Evaluation of field-testing protocols to diagnose insulin dysregulation in ponies using a Bayesian approach.

Field tests and their association with laminitis have not been evaluated in large cohorts. The objectives of this study were to evaluate the performance of basal insulin (BI), the oral sugar test (OST) and the insulin tolerance test (ITT) to diagnose ID and investigate their association with laminitis. Insulin dysregulation status was determined in 146 ponies using BI (insulin concentration >20 µIU/mL), an OST (insulin concentration >65 µIU/mL at 60 or 90 min after oral administration of 0.45 mL/kg corn syrup) and an ITT (< 50% reduction in glucose concentration 30 min after intravenous administration of 0.1 IU/kg insulin). Laminitis was identified using modified-Obel scores. A Bayesian approach was used to define the characteristics of the tests and receiver operating characteristic curves were used to assess their association with laminitis. All tests were well tolerated and laminitis was diagnosed in 9% of ponies. Insulin dysregulation was diagnosed in 15% of ponies using BI, 38% using the OST and 54% using the ITT with 11% of ponies positive for all three tests. The sensitivities and specificities of BI, the OST and the ITT to diagnose ID were 0.52 (95% confidence interval [CI], 0.35-0.79) and 0.97 (95% CI, 0.91 - 1.00), 0.84 (95% CI, 0.70 - 0.94) and 0.60 (95% CI, 0.49 - 0.71), and 0.85 (95% CI, 0.68-0.96) and 0.88 (95% CI, 0.75 - 0.97), respectively. Only BI and the OST were associated with laminitis (P = 0.003 and 0.015, respectively).

Prevalence and predictors of traditional medicine utilization among persons living with AIDS (PLWA) on antiretroviral (ARV) and prophylaxis treatment in both rural and urban areas in South Africa.

Previous studies have reported that majority of antiretroviral (ARV) treatment-naïve patients use traditional medicine (TM). Given that TM use is ubiquitous in South Africa especially for chronic conditions, there is a potential for ARV non-adherence and serious drug interactions among patients with HIV/AIDs who use TM. The motivating factors for TM use in HIV/AIDS patients on ARV and prophylaxis treatment have not been well defined in South Africa. This study aimed to investigate the prevalence, facilitators, predictors, and types of TM used among persons living with HIV/AIDS on antiretroviral treatment. The study was a cross-sectional survey which involved 100 participants enrolled at ARV clinics in two South African provinces. Univariate and bivariate analyses were performed to assess the relationships between variables and potential predictors of TM. Sixteen percent of participants on ARV reported TM use. Seventy-nine percent used TM prior to a diagnosis of HIV. Participants were more likely to use TM if they were from a rural province, female, older, unmarried, employed, had limited education, or were HIV-positive for less than five years. TM users reported utilizing herbal or medicinal mixtures that were claimed to heal all conditions. This study provides insights into the treatment modalities selected by patients with HIV/AIDS in South Africa who are receiving ARV. This study revealed that less than 20% of participants co-used TM and ARV. However, close to 80% of participants utilize TM before contracting HIV, which is in keeping with approximate estimates by the WHO.

Parental influence on gender and marital role attitudes: implications for intervention.

This study examined parental influence on specific attitudes toward marital and parental roles and nonspecific gender-role attitudes. Respondents included 173 young adults (127 women and 46 men) and their parents. Young adults' scores on the Role Orientation scale of the Marital Satisfaction Inventory and on masculinity and femininity scales from two self-report inventories (the MMPI-2 and the Personal Attributes Questionnaire) were correlated with their parents' scores on a subset of these same measures. Results indicated that (a) marital and parental role attitudes among young adults are strongly related to specific marital role attitudes of their parents but only modestly related (for women) or unrelated (for men) to nonspecific gender-role attitudes of their parents; (b) broader gender-role attitudes of young men are moderately correlated with their father's attitudes but generally uncorrelated with their mother's attitudes regarding gender roles; and (c) the relationship of young women's gender-role attitudes to both maternal and paternal attitudes is generally weaker and more complex than for young men. Clinical implications of these findings for assessment and intervention with distressed couples are discussed.

Effect of gonadotropin-releasing hormone at estrus on subsequent luteal function and fertility in lactating Holsteins during heat stress.

The experiment used lactating Holstein cows (n = 94) from three herds in Mississippi. The experiment was conducted during late summer when temperatures were hot enough to cause a measurable stress response in cows in order to determine the effect of GnRH administration during estrus on fertility and subsequent luteal function of cows under these conditions. The mean ranges for ambient temperature, relative humidity, and temperature-humidity index during the study were 21.4 to 32.8 degrees C, 67 to 95%, and 21.6 to 29.6, respectively. After injection of PGF2 alpha for synchronization of estrus, cows were alternately injected with 100 micrograms of either GnRH or saline (control) at detection of estrus, followed by AI 10 to 12 h later. From 14 treated cows and 14 control cows, blood samples were drawn by venipuncture just prior to treatment and at 5-d intervals until 30 d after treatment. Serum progesterone concentrations were determined by radioimmunoassay. Mean progesterone concentrations were higher for the cows treated with GnRH than for the controls. The pregnancy rate from first AI was 28.6% for all treated cows and 17.7% for control cows. On d 20, 42.8% of the treated cows and 57.1% of the control cows exhibited progesterone concentrations that were typical of pregnancy. When pregnancy was diagnosed in these cows after d 45, pregnancy rates had decreased significantly for controls but not for cows given GnRH at estrus, suggesting greater embryonic survival in treated cows. We concluded that GnRH treatment enhanced secretion of luteal progesterone and embryo survival.

Cutaneous leiomyosarcoma: case reports and review of the literature.

Two patients with leiomyosarcomas are described, both men with lesions located in the upper trunk. Both lesions were present for more than 10 years. The diagnosis was established after excisional biopsy. The diagnostic criteria are a tumor size greater than 2.5 cm in diameter; a mitotic rate of 1 or more per 5 high-power fields in cellular areas; and electron microscopic verification of smooth muscle cells. The literature is reviewed and the clinical presentation of the 2 patients is discussed. Wide excision is the treatment of choice after histological diagnosis.

The pathogenicity and cultural characteristics of virulent, intermediate and benign strains of Bacteroides nodosus causing ovine foot-rot.

The relationship between the cultural and biochemical characteristics of 22 strains of Bacteroides nodosus and their virulence for sheep was examined. Virulent, intermediate and benign strains were recognised. Although there was some relationship between virulence and colony morphology on hoof medium with 4% agar, colonies of one virulent and 4 intermediate strains resembled those of benign strains. However, on hoof medium with 2% agar and on blood Euonagar, colonies of this virulent and one intermediate strain differed from each other and the other 3 intermediate strains, which in turn differed from the benign. The degree of piliation, as assessed by electron microscopy, was not a reliable indicator of virulence in strains not possessing a beaded colony type. Together, the results of colony morphology and proteolytic tests such as zymogram, degrading proteinase and elastin-agar tests allowed better discrimination of virulent and benign strains. Intermediate strains generally possessed virulent protease activity. In strains with benign zymogram patterns, activity bands 2 and 3 were more labile than in strains with virulent patterns. The addition of CaCl2 to the culture medium resulted in greater stability of proteolytic activity, particularly with benign strains, and prevented the disappearance of protease activity in the band 5 position in virulent, intermediate and benign strains during prolonged incubation. There were slight differences in the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) patterns of outer membrane proteins of some benign strains but those of intermediate category resembled virulent strains. There was some relationship between the apparent Mr of the pilin monomer on SDS-PAGE gels and serogroup specificity.

Cross-protection from Bacteroides nodosus vaccines and the interaction of pili and adjuvants.

The effects of vaccination of Merino sheep with the purified pili or the whole cells of Bacteroides nodosus strain 198, either in oil or alum-oil adjuvant, on the severity of foot-rot induced with the homologous strain (198) and a heterologous strain (217) were determined in a field experiment, on flood irrigated pasture. The efficacy of the whole cell vaccines was comparable to that of purified pili vaccines, against homologous challenge, when both had a similar content of pilus antigen although the purified pili vaccines induced significantly greater homologous pilus agglutinating antibody titres than the whole cell vaccines. However, against heterologous challenge, the whole cell vaccines in oil (CO) or alum-oil (CAO) provided significantly greater protection than a purified pili-in-oil (PPO) vaccine, the number of severely affected feet in sheep vaccinated with PPO being similar to that of the unvaccinated group. The group vaccinated with purified pili in alum-oil (PPAO) was intermediate between these two extremes. The superior performance of the PPAO in comparison to the PPO vaccine, against heterologous challenge, was associated with significantly higher mean ELISA titres to the outer membrane complex. Western blot analyses implicated a role in cross-protection for outer membrane proteins, in particular a protein Mr 78,000. The PPO vaccine produced fewer, smaller and less persistent vaccination reactions at the inoculation sites than did the other vaccines. Bodyweight gains in the period prior to challenge were much lower for the groups vaccinated with CO and CAO than for the controls and those vaccinated with purified pili, due presumably to the larger vaccination reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

Studies into immunisation of cattle against interdigital necrobacillosis.

Calves were immunised with fractions of Fusobacterium necrophorum incorporated in mineral oil adjuvant and then each foot was experimentally exposed to interdigital necrobacillosis (foot abscess) by subcutaneous injection of homologous organisms through the interdigital skin. The number of cells from an 18 h liquid culture that might be expected to cause 50% of the feet of control calves to develop marked swellings following subcutaneous injection was shown to be approximately 2.2 X 10(8) cells. Immunity was shown to be associated with antigens that were located in the supernatant of the culture, and which may be identical with or closely associated with the exotoxins.

Expression of the pilin gene from Bacteroides nodosus in Escherichia coli.

Bacterial plasmids that direct the expression in Escherichia coli of the pilin of Bacteroides nodosus were constructed. The quantity of pilin produced was greater than that of the pilin synthesized by B. nodosus, but no surface structural pili were present; pilin was found associated with the inner membrane of E. coli. Vaccination of sheep with E. coli containing pilin elicited increases in agglutinating and enzyme-linked immunosorbent assay antibody titers, which in turn were lower than the titers in sheep immunized with pilin from B. nodosus. The E. coli-produced pilin vaccine initially appeared to delay the progression of infection in immunized sheep after a challenge with virulent homologous B. nodosus, but at a later time the severity of foot rot was similar to that in sheep vaccinated with a placebo.

The protection given by pilus and whole cell vaccines of Bacteroides nodosus strain 198 against ovine foot-rot induced by strains of different serogroups.

A highly purified pilus vaccine prepared from cells of Bacteroides nodosus strain 198 provided a high level of protection against homologous challenge and small, not statistically significant, levels of protection against challenge with 4 other strains each from different serogroups. In a second experiment, a partially purified pilus vaccine from strain 198 induced significant immunity to 1 of 4 heterologous strains which were different from those used in the first experiment. In a third experiment a strain 198 whole cell vaccine produced significant immunity against 3 of 6 heterologous strains used in the first 2 experiments. There was no obvious relationship between the colony type, degree of piliation and level of cross-protection obtained against a particular strain. The results provide further evidence that immunogens associated with, but distinct from, the pilus are involved in cross-protection and that cross-protective antigens are common to some, but not all, strains.

Differences between breeds of sheep in their responses to Bacteroides nodosus vaccines.

In a field experiment on irrigated pasture, sheep of several breeds were vaccinated twice, subcutaneously, in the upper neck, with Bacteroides nodosus vaccine containing either depiliated cells (DC vaccine), or whole, piliated cells (WC vaccine) and the responses were measured over the following 14 weeks. DC vaccine was as effective as WC vaccine in protecting against the development of foot-rot under conditions of moderate challenge, although the WC vaccine induced significantly higher pilus agglutinating antibody titres. Foot-rot developed in significantly more vaccinated Merinos (Peppin and Saxon strains) than in Romney Marsh, Dorset Horn or Border Leicester breeds. Agglutinating antibody titres after WC vaccination were significantly lower in the Peppin Merino than in the other sheep for the first 6 weeks, while after DC vaccination the titres remained elevated longer in the Border Leicester and Saxon Merino and were significantly higher from 6 weeks onwards. Reactions at the inoculation sites were generally larger in the British breeds than in the Merinos and among the former the reactions were largest, most numerous and most frequently discharged their contents in the Dorset Horn. Bodyweight gains in all vaccinated sheep were initially reduced, compared with controls, but the differences were no longer significant after the eighth week.

The role of Fusobacterium necrophorum and Bacteroides melaninogenicus in the aetiology of interdigital necrobacillosis in cattle.

When cultures of known pathogenic strains of Fusobacterium necrophorum, isolated either from cattle or sheep were injected through the interdigital skin of cattle typical lesions of interdigital necrobacillosis were produced. The inclusion of Bacteroides melaninogenicus in the inoculum did not appear to contribute to the development of lesions.

Cultural characteristics and virulence of strains of Fusobacterium necrophorum isolated from the feet of cattle and sheep.

Sixty-one isolates of Fusobacterium necrophorum were recovered for study. Thirty-one were obtained from lesions of foot abscess in cattle (25) and sheep (6), 28 were from interdigital lesions in cattle and 2 were from the normal interdigital skin of cattle. The majority of isolates from lesions of foot abscess were virulent, belonged to biotype AB (Fievez 1963), produced flat, irregular shaped, greyish colonies and haemolysis on blood agar, and grew as turbid filamentous suspensions in liquid media. They produced a soluble exotoxin, a leucocidin, and were pathogenic for cattle and mice. Virulent isolates also produced a haemolysin which most readily lysed bovine, equine and chicken erythrocytes; those from sheep were less susceptible while those of rabbit and pig were the most resistant. Isolates recovered from lesions of the feet not classified as foot abscess and from clinically normal feet were predominantly of the B biotype and caused few experimental lesions, produced convex, round, yellow colonies, flocculated and sedimented while growing in liquid medium and produced little or no haemolysin or leucocidin. Routine differentiation between virulent and non-virulent bovine isolates of F. necrophorum could be achieved by assessing the colour, morphology, and degree of haemolytic activity of colonies grown on blood agar.

Primary structure of pilin protein from Bacteroides nodosus strain 216: comparison with the corresponding protein from strain 198.

The amino acid sequence of pilin protein from Bacteroides nodosus strain 216 was determined. The protein had a calculated molecular weight of 15962 and contained the same number of amino acid residues (151) as the pilin from the previously sequenced strain 198. The sequence of the first 44 residues was common to both strains, including the unusual amino-terminal amino acid, N-methylphenylalanine. Of the remaining 107 residues, 37% of them differed between the two strains. Comparison of hydrophilicity profiles constructed from the sequence data indicated that a conserved region around residues 71-72 was probably the site of an antigenic determinant.

Biochemical and functional properties of a leucocidin produced by several strains of Fusobacterium necrophorum.

A soluble exotoxin (a leucocidin) which was lethal to peripheral blood leucocytes from cattle, sheep, rabbits and man (in order of decreasing sensitivity) was elaborated by a variety of isolates of Fusobacterium necrophorum when the majority of organisms were present as filaments in liquid culture. Maximum production of the leucocidin was achieved by concentrations of bacteria equivalent to between 4 X 10(7) and 4 X 10(8) short cells per ml of culture above which no further increase in titre was observed. The ability of different batches of medium to support production of leucocidin was reflected in their capacity to enable F. necrophorum to grow to this range of concentration. Prolonged culture of the organism, resulting in a decline to below 6 in the pH of the medium was associated with a depression in the titre of leucocidin, presumably due to its inactivation under these conditions. The leucocidin was stable at 4 degrees C for at least 30 days, to extremes of pH (4 to 9) for 1 h at room temperature, and showed maximum activity in assays conducted at 37 degrees C at pH 7 to 8. The exotoxin was inactivated by heating at 56 degrees C for 30 min and possessed a molecular weight around 250,000 to 300,000 as determined by gel filtration and membrane partition chromatography.

Differences between strains of Bacteroides nodosus in their effects on the severity of foot-rot, bodyweight and wool growth in Merino sheep.

The effects of 3 ovine and one bovine strains of Bacteroides nodosus on the severity of foot-rot, bodyweight and wool growth were compared in Merino sheep in a field experiment. Based on the severity of the induced foot lesions, one strain was classed as virulent (causing underrunning lesions in most feet), one was benign (causing lesions of the interdigital skin only), and 2, including the bovine strain, were of intermediate virulence (causing underrunning lesions in a small proportion of feet). Differences among strains in their effect on foot-rot severity and bodyweight were significant when compared over the whole experimental period, but were not significant at any single time of measurement, because of large differences between replicates. Bodyweight loss and severity of foot-rot caused by the virulent strain were significantly greater than that caused by the benign strain. The intermediate strains lay between these 2 extremes in terms of both bodyweight and foot-rot scores but were not significantly different from either in a statistical sense. Total greasy wool weight did not differ among groups over the whole experiment, but the rate of wool growth during the period when foot lesions were most prevalent and severe was reduced appreciably by the virulent strain and to a lesser extent by the intermediate strains.