Risk perception of NSAIDs in South Dakota in comparison with Slovakia and Greece.

AIM: Adverse effects (ADRs) of non-steroidal anti-inflammatory drugs (NSAIDs) represent a public health problem. To decrease the negative effect on the population, an improvement of risk awareness is crucial. We aimed to evaluate the risk perception and the use of NSAIDs in South Dakota in comparison with Slovakia and Greece. METHOD: A structured questionnaire evaluating NSAID use in 185 patients in a hospital in South Dakota. RESULTS: 95.7 % of respondents reported the use of analgesics. On 1-10 visual analogue scale, perceived risk of NSAIDs was 4.27±2.46, similar to Greece (4.36±2.41, p=0.360), but significantly higher than in Slovakia (3.8±1.9, p=0.038). Only 12.4 % were familiar with gastrointestinal ADRs and only 1.1 % were aware of cardiovascular risk. Although 57.8 % were informed about ADRs by their doctor or pharmacist, only 33.0 % were informed spontaneously, without actively asking. Providers in South Dakota were informing patients spontaneously more often than in Slovakia (15.9 %, p≤0.001) and on par with Greece (36.3 %, p=0.631). CONCLUSIONS: Public awareness about NSAID risk is dangerously low. Only a third of providers are informing patients about possible risks spontaneously (Tab. 6, Ref. 15) Keywords: non-steroidal anti-inflammatory drugs, risk perception, adverse effects, cardiovascular risk, gastrointestinal risk.

Molecular characterization of field isolates and vaccine strains of infectious bursal disease virus.

The present investigation was conducted to study the genetic heterogenicity and molecular polymorphism among the field isolates and vaccine strains of infectious bursal disease virus (IBDV). Samples of bursa of Fabricius from 15 suspected outbreaks of infectious bursal disease (IBD) were subjected to agar gel precipitation test (AGPT), virus isolation and reverse transcription-polymerase chain reaction (RT-PCR) combined with restriction fragment length polymorphism (RFLP). Nine out of 15 samples were found positive in AGPT while 14 were found positive both by virus isolation and RT-PCR. PCR amplified 474bp fragment from the variable region of VP2. Sac I, Stu I, Alu I, Ssp I and Mbo I restriction enzymes were used for characterization of all the 14 IBDV isolates and four reference vaccine strains. Sac I, Stu I, Alu I and Ssp I could differentiate classical virulent IBD (cvIBD) vaccine virus strains from very virulent IBD (vvIBD) field isolates by their varying restriction patterns. Based on above results two field isolates (VPL and VMK) were placed in cvIBD virus group and 12 field isolates were placed in vvIBD virus group. Virus neutralisation test (VNT) using rabbit raised Georgia strain anti-serum, however, could not differentiate between cvIBD virus and vvIBD virus. It was concluded that RT-PCR combined with RFLP assay using restriction enzymes Sac I, Stu I, Alu I and Ssp I can be used for rapid differentiation and classification of field isolates of IBDV.

Effect of fowl adenovirus-1 (IBH isolate) on humoral and cellular immune competency of broiler chicks.

Fowl adenovirus-1 (FAV-1), isolated from field outbreaks of inclusion body hepatitis (IBH), was administered orally to 3-week-old disease-free broiler chicks. Humoral immune competency was evaluated by determining the antibody response of infected chicks to sheep red blood cells (SRBC) and Brucella abortus. FAV-1 infection significantly decreased the antibody response of chicks to B. abortus (T-cell-independent antigen) by decreasing IgM responses, however, the decreased antibody response to SRBC (T-cell-dependent antigen) was statistically non-significant. Bursal index was also found lowered in infected chicks as compared to the control chicks. A significant decrease was seen in blastogenesis response of peripheral blood lymphocytes to phytohaemagglutinin (PHA-P) in FAV-1-infected chicks on 2 and 3 weeks post-infection (WPI). These results indicated that FAV-1 affects humoral as well as cellular immune competency of infected chicks.

Coronary endothelial dysfunction after heart transplantation predicts allograft vasculopathy and cardiac death.

BACKGROUND: Coronary endothelial dysfunction may be an early marker for cardiac allograft vasculopathy (CAV) in orthotopic heart transplant recipients. Using serial studies with intravascular ultrasound and Doppler flow-wire measurements, we have previously demonstrated that annual decrements in coronary endothelial function are associated with progressive intimal thickening. The present study tested whether endothelial dysfunction predicts subsequent clinical events, including cardiac death and CAV development. METHODS AND RESULTS: Seventy-three patients were studied yearly beginning at transplantation until a prespecified end point was reached. End points were angiographic evidence of CAV (>50% stenosis) or cardiac death (graft failure or sudden death). At each study, coronary endothelial function was measured with intracoronary infusions of adenosine (32-microgram bolus), acetylcholine (54 microgram over 2 minutes), and nitroglycerin (200 microgram) into the left anterior descending coronary artery; intravascular ultrasound images and Doppler velocities were recorded simultaneously. Of the 73 patients studied, 14 reached an end point during the study (6 CAV and 8 deaths, including 4 with known CAV, 1 graft failure, and 3 sudden). On the last study performed, the group with an end point had decreased epicardial (constriction of 11.1+/-2.9% versus dilation of 1.7+/-2.2%, P=0.01) and microvascular (flow increase of 75+/-20% versus 149+/-16%, P=0.03) endothelium-dependent responses to acetylcholine compared with the patients who did not reach an end point. Responses to adenosine and nitroglycerin did not differ significantly. CONCLUSIONS: Endothelial dysfunction, as detected by abnormal responses to acetylcholine, preceded the development of clinical end points. These data implicate endothelial dysfunction in the development of clinically significant vasculopathy and suggest that serial studies of endothelial function have clinical utility.

Detection of bovine herpesvirus-1 infection in breeding bull semen by virus isolation and polymerase chain reaction.

Serum samples from 51 apparently healthy breeding bulls were screened for bovine herpesvirus-1 (BHV-1) antibodies using an avidin-biotin enzyme-linked immunosorbent assay, revealing a sero-positive prevalence rate of 45.09%. Semen samples were then collected from 12 of the sero-positive and 12 of the sero-negative bulls and tested for BHV-1 antigen using both a virus isolation assay and a polymerase chain reaction (PCR) assay; PCR was applied to detect BHV-1 deoxyribonucleic acid by using primers selected from the relatively conserved sequence of the gl glycoprotein gene to amplify a 468 base pair fragment. The PCR-amplified products were confirmed as BHV-1 by restriction enzyme, Dde 1, which produced fragments of predictable sizes, namely 340 and 128 base pairs. Positive virus isolation test results, confirmed by virus neutralisation, found BHV-1 antigen in the semen of five sero-positive and six sero-negative bulls. In comparison, positive PCR results found BHV-1 genome in the semen of six sero-positive and eight sero-negative bulls. From the 24 semen samples tested, 14 were shown to be positive by PCR and 11 by virus isolation. The sensitivity and specificity of virus isolation were 57.14% and 70% respectively, and were significantly lower than PCR. In the semen samples taken from sero-negative bulls, BHV-1 was detected more often by PCR methods than by virus-isolation, suggesting that PCR is a more sensitive method for BHV-1 screening in bulls.

Cell mediated immune response of chicks following fowl adenovirus type-1 infection.

Cellular immune response of 6-week-old chickens infected with fowl adenovirus type-1 was evaluated by both in vivo and in vitro assay. In vitro assay, enumerating peripheral T lymphocyte by alphanaphthyl acetate esterase staining, revealed that cellular immunity appeared as early as 1 week post-infection and was maintained up to 5 weeks post-infection. In vivo assay by phytohaemagglutin skin test showed cellular immunity until 2 weeks post-infection. Thereafter no immunity was observed by this assay.

Comparative studies on the antigenicity of extra- and intracellular viruses of fowl pox.

The intra- and extracellular virus of three strains of fowl pox virus, when precipitated in succession with different saturation of ammonium sulphate revealed three antigens in gel diffusion test in the precipitates obtained at 25%, 50% and 75% of saturation. Further analysis of each positive antigen by dot ELISA revealed that the extracellular virus of FS-8 and HP 1 strains possessed excess antigenic protein at 50% saturation compared to their intracellular viruses. While no difference between extra- and intracellular viruses of FS-4 strain was observed.

An improved dot ELISA to detect fowl adenovirus type-1 antigen.

An improved dot immunobinding assay to detect fowl adenovirus type-1 is described. The method consists of spotting of antigen on nitrocellulose membrane sheet, blocking with either 5% acetic acid or with 5% defatted milk powder and fixation of either antigen or antigen-antibody complex, with either 50% methanol or 0.25% glutaraldehyde or 0.2% tannic acid. The results revealed that fixation of both antigen and antigen-antibody complex resulted in 4-fold increase in sensitivity when acetic acid was used as blocking agent. The use of two substrates simultaneously resulted in more colour intensity and clarity than using both substrates separately.

Comparative evaluation of cell culture-adapted and chicken embryo-adapted fowl pox vaccine strains.

Two types of vaccines, chicken embryo adapted (VacCE) and cell culture adapted (VacCC), were tested for their efficacy to elicite the immune response in birds vaccinated at 2 and 8 wk of age. The cell-mediated immune response studied by blastogenesis assay showed that birds vaccinated at the second week of age by both VacCE and VacCC vaccines had significant increase in T-lymphocyte count at 21 days postvaccination (PV) and 7 days postchallenge (PC), whereas in birds vaccinated at 8 wk of age, a significant increase was seen at 21 days PV and 7 days PC with the VacCC vaccine. The rise in passive hemagglutination titers was observed up to 21 days PV and 7 days PC in birds vaccinated at 2 wk of age. However, only the birds vaccinated with VacCC at 8 wk of age showed rise in titers at days 21 PV and 7 PC. Birds were challenged 90 days PV by scarification on the thigh region, and the birds vaccinated with VacCC showed 90% and 70% protection when vaccinated at 2 and 8 wk, respectively. The birds vaccinated with VacCE showed only 60% and 20% protection at the corresponding levels, respectively.

Isolation of A/Equi-2 virus during 1987 equine influenza epidemic in India.

Processing of nasal materials from clinical cases during the 1987 influenza epidemic in Northern and Central India resulted in the isolation of two haemagglutinating agents; one each from donkeys and horses at Bhiwani in Haryana State and Ludhiana in Punjab State, respectively. These were typed as Influenza A/Equi-2 viruses by haemagglutination inhibition test. The two isolates were designated as A/Equi-2/Bhiwani/1/87 and A/Equi-2/Ludhiana/1/87. The Bhiwani/87 isolate was confirmed to have H3N8 antigenic structure and was indistinguishable from the Miami/63 strain of A/Equi-2 virus. However, the A/Equi-2 Ludhiana/87 isolate was closely related to the Fontainebleau/79 strain of A/equi-2 virus.

Genomic diversity and prevalence of Rotavirus in cow and buffalo calves in northern India.

Faecal samples were collected from seventy-eight diarrhoeic cow and buffalo calves between November 1998 and February 1999 to study the genomic diversity and prevalence of Rotavirus infection by ribonucleic acid polyacrylamide gel electrophoresis (RNA-PAGE) and enzyme-linked immunosorbent assay (ELISA). In the organised dairy farm (where daily production and health records were maintained), the overall prevalence of infection with Rotavirus, recorded by RNA-PAGE and ELISA, was 27.02% (10/37) in both cow and buffalo calves. In unorganised dairy herds (where no production or health records were maintained), RNA-PAGE and ELISA detected infection with Rotavirus in 26.8% (11/41) of cow and 19.5% (8/41) of buffalo calves. Five distinct electropherotypes were found to circulate in cow and buffalo calves. All were short electropherotypes except the single long electropherotype observed in a buffalo calf in an unorganised dairy herd. Some differences in RNA migration pattern were observed when these electropherotypes were compared with the neonatal calf diarrhoea virus strain of Rotavirus. Some electropherotypes were restricted to one farm while others were found in both organised and unorganised dairy herds and in both cow and buffalo calves.

Epidemiology of inclusion body hepatitis in poultry in northern India from 1990 to 1994.

The epidemiology of inclusion body hepatitis (IBH) was studied in poultry in northern India, from April 1990 to March 1994, to evaluate the various factors responsible for causing and determining the severity of the disease. Broiler chicks and Japanese quail (Coturnix coturnix japonica) were the species examined. The factor observed to be most commonly associated with IBH was the presence of aflatoxins in the feed at higher than permissible levels, i.e. 20 parts per billion. Avian adenovirus-1 was isolated from the livers of affected birds. In the final year of the study, a number of outbreaks of IBH caused heavy mortalities among three to five-week-old broiler chicks, which displayed typical IBH lesions in addition to hydropericardium.

Pathogenicity of quail's inclusion body hepatitis virus (avian adenovirus-1) for Japanese quails and broiler chicks.

Quail (Coturnix coturnix japonica) and broiler (Gallus domesticus) chicks were inoculated experimentally with IBH virus (avian adenovirus-1) derived from quails to determine its pathogenicity. Quail chicks were inoculated by the intraperitoneal route at 3, 4, 5, 6 or 7 weeks of age. Lesions were encountered most frequently in the liver, kidneys and lungs. These included pale, swollen and mottled liver, swollen nephrotic kidneys, and congested and pneumonic lungs. The lesions were severe in birds inoculated at 5 weeks of age. Large basophilic intranuclear inclusion bodies were seen in hepatocytes and occasionally in the renal epithelium. The results showed that this isolate is pathogenic for quails above 3 weeks of age. Broiler chicks were inoculated at 4 weeks of age by the intraperitoneal route. The lesions produced in these chicks were similar to those of adenovirus-induced inclusion body hepatitis. Viral antigen was also demonstrated by dot-ELISA in suspension of liver tissue from both quail and broiler chicks.

The use of PCR combined with restriction enzyme analysis to characterize fowl adenovirus field isolates from northern India.

Ten fowl adenoviruses (FAVs), isolated from suspected cases of inclusion body hepatitis (IBH) in quails and broilers, were characterized by a hexon-based polymerase chain reaction (PCR) combined with restriction enzyme analysis (REA) of the amplified DNA fragments. All the isolates could be detected using H1/H2 and H3/H4 primer sets. Amplification of DNA with H1/H2 and H3/H4 primer sets resulted in fragments of approximately 1219 bp and 1319 bp, respectively. HaeII digestion of the H1/ H2 PCR products and HpaII digestion of the H3/H4 PCR products characterized all the isolates in FAV groups, known from genomic typing using the whole DNA. For some of the isolates, neutralization tests were used to confirm these results. The results revealed that, as well as FAV serotype 1, which is the sole member of DNA group A, FAVs of DNA group E are also associated with IBH in poultry in northern India. The FAV specific PCR combined with REA was found to be very useful in investigating the epidemiological situation in the field. It was even possible to define mixed infections with more than one FAV.

Neutralising antibody and challenge response to live and inactivated avian adenovirus-1 in broilers.

Immune responses to live and inactivated avian adenovirus-1 were evaluated in broilers by neutralising antibody response and challenge reaction. The neutralising antibody titre was 1:256, 1:64 and 1:32 in live virus, inactivated virus and uninoculated control birds respectively at 3 weeks post inoculation when they were challenged. At one week post challenge the antibody titres were 1:941, 1:247 and 1:375 in live virus, inactivated virus and control birds, respectively. There was a booster effect of challenge in the live virus inoculated group up to 3 weeks post challenge. The post challenge histopathological evaluation of liver, kidneys, lungs and bursa revealed less severe changes in the live virus inoculated group as compared to other groups.