The role of flow cytometry in the diagnosis of non- Hodgkin's lymphoma, Hodgkin's lymphoma, granulomatous inflammation and reactive lymph node specimens.

PURPOSE: In this study we aimed to compare the flow cytometry (FC) results of patients with B cell lymphoma, T cell lymphoma, Hodgkin's lymphoma, granulomatous inflammation and reactive lymph node and investigate the role of FC in malignant or non malignant conditions. METHODS: Ninety patients were divided into 5 groups according to histopathology results. Patients were compared according to cytokeratin and positivity percentage of the following surface markers: CD45, CD19, CD5, CD19-CD5, CD4, CD8, CD3,CD16-CD56, CD10, CD10-CD19, CD23, CD20, CD4-CD8, CD3-CD16-56, CD30, CD38, kappa and lambda light chains, CD20-CD23. Patients were also compared according to the intensity of the expression (exp) of same markers. ROC curve analysis was performed for CD19+ cell percentage, CD38 exp, kappa/lambda and lambda/kappa ratios. RESULTS: 1) Kappa/lambda and lambda/kappa ratios can distinguish B cell lymphoma from T cell lymphoma, Hodgkin's lymphoma, granulomatous inflammation and reactive lymph node; 2) CD19+ cell percentage can distinguish T cell lymphoma from Hodgkin's lymphoma, granulomatous inflammation and reactive lymph node; 3) CD38 exp can partly distinguish B cell lymphoma from T cell lymphoma, Hodgkin's lymphoma, granulomatous inflammation and reactive lymph node and T cell lymphoma from granulomatous inflammation, T cell lymphoma from reactive lymph node, Hodgkin's lymphoma from reactive lymph node. CONCLUSION: Flow cytometry has a role in distinguishing lymphomas from non malignant lesions.

Apoptosis patterns in eutopic and ectopic endometrium, adhesions and normal-looking peritoneum from women with or without endometriosis.

OBJECTIVE: To assess the apoptosis rate in eutopic and ectopic endometrial stromal and glandular cells, normal peritoneum and adhesions in women with endometriosis. METHODS: A total number of 97 women with (n:60) and without (n:37) histopathologically confirmed endometriosis who underwent laparoscopy or laparotomy in the early follicular phase of the menstrual cycles for pain and infertility were included in this study. Stage I/II and stage III/IV were categorized as early staged and late-staged endometriosis. The endometrial samples were obtained with a Novack cannula from the corpus of the uterus. Normal-looking peritoneum, peritoneal implants and adhesions were sampled and fixed in formaldehyde for immunohistochemical staining with Bcl-2 and Bax. Tissue samples were fixed in formaldehyde for the assessment of apoptosis via terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and M30 cytoDEATH antibody. RESULTS: The intensity of Bax staining of normal-looking peritoneum in early staged endometriosis was higher, compared to women with late-staged and women without endometriosis (P = 0.03). However, degree of Bcl-2 staining did not differ among early and late-staged endometriosis and women without endometriosis (P = 0.1). In terms of Bcl-2 and Bax staining in the stromal and glandular parts of the eutopic endometria, no significant differences were detected among three groups. In cases with early- and late-staged endometriosis the intensity of Bax and Bcl-2 stainings did not differ in both stromal and glandular parts of ectopic endometria. Number of cells with positive apoptotic signals assessed via TUNEL (P = 1.0) and M30 cytoDEATH antibody (P = 0.59) in normal-looking peritoneum did not differ between three groups. In addition, no difference in term of numbers of apoptotic cells obtained from adhesions was observed between three groups (for TUNEL, P = 0.29, for M30, P = 0.19). CONCLUSIONS: Apoptosis patterns did not differ in the eutopic and ectopic endometria as well as adhesions of women with or without endometriosis.

Cytokine and immune cell levels in peritoneal fluid and peripheral blood of women with early- and late-staged endometriosis.

OBJECTIVE: To investigate the level of cytokines and immune cells in the peripheral blood (PB) and peritoneal fluid (PF) of different stages of endometriosis. METHODS: A prospective study was conducted to include 97 women with (n 60) and without (n 37) histopathologically confirmed endometriosis. Based on rASRM classification, stage I/II and stage III/IV were categorized as early-and late-staged endometriosis. Prior to surgery, 10 ml of blood was withdrawn from antecubital vein and serum was obtained. Aliquots were made and stored at -70 degrees C until assayed for cytokines. PF was aspirated from the pouch of Douglas. Peripheral and PF samples were analyzed by ELISA in terms of IL-2, IL-4, IL-10 and IFN-gamma. Determinations of T helper, T suppressor, NK, and B cells were assessed by using cluster determinant-3 (CD-3), CD4, CD8, CD25, CD28, CD45, CD16, CD23 and antibodies against early T cell activation antigens such as CD45RA/CD45RO, CD-69 and late activation antigens such as HLA-DR. A multiparameter flow cytometry was applied to detect the cell activation antigen expression. RESULTS: In terms of cytokine levels in PB and PF's of control group and early- and late-staged endometriosis cases, no significant difference was depicted in the cytokine levels (p > 0.05). Levels of immune cells did not differ between three groups (p > 0.05). CONCLUSIONS: The result of this study did not show any significant difference in PB and PF cytokine and lymphocyte subgroups between normal and early- and late-staged endometriosis.

Effect of smoking on semen parameters of men attending an infertility clinic.

OBJECTIVE: To assess the effects of smoking on total sperm count, progressive sperm motility and sperm morphology among couples attending an infertility clinic. METHODS: A total of 223 sperm samples (126 smokers and 97 nonsmokers) from men attending an infertility clinic for routine infertility workup were compared on the basis of standard semen analysis. RESULTS: Cigarette smoking is negatively correlated with progressive motile sperm count (r = -0.1464, p = 0.042), but not with sperm concentration (p = 0.961), total motile sperm count (p = 0.890) and sperm morphology (p = 0.838). Furthermore, packages/ year (cumulative dose of cigarettes) did not correlate with any of the sperm parameters including sperm density (p = 0.976), total (p = 0.559) and progressive (p = 0.406) motile sperm count and sperm morphology (p = 0.449). CONCLUSIONS: Although the effect of smoking on male infertility remains inconclusive, smoking had an adverse effect on the progressive sperm motility, irrespective of total amount of cigarettes smoked per day.