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Biomedical subjects

J L Duncan

Publications and source records attributed to J L Duncan.

At least 73 records · Page 4Linked to original sources

A comparison of the responses to repeated experimental infections with Haemonchus contortus among Scottish Blackface lambs.

Twenty helminth-naive Scottish Blackface lambs were given three infections with 10,000 infective larvae of Haemonchus contortus at 8 week intervals. An additional six lambs served as uninfected controls and eight lambs were infectivity controls. The lambs were 7 months old at the start of the infection. Four of the 20 lambs developed severe haemonchosis and were put down during the experiment. The remaining 16 lambs plus uninfected controls were necropsied 8 weeks after the third infection. The mean faecal egg count peaked 6-8 weeks after the first infection, gave a second smaller peak 6-8 weeks after the second infection but produced no peak after the third infection. Mean red blood cell counts fell rapidly during the first infection, then rose gradually during the second and third infections. The mean values suggested that two infections were sufficient to produce effective immunity in the sheep population but they masked considerable individual variation. Eleven animals appeared relatively resistant following the first infection, while two animals were relatively susceptible to even the third infection. The repeatability of mean faecal egg counts or mean red blood cell counts for each animal during the replicate infections were very high, because the rankings of the individual sheep remained remarkably stable. Faecal egg counts were very strongly correlated with red blood cell counts. Multiple regression analysis showed that four factors--faecal egg counts, red blood cell counts, weight and sex--accounted for essentially all of the observed variation in worm burdens among the lambs. Therefore, under these controlled experimental conditions, additional markers appear unnecessary for the detection of resistance status.

Analysis of Variance↗

World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) second edition of guidelines for evaluating the efficacy of anthelmintics in ruminants (bovine, ovine, caprine)

The first edition of the W.A.A.V.P. anthelmintic guidelines for ruminants was published in 1982. Since then improved parasitological procedures have been developed, new therapeutic and prophylactic products have appeared requiring different test methods, and registration authorities are requesting more detailed record keeping and data validation. This second edition addresses these developments and fulfills the original goal of publishing guidelines for high quality, scientifically valid testing standards for trials that would be accepted as proof of efficacy by registration authorities regardless of country of origin. This second edition includes updated guidance on standard parasitological procedures, dose titration, dose confirmation and clinical trials, and provides guidelines for evaluating products for efficacy against anthelmintic resistant parasites, persistence of activity and prophylactic activity. Tests for efficacy against nematodes, trematodes and cestodes are included.

Animals↗

Platelet count and the outcome of operation for ruptured abdominal aortic aneurysm.

PURPOSE: The purpose of this study was to determine the relationship between admission and postoperative platelet count (PC) and mortality and morbidity rates after emergency surgery for ruptured abdominal aortic aneurysm (AAA). METHODS: We performed a case record review of 65 consecutive patients admitted with ruptured AAA. RESULTS: Five patients did not undergo operation because of poor clinical condition, 12 patients died at operation, 13 patients died after operation, and 35 patients survived (operative mortality rate of 58%). Fifteen patients had an admission PC of less than 150 x 10(9)/L, of whom 14 (93%) died; 27 patients had an admission PC of 150 to 250 x 10(9)/L, of whom 8 (30%) died; and 18 patients had an admission PC of greater than 250 x 10(9)/L of whom 3 (17%) died. At the end of operation 29 patients had a PC of less than 100 x 10(9)/L, of whom 13 (45%) died, 20 (69%) had development of multiorgan failure (MOF), and 6 (21%) required relaparotomy for bleeding complications. By contrast, all 19 patients having a PC of 100 x 10(9)/L or greater at the end of the operation survived, three (16%) had development of MOF, and none required further surgery for bleeding. There was a significant inverse correlation between PC at the end of the operation and the number of postoperative days spent on a ventilator, in the intensive therapy unit, and in hospital. After operation, 15 patients had development of thrombocytosis (PC > 400 x 10(9)/L), of whom 10 had development of deep venous thrombosis and 8 had development of pulmonary embolism. There were no deep venous thromboses or pulmonary embolisms in patients who did not have development of a postoperative thrombocytosis. CONCLUSIONS: There is a direct correlation between PC on admission to the hospital and death after emergency repair of ruptured AAA. Thrombocytopenia at the end of the operation is associated with a high risk of morbidity from continuing hemorrhage or MOF. The development of postoperative thrombocytosis is associated with a high risk of thromboembolic complications. PC provides a simple marker of outcome in patients diagnosed with and undergoing operation for ruptured AAA.

Aged↗

An immunodominant antigen on the Dictyocaulus viviparus L3 sheath surface coat and a related molecule in other strongylid nematodes.

Monoclonal antibody 2A6 binds to the surface of the Dictyocaulus viviparus L3 sheath and identifies a molecule which is highly antigenic in both infected and vaccinated cattle. Immunogold electron microscopy reveals that the antibody binds to a highly stable surface coat overlying the epicuticle of the L3 sheath. The binding to the cuticular surface is stage specific, being restricted to the surface of the L3 sheath, although the antigen can be detected on Western blots of parasite homogenates from the L1 stage. Immunofluorescence on live parasites demonstrates that the antibodies also bind to the L3 sheath surface of many other strongylid nematodes. The antigen detected by 2A6 differs in molecular weight between the species; in D. viviparus the antigen is poorly resolved between 29 and 40 kDa by SDS-PAGE whereas in nematodes from the family Trichostrongyloidea a single band or doublet of approximately 20 kDa is detected, suggesting that a related molecule is present in these species. Although infection and vaccination with D. viviparus elicits a strong immune response to the L3 sheath antigen this does not appear to be the case following infection with other strongylid nematodes.

Animals↗

The distribution of faecal nematode egg counts in Scottish Blackface lambs following natural, predominantly Ostertagia circumcincta infection.

Faecal samples were taken in 2 consecutive years from Scottish Blackface lambs on a commercial farm in central Scotland to examine variation among lambs in the number of nematode eggs in their faeces. Samples were taken at intervals of 4 weeks from 1 to 5 months of age. Lambs were treated with a broad-spectrum anthelmintic at each sample date. Ewes and lambs were naturally infected by grazing pasture contaminated with mixed, predominantly Ostertagia circumcincta, nematode species. The ewes were removed from the lambs when the lambs were three months old. Only 3 of the 6 sets of samples taken prior to removal of the ewes fitted a negative binomial distribution but all sets of samples taken after this did so. The mean and the variance differed with the month and year of sampling. The amount of variation among lambs increased as the lambs grew from 3 to 5 months of age, possibly because of heterogeneity in the rate of development of protective responses. Counts taken in August and September, when the lambs were 4 and 5 months of age, were positively correlated with each other. In contrast, counts taken at 1 or 2 months of age were negatively correlated with counts taken when the lambs were 3-5 months old. Sex, date of birth, sire and dam were all associated with variation in faecal egg counts but the size and significance of their influence was dependent upon the date when faecal samples were taken. Four factors: sire, dam, sex and year accounted for over two-thirds of the variation among older lambs in faecal egg counts.

Animal Husbandry↗

Effect of vaginal fluid on adherence of type 1 piliated Escherichia coli to epithelial cells.

Colonization of the vaginal mucosa with uropathogens from fecal flora is an important step in ascending urinary tract infections (UTIs) in women. Colonization is influenced by interactions between uropathogens, vaginal fluid, and epithelial cells. In this study, vaginal fluid from 21 women was assessed for effects on adherence of type 1 piliated Escherichia coli to the A431 cell line. Adherence to cells was enhanced by all fluid specimens when tested at low fluid protein concentrations in an in vitro assay. At higher concentrations, certain specimens maintained enhanced binding whereas others resulted in diminished binding. Increases in adherence were associated with increased binding of E. coli to vaginal fluid in vitro and with higher vaginal fluid pH. These results demonstrate that vaginal fluid significantly alters the adherence of type 1 piliated E. coli to epithelial cells in vitro and, therefore, should be studied as a potential modifier in bacterial colonization and UTIs in vivo.

Adult↗

Regulation of egg production, worm burden, worm length and worm fecundity by host responses in sheep infected with Ostertagia circumcincta.

Following infection with Ostertagia circumcincta there was considerable variation in worm burdens, worm size and number of inhibited larvae even among sheep matched for age, sex, breed, farm of origin and history of parasite exposure. There was also substantial variation among sheep in the concentration of mast cells, globule leucocytes, eosinophils, IgA-positive plasma cells and parasite-specific IgA in the abomasal mucosa. With the exception of faecal egg counts over time, the parasitological and immunological traits were all continually distributed among animals and sheep did not fall into discrete high and low-responder categories. The responses were correlated. Sheep with more mast cells also had more globule leucocytes, more eosinophils, more IgA plasma cells and greater amounts of parasite-specific IgA in the abomasal mucosa. Female worm length was strongly and positively correlated with the number of eggs in utero. Faecal egg counts were associated with variation in worm number and with variation in the number of eggs in utero. The worm burden was negatively correlated with the number of globule leucocytes in the abomasal mucosa, suggesting that worm numbers are regulated by immediate hypersensitivity reactions. Decreased female worm length was associated with an increased local IgA response to fourth stage larvae. The number of inhibited larvae was positively associated with the size of the local IgA response and positively associated with the size of the worm burden. The results suggest that variation among mature sheep in faecal egg counts is due, at least in part, to variation in local IgA responses which regulate worm fecundity and to variation in local immediate hypersensitivity reactions which regulate worm burdens.

Animals↗

Binding of type 1-piliated Escherichia coli to vaginal mucus.

To better understand the interactions involved in bacterial adherence and the role of mucus in the pathogenesis of urinary tract infections, we developed a system to study the binding of a recombinant Escherichia coli strain, HB101/pWRS1-17, expressing type 1 pili, to vaginal mucus collected from 28 women. Bacteria bound to differing extents to all specimens examined, and preincubation of bacteria with mannose inhibited binding by 50 to 89%. Additionally, all mucus samples showed reactivity with anti-mannose antibody, and the levels of reactivity correlated with the levels of bacterial binding, suggesting that the mannose-terminal saccharides present on these glycoproteins are the receptors for the binding of type 1-piliated bacteria. Mucus specimens collected over periods of 5 days and 12 weeks exhibited significant variation in bacterial binding, indicating temporal differences in the ability of vaginal mucus to act as a receptor for type 1-piliated E. coli. The results show that vaginal mucus can bind bacteria and may thus influence the initial attachment and subsequent colonization of the vaginal and urinary tract epithelium by E. coli.

Bacterial Adhesion↗

Analysis of the fimB promoter region involved in type 1 pilus phase variation in Escherichia coli.

Many strains of Escherichia coli express type 1 pili which undergo phase variation during growth in broth, but become uniformly nonpiliated when passaged on agar media. In a previous analysis of these agar phase locked strains, we demonstrated that fimB and fimE, which mediate the site-specific DNA rearrangement involved in phase variation, are differentially transcribed under the two growth conditions. In this study the fimB promoter region was sequenced and characterized in agar phase locked strain J96. Primer extension analysis of the fimB gene identified three putative transcription initiation sites. Transcription starting from two of the sites (P1 and P2) would produce an mRNA that approximates the transcript size of fimB previously detected by Northern blot hybridizations. Gel mobility shift studies revealed the presence of promoter binding activity in broth-grown cell extracts, but not in extracts from agar-grown cells, that reacted specifically with DNA fragments upstream of the P1 and P2 promoter regions. This putative binding protein may be involved in the regulation of fimB, perhaps by acting as a transcriptional activator.

Bacterial Proteins↗

The immunogenicity of the acetylcholinesterases of the cattle lungworm Dictyocaulus viviparus.

Somatic extracts and excretory/secretory (ES) products of the adult stage of the cattle nematode, Dictyocaulus viviparus, were examined for acetylcholinesterase (AChE) activity. Both were found to contain activity which had an optimum pH of 9.5, however, the adult ES products contained over 200 times more AChE activity per unit protein. Gel electrophoresis and specific enzyme staining revealed 5 migratory isoforms of AChE which were common to adult ES products and adult homogenates. Comparison of L3 with L4 and adult extracts indicated that the AChE were only produced by later developmental stages of this parasite. The antigenicity of D. viviparus AChE was demonstrated by binding to serum IgG from naturally and experimentally infected calves but the enzymes were not recognized by calves vaccinated twice with 400 Gy-irradiated larvae. This is the first report of helminth AChE release by a parasitic nematode in a pulmonary location. The presence of these enzymes in such high amounts in the ES products, along with their immunogenicity, suggests that they might have an important role to play in the immunobiology of D. viviparus in the lungs.

Acetylcholinesterase↗

Variation of blood group antigen expression on vaginal cells and mucus in secretor and nonsecretor women.

The expression of blood group antigens on the surface of urothelial cells and in mucus is controlled partly by the blood type and secretor status of the individual. To our knowledge, the possibility that the levels of these antigens vary with time has not been previously assessed. We determine if the pattern and/or intensity of blood group antigen expression on vaginal epithelial cells and mucus changed with time. Cell and mucus specimens were collected weekly for 3 months from 10 women: 5 (2 secretors and 3 nonsecretors) with and 5 (3 secretors and 2 nonsecretors) without a history of urinary tract infections. In addition, samples were collected on 5 consecutive days from 5 of these individuals. The cell and mucus samples were assayed for ABH and Lewis blood group antigens using monoclonal antibodies in cell concentration immunofluorescence and enzyme-linked fluorogenic assays, respectively. Although the pattern of antigen expression in the vaginal cell and mucus samples was consistent with the blood type and secretor status of an individual, in all women the level of antigen expression changed significantly and rapidly during the 3-month and 5-day periods. The results show a previously unrecognized phenomenon and demonstrate that the expression of blood group antigens on vaginal cells and in mucus is a dynamic process.

ABO Blood-Group System↗

Blood group antigen expression on vaginal cells and mucus in women with and without a history of urinary tract infections.

Adherence of bacteria to carbohydrate receptors on the surface of vaginal epithelial cells is a critical event that precedes bacterial colonization of the vaginal mucosa and subsequent urinary tract infection. Blood group antigens, carbohydrate structures bound to lipids or proteins, constitute an important component of the uroepithelial cell membrane and may affect susceptibility to urinary tract infections. To determine if the ABH and Lewis antigen expression in women with a history of urinary tract infections differed from that of women without such a history, vaginal specimens from 87 women (42 patients and 45 controls) were analyzed for the detection of these antigenic determinants using monoclonal antibodies in quantitative immunoassays. The profile of ABH antigen expression was generally consistent with the ABO phenotype of the individual and appeared to be influenced by the secretor status. Secretors expressed higher levels of A, B and H determinants than nonsecretors. In addition, Lewis antigens were detected on vaginal cells and in mucus. Samples from nonsecretors expressed higher levels of Le(a) and Le(x) antigens, whereas secretors expressed higher levels of Le(b) and Le(y) antigens. The levels of antigen expression varied widely among individuals with the same blood type and secretor status. Comparisons between patient and control groups showed no significant differences in ABH or Lewis antigen expression overall, or when controlling for ABO or secretor phenotypes, respectively. These findings confirm our previous observations on healthy women, and document the heterogeneity of blood group antigen expression on vaginal epithelial cells and mucus from women with a history of urinary tract infections.

ABO Blood-Group System↗

Genetic control of the antibody repertoire against excretory/secretory products and acetylcholinesterases of Dictyocaulus viviparus.

Outbred Dunkin-Hartley and inbred strain 2 and strain 13 guinea pigs were immunized with Dictyocaulus viviparus adult ES products prior to challenge with third stage larvae. Antibody responses of the three strains to adult ES products and the acetylcholinesterase (AChE) isoforms which they contain were examined. Using immunoprecipitation and ELISA, it was observed that responses in the three strains to adult ES products were distinct: considerable heterogeneity in the antibody repertoire was observed between outbred Dunkin-Hartley animals, with only slight variation occurring amongst the inbred individuals. Responses to the AChE isoforms were heterogeneous amongst individual outbred guinea pigs but were more consistent in inbred strain 2 and 13 animals in which strain-specific patterns of recognition were observed. Previous studies with nematode infections have indicated a role for the major histocompatibility complex in determining the nature and level of the immune response. As the inbred strains bear different alleles at the Class II region but are identical at the Class I region, the differences observed are likely to be due to genes mapping to the Class II locus. This is therefore the first report of genetic restriction of the antibody repertoire to secreted AChEs of a parasitic nematode.

Acetylcholinesterase↗

Effect of Lewis blood group antigen expression on bacterial adherence to COS-1 cells.

Epithelial cells from secretor individuals demonstrate decreased bacterial adherence compared with cells from nonsecretors. Lewis blood group antigen expression is one component of the secretor/nonsecretor phenotype and several epidemiologic studies have suggested a link between Lewis blood group antigen phenotype and susceptibility to urinary tract infections. In this study, we examined the possibility that the expression of the difucosylated Lewis blood group determinants, Leb and Ley (associated with the secretor phenotype), made cells less susceptible to Escherichia coli adherence by masking receptors for pili. COS-1 cells, which do not produce Lewis (Lea, Leb, Le(x), and Ley) blood group antigens, were used as target cells for bacterial adherence. The surface blood group antigen expression pattern of the cells was then modified by cotransfection with plasmids containing DNA inserts encoding alpha (1,2)-fucosyltransferase and alpha (1,3)- and alpha (1,4)-fucosyltransferases, resulting in the expression of Leb and Ley. E. coli HB101 expressing various adhesins (type 1, PapJ96, PapIA2, PapAD110, Prs, and S) from recombinant plasmids bound equally well to untransfected cells and transfected cells expressing Lea and Le(x) (nonsecretor phenotype) and Leb and Ley (secretor phenotype) antigens. We conclude that the presence of Leb and Ley antigens on cells from secretors does not alone mask receptors for E. coli pili or hinder bacterial adherence.

Animals↗

Efficacy of sliding-scale insulin therapy: a comparison with prospective regimens.

OBJECTIVE: To compare the efficacy of retroactive sliding-scale insulin therapy, proactive therapy, and a combination of the two methods in establishing glycemic control in hospitalized diabetic patients. METHODS: Medical records of 47 diabetic ketoacidosis inpatients were reviewed retrospectively. RESULTS: The sliding-scale insulin therapy group's glucose deviation score (167.4) was significantly higher than the deviation for the proactive (112.9) and combination (121.3) groups. The sliding-scale insulin therapy group also had a significantly higher median glucose value (262.5) than the proactive (199.9) and combination (221.2) groups as well as a significantly higher number of nursing shifts (0.70) in which a glucose of 250 mg/dl or greater was recorded than in the proactive (0.37) and combination (0.40) groups. The proactive group was on their treatment regimen significantly less time than the combination group (5.5 vs 10.4 nursing shifts, respectively). The proactive group was hospitalized significantly fewer days (4.4) than the combination (6.3) and sliding-scale insulin therapy (6.3) groups. CONCLUSIONS: The present study lends support to previous concerns that sliding-scale insulin therapy is less effective than preventive therapy in the management of hospitalized diabetic patients.

Acid-Base Equilibrium↗

Survival and development of ivermectin-resistant or susceptible strains of Haemonchus contortus under field and laboratory conditions.

The free-living development of three strains of Haemonchus contortus was studied in two experiments. Day 21 faecal samples were collected from lambs infected with either a susceptible strain, a laboratory-selected ivermectin (IVM) resistant strain or a South African field strain showing multiple anthelmintic resistance, which included IVM. No eggs hatched in samples cultured at 4 or 10 degrees C. At 22 degrees C the laboratory-selected strain showed the highest rate of development while at 27 degrees C the susceptible strain produced the highest yield of third stage larvae (L3): at both temperatures the field strain showed the lowest percentage development to L3. The second experiment was a field study carried out in southern Brazil. Faeces containing either an IVM-susceptible or an IVM-resistant strain of H contortus were placed in two series of grass plots during each of three summer months. Soil subsequently yielded more larvae than did grass suggesting migration or mechanical transport into the soil. For plots contaminated during the first two months there was no significant difference in recovery rate between the two strains (P > 0.05). When contamination occurred during the third month, the IVM-resistant strain produced significantly higher recovery rates (P < 0.05) from both pasture and soil.

Animals↗