Multi-site DNA polymorphism analyses of Leishmania isolates define their genotypes predicting clinical epidemiology of leishmaniasis in a specific region.

Leishmania isolates from 57 cases of human cutaneous (CL), human visceral (VL), and canine visceral (CVL) leishmaniasis in Turkey were grouped by multi-site DNA polymorphism analyses into five genotypes. The initial grouping was based on DNA heterogeneity of the faster-evolving mitochondrion (kinetoplast) minicircles and the intergenic regions of two nuclear repetitive genes. Taxonomic affiliation and phylogenetic relationships of the five genotypes were inferred by comparing them with reference species for sequence heterogeneity in a approximately 1.4 kb conserved single-copy gene, encoding N-acetylglucosamine-1-phosphate transferase (NAGT). Alignment of the available sequences revealed no gap, but up to 7% scattered base substitutions, suggesting that this functionally important gene is a suitable marker. Three genotypes are completely identical to the NAGTs of the reference species, identifying them as L. infantum, L. tropica. and L. major, respectively. The remaining two are recognized as L. major NAGT variants with one and four base substitutions, respectively. As expected, Maximum Likelihood analysis of the NAGT sequences separates them into three clades, corresponding to the three species. The majority of the isolates obtained are L. infantum and L. tropica, which have been known to cause infantile VL and anthroponotic CL in western and southeastern Turkey, respectively. Unexpected is the finding of Leishmania major variants and their dispersal, possibly as previously unrecognized clinico-epidemiologic entities of CL and VL.

Osteoarthricular involvement of brucellosis in Turkey.

The aim of this study was to determine the rates, types, clinical features and treatment of osteoarthricular involvement of brucellosis in Turkey. In a restrospective study in adults, we investigated 238 patients diagnosed with brucellosis over a period of 6 years. A diagnosis of brucellosis was established by isolation of Brucella species in blood or by a compatible clinical picture together with a standard tube agglutination titre of > or = 1/160 of antibodies for brucellosis and/or demonstration of an at least four-fold rise in antibody titre in serum specimens taken over 2 or 3 weeks. Osteoarthricular involvement was defined by inflammatory signs in peripheral joints or by unrelieved pain at rest together with radiological alterations and/or radionuclide uptake in any deep joint. Eighty-seven patients (36.5%) had osteoarthricular involvement (58.6% female, 41.4% male), 47 (54.1%) of whom were reported to consume unpasteurised dairy products. The mean age was 32.3 +/- 16 years. Sacroiliitis was the most common involvement (n = 53, 60.9%) followed by peripheral arthritis (n = 17, 19.5%), spondylitis (n = 12, 13.8%) and bursitis (n = 5, 5.7%). During the observation period, 60 (69%) patients with osteoarthricular involvement and radiographic abnormalities. A bone scan was positive in 15 patients with no radiographic abnormalities. All patients received merely medical treatment and relapse occurred in five (5.7%) patients. Sacroiliitis has been determined as the most frequently observed type of osteoarthricular involvement in brucellosis in Turkey.

Serodiagnostic value of ELISA in pulmonary tuberculosis in Turkey where tuberculosis is highly prevalent.

We studied the diagnostic utility of ELISA in pulmonary tuberculosis (TBC) using serologically active glycolipid antigens. Twenty-seven patients who were smear positive, and 30 patients who were smear negative, but with evidence of active TBC (sputum culture positive in 10, response to anti-TBC chemotherapy in 20) were included in the study group. Twenty cured patients who had been free of TBC for at least 1 year, 50 TBC-free persons with PPD results of 0-10 mm and more than 10 mm, and 21 patients with active inflammatory diseases other than TBC formed the control groups. Sensitivity and specificity were 96 and 91%, respectively. We think that serologically active glycolipid antigens are as sensitive and specific as other purified antigens, and even superior to them from the point of view of their production: their extraction is quicker and easier.

Serodiagnosis of Asian leishmaniasis with a recombinant antigen from the repetitive domain of a Leishmania kinesin.

rK39 is a recombinant product of the 39 amino acid repeats found in a kinesin-like gene of visceral Leishmania spp. This and other antigens were compared for immunodiagnostic potential by enzyme-linked immunosorbent assay with sera from confirmed cases of Asian cutaneous and visceral leishmaniasis. In preliminary trials, rK39 proved superior to 2 purified Leishmania antigens, a cytosolic protein (p36) and a membrane protein (gp63), for immunodiagnosis of visceral leishmaniasis. Of the 53 visceral cases from China and Pakistan assayed, 52 were seropositive (98%) at a 10(-1) dilution with 36 ng of rK39. End point titrations of 27 highly positive samples yielded anti-rK39 antibody titres ranging from c. 10(-3) to beyond 10(-4). Antigen titrations with one positive serum further revealed that rK39 was 25-fold more sensitive than Leishmania whole cell soluble lysates. 31 cutaneous leishmaniasis cases from Turkey assayed for anti-rK39 antibody gave reactions ranging from negative or marginally positive to positive. In Brazil, all cutaneous and mucocutaneous leishmaniasis cases gave negative results in this assay.

The prevalence of anti-HCV positivity in patients undergoing haemodialysis or with malignant disease.

We studied the occurrence of anti-hepatitis C (HCV) antibodies in patients with malignant disease (53), in patients undergoing haemodialysis (56), and in blood donors (204) as healthy population controls. The study was carried out using the second-generation EIA test. Anti-HCV positivity was 23.2% in haemodialysis (HD) patients, 0.5% in blood donors, and 0% of the patients with malignant disease (MD). There was no association between anti-HCV positivity and the results of AST, ALT and HBsAg tests in patients and controls. But there was significant association of blood transfusion frequency and duration of HD with anti-HCV positivity in patients undergoing HD, and conversely an absence of this association in patients with MD. However, two of the anti-HCV-positive HD patients did not have any blood transfusion history. One HCV-positive blood donor had a history of surgical operation. Nosocomial transmission of HCV infection has replaced blood transfusion as the main risk factor in HD patients, and preventive measures should be performed in this direction to control infection.

Plasma levels of tumor necrosis factor alpha during hemodialysis.

Tumor necrosis factor alpha (TNF-alpha) levels were measured by immunoradiometric assay in chronically hemodialyzed patients consisting of 9 males and 3 females aged between 14 and 60 years. TNF-alpha levels were 5.47-15.1 pg/ml (mean 9.76 +/- 6.63) before hemodialysis (HD), and 5.75-58.55 pg/ml (mean 22.15 +/- 15.14) after HD. According to these results TNF-alpha levels in chronically hemodialyzed patients were within normal limits, but after 4 h of HD, higher levels of TNF-alpha were found.