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R A Wilson

Publications and source records attributed to R A Wilson.

At least 37 records · Page 2Linked to original sources
PubMed2000

Expression of virulence factors among Escherichia coli isolated from the periurethra and urine of children with neurogenic bladder on intermittent catheterization.

BACKGROUND: Patients with neurogenic bladder caused by spinal cord injury or myelomeningocele empty their bladder several times a day by intermittent catheterization. Bacteriuria without symptoms of infection is frequently present in these patients. Occasionally a clone of Escherichia coli that has been carried for weeks without symptoms causes a symptomatic urinary tract infection. Virulence factors are commonly expressed among E. coli causing infection in patients with normal urinary tracts. However, it is unknown whether expression of virulence factors by an E. coli clone colonizing the neurogenic bladder increases the risk of subsequent infection. In this study we examined the prevalence of virulence factor expression among E. coli isolated from the periurethra and urine of patients with neurogenic bladder. METHODS: The prevalence of virulence factors was examined among E. coli isolated from the periurethra and urine in patients with neurogenic bladder who received intermittent catheterization and were followed for 6 months. Representative isolates from the 37 clonal types of E. coli detected in the periurethra and urine of children with neurogenic bladder were assessed for O antigen, hemolysin, aerobactin, serum resistance and type I and P-adhesin. RESULTS: All clones were serum-resistant and expressed type I adhesin, none expressed aerobactin and two expressed hemolysin. The presence of P-adhesin was not unique to clones associated with symptomatic infection. The presence of P-adhesin carried for weeks in a clone did not predict subsequent infection in the neurogenic bladder. CONCLUSION: Bacterial virulence factors did not predict infection of the neurogenic bladder.

Adolescent↗
PubMed2000

Tetanus toxin fragment C expressed in live Salmonella vaccines enhances antibody responses to its fusion partner Schistosoma haematobium glutathione S-transferase.

Tetanus toxoid has been used widely as an adjuvant. The atoxic fragment C from tetanus toxin (TetC) is potently immunogenic when expressed in Salmonella vaccine strains and has been used as a fusion partner for antigens (Ag). However, there has been no formal comparison of the immunomodulatory impact of TetC on its fusion partners. In this study, we have addressed this important issue. The protective 28-kDa glutathione S-transferase (GST) from Schistosoma haematobium (Sh28GST) was expressed either as a fusion to TetC or as the full-length Sh28GST alone in a nonvirulent aroA-attenuated strain of Salmonella enterica serovar Typhimurium. The Sh28GST proteins were soluble and stably expressed in Salmonella, as evaluated by Western blotting with TetC and/or Sh28GST antisera. Mice were immunized orally with a single dose of the live recombinant Salmonella. The constructs were stable in mice but, dramatically, only the strain expressing the TetC-Sh28GST fusion elicited significant antibody (Ab) responses directed against Sh28GST as determined by enzyme-linked immunosorbent assay. An analysis of the isotype profiles showed that these mice also produced anti-Sh28GST immunoglobulin A and GST-neutralizing assays revealed high levels of neutralizing Abs in sera. These are important correlates of protection in schistosomiasis. In addition, stimulation of spleen cells from immunized mice with Sh28GST Ag showed that both strains, expressing Sh28GST alone or the TetC-Sh28GST fusion, were able to stimulate the secretion of Th1-related cytokines (gamma interferon and interleukin 2) to comparable levels. Thus, TetC has modulated the immune responses generated against its fusion partner, Sh28GST, by markedly enhancing the Ab responses elicited. These results have important implications in the rational development of live vaccines.

Adjuvants, Immunologic↗
PubMed2000

Role of bacterial association and penetration on destruction of Escherichia coli O157:H7 in beef tissue by high pH.

This study was undertaken to determine if association with collagen enables Escherichia coli O157:H7 to resist high-pH treatments and to determine the effects of high pH on the survival of E. coli O157:H7 within different layers of beef tissue. E. coli O157:H7 was inoculated onto purified bovine type I collagen on 12-mm2 circular glass coverslips, plain 12-mm2 circular glass coverslips (control), and 12-mm2 irradiated (cobalt-60) lean beef tissue. The rates of destruction of E. coli O157:H7 inoculated on coverslips in pH 10.5 NaHCO3-NaOH buffer at 35 degrees C were determined at various sampling times. E. coli O157:H7 cells associated with collagen and treated in the same manner were also examined using scanning electron microscopy to determine if association with collagen enabled the organism to resist high-pH treatments. The inoculated tissue was treated in pH 13.0 NaHCO3-NaOH buffer at 25 degrees C, and penetrating cells of E. coli O157:H7 were recovered using a cryostat technique. There was no significant difference (P < 0.05) between the rates of destruction of collagen-associated E. coli O157:H7 and non-collagen-associated E. coli O157:H7 following exposure to high-pH treatments. Scanning electron micrographs showed that collagen-associated E. coli O157:H7 cells appeared physically damaged by exposure to high-pH treatments, and association of E. coli O157:H7 to collagen did not increase the resistance of the organism to destruction by high-pH rinses. No significant differences were seen between 20 ml of NaHCO3-NaOH buffer at pH 13.0 (treatment) and 20 ml of distilled water at pH 7.0 (control) when E. coli O157:H7 cells were recovered in beef tissue at depths of up to 2,000 microm (P < 0.05). The ability of E. coli O157:H7 to penetrate beef tissue may be an important factor in reducing the effectiveness of high-pH treatments in killing this organism on beef tissue. This finding should be considered in the future when designing treatments to decontaminate beef carcasses.

Animals↗
PubMed2000

Proximal femoral fracture in Northern Ireland between 1985-1997--trends and future projections.

The aims of this study were to identify changing trends with time of the incidence of proximal femoral fracture and to enable future number of hip fractures to be projected. Hospital theatre records in Northern Ireland were surveyed in 1985, 1991, 1994 and 1997 to establish the number of surgical procedures for proximal femoral fracture. The age and sex specific rates for males and females aged 50+ years were calculated. Analysis of age and sex specific incidence rates was undertaken using linear regression and Poisson regression. A 1.6% increase per annum (95% CI 1.0-2.2) was noted in males and females from 1985 to 1997. Projected increases in rates of proximal femoral fracture were calculated using population projection for 2001, 2006, 2011 and 2016. Modelling (a) assuming the age standardised rates in 1997 remain static and continue predicted a 55% fracture increase in males and a 29% increase in females by 2016, (b) assuming the secular increases continued predicted a 93% fracture increase in males and a 67% increase in females and (c) assuming further linear growth on a log-scale predicted a fracture increase in males of 135% and 99% in females. The number of proximal femoral fractures in Northern Ireland is increasing faster than that anticipated due to demographic changes alone, supporting a secular increase which was evident throughout the period of time studied, in contrast to that reported from other regions in the United Kingdom.

Age of Onset↗
PubMed1999

TNF is essential for the cell-mediated protective immunity induced by the radiation-attenuated schistosome vaccine.

C57BL/6 mice exposed to the radiation-attenuated schistosome vaccine exhibit high levels of protective immunity. The cell-mediated pulmonary effector mechanism involves IFN-gamma-producing CD4+ T cells in a focal response around challenge larvae. IFN-gamma can promote production of TNF and can synergize with this cytokine in its actions on responder cells. We have examined whether TNF plays a role in lung phase immunity to schistosomes using mice with a disrupted gene for TNFRI (TNFRI-/-). The most dramatic finding was that the schistosome vaccine elicited no protection whatsoever in these mice. However, this could not be attributed to a lack of responder cells, because more lymphocytes were lavaged from the airways of TNFRI-/- than wild-type mice. Furthermore, CD4+ T cells were equally represented in airway populations from the two groups and produced IFN-gamma upon Ag stimulation in vitro. In contrast, pulmonary macrophage function was defective in TNFRI-/- mice, as indicated by a failure to up-regulate inducible NO synthase mRNA. Histopathological analysis revealed that focal infiltrates were of similar size and cell composition in the two groups but that more parasites were free of foci in the TNFRI-/- mice. These animals had a greatly impaired IgG response to schistosomes, which may explain their lack of residual protection due to Ab in a situation where cell-mediated immunity is disabled. We suggest that the absence of protective immunity could result from a retarded build-up of leukocytes around migrating lung worms and/or a deficit in accessory cell function within a focus, both of which would permit parasite escape.

Animals↗
PubMed1999

Immune responses to the radiation-attenuated schistosome vaccine: what can we learn from knock-out mice?

The goal of an effective schistosomiasis vaccine has proved elusive but the protective immunity induced in mice by radiation-attenuated cercaria larvae provides an appropriate model from which such a vaccine might be developed. Using gene-disrupted mice, we have analysed the process of immune priming by attenuated larvae of Schistosoma mansoni and the nature of the pulmonary effector response directed against a challenge infection. Vaccination stimulates expansion of IFNgamma-producing T-helper cells in the skin-draining lymph nodes. IL-12 is crucial in determining the Thl direction of this initial response but the cells of origin and the parasite components which stimulate its production are unknown. In the effector response, focal aggregates comprising mainly mononuclear cells accumulate around challenge larvae in the lungs, a process orchestrated by IFNgamma. This cytokine up-regulates nitric oxide synthase activity but we were unable to implicate nitric oxide as a cytotoxic agent causing challenge parasite elimination. An alternative action for IFNgamma may be to up-regulate adhesion molecule expression, increasing the cohesiveness of effector foci the better to block parasite migration, but the adhesive interactions so far examined do not appear relevant. In contrast, TNF induction is essential to protection, and we are currently testing the hypothesis that it determines the speed of the effector response following arrival of challenge larvae in the lungs.

Animals↗
PubMed1999

Characterization of eae+ Escherichia coli isolated from healthy and diarrheic calves.

Strains of Escherichia coli from 101 healthy and 114 diarrheic calves were screened by PCR for the eae (intimin) gene and Shiga toxin genes (stx). Each eae+ and eae/stx+ strain was examined for antimicrobial susceptibility, enterohemolysin activity, and the somatic O antigen was determined. An immunoassay was used to detect Shiga toxin antigens for the eae/stx+ E. coli. Significantly more (p = 0.005) of the healthy calves carried eae+ and eae/stx+ E. coli in their feces when compared to strains from diarrheic calves. Moreover, Shiga toxin antigens were detected significantly more (p = 0.001) often among the eae/stx+ strains from healthy calves when compared to eae/stx+ strains from diarrheic calves. However, significantly more (p = 0.001) of the eae+ and eae/stx+ strains from diarrheic calves were resistant to at least one of the antimicrobials tested, and the strains from diarrheic calves had a significantly (p = 0.05) higher rate of antimicrobial resistance to at least two different antimicrobial classes. No significant difference (p> or =0.05) was detected among the eae+ and eae/stx+ strains from healthy and diarrheic calves for enterohemolysin production. Serogroups O-negative, O5, O26, and O111 were predominate among both healthy and diarrheic calves.

Adhesins, Bacterial↗
PubMed1999

Characterization of Escherichia coli isolated from healthy dogs.

Five month old dogs from a Midwestern research kennel occasionally developed bloody diarrhea after shipment to other facilities. As previous diagnostic efforts failed to reveal any potential pathogens in feces from normal and diarrheic dogs, Escherichia coli was investigated for select virulence properties that may contribute to the occurrence of bloody diarrhea. Fecal swabs from 52 healthy dogs were examined for E. coli. Two hundred and sixty E. coli-like colonies were screened by PCR for the attaching and effacing (eae) gene, Shiga toxin (stx) genes, and the heat-stable enterotoxin type A (sta) gene. One hundred forty two of the 260 E. coli-like colonies (54.6%) from 43 dogs were eae or sta positive; and 60 of the eae and/or sta positive isolates were examined further. Among the 60 isolates, 23 (38.3%) possessed the eae gene, 32 (53.3%) possessed the sta gene, and five (8.3%) possessed both eae and sta genes (eae+/sta+). Of the 60 isolates, six sta+ and one eae+/sta+ isolates were hemolytic. When examined in the suckling mouse assay, five of six sta+ isolates and three of four eae+/sta+ isolates gave gut-to-remaining carcass ratios > or =0.083, indicating expression of heat-stable enterotoxin. These enterotoxin-producing isolates belonged to serogroups O42, O170, and O-negative.

Animals↗
PubMed1999

Characterization, cloning and immunogenicity of antigens released by lung-stage larvae of Schistosoma mansoni.

Lung-stage schistosomula are the target of protective immunity in mice vaccinated with attenuated cercariae of Schistosoma mansoni. Therefore, proteins present at this developmental stage, and in particular those which are secreted, are a potential source of novel vaccine candidates. However, little information is available about such molecules. Here we describe the cDNA clones identified by screening expression libraries with serum raised against proteins released by lung-stage schistosomula. In total, 11 different cDNA species were identified, 6 of which have been described previously in S. mansoni; these included fructose 1,6-bisphosphate aldolase and Sm21.7 which together accounted for two-thirds of all positive clones. Of the 5 newly described schistosome genes, 1 cDNA had a high degree of homology to the s5a subunit of 26S proteasomes, most significant being with the human protein. The remaining 4 clones showed no significant homologies to any genes sequenced previously. Fructose 1,6-bisphosphate aldolase, Sm21.7, the proteasome homologue and 1 unknown clone (A26) have been expressed in a bacterial expression system and serum produced against each recombinant protein. Immunolocalization showed fructose 1,6-bisphosphate aldolase, Sm21.7 and the proteasome homologue to be most abundant in muscle cells whilst clone A26 was distributed throughout many tissues, but was most abundant in the tegument. Analysis of the cellular immune responses of vaccinated mice showed 3 of the 4 expressed clones to be highly immunogenic, inducing the secretion of large quantities of the Th1-type cytokine interferon gamma.

Amino Acid Sequence↗
PubMed1999

The radiation-attenuated schistosome vaccine induces high levels of protective immunity in the absence of B cells.

Radiation-attenuated cercariae of Schistosoma mansoni elicit consistently high levels of protective immunity in mice. The cell-mediated pulmonary effector mechanisms have been well characterized but the role of B cells and antibodies remains ill defined. We have compared the immune responses of B-cell-deficient (muMT) mice and their wild-type (WT) counterparts following exposure to the attenuated vaccine. Both groups mounted a T helper type 1 (Th1)-biased response in the skin-draining lymph nodes after vaccination. Interferon-gamma was the dominant cytokine secreted by airway leucocytes after challenge in both muMT and WT mice, but there was a somewhat greater Th2 component in the former animals. The cellular infiltrates observed in the airways, and the pulmonary effector foci, were of similar composition in the two groups although some large foci were present in the muMT mice. There was a marked dichotomy in the protection induced in muMT animals by a single vaccination, with two-thirds showing levels similar to their WT counterparts, demonstrating that cell-mediated mechanisms alone can provide adequate protection. The remaining muMT mice had a mean worm burden identical to that of their challenge controls. A possible explanation is that a proportion of the muMT animals have a genetic defect closely associated with the mu-heavy-chain locus on chromosome 12, which affects their ability to mount a protective cell-mediated response. Three vaccinations enhanced the immunity of WT animals, most likely by augmenting antibody-mediated mechanisms. In contrast, no enhancement was seen in muMT mice, suggesting that the cell-mediated response is not boosted by multiple exposures to attenuated larvae.

Animals↗
PubMed1999

Interleukin-12 can directly induce T-helper 1 responses in interferon-gamma (IFN-gamma) receptor-deficient mice, but requires IFN-gamma signalling to downregulate T-helper 2 responses.

An in vivo model of pulmonary granuloma formation around embolized schistosome eggs was investigated as an environment in which to analyse a role for interleukin-12 (IL-12) in the differentiation of T-helper 1 (Th1) and Th2 subsets. Specifically, mice deficient for the interferon-gamma receptor (IFN-gammaR-/-) were used to determine the role for IL-12 in the absence of IFN-gamma-mediated signalling. We show that recombinant IL-12 administered to IFN-gammaR-/- mice caused the up-regulation of mRNA for IFN-gamma in lung tissue, and the secretion of abundant IFN-gamma by in vitro-cultured lymph node cells in response to egg antigens. This indicates that IL-12 can act independently of IFN-gamma to induce the development of Th1 cells. Administration of rIL-12 to wild-type mice markedly reduced the secretion of Th2-associated cytokines, IL-4 and IL-5. However, these cytokines were not dramatically reduced in IFN-gammaR-/- mice treated with IL-12. We conclude that inhibition of these cytokines by IL-12 is primarily dependent upon effective IFN-gamma signalling, although abrogation of T-cell derived IL-10 appeared to be dependent upon IL-12. We also show that increases in mRNA for the beta2 subunit of the IL-12 receptor and the p40 subunit of IL-12 after rIL-12 treatment were lower in IFN-gammaR-/- mice, compared to wild-type mice, indicating that their expression was primarily dependent upon IFN-gamma with only a minor role for IL-12.

Animals↗
PubMed1999

Correlation of periurethral bacterial flora with bacteriuria and urinary tract infection in children with neurogenic bladder receiving intermittent catheterization.

Periurethral bacteria are inoculated daily into the urine of children with neurogenic bladder during clean intermittent catheterization (CIC). We examined how frequently periurethral bacterial species produced bacteriuria in children followed longitudinally. When Escherichia coli was detected on the periurethra, bacteriuria was also present 93% of the time. When Klebsiella, Pseudomonas, or Enterococcus species or nonpathogens were detected on the periurethra, bacteriuria was present 80%, 40%, 40%, and 25% of the time, respectively. Clonal typing of multiple colonies of E. coli from each periurethral and urine culture revealed that children carried only one or two E. coli clones in their urinary tracts over months of surveillance. When E. coli was detected in the urine, the identical clone was on the periurethra. E. coli persisted for weeks in the urine without causing symptoms. Occasionally the same E. coli clone carried for weeks caused a urinary tract infection. Bacteriuria frequently occurs after inoculation of periurethral E. coli into the urine during CIC.

Bacteriuria↗
PubMed1999

Genetic diversity among Escherichia coli isolates carrying f18 genes from pigs with porcine postweaning diarrhea and edema disease.

Multilocus enzyme electrophoresis was applied to detect allelic variation and multilocus genotypes (electrophoretic types [ETs]) among 43 Escherichia coli isolates from weaned pigs suffering from edema disease or from diarrhea. ETs were analyzed in relation to O serogroups and virulence genes (sta, stb, lt, stx2, and f18) by DNA hybridization. Genomic diversity was the lowest in serogroup O138, while virulence genes (stx2 and f18) were the most uniform in serogroup O139. In general, the serogroups or toxin and F18 fimbria types were not related to selected ETs, suggesting that the toxin and f18 fimbria genes in E. coli isolates from pigs with postweaning diarrhea or edema disease occur in a variety of chromosomal backgrounds.

Animals↗
PubMed1999

Formation of multinucleated giant cells in the mouse lung is promoted in the absence of interleukin-12.

The formation of multinucleated giant cells (MGCs) in an in vivo model of pulmonary inflammation was investigated to determine whether these cells are the result of a dominant T helper (Th) 1 or Th2 cytokine environment. We report that knockout (KO) mice with a disrupted interleukin (IL)-12 p40 gene exposed to the helminth Schistosoma mansoni had abundant and very large MGCs (> 50 microm) in their lungs concurrent with extensive eosinophilia and a population of large macrophages. Many of the MGCs and macrophages appeared to have phagocytosed eosinophils as part of a clearance process. The KO mice also had a strongly polarized Th2 immune response as judged by elevated levels in the lungs of messenger RNA (mRNA) transcripts for IL-4, IL-5, IL-6, and IL-13, but decreased levels of mRNA for interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). In addition, cells recovered by bronchoalveolar lavage from the airways of these mice secreted a Th2-biased profile of cytokines upon restimulation in vitro with parasite antigen. In contrast, wild-type C57BL/6 or KO mice treated with recombinant IL-12 had a polarized Th1 phenotype with elevated levels of IFN-gamma and TNF-alpha mRNA in the lungs, and an airway cell population that secreted abundant IFN-gamma. Very few MGCs were detected in these mice, and there was an absence of pulmonary eosinophilia. We conclude that the formation of MGCs in our model is promoted in the absence of IL-12 and is linked instead to the abundance of Th2 cytokines, notably IL-4 and IL-13.

Animals↗
PubMed1999

Factors in cardiac risk stratification of candidates for renal transplant.

BACKGROUND: Renal transplant candidates are at high risk of fatal and nonfatal cardiac events. METHODS: This study evaluated five clinical risk factors--age at least 50 years, insulin-requiring diabetes mellitus, angina, congestive heart failure and an abnormal electrocardiogram (ECG) (excluding left ventricular hypertrophy)--that had been used in the first tier of a two-tiered prospectively applied risk stratification algorithm. RESULTS: Using multiple logistic regression analysis, age at least 50 years, abnormal ECG, and diabetes mellitus were independently predictive of cardiac death. Of the two remaining clinical risk factors, the presence of angina had independent predictive value for nonfatal cardiac events (myocardial infarction, coronary angioplasty, bypass surgery, and unstable angina). The independent predictive value of congestive heart failure approached statistical significance. CONCLUSION: Clinical risk-factor analysis is helpful in identifying renal transplant candidates at high risk for fatal or nonfatal cardiac events.

Algorithms↗
PubMed1999

Escherichia coli O157:H7 in retail ground beef in Seattle: results of a one-year prospective study.

Escherichia coli O157:H7 was sought systematically in 1,400 samples of retail ground beef in Seattle in a 1-year prospective study. Sorbitol-nonfermenting, lactose-fermenting, indole-positive colonies isolated after enrichment culture were probed for the presence of Shiga toxin genes. Totals of 67,040 sorbitol-nonfermenting and 66,705 sorbitol-fermenting colonies were characterized, but E. coli O157:H7 was not identified. The sensitivity of this technique was usually sufficient to detect E. coli O157:H7 at a concentration below 1 CFU/g of meat. These data demonstrate that retail ground beef in Seattle is neither frequently nor heavily contaminated with E. coli O157:H7.

Animals↗
PubMed1999

Comparison of laparoscopic and open ventral herniorrhaphy.

The repair of large and/or recurrent ventral hernias is associated with significant complications and a recurrence rate that can be more than 50 per cent. Laparoscopic ventral herniorrhaphy, a recent development, has been shown to be safe and effective in the repair of ventral hernias. This study retrospectively reviews all ventral hernia repairs over a 3-year period, November 1995 through December 1998, at a community-based teaching hospital. The purpose of the study was to compare open and laparoscopic repairs. A total of 253 ventral hernia repairs were performed during this time, 174 open and 79 laparoscopic. The age, weight, and sex distribution was similar for each group. The hernias in the open group averaged 34.1 cm2 in size, and mesh used averaged 47.3 cm2. In the laparoscopic group, the hernia defect averaged 73.0 cm2, and the mesh size averaged 287.4 cm2. Operative time was longer in the open group, 82.0 versus 58.0 minutes. In the open group, there were 38 (21.8%) minor and 8 (4.6%) major complications, compared with 13 (16.5%) minor and 2 (2.5%) major complications in the laparoscopic group. Hospital stay was shorter for the laparoscopic group, 1.7 versus 2.8 days. At an average follow-up of 21 months (range, 2-40 months), there have been 36 recurrences in the open group (20.7%) compared with 2 recurrences in the laparoscopic group (2.5%). In this series, laparoscopic ventral herniorrhaphy compares favorably to open ventral herniorrhaphy with respect to wound complications, hospital stay, operative time, and recurrence rate.

Abdomen↗