Investigation of the relationship between lymphocyte subsets and intestinal parasites.

We analyzed the peripheral blood lymphocyte subsets of cancer patients infected with intestinal parasites, with an aim to find out the relationship between the levels of different types of lymphocytes with the prognosis of patients. 201 cancer patients aged 18 and over were included. Stool samples of the patients were examined using native-lugol, trichrome, modified trichrome (Weber's Trichrome stain), and modified Ziehl-Neelsen staining methods. Microsporidia and Cryptosporidium parvum were investigated at the genus and species levels using PCR. Lymphocyte subsets were determined by flow cytometry in blood samples. One or more parasite species were detected in 115 (56.7%) patients. The most common parasite species were Microsporidia, Blastocystis and Entamoeba coli, respectively. The frequency of parasites was high in patients with low lymphocyte percentage, CD3+ T cell and CD3+ CD4+ T (Th) cell levels in blood samples studied by flow cytometry. Microsporidia infection was significantly higher in patients with low lymphocyte percentage and Th cell levels. Similarly, C. parvum infection was found to be significantly higher in patients with low T lymphocyte percentage and Th cell level. Finally, Blastocystis infection was significantly higher in patients with low lymphocyte percentage and CD4/CD8 ratio higher than 1. The decrease in lymphocyte percentage, CD3+ T cell and Th cell count, and low CD4/CD8 ratio in cancer patients increase the frequency of intestinal parasitic infections. Based on these results, lymphocyte subsets may help identify cancer patients at high risk of opportunistic parasites. We suggest that opportunistic parasitic infections affecting the clinical course of the disease should be considered by clinicians during the follow-up and treatment of patients.

Application of Flow Cytometry in the Early Diagnosis of Neonatal Sepsis.

GOALS: To raise awareness of the utility of flow cytometric detection of inflammatory markers in the early diagnosis of neonatal sepsis. PROCEDURES: In accordance with the Töllner scoring system, cases with ≥10 points are accepted as having "clinical sepsis" and cases with 0-4 points as having "no sepsis". The study group consisted of 50 newborns with clinical sepsis as well as a control group of 50 newborns without sepsis. In all cases, blood counts, C-reactive protein (CRP) levels, and procalcitonin (PCT) levels were recorded. Additionally, the "cluster of differentiation" (CD)64, CD11b, and CD62L adhesion molecules and the presence of the human leukocyte antigen HLA-DR on monocyte and neutrophil surfaces were examined by flow cytometry. RESULTS: The levels of acute-phase reactants CRP and PCT were significantly higher in the study group than in the control group (p<0.05). The cell adhesion molecules CD11b and CD64 and the human leukocyte antigen HLA-DR were significantly higher in the study group (p<0.05); CD62L levels were similar to those in the control group (p>0.05). Furthermore, receiver operating characteristic analysis indicated that neutrophil CD11b (nCD11b) is a diagnostic marker for neonatal sepsis (area under the curve [AUC]: 0.72, 95% confidence interval [CI]: 0.62-0.82, p<0.001). The sensitivity, specificity, and positive predictive value (PPV) for nCD11b were 72%, 68%, and 58.4%, respectively. Similarly, monocyte CD11b (mCD11b) positivity was found to be diagnostic (AUC 0.77, 95% CI: 0.68-0.87, p<0.001). The sensitivity and specificity for mCD11b were 72% and 68%, respectively. In addition, the sensitivity, specificity, and PPV for nHLA-DR were 62%, 60%, and 60.8%, respectively. CONCLUSION: In addition to acute-phase proteins, cell surface antigens such as CD11b, CD64, and HLA-DR should be used in routine investigations for the early diagnosis of neonatal sepsis. Such usage in combination with acute-phase reactants may enhance diagnostic accuracy.

Lymphocyte subpopulations in pulmonary tuberculosis patients.

Protection against Mycobacterium tuberculosis is based on cell-mediated immunity, most importantly involving CD4(+) and CD8(+) T-cell subsets. The aim of this study was to evaluate CD4(+) and CD8(+) T-cell profiles and CD19(+) and CD3(+)CD(16 + 56)(+) populations in patients with pulmonary tuberculosis. CD4(+) and CD8(+) T cells, B-lymphocytes, and natural killer (NK) cells were evaluated in 75 active (APTB) and 25 inactive (IPTB) pulmonary tuberculosis cases and 20 healthy subjects (HCs). The results were compared at different stages of antituberculosis treatment in the APTB patients and also according to X-ray findings in the newly diagnosed APTB patients. The percentages of CD4(+) T cells were significantly lower (P < .01) and those of CD3(+)CD(16+56)(+) cells were significantly higher (P < .01) in APTB patients than in HCs. CD8(+) T cells were significantly decreased (P < .05), and CD3(-)CD(16+56)(+) cells were significantly increased (P < .01), in IPTB patients compared to HCs. The percentages of CD4(+), CD8(+), CD3(-)CD19(+), and CD3(-)CD(16+56)(+) cells showed no differences at different times of the antituberculosis regimen, and different stages of newly diagnosed APTB patients. APTB patients have a reduced percentage of circulating CD4(+) T cells and an increased percentage of NK cells compared with healthy individuals. These cells could play important roles in the immune response to M tuberculosis infection.

Changes in serum cytokine levels in active tuberculosis with treatment.

It has been reported that IFN-gamma, TNF-alpha, and IL-12 stimulate, and that IL-10, TGF-beta, and IL-4 suppress the protective immune response against tuberculosis. We aim to evaluate changes in the serum levels of pro and antiinflammatory cytokines in active pulmonary tuberculosis (APTB) and the possible effects of treatment on these changes. Serum IL-12p40, IL-4, IL-10, TNF-alpha, IFN-gamma, and TGF-beta1 levels were determined in 20 APTB cases (group 1) before and 2, 4, and 6 months after therapy. The same parameters were also determined in 9 inactive pulmonary tuberculosis (IPTB) cases (group 2) and 9 healthy controls (HC, group 3). Before treatment, the mean serum IFN-gamma, TNF-alpha, and IL-10 levels in group 1 were statistically higher than those in group 2 (P=.001, P=.024, P=.016, resp) or group 3 (P=.003, P=.002, P=.011, resp). The levels in group 1 decreased significantly after treatment (P=.001 for IFN-gamma, P=.004 for TNF-alpha, P=.000 for IL-10). The serum levels of IL-12p40 were significantly higher in group 1 than in group 3 (P=.012) and decreased insignificantly after treatment. There was no difference in serum IL-4 and TGF-beta1 levels among the groups (P>.05). Because the serum IL-12p40, IL-10, TNF-alpha, and IFN-gamma levels were high in APTB, we believe that these cytokines have important roles in the immune response to Mycobacterium tuberculosis (M tuberculosis). These parameters could be used in follow-up as indicators of the success of APTB therapy.

Serum neopterin levels in patients with brucellosis.

OBJECTIVES: The aim of this study was to evaluate the importance of neopterin levels in brucellosis and its alterations with treatment. METHODS: Thirty patients who had positive clinical findings, Brucella standard tube agglutination tests and/or positive blood culture were included in the study. The control group was composed of 30 healthy subjects. Neopterin (NPT) levels were measured by ELISA according to the protocol of manufacturer. RESULTS: Mean NPT levels were 54.15+/-33.7 nmol/l in study groups and 8.75+/-2.48 nmol/l in controls (P<0.001). It was determined that NPT levels measured at the time of admission of the patients had a linear correlation between the duration of complaint (r=0.560, P=0.001). While the mean NPT levels in patients with complaints of over 30 days were 79.07+/-34.9 nmol/l and it was 39.71+/-23.4 nmol/l in the patients with complaints of less than or equal to 30 days (P=0.002). NPT values measured after treatment (20.48+/-25.6) was determined lower by statistical means than the values measured before treatment (54.15+/-33.7, P=0.0001). The NPT levels of patients with positive blood culture (75.56+/-33.6) were higher than those of with negative blood cultures (32.72+/-15.2, P=0.001). CONCLUSION: It was concluded that the NPT levels were higher in patients with brucellosis. In addition, it was found that the levels of NPT were higher in non-responding patients than responded to the brucellosis treatment. It was thought that monitoring NPT levels could be useful for the extent and activity of the disease.

Protective effects of different antioxidants and amrinone on vancomycin-induced nephrotoxicity.

We have studied the effects of three antioxidants and amrinone, an inotropic agent, against vancomycin-induced nephrotoxicity in rats by investigating renal function and morphology. Thirty adult female Sprague Dawley rats (168-234 g) were divided into six groups. A saline-treated group served as control. The other five groups were treated for 7 days with vancomycin alone or in combination with alpha-lipoic acid, Ginkgo biloba extract 761, melatonin or amrinone. On day 8, all the rats were sacrificed by decapitation, kidney tissues were excised immediately and blood and kidney samples were collected. Blood urea and creatinine, kidney tissue malondialdehyde levels, and kidney superoxide dismutase and glutathione (GSH) peroxidase activities were measured. The kidneys were also examined for histological changes. Vancomycin administration led to increased urea, creatinine and malondialdehyde levels and decreased superoxide dismutase and GSH peroxidase activities. Co-administration of alpha-lipoic acid, Ginkgo biloba extract, melatonin or amrinone with vancomycin prevented the increases in the urea, creatinine and melondialdehyde levels and also resulted in higher superoxide dismutase and GSH peroxidase activities. The antioxidants and AMR improved the renal pathology compared to rats treated with vancomycin alone (P<0.05). These results indicate that the three antioxidants and amrinone have potential protective effects against vancomycin-induced nephrotoxicity, which might in part be due to inhibition of free oxygen radical production. Amrinone was the most effective drug as judged on the basis of the pathological findings.

Determination of intracellular cytokines produced by Th1 and Th2 cells using flow cytometry in patients with brucellosis.

The aim of the study was to evaluate intracellular interferon-gamma (IFN-gamma), and interleukin-4 (IL-4) levels in pre- and post-treatment periods of brucellosis patients and to determine the relationship between these parameters and patients' clinical findings. Twenty-five patients diagnosed as brucellosis and 11 aged-matched healthy volunteers were included in the study. CD3+CD4+ T lymphocytes levels were significantly lower in patients with brucellosis as compared to the control group. CD3+CD8+ T lymphocytes and CD3+IFN-gamma+ levels were increased in brucellosis patients compared with the control group. CD4+IFN-gamma+ and CD4+IL-4+ levels were no different between patients and healthy individuals. CD3+IL-4+ levels decreased in patients compared with healthy controls. Pre-treatment CD3+IFN-gamma+ levels dramatically increased in patients responsive to management compared with the unresponsive ones. In responsive cases, CD3+IFN-gamma+ levels decreased statistically after the treatment while in unresponsive cases no meaningful change was observed with respect to treatment. Adding IFN-gamma to the treatment for improving the depleted levels of IFN-gamma can be beneficial in patients with brucellosis who shows a tendency to chronicity or patients who do not respond to the treatment.