Differential flow cytometric detection of intracellular cytokines in peripheral and peritoneal mononuclear cells of women with endometriosis

OBJECTIVE: The pathogenesis of endometriosis is related to functional changes in CD3+ and CD14+ cells observed both at the local and systemic level. Here we investigated whether, and if so, how the body compartment influences cytokine expression in stimulated peritoneal and peripheral CD3+ and CD14+ cells of women with endometriosis. STUDY DESIGN: Isolated peripheral blood (PB) and peritoneal fluid (PF) mononuclear cells from women with endometriosis were cultured under non-adherent conditions and stimulated with PMA and ionomycin for 6h to induce intracellular cytokine synthesis of TNF-alpha, IFN-gamma, and IL-8 by CD3+ cells or with LPS for 9h to produce TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 by CD14+ cells. RESULTS: The percentages of positive CD3+ cells stained for TNF-alpha and IFN-gamma were significantly higher and those stained for IL-8 were significantly lower in PF compared with PB, this being independent of the stage of endometriosis. In contrast, the percentages of CD14+ cells producing TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 were significantly higher in PB than PF of women with endometriosis. CONCLUSIONS: Monocytes/macrophages and lymphocytes derived from the peripheral and peritoneal compartments of women with endometriosis differentially respond to stimulated cytokine synthesis induction. However, it is difficult to state whether the observed phenomenon is more related to body compartment influence per se or to the presence of endometriosis.     

Über Immunisierung Gegen Diphtherie in Warschau

Zusammenfassung Es wird darauf aufmerksam gemacht, daß bei Krankheiten, gegen welche sich gleichsam spontan im Alter eine Immunität ausbildet und bei welchen die betreffenden Erreger eine Durchseuchung bewirken, Unterschiede in der Immunisierbarkeit vorhanden sein müssen, die vom Alter und Durchseuchungsgrad abhängig sind. Da die Durchseuchung in verschiedenen epidemischen Perioden verschieden, geringer auf dem Lande als in der Stadt und abhängig von der Kontaktmöglichkeit und anderen Faktoren ist, so kann auch der Erfolg der Immunisierung bei quantitativer Betrachtung von Fall zu Fall und von Ort zu Ort verschieden sein.Da die Diphtherieepidemien in Form von Wellen verlaufen, so kann man nach dem bloßen Verschwinden einer Epidemie nach der Impfung noch nicht ohne weiteres auf den Erfolg der Impfung schließen.Bei der statistischen Bearbeitung des Impferfolges soll der verschiedenen Expositionszeit bei den Geimpften und Ungeimpften Rechnung getragen werden. Man muß auch die Anzahl der geimpften Kinder von der Anzahl der Kinder zwischen o und 15 Jahren, die sich im Zustande der erworbenen Immunität befinden, unterscheiden. Es wurde auf verschiedene Fehler bei der Berechnung der Morbidität bei Geimpften und Ungeimpften aufmerksam gemacht. In den Jahren 1931–1934 wurden in Warschau 94920 Kinder geimpft. Im Jahre 1934 waren 86602 der Kinder zwischen 0 und 15 Jahren=31,2% der kindlichen Bevölkerung im Zustande der gegen Diphtherie erworbenen Immunität. Verwendet wurde lediglich das Anatoxin vonRamon, und zwar durchschnittlich 60–90 Flockungseinheiten.Es wurde eine 4 fache Abnahme der Morbidität und eine 8–10 fache Abnahme der Mortalität bei Geimpften festgestellt. Die Diphtherieimpfung wurde daher auf breiter Basis empfohlen.E. Grodzki, J. Jakubkiewicz, W. Mazurek und M. Szeynman führten die Impfungen durch. Die statistische Bearbeitung lag in den Händen von E. Grzegorzewski.     

Event-free survival in patients after an acute coronary event with exercise-induced normalization of the T-wave

BACKGROUND: Risk stratification of patients with unstable angina or non-Q-wave myocardial infarction (MI) is an unresolved clinical problem. The prognostic value of T-wave normalization (TWN) during exercise has not been studied in this group of patients. HYPOTHESIS: Event-free survival in clinically stable patients after an acute coronary event without ST-segment elevation can be predicted by the presence of exercise-induced TWN. METHODS: Sixty-five patients (43 men and 22 women, mean age 62+/-10 years) entered the study. The diagnosis of unstable angina and non-Q-wave MI was made in 40 and 25 patients, respectively. A treadmill exercise test was performed in all patients after clinical stabilization. The patients were divided into three groups: those with negative baseline T waves and exercise-induced TWN (Group 1); those with negative baseline T waves, but without TWN (Group 2); and those with positive baseline T waves (Group 3). The patients were followed up for 6 months. RESULTS: During follow-up, serious cardiovascular complications occurred in 15 (23%) patients. These included exacerbation of ischemic heart disease (14 patients) and acute MI (1 patient). Event-free survival was greater in patients in Group 1 (95%) than in those in Group 2 (68%, p < 0.034) or Group 3 (71%, NS). Among all patients studied, exercise-induced TWN was predictive of event-free survival with a sensitivity of 38% and a specificity of 93%. CONCLUSIONS: In clinically stable patients after an acute coronary event without ST-segment elevation, exercise-induced TWN is a specific but n ot sensitive predictor of event-free survival after 6 months.     

Preoperative evaluation of chronic alcoholics assessed for surgery of the upper digestive tract

BACKGROUND: Alcoholics are at risk of developing major complications in the postoperative period. Adequate prophylactic treatment, as well as preoperative abstinence, can significantly decrease the rate of complications. However, the preoperative diagnosis of alcoholism is difficult to establish. The purpose of this study was to assess whether three preoperative visits, an alcohol-related questionnaire (CAGE), and the laboratory markers carbohydrate-deficient transferrin (CDT) and gamma-glutamyltransferase (GGT) would increase the rate of detection of chronic alcoholics. METHODS: The study included the Departments of ENT, Facial and Maxillofacial Surgery, and General Surgery of a university hospital; 705 male patients were assessed for tumor surgery of the upper digestive tract and were allocated to 5 different groups. All patients were seen three times, and five different strategies were used to detect chronic alcoholics. The gold standard was the diagnosis of alcohol misuse made by an experienced (blinded) investigator according to the DSM-III-R. The main outcome measurements were the detection rates of the different test strategies. RESULTS: By clinical routine alone, only 16% were detected during the first visit and 34% after three visits. If the CAGE questionnaire was added, sensitivity increased to 64%. The further addition of GGT or CDT led to 80 and 85% detections, respectively. A combination of all tests had a sensitivity of 91%. CONCLUSIONS: To detect more alcoholic patients at risk for major complications, patients should be seen more often, and additional diagnostic tools such as the CAGE, CDT, and GGT should be used before surgery.     

Evaluation of angiogenesis, p-53 tissue protein expression and serum VEGF in patients with endometrial cancer

Endometrial carcinoma occurs mostly in post-menopausal women. Classical methods of prognostication, as FIGO stage and histopathologic grade, could be improved by applying additional techniques, utilizing molecular biology and immunochemistry. p-53 tumor suppressor gene, the most commonly mutated gene in human cancers has been shown to play an important role in the biology of gynecologic carcinomas. Angiogenesis, a process of formation of new vessels, being connected to tumors progression and metastatic potential was shown to be linked with tumor suppressor genes expression. The aim of the study was to evaluate relationships between intensity of tumor angiogenesis, serum levels of Vascular Endothelial Growth Factor (VEGF) and tissue p-53 protein expression in endometrial adenocarcinoma. Angiogenic Point's Density (APD) was calculated in hot spots areas using the morphometric appliance. For detection of p53 protein in tumor samples, LSAB + Kit Alkaline Phosphatase (DAKO) was used. VEGF levels were assessed in patient's blood sampled before the operation. Overexpression of p53 protein was found in tumor tissue in 35.2% of cases and mean angiogenic points density was greater in p53 positive cases. Serum levels of VEGF were above the cut off level in 54.5% of patients, in those cases angiogenesis was also elevated. In cases of p53 overexpression, VEGF levels tended to be greater as compared with p53 negative cases. In conclusion, our study demonstrated that angiogenesis was more intensive in p53 positive cases, confirming the hypothesis of tumor suppressor-gene regulation of the process of neovascularization. Serum levels of VEGF were borderline-significantly higher in cases of p53 overexpression, they were also correlated to the angiogenesis. Joint assessment of angiogenesis and tumor suppressor genes expression may contribute to reliable evaluation of the biology of endometrial carcinoma.     

Effect of Asthma Call-back Survey methodology changes on work-related asthma estimates, 19 states, 2007–2012

Objective: Asthma Call-back Survey methodology has been changed recently, as a new sampling design, weights calculation (2011–2012), and revised work-related asthma (WRA) section (2012) were implemented. To assess the effect of these changes on the WRA and possible WRA estimates among ever-employed adults with current asthma, we analyzed 2007–2012 data for 37?505 ever-employed adults (≥18 years) collected from 19 US states (representing an estimated 10 million adults each year). Methods: Using data from landline telephone (LLP) households, we calculated estimates applying poststratification weights (2007–2010) and “raking” weights (2011–2012). Also, using data from LLP/cellular telephone (CP) households combined, we calculated estimates applying “raking” weights (2012). Results: Based on LLP household data, the WRA estimates ranged from 7.8% to 9.7% during 2007–2010, was 9.1% in 2011 and 15.4% in 2012. Possible WRA estimates ranged from 35.1% to 38.1% during 2007–2010, was 38.1% in 2011 and 39.8% in 2012. Using the 2012 LLP/CP household data, the WRA and possible WRA estimates were 15.4% and 38.9%, respectively. Conclusions: Implementation of “raking” weights did not substantially change the WRA or possible WRA estimates among ever-employed adults with current asthma. The WRA and possible WRA estimates based on LLP and LLP/CP samples in 2012 were comparable, as CP users are younger and less likely to have WRA. The substantial upward shift in the 2012 WRA estimates likely was associated with the revision to the WRA section.     

Restoration of Ca2+ influx and degranulation capacity of variant RBL-2H3 cells upon implantation of isolated cromolyn binding protein

Recently, variants of the rat basophilic leukemia cells (RBL-2H3), deficient in their binding capacity for the antiallergic drug cromolyn but displaying unimpaired ability to bind IgE, were selected and cloned [Mazurek, N., Bashkin, P., Petrank, A. & Pecht, I. (1983) Nature (London) 303, 528-530]. Although the histamine content and the number of IgE receptors in these variants are similar to those of the parental cells, they cannot be stimulated immunologically to allow Ca(2+) influx and to degranulate. In contrast, the Ca(2+) ionophore A23187 causes these variants to degranulate, indicating that the mechanism distal to the Ca(2+) gating is intact in the variants. The cromolyn binding protein (CBP), present in the membranes of RBL-2H3 cells, has recently been isolated by affinity chromatography under nondenaturing conditions. In the current study we have used Sendaivirus envelopes as fusogenic carriers to implant the purified CBP into the membrane of variant basophils that were defective in it. This fusion leads to the restoration of Ca(2+) uptake and degranulation capacity of the variants after IgE-mediated stimulation. These restored activities seem to show a sigmoidal dependence on the amount of incorporated CBP. Saturation values comparable to those of the parental line are reached when the level of implanted CBP approaches its density on the latter line. The restored capacity is due to the implanted CBP, because the reinstated immunological response can be blocked by the inhibitory drug cromolyn and by monoclonal antibodies specific to CBP, both shown to prevent Ca(2+) uptake and degranulation in mast cells and parental RBL-2H3 cells. These results point out that CBP plays an important role in the Ca(2+) gating process resulting in degranulation.