Legionnaire's disease: a nosocomial outbreak in Turkey.

Six nosocomial cases of Legionella pneumophila occurred over a two-week period, with one further case being diagnosed retrospectively after 30 days. Strains isolated from the hospital water system were clonally related to a single sputum isolate. A sero-epidemiological investigation into legionella exposure amongst staff and inpatients was undertaken at the eight-year-old Inonu University Medical Centre in Turkey, which has 600 beds and central air conditioning. There is no disinfection programme for the hospital water system. A total of 500 serum samples (400 hospital staff and 100 inpatients) were screened for antibody to L. pneumophila by enzyme-linked immunosorbent assay (ELISA). Seroreactive cases were confirmed by a four-fold antibody rise in ELISA, a high indirect immunofluorescent assay (IFA) antibody titre or a positive urinary antigen test. ELISA showed that 24 (6%) of the 400 hospital staff and seven (7%) of the 100 inpatients had antibody titres higher than the cut-off value. ELISA-seroreactive cases were followed for two to four weeks. Of these subjects, seven (three patients and four staff) showed a four-fold rise in antibody titre by ELISA, six (three patients and three staff) had a high IFA titre, three patients with pneumonia had a positive urinary antigen test, and one of these patients also had a positive sputum culture. In addition, 22 water distribution systems were screened for the presence of L. pneumophila by culture. L. pneumophila was isolated from 15 sites. Pulsed-field gel electrophoresis typing indicated that all strains isolated from water systems were identical and clonally related to the strain isolated from sputum. Superheating and flushing of water systems were undertaken with legionella being re-isolated from four sites. Repeated superheating and flushing eliminated legionella completely. This study demonstrated that rapid detection of L. pneumophila and adequate superheating and flushing of water systems are effective for elimination and reduction of spread of this organism.

Disulfiram, as a candidate NF-κB and proteasome inhibitor, prevents endometriotic implant growing in a rat model of endometriosis.

OBJECTIVE: Disulfiram (DSF) exerts its therapeutic effects through oxidative, proteasome, and nuclear factor kappa beta (NF-κB) pathways. The study was planned to test the impact of DSF on growing of endometriotic implants in rats with experimentally induced endometriosis. PATIENTS AND METHODS: Thirty rats were labeled as the control (n = 8), sham (n = 6), GnRH-agonist (n = 8) and the DSF (n = 8) groups. The rats in the group 3 exposed to single dose leuprolide acetate. The rats in group 4 were treated with DSF for 21 days. The serum activity of oxidant and antioxidant markers, total oxidant status (TOS), total antioxidant status (TAS), interleukin-1ß, and tumor necrosis factor-α (TNF-α) were determined. Implants were processed for NF-κB, PCNA, and CD34 immunostaining. RESULTS: The serum concentration of malondialdehyde in the DSF group was significantly higher than those in other groups. The concentration of TAS, TNF-α, and interleukin-1ß in the DSF group considerably decreased compared to control group. Following treatment with DSF while the percentage of Grade 1 and 2 implants increased the percentage of Grade 3 and 4 implants decreased. The implants disappeared totally in two cases in the DSF group and one case in the GnRH-agonist group. The mean H-Scores of implant NF-κB and PCNA in DSF treated animals were found to significantly lower than those of the control group. CONCLUSIONS: By decreasing NF-κB expression, angiogenesis, and cell proliferation DSF prevents the growth of endometriotic implants.

Restoration of T and NK cell function in GM-CSF mobilized stem cell products from breast cancer patients by monocyte depletion.

Rapid immune reconstitution is observed following autologous peripheral blood stem cell transplantation (PSCT) as compared to autologous bone marrow transplantation (ABMT), although it is depressed compared to that observed in normal individuals. The immune dysfunction occurs despite the restoration of normal lymphoid cell numbers and may be associated with the immunologic characteristics of the infused peripheral blood stem cell (PSC) product. We report herein that the in vitro T cell proliferation and NK activity in PSC products of breast cancer patients are significantly increased following the removal of CD14+ monocytes (33 +/- 2% of the PSC product) by carbonyl iron magnetic cell isolation (CI). In vitro expansion of PSC cells cultured for 7-21 days in the presence of interleukin-2 (IL-2) is also significantly increased by depletion of the phagocytic cells. The PHA and IL-2 mitogenic responses, as well as NK activity of the expanded cells, was also significantly increased by the depletion of the phagocytes. In summary, the depletion of phagocytic monocytes from PSC products restores the proliferative and functional properties of T and NK lymphocytes and may facilitate adoptive cellular therapy, as well as rapid immunologic reconstitution post-PSCT.

Molecular epidemiology of tuberculosis in Turkey.

Our objective was to determine the extent of fingerprint pattern diversity of Mycobacterium tuberculosis isolates from Turkey. Of the 320 patient isolates, 81 (25.3%) carried

Effect of nitric oxide synthase inhibition in experimental Pseudomonas keratitis in rabbits.

PURPOSE: The aim of this study was to assess the efficacy of the nitric oxide synthase inhibitor L-NG-nitro-arginine-methyleser (L-NAME) in the treatment of experimental Pseudomonas keratitis. METHODS: Twelve young New Zealand white rabbits were given intrastromal injections of Pseudomonas aeruginosa strains. Twenty-four hours later, the rabbits were randomly divided into four groups: group 1 was treated with topical 0.3% ciprofloxacin drops and a single subconjunctival injection of L-NAME (150 mg/kg); group 2 received topical 0.3% ciprofloxacin drops alone; group 3 received a single subconjunctival injection of L-NAME alone; group 4, the control group, was treated with topical balanced salt solution (BSS) drops. One drop of either ciprofloxacin of BSS was applied at each treatment interval. Twenty-six hours after the start of treatment, the eyes were examined by slit lamp to assess inflammation. Aqueous humor specimens were collected from each eye to measure the nitric oxide concentration. Corneas were removed to count bacteria results. RESULTS: Slit lamp examination cell scores were significantly lower for groups 1 and 3 than groups 2 and 4 eyes (p = 0.002 to p = 0.004). No viable bacteria were detected in the corneas of groups 1 and 2. Group 3 corneas had significantly fewer bacteria (6.33 +/- 0.42 x 10(3)) than group 4 (5.94 +/- 0.16 x 10(4)) (p < 0.05). Nitric oxide levels in the aqueous humor were significantly lower for group 1 eyes than for groups 2, 3, or 4 (p = 0.02, p = 0.01, and p = 0.003, respectively). CONCLUSIONS: We conclude that nitric oxide synthase inhibitors may be a useful adjunct but are not a replacement for traditional antibiotic drop therapy.

Tularemia: a case transmitted from a sheep.

Tularemia is an arthropod-borne infectious disease caused by Francisella tularensis, a gram-negative microorganism that normally resides in a wide range of wild and domestic animals. The disease is characterized by a sudden onset with high fever, headache, malaise, chills, myalgia, and arthralgia. A short time after exposure, an inflamed and ulcerated lesion rapidly appears at the site of entry. A regional lymphadenopathy follows the cutaneous presentation. Cultures from the lesions or blood generally give negative results. Histopathologic examination reveals either a nonspecific inflammatory infiltrate or an infectious granuloma. The most useful laboratory procedure in the diagnosis of tularemia is serologic tests. Streptomycin, gentamicin, and tetracycline are the drugs of choice in the treatment. Quinolones are also effective. Tularemia is fairly rare in Turkey. We present a typical case of ulceroglandular tularemia transmitted from a sheep to a young man.

Detection of Alloiococcus otitidis in the nasopharynx and in the outer ear canal.

Alloiococcus otitidis has been recovered from the middle ear of children with otitis media with effusion, but its natural habitat is not known. To determine whether the nasopharynx and the outer ear canals are the natural habitats of A. otitidis, 145 swabs (50, nasopharynx; 95 outer ear canal) collected from 50 children were screened by polymerase chain reaction. A. otitidis DNA was detected in seven (4.8%) of the 145 specimens, of which four were nasopharynx, and three outer ear canal. These results indicate that the nasopharynx and outer ear canal may be the body sites for localization of A. otitidis.

[Incidence of rubella IgG antibodies in young girls]. / Genç kizlarda rubella IgG antikoru insidansi.

Rubella IgG antibodies were searched in 222 sera from female students of Selçuk University School of Medicine and from Institute of Higher Education for Nurses aged between 18-20 years. Of these 31 (13.96%) were found seronegative by ELISA. We discussed the risk of infection of these young girls.

Longstanding skin ulcers due to Mycobacterium tuberculosis in a healthy man.

Cutaneous tuberculosis, a rare form of extrapulmonary tuberculosis, has a wide variety of clinical presentations and continues to be one of the most important dermatological diseases in developing countries.The sites of predilection are neck, supraclavicular region, axilla, and groin. Single or multiple cutaneous and subcutaneous nodules first appear and breakdown later resulting in undermined ulcers with a purulent discharge, sinuses, and disfiguring scars. We report a multifocal case of scrofuloderma in a 47-year-old immunocompetent man treated successfully with four antituberculotic drugs, albeit lately due to the loss of awareness to the disease.

Comparison of the Tzanck test and polymerase chain reaction in the diagnosis of cutaneous herpes simplex and varicella zoster virus infections.

BACKGROUND: Although the diagnosis of herpes simplex virus (HSV) and varicella zoster virus (VZV) infections is usually made clinically, the Tzanck test, electron microscopy, viral culture, polymerase chain reaction (PCR), and serologic tests can be utilized to verify the diagnosis. METHODS: We conducted a study on a total of 98 patients (77 patients with recurrent herpes simplex and 21 patients with herpes zoster) to evaluate the reliability and reproducibility of the Tzanck test in comparison with PCR. RESULTS: In herpes virus infections, the general positivity rates of the Tzanck test and PCR were 61.2% and 79.6%, respectively. The difference between the positivity rates of the two tests was statistically significant. The positivity rates of the tests differed according to the type and duration of the lesions. CONCLUSIONS: Although PCR was superior to the Tzanck test, the Tzanck test has also been proven to be a reliable diagnostic method, with a sensitivity of 76.9% and a specificity of 100%. We recommend the use of this easy, quick, reproducible, and inexpensive diagnostic test more often in dermatologic practice, especially in cutaneous herpes virus infections.

The effect of Na+-K+ ATPase inhibition by ouabain on histamine release from human cutaneous mast cells.

BACKGROUND: There are controversial reports on the effect of sodium-potassium adenosine triphosphatase (Na+-K+ ATPase) inhibition on mast cell mediator release. Some of them have indicated that ouabain (strophanthin G), a specific Na+-K+ ATPase inhibitor, inhibited the release, whereas the others have shown that ouabain had no effect or even had a stimulatory effect on the mediator secretion. Most of these studies have utilized animal-derived mast cells. The aim of this study was to determine the effect of Na+-K+ ATPase inhibition on human skin mast cells. METHODS: Unpurified and purified mast cells were obtained from newborn foreskins and stimulated by calcium ionophore A23187 (1 microM) for 30 min following a 1 hr incubation with various concentrations (10(-4) to 10(-8) M) of ouabain. Histamine release was assayed by enzyme-linked immunosorbent assay (ELISA). RESULTS: The results indicated that ouabain had no significant effect on the non-immunologic histamine release from human skin mast cells, in vitro. CONCLUSIONS: Na+-K+ ATPase inhibition by ouabain had no significant effect on the non-immunologic histamine release from human cutaneous mast cells and suggested differences between human and animal mast cells.

A case of sporotrichosis treated with itraconazole.

Lymphocutaneous sporotrichosis is caused by Sporothrix schenckii, a dimorphic fungus commonly existing on decaying plants and in the soil. The fungus has a worldwide distribution but is more prevalent in temperate and tropical climates. Infection may result from traumatic inoculation of contaminated material such as soil, hay, moss, plant debris, splinters, thorns and barbs. Inoculation is also possible through some animal scratches and bites. We describe a typical case of lymphocutaneous sporotrichosis, which was confirmed by clinical, histological and microbiological features. The patient was successfully treated with 400 mg daily systemic itraconazole for 4 months.

False-positive reaction between syphilis and hepatitis C infection.

There are limited data about false-positive reactions against hepatitis C virus (HCV) in syphilitic patients and false-positive reactions against syphilis in the patients with HCV infection. The aim of this study was to demonstrate the false-positivity of syphilis in patients with HCV infection, the false-positivity of anti-HCV in patients with syphilis and the validity of the serological tests in such patients. Fifty patients with positive anti-HCV, 21 patients with positive VDRL and 50 healthy subjects were studied. Syphilis serology was determined by the Venereal Disease Research Laboratory (VDRL) test and microhemagglutination for T. pallidum (MHA-TP) test. Hepatitis C serology was determined by a second generation ELISA (Ortho Diagnostics) test for HCV antibody, and anti-HCV positive patients were tested for HCV RNA by polymerase chain reaction (PCR). All assays were performed on all subjects. Not only the false-positive VDRL reaction in the patients with HCV infection but also false-positive anti-HCV tests in syphilitic patients have been observed. Four patients with syphilis had positive anti-HCV and negative HCV-RNA, whereas 10% (5 of 50) of patients with hepatitis C infection had positive VDRL and these patients were negative for MHA-TP test. The rates of false-positivity of VDRL and anti-HCV were higher than within the control group (p < 0.05). According to these data, positive anti-HCV in syphilitic patients and positive VDRL in chronic hepatitis C may be false-positive results with regard to the reaginic tests. Therefore, therapeutic measures should not be initiated without confirmation with a treponemal test or PCR. VDRL and HCV-ELISA tests may be interacted with IgM or IgG antibodies. This relationship should be investigated in further studies.

Impaired T and NK cell response of bone marrow and peripheral blood stem cell products to interleukin (IL)-2.

The function of steady-state and interleukin (IL)-2-co-cultured mononuclear cells differs significantly between bone marrow (BM) products, growth factor-mobilized peripheral blood stem cell (PSC) products and normal peripheral blood mononuclear cells (PBMC). The natural killer (NK) cell activity and T cell proliferative response of PSC products from non-Hodgkin's lymphoma (NHL) patients are significantly higher than that of BM products and similar to normal PBMC. However, following a five-day co-culture with IL-2 (100 IU/ml), the NK activity of PSC, PBMC, and BM products (lytic units) was increased 176-, 40-, and 14-fold, respectively, compared to that observed prior to IL-2 culture. In contrast, lymphokine activated killer (LAK) cytotoxicity prior to IL-2 culture was low in PSC and BM products and normal PBMC, but was significantly increased in PSC products and PBMC following IL-2 co-culture. The proliferative response of PSC and BM products to the T cell mitogen phytohemagglutinin (PHA) was significantly lower than that observed with normal PBMC; however, PSC had a significantly higher response than cells from BM products. Similar patterns of T cell PHA mitogenic response were observed after IL-2 co-culture. In addition, the IL-2 mitogenic responses of IL-2-co-cultured PSC and BM products were also significantly lower than that observed with PBMC co-cultured with IL-2. The IL-2 mitogenic response of PBMC was also significantly increased compared to prior to IL-2 co-culture; whereas, the IL-2 mitogenic responses from PSC and BM cells were not. In summary, co-culture with IL-2 can increase the NK and LAK cell cytotoxicity of PSC and BM products from NHL patients, but IL-2 co-culture does not improve T cell function within either BM or PSC products.

Vascular endothelial growth factor levels are increased and associated with disease activity in patients with Behçet's syndrome.

BACKGROUND/AIMS: Vascular endothelial growth factor (VEGF) is a cytokine participating in inflammation with potent endothelial cell effects. It is produced by macrophages, neutrophils and vascular endothelial cells and can alter vessel permeability. Behçet's syndrome is a systemic inflammatory disorder with unknown etiology. Vascular endothelial dysfunction is one of the prominent features of the disease. We previously demonstrated the possible involvement of proinflammatory cytokines [tumor necrosis factor (TNF)-alpha, soluble interleukin-2 receptor (sIL-2R), interleukin (IL)-6 and IL-8], nitric oxide (NO) and adrenomedullin in the etiopathogenesis of Behçet's syndrome. Since VEGF expression is induced by these cytokines and VEGF itself is a potent stimulator of NO production with endothelial cell effects, this study aimed to investigate whether VEGF was affected during the course of Behçet's syndrome. We also assessed the possible involvement of VEGF in ocular Behçet's syndrome or in disease activity. METHODS: This multicenter case-control study included a total of 39 patients with active (n = 22) or inactive (n = 17) Behçet's syndrome (mean age, 38.1 +/- 10.4 years; 21 men and 18 women) satisfying International Study Group criteria, and 15 healthy hospital-based control volunteers (mean age, 39.2 +/- 9.3 years; eight men and seven women) matched for age and gender from a similar ethnic background. Patients were examined by a dermatologist and an ophthalmologist with an interest in Behçet's syndrome. Plasma VEGF concentrations were measured using a newly established enzyme-linked immunosorbent assay. Clinical findings and acute-phase reactant parameters such as erythrocyte sedimentation rate, alpha1-antitrypsin, alpha2-macroglobulin, and neutrophil count were used to classify the disease in Behçet's patients as active or inactive. The Wilcoxon test or the Mann-Whitney U-test was used for statistical analysis as indicated and the results were expressed as mean +/- SD, with range. RESULTS: The mean plasma VEGF level in patients with Behçet's syndrome (291.9 +/- 97.1 pg/mL; range 121-532 pg/mL) was higher than that in control subjects (103.0 +/- 43.6 pg/mL; range 25-187 pg/mL) and the difference was significant (P < 0.001). Patients with active disease had significantly (P < 0.001) higher VEGF levels than patients with inactive disease (347.6 +/- 87.1 vs. 219.9 +/- 51.6 pg/mL). In addition, ocular Behçet's patients (n = 23) had higher VEGF levels (315.7 +/- 92.1 pg/mL) than nonocular patients (n = 16, 257.8 +/- 96.6 pg/mL) and the difference was of borderline significance (P = 0.041). The levels of all acute-phase reactant parameters were significantly higher in the active stage than in the inactive stage (for each, P < 0.01) or in control subjects (for each, P < 0.001). CONCLUSIONS: VEGF may participate in the course of Behçet's syndrome, especially in the active stage, and elevated levels of VEGF may be an additional risk factor for the development of ocular disease, contributing to poor visual outcome.

IL-2 expansion of T and NK cells from growth factor-mobilized peripheral blood stem cell products: monocyte inhibition.

The expansion of T and natural killer (NK) cells in growth factor-mobilized peripheral blood stem cell (PSC) products with interleukin-2 (IL-2) requires a reduction in monocyte frequency. Monocytes are enriched with stem cells during apheresis and, in this series of growth factor-mobilized PSC products from breast cancer patients, represented 36 +/- 6% of the cells in the product. Immunophenotyping studies revealed that monocytes inhibited the proliferation of NK cells (CD56+ and CD3- CD8+ CD56+ cells) and T cells (CD3+, CD4+, and CD8+ cells) during IL-2 co-culture for 7, 14, or 21 days. A reduction in monocytes resulted in 61-fold expansion of CD3- CD8+ CD56+ cells compared with a 3.7-fold increase of CD3+ cells by day 21. In addition, following IL-2 co-culture, cells from PSC products with a reduced frequency of monocytes had a significantly increased T cell mitogenic response and NK cell activity in PSC products compared with intact products. We suggest that monocytes inhibit the IL-2-dependent proliferation and augmented function of NK and T cells from growth factor-mobilized PSC products.