Cell-type-specific determination of reactive oxygen species by flow cytometry.

BACKGROUND: Seminal leukocyte-generated reactive oxygen species may have a significant impact on sperm intracellular reactive oxygen species levels, therefore contributing to oxidative damage and consequent functional impairment of spermatozoa. This relationship may be utilized for male urogenital inflammation-driven oxidative stress diagnostics. OBJECTIVE: To obtain seminal cell-specific, reactive oxygen species-related fluorescence intensity cut-off values to differentiate leukocytospermic samples displaying reactive oxygen species overproduction (oxidative burst) from normozoospermic seminal samples. MATERIAL AND METHODS: Ejaculates gained by masturbation were obtained from patients in the framework of andrology consultations. The results published in this paper were generated from samples for which the attending physician requested spermatograms and seminal reactive oxygen species laboratory tests. Routine seminal analyses were performed according to World Health Organization guidelines. Samples were divided into normozoospermic "non-inflamed," and leukocytospermic groups. The semen was stained by 2',7'-dichlorodihydrofluorescein diacetate and the reactive oxygen species-related fluorescence signal and the percentage of reactive oxygen species-positive spermatozoa within the living population were quantified by flow cytometry. RESULTS: Reactive oxygen species-related mean fluorescence intensity was higher in both spermatozoa and leukocytes from leukocytospermic samples than in those from normozoospermic samples. Mean fluorescence intensity in spermatozoa was positively and linearly correlated with mean fluorescence intensity measured in leukocytes in both groups. DISCUSSION: The capacity of spermatozoa to generate reactive oxygen species is at least three log lower than that of granulocytes. The question is whether the reactive oxygen species-producing machinery of spermatozoa is capable of causing autologous oxidative stress or whether leukocytes are the predominant source of seminal oxidative stress. Based on our observations, the reactive oxygen species production of leukocytes may have a significant impact on the overall reactive oxygen species levels measured in spermatozoa. CONCLUSION: Reactive oxygen species-overproducing leukocytospermic and normozoospermic seminal samples can reliably be differentiated based on reactive oxygen species mean fluorescence intensity measurement.

Preclinical characterization of CPL302-253, a selective inhibitor of PI3Kδ, as the candidate for the inhalatory treatment and prevention of Asthma.

Asthma is a common chronic inflammatory disease. Although effective asthma therapies are available, part of asthmatic population do not respond to these treatment options. In this work we present the result of development of CPL302-253 molecule, a selective PI3Kδ inhibitor. This molecule is intended to be a preclinical candidate for dry powder inhalation in asthma treatment. Studies we performed showed that this molecule is safe and effective PI3Kδ inhibitor that can impact many immune functions. We developed a short, 15-day HDM induced asthma mouse model, in which we showed that CPL302-253 is able to block inflammatory processes leading to asthma development in vivo.

[Sleep disturbances and nightmares as risk factors for suicidal behavior among men and women]. / Alvászavarok és rémálmok mint az öngyilkossági magatartás férfiak és nok körében.

INTRODUCTION: Today, more and more international studies analyze the connection between sleep disturbances, including nightmares, and the risk of suicide. The majority of these studies focuses on patients with depression and PTSD; a few studies use community samples. The present paper is based on a community sample research; it examines gender differences in the relationship between nightmares and suicidal behavior (ideation and attempt) during the three years preceding the study. METHODS: The database was derived from the data of the national representative survey Hungaro study 2006, the objective of which was the health status of the Hungarian adult population (N=4642). The five-item version of Athens Insomnia Scale was used for the examination of sleep disturbances; depression was measured with the shortened version of the Beck depression questionnaire. The questions dealing with nightmares were part of the Sleep Quality Questionnaire. Sleep problems, nightmares and depression were considered as risk factors for both suicidal ideation and suicide attempts. Associations were analyzed by logistic regression. The data were corrected for age, gender, education, and financial situation. RESULTS: Our analysis verified the connections between suicidal thoughts and attempts, and nightmares. The results show that the frequent occurrence of nightmares is associated with a nearly four times higher risk of suicide attempts among men (OR=3,89) and a threefold increase among women (OR=1,74). For suicidal thoughts, nightmares correlate with a three times higher risk for men (OR=2,97) and one and the half higher risk for women (OR=1,58). In our sample, frequent nightmares and sleep disorders increase the risk of suicidal behavior more than depression. CONCLUSION: After clinical studies proved the influence of sleep problems in various patient groups, our epidemiological study seem to confirm that nightmares have an independent role in the formation of suicidal behavior.

[Knowledge about and attitudes toward suicide in the Hungarian general population and in the helping professions]. / Ongyilkossággal kapcsolatos ismeretek, attitüdök a magyar lakosság és a segítô foglalkozású szakemberek körében.

In the framework of an international suicide prevention program, we examined suicide-related attitudes and knowledge using the questionnaire constructed by Hubbard-McIntosh in the Hungarian population (N=4238) and among 231 helping professional. Our results show that proper information and false assumptions as well as attitudes toward suicide (refusal or acceptance of the right to commit suicide in general and exceptional situations in life) are closely related to several characteristics of the individual's socio-demographic position, to geographical location, to depression, and to the personal involvement in suicide-related events. In line with our expectations, the level of knowledge of helping professionals is higher than that of the general population. Compared to the general population, a significantly higher proportion of helping professionals deny the right to commit suicide; however, in relation to a particular situation of life - e.g. terminal illness - their opinion is similar to that of the general population.

Nitric oxide involvement in the delayed antiarrhythmic effect of treadmill exercise in dogs.

We have shown previously that a single period of treadmill exercise in dogs protects the heart against the severe ventricular arrhythmias that arise when a major (anterior descending) branch of the left coronary artery is occluded following anaesthesia 24 h later. This protection is aminoguanidine sensitive, suggesting a role for nitric oxide (NO) in this exercise-induced delayed antiarrhythmic effect. The present study has further examined the possible role of NO as a mediator and/or as a trigger using the selective induced (iNOS) inhibitor S-(2-aminoethyl)-methyl-isothiourea (AEST) and the specific but not selective nitric oxide synthase inhibitor N(omega)-nitro-L-arginine-methyl-ester (L-NAME). Exercise markedly reduced the severity of ischaemia and reperfusion-induced ventricular arrhythmias 24 h later. Thus, only one of the dogs (8%) so exercised fibrillated on occlusion (contrast 46% in the control, non-exercised dogs; P<0.05) and the marked changes in the inhomogeneity of electrical activation that occur in the ischaemic region following occlusion were much reduced (P<0.05 compared to controls). This delayed exercise-induced cardioprotection was significantly attenuated by the nitric oxide synthase (NOS) inhibitors L-NAME, given prior to the exercise protocol and by AEST given prior to the coronary artery occlusion. For example, survival from the ischaemia-reperfusion insult was 54% in the exercise dogs, 0% in the controls and 14% in those dogs given a NOS inhibitor. We conclude that nitric oxide (NO) is both the trigger and the mediator of this delayed protection against ischaemia and reperfusion-induced arrhythmias.

Delayed exercise-induced protection against arrhythmias in dogs--effect of celecoxib.

A 20-min period of treadmill exercise in dogs, sufficient to increase heart rate by 90-100 beats min(-1), markedly decreases the severity of the life-threatening ventricular arrhythmias that result when, following anaesthesia 24 h later, the left coronary artery is occluded. This protection was unaffected by the administration of the selective cyclooxygenase-2 inhibitor celecoxib (two times 3 mg kg(-1) given intravenously). These results show that prostacyclin, derived from cyclooxygenase-2, plays no role in this pronounced delayed cardioprotection.

Sildenafil (Viagra) reduces arrhythmia severity during ischaemia 24 h after oral administration in dogs.

Sildenafil (Viagra) prolongs repolarisation in cardiac muscle, an effect that could lead to ventricular fibrillation (VF). Sildenafil (2 mg kg(-1)) was given by mouth to 12 mongrel dogs and, 24 h later, these dogs were anaesthetised, thoracotomised and subjected to a 25 min occlusion of the anterior descending coronary artery. Haemodynamic parameters were similar in this and the control group, but there were fewer and less serious ventricular arrhythmias during occlusion in the sildenafil group (VF 17 vs 60%; ventricular premature beats 140+/-52 vs 437+/-127% and episodes of ventricular tachycardia 4.0+/-3.2 vs 19.3+/-7.7%, all P<0.05). However, reperfusion VF and indices of ischaemia severity (epicardial ST-segment mapping, inhomogeneity) were not modified by the drug. Sildenafil increased the QT interval, especially during ischaemia. Our conclusion is that ischaemia-induced ventricular arrhythmias are reduced by sildenafil, but this protection is less pronounced than that following cardiac pacing or exercise.