol wires means that on unloading they return to their original shape following deformation. These properties are of interest in endodontology as they allow construction of root canal instruments that utilize these favourable characteristics to provide an advantage when preparing curved canals. This review aims to provide an overview of Nitinol alloys used in dentistry in order for its unique characteristics to be appreciated.

Alloys↗
PubMed2000

Analysis of iceA1 transcription in Helicobacter pylori.

BACKGROUND: Transcription of the Helicobacter pylori iceA1 gene is induced following adherence of the bacterium to gastric epithelial cells in vitro, suggesting that this gene might be involved in H. pylori pathogenesis. Consequently, the current studies were undertaken to characterize iceA1 transcription and to define the structure of iceA1-containing transcripts to evaluate the potential of this gene to encode functional proteins. MATERIALS AND METHODS: Northern blots and primer extension of RNA isolated from broth-grown cultures of various H. pylori strains was done to analyze iceA1-specific gene transcription. Reverse transcriptase (RT)-PCR was used to determine the levels of iceA1 transcripts derived from readthrough transcription that was initiated upstream of iceA1 within the 5'-flanking cysE gene. RESULTS: Three major transcripts were detected and each was initiated from a common promoter, designated PI. Two of these transcripts were comprised of iceA1 sequence, while a third transcript was dicistronic and included the downstream gene, hpyIM. In addition, 10-fold lower levels of iceA1 transcripts were initiated upstream of PI, either within or immediately downstream of cysE. CONCLUSIONS: The present analysis suggests that iceA1 does not encode a functional protein in the majority of H. pylori strains. However, transcription of hpyIM, which encodes a highly conserved DNA adenine methyltransferase, is linked to iceA1 transcription. Therefore, iceA1 may affect H. pylori virulence in vivo through transcriptional regulation of hpyIM expression levels, which may result in specific variations in DNA methylation patterns leading to alteration in the expression of genes involved in virulence or pathogenesis.

Acetyltransferases↗
PubMed2000

Quasispecies development of Helicobacter pylori observed in paired isolates obtained years apart from the same host.

Helicobacter pylori isolates show greater genetic diversity than other bacterial species studied, but the basis for this phenomenon is unknown. Whether detectable genomic mutation appears within an H. pylori population during persistent colonization was investigated. Paired H. pylori populations obtained across 7- to 10-year intervals from 13 patients were characterized by use of methods including polymerase chain reaction (PCR) genotyping for cagA, vacA, iceA, recA, and IS605; random arbitrarily primed DNA (RAPD)-PCR and amplified fragment length polymorphism (AFLP) analysis; and ELISA, to determine Lewis phenotypes. Genotyping, including recA sequence analysis, revealed that initial and follow-up populations represented the same population in 11 patients (85%). Nevertheless, distinct dissimilarities were shown within each of these 11 pairs by both RAPD-PCR and AFLP analyses. During follow-up, Lewis-y levels, but not Lewis-x levels, decreased significantly. The changes detected by RAPD-PCR and AFLP indicate that genetic drift occurs within H. pylori populations over the course of years of colonization of a single host.

Adult↗
PubMed2000

Modulation of glutathione and glutamate-L-cysteine ligase by methylmercury during mouse development.

The antioxidant tripeptide glutathione has been proposed to be important in defense against oxidative stress and heavy metal toxicity. We evaluated alterations in glutathione regulation and synthesis associated with low-level chronic methylmercury (MeHg) exposure in the developing mouse fetus. Female C57Bl/6 mice were given 0, 3, or 10 ppm MeHg in the drinking water for 2 weeks prior to breeding and throughout pregnancy. Fetuses were collected on gestational days (gd) 12 and 16. Total glutathione, reduced glutathione (GSH), oxidized glutathione (GSSR), and glutamate-L-cysteine ligase (Glcl) activity were assessed in yolk sacs and fetuses at gd 16. Western and Northern blots for Glcl-catalytic (Glclc) and Glcl-regulatory (Glclr) subunits were performed on gd 12 and gd 16 fetuses. There were no changes in total glutathione in gd 16 mouse fetuses with exposure, but there were dose-related decreases in GSH and increases in GSSR. In contrast, visceral yolk sacs exhibited an increase in total glutathione in the low-dose groups, but no changes in the high-dose group. There were no changes in Glcl activity in fetuses, but there was a 2-fold increase in Glcl activity in yolk sacs from both low-dose and high-dose groups. There was a 2-fold induction in GLCLC: mRNA and protein in the gd 16 yolk sacs at both 3 and 10 ppm MeHg. No treatment-related changes in Glclr protein in either gd 12 or gd 16 yolk sacs or fetuses were found. Thus, the yolk sac is capable of up-regulating Glclc and GSH synthetic capacity in response to MeHg exposure. This increase appears to be sufficient to resist MeHg-induced GSH depletion in the yolk sac; however fetal glutathione redox status is compromised with exposure to 10 ppm MeHg.

Animals↗
PubMed2000

Campylobacter fetus sap inversion occurs in the absence of RecA function.

Phase variation of Campylobacter fetus surface layer proteins (SLPs) occurs by inversion of a 6.2-kb DNA segment containing the unique sap promoter, permitting expression of a single SLP-encoding gene. Previous work has shown that the C. fetus sap inversion system is RecA dependent. When we challenged a pregnant ewe with a recA mutant of wild-type C. fetus (strain 97-211) that expressed the 97-kDa SLP, 15 of the 16 ovine-passaged isolates expressed the 97-kDa protein. However, one strain (97-209) expressed a 127-kDa SLP, suggesting that chromosomal rearrangement may have occurred to enable SLP switching. Lack of RecA function in strains 97-211 and 97-209 was confirmed by their sensitivity to the DNA-damaging agent methyl methanesulfonate. Southern hybridization and PCR of these strains indicated that the aphA insertion into recA was stably present. However, Southern hybridizations demonstrated that in strain 97-209 inversion had occurred in the sap locus. PCR data confirmed inversion of the 6.2-kb DNA element and indicated that in these recA mutants the sap inversion frequency is reduced by 2 to 3 log(10) units compared to that in the wild type. Thus, although the major sap inversion pathway in C. fetus is RecA dependent, alternative lower-frequency, RecA-independent inversion mechanisms exist.

Animals↗
PubMed2000

Helping Wisconsin women quit smoking: a successful collaboration.

The cost of treatments for tobacco dependence frequently presents a financial barrier to their use. To overcome such barriers, the Wisconsin Women's Health Foundation, the Wisconsin Bureau of Public Health, the McNeil Consumer Healthcare, and the University of Wisconsin Center for Tobacco Research and Intervention collaborated in an initiative to distribute nicotine patches to Wisconsin women at no cost. As a result of this collaborative effort, approximately 19,000 women received a 6-week course of Nicotrol Patches. To evaluate the effectiveness of this initiative, a sample of 500 recipients were contacted and surveyed by telephone 6 months after receiving their patches. Approximately 22% of these women reported total abstinence at 6 months, and another 77% reported they had reduced their smoking. At follow-up, women who had successfully quit rated their health status significantly better than women who were still smoking. More than 99% of respondents recommended that the program be repeated. Extrapolating the observed abstinence rate to the 19,000 patch recipients, an estimated 4000 Wisconsin women successfully quit smoking as a result of this program.

Administration, Cutaneous↗
PubMed2000

Retrospective analysis of an outbreak of B virus infection in a colony of DeBrazza's monkeys (Cercopithecus neglectus).

In 1981, an outbreak of herpetic disease developed in a colony of DeBrazza's monkeys (Cercopithecus neglectus). In seven of eight infected animals, clinical signs of infection included vesicular and ulcerative lesions on the lips, tongue, and/or palate. Histologic examination of lesions revealed intranuclear inclusion bodies, and electron microscopy revealed nucleocapsids and virions with typical herpesvirus morphology. Although a virus was isolated that appeared similar to monkey B virus, techniques available at the time did not allow precise identification of the virus. Analysis of serum from one surviving monkey collected 12 years after the outbreak revealed a pattern of reactivity characteristic of B virus-positive serum on the basis of results of ELISA and western immunoblot analysis. Polymerase chain reaction analysis of archived paraffin-embedded tissue specimens and molecular analysis of the one viral isolate obtained from a DeBrazza's monkey indicated that the virus responsible for the outbreak was a new genotype of B virus. Testing of sera from lion-tailed macaques (Macaca silenus) housed in an adjacent cage at the same zoo indicated that these animals harbored this virus and, thus, were the likely source of the virus that infected the DeBrazza's monkeys. This study documents usefulness of archiving samples from disease outbreaks for later analysis. In addition, this incident underscores the importance of considering herpes B virus infection when outbreaks of disease having characteristics of herpetic infections develop in nonhuman primates kept at institutions that also house macaques.

Animals↗