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The interaction between the bacteria and the host is a key factor determining the clinical consequences of H. pylori infection. The immune system plays an important role in either promoting or preventing the disease. The mucosal production of TNF-a, IL-6, IL-8 and IL-10 and the CagA status were investigated in H. pylori-positive patients with duodenal ulcer (DU). The concentrations of these cytokines in gastric antral mucosal specimens from patients infected with H. pylori (n = 40) were determined by ELISA and compared with data on mucosal specimens from H. pylori-negative patients (n = 12). The local TNF-a, IL-6 and IL-8 concentrations in the antral biopsy samples were significantly higher (p < 0.001) in the patients infected with H. pylori than in the samples from the H. pylori-negative subjects. CagA positivity was demonstrated in 39 (97.5%) of the 40 patients with DU, and in 41 (70.7%) of H. pylori-positive (58 of 100) healthy blood donors. In complementary studies focusing on extragastric disease, it was found that 57% of patients with ischaemic heart disease were seropositive as concerns H. pylori, and 91% of them had antibodies against human heat shock protein 60, too. This study suggests that, besides the bacterial virulence factor, the host response of an increased mucosal production of inflammatory cytokines can be relevant to the gastric pathophysiology in H. pylori-induced DU. At the same time, in ischaemic heart diseases the role of autoimmune processes induced by H. pylori cannot be excluded.
Hepatitis B virus (HBV) transmission via blood and other body fluids from infected individuals to healthy people has been largely demonstrated. However, in the current literature, there is little information available on the potential role of cerumen in HBV transmission.Cerumen and blood were collected from 70 patients infected with HBV and 70 volunteer healthy people were selected as the control group, and the samples were evaluated by ELISA and Real-time PCR.All the patients proved positive for HBsAg and anti HBc total. Sixty-one of the 70 cerumen samples of cases (82.1%) and 5 (7%) of controls were positive for HBV DNA with ranges from 1.53 × 102 to 2.9 × 108 and 1.3 × 102–2.6 × 105/ml, respectively. In three patients, the level of HBV DNA in cerumen was higher than that in the serums. The patients who were positive for HBeAg showed a higher rate of HBVDNA in the serum and cerumen.The results of this study showed the level of HBV DNA as a probably indicator of high risk transmission factor, which was present in the cerumen of chronic hepatitis B patients in west of Iran.
The presence of WNV in Europe has been well known for decades, although the first human infections and avian outbreaks were diagnosed in Hungary only in 2003. An annual average of 6–8 cases of the neuroinvasive form of WNV infection has been detected in the region since then, but a higher number (17) of WNV associated neuroinvasive disease occurred in 2008.In 2004, a surveillance system was established for monitoring WNV-associated meningo-encephalitis cases in Hungary, but a milder type of illness (with fever, rash and/or influenza like symptoms) is not followed. Fifty-two sera of 45 patients with mild clinical symptoms (fever, exanthema) were tested for anti-WNV antibodies in 2008 in a retrospective study by immunofluorescence test and ELISA. Seven patients had antibodies against WNV, serologic evidence of recent WNV infection was found in 4 out of the 7 patients. Infections could be acquired predominantly in August and in September, which seems to be a risk period for WNV in Hungary.The possibility of a recent WNV infection should be taken into consideration in the occurrence of fever and rush at late summer. Differential diagnosis of exanthematous patients should include WNV serology tests and should be done routinely.
The aim of this study was to investigate the effects on the immune response of levamisole alone and in conjunction with Candida albicans stimulation in human macrophage cell culture by determining the alterations in the levels of cytokine release.Levamisole treatment was performed before, during and after infecting U-937 human macrophage cells with C. albicans. In cell supernatants, interleukin (IL)-1b, IL-12, IL-18, tumour necrosis factor alpha (TNF-α) levels were measured by ELISA.In vitro levamisole treatment accompanied by C. albicans stimulation significantly increased IL-12, IL-1β and IL-18 production in macrophage cells (p < 0.05). It was observed that when administered before C. albicans infection, levamisole significantly increased IL-12 and IL-1β production in macrophage cells (p < 0.05). Another finding was that when applied to macrophage cells simultaneously with C. albicans infection, or before infection with C. albicans, levamisole suppressed the TNF-β production stimulating effect of C. albicans (p < 0.05).These results indicated that levamisole could be useful in treating patients infected with C. albicans or in protecting individuals under the risk of being infected with this pathogen. There is a need for further experimental and clinical studies on this hypothesis.
Many lines of evidence propose that psoriasis is a T cell-mediated disease where T cell activation is followed by secretion of inflammatory cytokines. To elucidate the functional state of T cells in guttate psoriasis, we analysed mRNA expression levels of T-bet and GATA-3 for Th1 and Th2 differentiation, respectively together with Th1 (IFN-γ) and Th2 (IL-4) cytokine mRNA expression. Relative quantification of T-bet, GATA-3, IFN-γ and Th2, and IL-4 transcripts in peripheral blood leukocytes (PBL) was conducted by real-time reverse transcriptase PCR (RT-PCR). Serum levels of IFN-γ and Th2 and IL-4 were also determined by ELISA. GATA-3 and IL-4 mRNA expression levels were lower in psoriatic patients as compared to normal healthy controls. The expression levels of T-bet and IFN-γ and Th2 genes were relatively similar in the patients and controls. In addition, a marked decrease in plasma IL-4 levels was observed in the psoriasis group, while no differences were observed with regard to levels of IFN-γ and Th2 between patients and normal subjects. Furthermore, a clear correlation between decreased IL-4 mRNA expression and IL-4 (P < 0.05) was revealed. These results suggested that altered balance between Th1 and Th2 cells transcription factor genes and they products may be implicated in the pathogenesis of psoriasis.
The aim of this study was to determine the clinical relevance of cardiac biomarkers [troponin I and T, creatine kinase-MB fraction (CK-MB) and lactate dehydrogenase (LDH)] in premature calves with respiratory distress syndrome. Seventy premature calves were admitted to the clinic within 24 h after birth. Respiratory distress syndrome was diagnosed in premature calves by clinical examination and venous blood gas analysis. Ten healthy calves, aged 5 days, were used as control. Cardiac troponin I and T were analysed using ELISA and ELFA, respectively. Serum CK-MB and LDH were also analysed in an automatic analyser. The calves had low venous pH, pO2, O2 saturation and high pCO2 values consistent with dyspnoea, hypoxaemia, and inadequate oxygen delivery. Mean serum troponin I, troponin T, CK-MB and LDH levels were increased in the premature calves compared to the control group. In conclusion, the results in this study demonstrated that serum CK-MB, troponin I and troponin T concentrations could be used for evaluating myocardial injury in premature calves with respiratory distress syndrome.
C-reactive protein (CRP) and haptoglobin (Hp) are well-known acute phase proteins in the dog. Currently, a commercial ELISA and a colorimetric assay are the methods of choice for measuring CRP and Hp, respectively; however, these assays showed interference when using haemolysed, lipaemic or hyperbilirubinaemic samples. Recently, time-resolved immunofluorometric assays (TRIFMAs) have been developed for measuring canine CRP and Hp. The aim of the present study was to evaluate the effect of increasing concentrations of haemoglobin, lipids and bilirubin in CRP and Hp serum measurements using these new fluoroimmunoassays. Haemolysis was produced by freezing blood cells at −20°C. The haemolysate was added to pooled sera at final concentrations of 0, 2.5, 5, 10, 20 and 40 g/L. A commercial emulsion of triglycerides was added to homologous pooled sera at 0, 0.35, 0.7, 1.4, 2.8, 5.6 and 11.2 mmol/L. Bilirubin, initially dissolved in dimethyl sulphoxide, was added to pooled sera at 0, 64.2, 128.4, 256.8, 513.7 and 1027.4 μmol/L. Addition of fresh haemolysate, triglycerides or bilirubin to serum samples did not affect either CRP or Hp concentrations (P ≥ 0.18), so the TR-IFMAs could be an alternative to the traditional tests for measuring canine CRP and Hp in those laboratories where immunofluorometric assays are available.
Serum amyloid A (SAA) is of interest as the circulating precursor of amyloid A protein, the fibrillar component of AA (secondary) amyloid deposits, and also as an extremely sensitive and rapid major acute phase protein. Serum concentrations of acute phase proteins (APPs) provide valuable information about the diagnosis and prognosis of various diseases, and thus the relevance of APPs for monitoring the health status of domestic animals is widely accepted. More importantly, the measurement of SAA concentration assists in assessing the prognosis in secondary amyloidosis, which is a common disease of geese, affecting an increasing number of animals. In the present study we introduce a highly sensitive goose-specific ELISA method for measuring SAA concentration in goose serum or plasma samples. Samples were taken from geese of the Landes Grey and Hungarian White breeds, which were stimulated for an acute phase reaction by administration of a commercially available fowl cholera vaccine containing inactivated Pasteurella multocida . Strong and characteristically rapid acute phase responses were measured in both breeds, peaking at approximately 24 h after inoculation. The maximum SAA concentration was 1200 μg/ml. At 72 h post-inoculation, the concentrations returned to pre-inoculation values. There was significantly (p = 0.004) less intense response in the control groups; however, a very mild increase of SAA levels was detected due to the stress inevitably caused by the sampling procedure.
In order to evaluate the seroconversion of horses to Babesia caballi and B. canis in Hungary, blood samples were collected from 371 animals on 23 different locations of the country. The presence of antibodies to B. caballi was screened with a competitive ELISA. All 29 positive samples came from one region (the Hortobágy). The prevalence of infection did not show correlation with sexes, and reached 100% in the age group of 2–5 years. Babesia canis -specific antibodies were demonstrated by IFAT in 6.74% of animals kept in 7 regions. The titres were low or medium level (1:40 to 1:160), indicating that the horses had previously been exposed to this piroplasm, but their infection must have been limited. The highest seropositivity rate was observed in the age group of 3–4 years, and males (stallions and geldings) were significantly more frequently infected than females. However, neither B. caballi nor B. canis could be identified in the peripheral blood samples of infected horses by PCR. Since most of the B. caballi -positive horses remained negative in the B. canis IFAT, whereas seroconversion solely to B. canis was detected in several regions of the country, serological cross-reaction between the two species can be discounted. This is the first serological evidence of horses being naturally infected with B. canis , supporting the view that piroplasms are less host specific than previously thought.
During a six-month period a region of Northern Sardinia was monitored to check the presence of mycobacterial infections in wild boars. Forty-eight serum and 229 biopsy samples were collected from different animals and examined by both traditional diagnostic techniques (culture, bacterioscopic and molecular tests) and enzyme-linked immunosorbent assay (ELISA). The latter was used to determine the antibody response against both methylated and nonmethylated Heparin-Binding Haemagglutinin (HBHA) protein. Nine mycobacterial strains were isolated: three M. avium ssp. paratuberculosis (Map), three M. avium , one M. interjectum and two M. scrofulaceum strains. By PCR, only one animal was positive for M. bovis , whereas 10 animals were positive for Map. Out of the 48 sera tested, 19 showed a good humoral response to methylated HBHA and 17 to nonmethylated HBHA. Our data provide new information on the prevalence of mycobacterial infection among wild boars in Northern Sardinia and suggest that a more effective program should be developed to monitor mycobacterial infections in the wild animal population.