Search

Additional file 1: Figure S1. Gating strategy to determine PD-1 and PD-L1 expression by DPSCs. DPSCs were identified by FSC and SSC (1) then live cells were gated by SSC/dead staining dot plot (2). Afterwards CD3neg CD4neg cells were selected (3) and fluorescence intensities in FITC and PECy7 channels shown by histograms. The mean percentage of adherent DPSCs in the FSC/SSC gating was 93.5% �� 2.2%. Figure S2. Gating strategy to determine PD-1 and PD-L1 expression by PBMCs pre-activated by CD3/CD28 linking. PBMCs were identified by FSC and SSC (1) then live cells were gated by SSC/dead staining dot plot (2). Afterwards CD3+CD56neg cells (T lymphocytes) were selected (3). CD4+ and CD4neg cells were further gated (4) and fluorescence intensities in FITC and PECy7 channels shown by histograms. Figure S3. Real Time PCR analysis of caspase 3 in PBMCs. Caspase 3 mRNA levels were evaluated in rPBMCs and aPBMCs alone, aPBMCs after DPSCs co-culture with and without PD-L1 inhibitor. Statistically significant increase of mRNA levels of caspase 3 was detected in aPBMCs after DPSCs co-culture with and without the addition of PD-L1 inhibitor (��P < 0.05 vs aPBMCs).

Введение. Стероидогенез представляет собой сложный ферментативный процесс, в котором циклический аденозин-монофосфат (цАМФ) и гуанозин-монофосфат (цГМФ) играют важную роль. Ингибиторы фосфодиэстеразы 5 (PDE5i) увеличивают цГМФ, увеличивая таким образом доступность NO. Цель: исследовать как длительное применение ингибитора фосфодиэстеразы 5 типа Варденафила влияет на надпочечниковый и тестикулярный стероидогенез у больных сахарным диабетом, с применением жидкостной хроматографии-масс-спектрометрии (LC-MS / MS). Дизайн. Длительное, проспективное, рандомизированное, плацебо-контролируемое, двойное слепое клиническое исследование. Материалы и методы. В исследование включены 54 пациента мужского пола, страдающие СД2, который был диагностирован в течение последних 5 лет. 26 и 28 пациентов были отнесены к группе, получающей исследуемый препарат и плацебо, соответственно. Вмешательства. Исследование состояло из фазы регистрации, фазы лечения (24 недели) (Варденафил 10 мг / плацебо дважды в день), и фазы наблюдения (24 недели). Контролируемые параметры: прогестерон (Р), 17-гидроксипрогестерон (17OHP), андростендион (А), тестостерон (Т), дегидроэпиандростерон (ДГЭА), ДГЭА сульфат (ДГЭАС), кортикостерон, 11-дезоксикортизола и кортизол (С) были оценены с использованием жидкостной хроматографии-масс-спектрометрии. Результаты. Никаких различий не было отмечено в уровне тестикулярных андрогенов в исследуемой и контрольной группой. Для надпочечников стероиды были оценены в соответствии с зоной, в которой они синтезируются. При анализе андрогенов пучковой зоны не было выявлено существенных различий в уровнях 11-дезоксикортизола как в исследуемой, так и в контрольной группе. В сетчатой зоне уровень ДГЭА значительно снизился во время лечения только в исследуемой группе (р = 0,007). При ретроспективном анализе более высокие уровни ДГЭА были зафиксированы на визитах 2 и 8. Отношение ДГЭАС / ДГЭА значительно увеличилось во время лечения только в группе, получающей препарат. Что касается клубочковой зоны надпочечников, уровень кортикостерона существенно изменился в ходе визитов как в исследуемой, так и в контрольной группе (р, Background. Steroidogenesis is a complex enzymatic process in which cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) play an important role. Phosphodiesterase-5 inhibitors(PDE5i) increase cGMP, improving NO availability. Objective. to investigate whether long-term, chronic treatment with the PDE5i Vardenafil affects adrenal and testicular steroidogenesis in diabetic men, using liquid chromatography-mass spectrometry (LC-MS/MS). Design. A longitudinal, prospective, investigator-started, randomized, placebo-controlled, double-blind, clinical-trial was carried out. Setting and Participants. 54 male patients affected by T2DM diagnosed within the last 5 years were enrolled. 26 and 28 patients were assigned to the verum and placebo-group, respectively. Interventions. The study consisted of an enrolment phase, a treatment phase (24weeks) (Vardenafil/placebo 10mg twice-daily), and a follow-up phase (24weeks). Outcome measurements: progesterone (P), 17-hydroxyprogesterone (17OHP), androstenedione (A), testosterone (T), dehydroepiandrosterone (DHEA), DHEA sulphate (DHEAS), corticosterone, 11-deoxycortisol and cortisol (C), were evaluated using LC-MS/MS. Results. No differences were seen in sex testicular steroids between study and control group. For the adrenal gland, steroids were considered according to the zona in which they are produced. Considering steroids produced in the zona fasciculata, no significant differences were seen in 11-deoxycortisol and C among visits, both in the study and in the control group. For the zona reticularis, DHEA significantly decreased during treatment only in the study group (p=0.007). At post-hoc test DHEA showed higher levels at visit 2 and 8 than in other visits. The DHEAS/DHEAS ratio significantly increased during treatment only in the verum group. Considering the adrenal zona glomerulosa, corticosterone significantly changed among visits both in the study and in the control group (p

Central hypogonadotropic hypogonadism (CHH) is an emerging pathological condition frequently associated with overweight, metabolic syndrome, diabetes, and midline defects. The genetic mechanisms involve mutations in at least twenty-four genes regulating GnRH neuronal migration, secretion, and activity. So far, the mechanisms underlying CHH, both in prepubertal and in adulthood onset forms, remain unknown in most of the cases. Indeed, all detected gene variants may explain a small proportion of the affected patients (43%), indicating that other genes or epigenetic mechanisms are involved in the onset of CHH. The aim of this review is to summarize the current knowledge on genetic background of CHH, organizing the large amount of data present in the literature in a clear and concise manner, to produce a useful guide available for researchers and clinicians.