Secretory immunoglobulin A concentration in non-stimulated and stimulated saliva in relation to the status of smoking.

Introduction: Secretory immunoglobulin A (sIgA) is a dominant immunoglobulin in the saliva. It is the first line of defense against microorganisms. Aim of the study: Analysis of secretory immunoglobulin A concentration in non-stimulated and stimulated saliva. Assessment of sIgA concentration in relation to the status of cigarette smoking of the investigated. Material and Methods: Survey and biochemical studies of saliva were conducted in the group of 109 people (smokers and non-smokers) aged 20-54. The smokers smoked from 5 to more than 20 cigarettes daily. The investigation material was nonstimulated and stimulated saliva collected from patients on the same day between 9.30 and 11.30 a.m, 1.5-2h after meal. Directly after non-stimulated and stimulated saliva collection Salivette tubes were placed in the ice container with the temperature of 4oC, then centrifuged at the temperature of 4oC for 12 minutes at 3000 r/min. The obtained supernatant was stored at the temperature of -75ºC until the assays were performed. sIgA concentration was determined using sIgA ELISA Kit (Immunodiagnostik AG, Germany). Statistical analysis was conducted with the use of Mann ­ Whitney test. While investigating the influence of age on the studied parameters Spearman correlation coefficient and its significance were used. Statistically significant test values were those of p<0.05. Results: In non-stimulated saliva sIgA concentration was significantly higher compared to stimulated saliva (Z = 4.00, p<0.001). No significant differences were stated in sIgA concentration in non-stimulated saliva between smokers and non-smokers (Z = 0.26, p>0.05). No essential differences were stated in sIgA concentration in stimulated saliva between non-smokers and smokers (Z = 0.23, p>0.05). Essential differences were stated between the groups. In men sIgA concentration in stimulated saliva was significantly higher compared to women (Z = 2.25, p<0.05). Conclusions: sIgA concentration in non-stimulated saliva is significantly higher in comparison to stimulated saliva. The status of cigarette smoking does not influence the essential differences in sIgA concentration in nonstimulated and stimulated saliva. In men sIgA concentration in stimulated saliva is significantly higher in comparison to women.

Stimulated and non-stimulated saliva as biological material in the assessment of cotinine concentration.

INTRODUCTION: A biomarker of the exposure to the tobacco smoke is cotinine - a nicotine metabolite with a half-life about 16 hours. Analysis of cotinine concentration in biological material allows objective assessment of the cigarette smoking status and environmental exposure to the tobacco smoke. AIM OF THE STUDY: The aim of the study was evaluation of the influence of method of saliva collection: stimulated or non-stimulated one by chewing a paraffin cube on cotinine concentration in the saliva as well as the analysis of the obtained cotinine values depending on the status of cigarette smoking. MATERIAL AND METHODS: Survey with the use of authorial survey questionnaire and biochemical survey of saliva were conducted among 115 people aged 20-50 who reported for treatment at the Department of Conservative Dentistry with Endodontics of the Medical University of Lublin. In the examined group 66 people declared cigarette smoking, 49 reported they had never smoked cigarettes and had never tried to smoke. Cotinine concentration in the saliva was assayed with the use of Cotinine test (Calbiotech). Obtained study results were submitted to statistical analysis using Ch2 test. Statistically essential test values were those of p<0.05. While investigating the influence of age on the examined parameters Spearman correlation coefficient and the test of its relevance were employed. RESULTS: Mean cotinine concentra- tion in non-stimulated saliva in smokers was 310.36 ng/ml, in stimulated saliva 305.61 ng/ml. No statistically essential differences were stated between stimulated and non-stimulated saliva (Z=0.36, p>0.05). Cotinine concentration in non-stimulated and in stimulated saliva was increasing with age of the investigated. CONCLUSIONS: Cotinine concentration in both in stimulated and non-stimulated saliva obtains similar values. Cotinine concentration both in stimulated and non-stimulated saliva increases with age of the investigated.

[Cotinine concentration in the saliva in relation to oral hygiene procedures]. / Stezenie kotyniny w slinie w relacji do przeprowadzania zabiegów higienicznych w jamie ustnej.

Cotinine is a biomarker of the exposure to the tobacco smoke, nicotine metabolite with half-life in the saliva which is 17 hours. Assessment of cotinine concentration enables among others verification of the questionnaire data as well as evaluation of both smokers and non-smokers exposure to the tobacco smoke. Practicing proper oral hygiene procedures is an essential factor of the prophylaxis of dental caries and periodontal diseases which influence general health state. The removal of dental calculus is achieved by proper teeth brushing and the use of additional oral aids. The aim of the study was evaluation of cotinine concentration in non-stimulated saliva in order to verify questionnaire data (smoker/non-smoker) and analysis of practicing oral hygiene procedures in relation to the status of cigarette smoking. Questionnaire and biochemical studies were conducted in the group of 116 people aged 20-54. In questionnaire survey 53 people (45.69%) confirmed cigarette smoking, 63 (54.31%) declared they had never smoked and never tried to smoke. Non-stimulated saliva was collected between 9(30) and 11(30), 1,5-2 hours after meal. Cotinine concentration was assayed with the use of Cotinine ELISA (Calbiotech, USA). Obtained study results were submitted to statistic analysis with the use of Chi2. Statistically essential test values were those with p<0,05. In the study group the mean value of cotinine concentration was 155.76 ng/ml. Brushing teeth once a day or less frequently was reported by 26.92% smokers and 4.76% non-smokers, brushing teeth at least twice a day was reported subsequently by 73.08% and 95.24% participants. Non-smokers in comparison with smokers considerably more frequently brushed their teeth, at least twice a day (XZ=11.11, p<0.001). Smokers used a toothbrush with medium hardness bristle (X2=6.05, p<0.05) as well as toothpicks to maintain hygiene of interdental spaces and teeth contact surfaces (X2=21.34, p<0.001) whereas they used dental floss less frequently (X2=10.64, p<0.01). Smokers more fre. quently brushed their teeth improperly (X2=1 3.41, p<0.001). Smokers in comparison with non-smokers did not practice proper oral hygiene which is an essential risk factor of the oral health. It is crucial for dental surgeons to conduct oral hygiene instructions in smokers as well as realization of health threats resulting from cigarette smoking.

[Lymphocyte activation markers in patients with ovarian cancer]. / Markery aktywacji limfocytów u pacjentek z rakiem jajnika.

INTRODUCTION: The process of T cells activation is necessary for the performance of the defense functions. Successful activation depends on direct lymphocyte - antigen-presenting cell contact and signal transmission to the lymphocyte. Activated T cells exhibit surface expression of molecules such as CD69, CD25 and HLA-DR. The effect of cell activation is a cascade of molecular events leading to proliferation and clonal expansion of antigen-specific T cells. OBJECTIVES: The aim of this study was to evaluate the phenotype and T cell activation markers: CD69, CD25 and HLA - DR in the peripheral blood and tumor tissue of ovarian cancer patients. MATERIAL AND METHODS: The study group consisted of 26 patients operated due to ovarian cancer (FIGO Ilb - IV). Mononuclear immune cells were isolated from peripheral blood and ovarian cancer tissue. To obtain peripheral blood lymphocytes, blood was collected into heparinized tubes and diluted 1:1 with PBS, then layered on Gradisol L and centrifuged 20 minutes at 2800 rpm. Mononuclear cells were washed twice with PBS and labeled with monoclonal antibodies. A small piece of tumor tissue (about 1cm3) was fragmented with a surgical blade. Minced tissue was suspended in PBS and layered on Gradisol L for mononuclear cells isolation. To assess the phenotype and activation status of peripheral blood and tumor infiltrating lymphocytes, we used FACS Canto cytometer and monoclonal antibodies conjugated with fluorochromes: anti-CD3-FITC, anti-CD4-PE-Cy5, anti-CD8-APC, anti-CD25-PE, anti-CD69-PE-Cy7, anti-HLA-DR-PE-Cy7. Statistical analysis was performed using the Statistica 5.0 and Wilcoxon test. RESULTS: In all cases we detected T helper CD3+CD4+ and cytotoxic CD3+CD8+. T lymphocytes in both blood samples and tumor tissues. We observed no statistically significant difference in the percentage of CD3+ CD4+ cells among the mononuclear cells present in peripheral blood and tumor tissue. The percentage of CD3+CD8+ cytotoxic T lymphocytes was higher among mononuclear cells isolated from the tumor tissue. The percentage of CD3+ lymphocytes expressing the very early activation marker CD69 was significantly higher among tumor infiltrating lymphocytes compared with peripheral blood lymphocytes. Similarly the percentages of CD3+CD4+CD69+ T helper lymphocytes and CD3+CD8+CD69+ cytotoxic T lymphocytes were significantly higher on lymphocytes isolated from tumor tissue when compared to blood. The expression of an early activation marker - CD25 was significantly higher on the CD3+ and CD3+CD8+ peripheral blood lymphocytes compared to CD3+ and CD3+CD8+ tumor infiltrating lymphocytes. There were no statistically important differences between the percentages of, isolated from blood and tissue, CD3+CD4+ T helper lymphocytes. The expression of the late activation marker - HLA-DR was significantly higher on CD3+ lymphocytes isolated from tumor tissue compared with peripheral blood. Similarly the percentages of CD3+CD4+ lymphocytes and CD3+CD8+ cytotoxic T cells expressing HLA-DR were significantly higher among tumor infiltrating lymphocytes when compared to peripheral blood ones. CONCLUSIONS: T cells obtained from ovarian cancer tissues are activated cells. The state of T cell activation may be the result of direct contact of these cells with tumor antigens. The low expression of CD25 may suggest abnormal clonal expansion of antigen-specific lymphocytes.