Cytokines in the male reproductive tract and their role in infertility disorders.

Summary
Chronic genitourinary inflammation results in Leukocytospermia (LCS), an elevated number of white blood cells (WBCs) in semen, which, in association with oxidative stress, may suppress sperm function, and manifest as male factor infertility. The current clinical diagnosis of LCS employs manual enumeration of WBCs and requires complex staining and laboratory skills or measurement of inflammatory cytokines and chemokines levels. Many patients with idiopathic infertility are asymptomatic. In search of better inflammatory markers for LCS, we evaluated expression of toll-like receptors 2 and 4 (TLR-2/4), cyclooxygenase-2 (COX-2), and nuclear factor (erythroid-derived 2)-like 2 (Nrf-2) in semen samples of age-matched infertile patients with and without LCS. We employed the usage of specific Western blot evaluation, cytokine array; immunofluorescence microscopy (IFM) followed by computer-based analysis, and other molecular approaches. As compared with non-LCS patients (n = 38), semen samples from LCS patients (n = 47) displayed significantly lower total sperm count (p < 0.01), motility (p < 0.0001), normal head count (p < 0.0001), and a significantly higher white blood cell count (p < 0.0001). Differential cytokine profiling of seminal plasma by antibody array revealed up-regulation of several pro-inflammatory chemokines in LCS samples. Western blot analysis of LCS seminal plasma (n = 15) also showed a significant increase in expression of TLR-2 (p < 0.001) and 4 (p < 0.01), COX-2 (p < 0.001), and Nrf-2 (p < 0.001) as compared with semen samples from non-LCS patients (n = 15). Computer-based objective IFM analysis of spermatozoa from LCS patients showed increased expression of TLR-4 (p < 0.001), Cox-2 (p < 0.01), and (Nrf-2) (p < 0.01). Significant differences in the subcellular localization of these proteins were evident in the sperm head and tail segments of LCS samples. Altogether, these observations suggest that TLR-2/4, COX-2, and Nrf-2 can serve as novel biomarkers of inflammation and oxidative stress. Therefore, developing a rapid assay for these biomarkers may facilitate early diagnosis and management of LCS especially in idiopathic and asymptomatic male infertility patients.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/andr.12074

Differential expression of novel biomarkers (TLR-2, TLR-4, COX-2, and Nrf-2) of inflammation and oxidative stress in semen of leukocytospermia patients.

Changes in serum obestatin, preptin and ghrelins in patients with Gestational Diabetes Mellitus.

Aims
To compare the incidence of large-for-gestational-age (LGA) infants in women diagnosed with gestational diabetes mellitus (GDM) early and late in pregnancy, and evaluate associated factors.

Methods
A total of 284 women with GDM who commenced antenatal care before 20 weeks of gestation were enrolled: 142 were diagnosed before 20 weeks (early GDM) and 142 were diagnosed after 20 weeks of gestation after normal initial screening tests (late GDM). Incidence of LGA infants were compared. Factors associated with LGA and pregnancy outcomes were evaluated.

Results
Both groups had comparable baseline characteristics. The late GDM group were more likely to gain weight greater than recommended (P = 0.009) and less likely to have optimal glycemic control (P = 0.035). Incidences of maternal and neonatal complications, including LGA, were not significantly different between the groups. Logistic regression analysis demonstrated that the timing of GDM diagnosis was not significantly associated with LGA. Less gestational weight gain than recommended decreased the risk of LGA by 76% (adjusted odds ratio [OR] 0.24, 95% confidence interval [CI] 0.09-0.67, P = 0.007), while gestational weight gain greater than recommended doubled the risk of LGA (adjusted OR 1.99, 95% CI 1.03-3.87, P = 0.041). Good glycemic control also reduced the risk of LGA by 66% (adjusted OR 0.34, 95% CI 0.16-0.71, P = 0.886).

Conclusion
Gestational weight gain and glycemic control, but not timing of diagnosis, were independently associated with LGA in women with GDM. Women diagnosed with GDM early had similar rates of LGA infants as women diagnosed late.

Incidence of large for gestational age infants when gestational diabetes mellitus is diagnosed early and late in pregnancy.

In order to explore the anti-inflammatory effects of Nodosin from Isodon serra, a traditional Chinese herb medicine, mouse T lymphocytes were incubated with Nodosin. In the current study, Nodosin suppressed the overproduction of the T lymphocytes; moreover, cell mitosis cycle was modulated by interfering with DNA replication in G1 stages via inhibition of IL-2 cytokine secretion at the mRNA level by Nodosin. Interestingly, Xylene-induced mouse tumescence model results suggested Nodosin depressed the murine ear-swelling extent and the level of IL-2 in the blood serum. Finally, Nodosin possessed significant anti-inflammatory effects and is a potential candidate for further clinical trial.

Anti-inflammatory function of Nodosin via inhibition of IL-2.

Inflammation of male accessory glands constitutes one of the causes of infertility and in the same time there is no accurate method to diagnose a silent inflammation. The study was aimed in the evaluation of the levels of interleukins (IL-1β) and (IL-18) in the seminal plasma and their significance in the diagnostics of semen pathology. 107 men diagnosed in the Andrology Outpatient were included in the study. Patients were divided in the 4 subgroups according to the semen parameters (normospermia, asthenoteratozoospermia, oligoasthenoteratozoospermia, and azoospermia). Next they were divided according to the incidence of leukocytospermia. The levels of proinflammatory cytokines were measured in the seminal plasma with the ELISA method. The significant correlation between IL-1β and IL-18 and the rate of abnormal spermatozoa was observed in the oligoasthenoteratozoospermia. The increased levels of IL-1β and IL-18 were found in patients with an elevated leukocyte count in the semen. In addition, the levels of IL-1β and IL-18 were shown to correlate with the number of leukocytes in semen. Our results suggest the role of IL-1β and IL-18 in the genesis of semen pathology that might be responsible for defects of spermiogenesis manifested with impaired morphology of spermatozoa. Moreover, it seems that the increased levels of IL-1β and IL-18 might be indicators of leukocytospermia and can be useful in diagnostics of male accessory gland inflammation.

/pdf/interleukin-1b-and-interleukin-18-and-their-connection-with-cf84oeml2r.pdf

Interleukin 1β and interleukin 18 and their connection with leukocytospermia in human semen

Male infertility is a serious diagnostic problem and in many cases the exact cause of failure to reproduce remains unknown. New diagnostic methods are being evaluated in search of more precise diagnosis and possibility of casual treatment. Measuring the level of cytokines, both in seminal plasma and serum, does not only expand the diagnostic options, but also, through the growing knowledge of immune processes, can give rise to new therapeutic methods of improving the quality of semen and increasing the chance to reproduce. The article reviews the role of proinflammatory cytokines in its pathogenesis.

Role of inflammatory cytokines in male infertility

https://www.termedia.pl/Journal/-10/pdf-15412-80?filename=Interleukin 1b and interleukin 18.pdf

Clinical immunology Interleukin 1β and interleukin 18 and their connection with leukocytospermia in human semen

Male infertility is a major diagnostic and therapeutic problem. Inflammatory process seems to be one of the most important factor influencing the parameters of seminogram. As currently used methods in above pathologies are still insufficient it has been postulated that novel parameters are necessary to make the diagnostics more accurate. Proinflammatory cytokines i.e. interleukins 6 (IL-6) and 8 (IL-8) seem to play an important role in inflammatory processes in male genital tract. The aim of present study was to evaluate their significance in semen diagnostic in correlation to seminogram parameters as well as to well established inflammatory signs i.e. leukocytospermia. We evaluated the correlation between the level of IL-6 and the rate of immobile spermatozoa in men with asthenoteratozoospermia that suggests its influence on impaired sperm motility. This observation was reinforced by the fact, that IL-6 level negatively correlated with slow progressive type of movement in this group of men. The increased levels of IL-6 and IL-8 in seminal plasma of men with leukocytospermia and correlation between IL-6 level and number of leukocytes in semen in this group of men suggested relation of IL-8 and 6 with mobilization of inflammatory infiltrate in male reproductive system.

https://www.termedia.pl/Journal/-10/pdf-15413-90?filename=Interleukin 6 and interleukin 8.pdf

Clinical immunology Interleukin 6 and interleukin 8 concentrations in seminal plasma of male with seminogram abnormalities

Summary Objectives: Gestational diabetes mellitus (GDM) is defined as any degree of glucose intolerance, first time detected in pregnancy. Early diagnosis of the disease may reduce fetal exposure to maternal hyperglycemia and decrease the risk of LGA. The aim of the study was to examine the influence of time and diagnostic method of GDM on the prevalence of LGA and pregnancy outcome among patients with gestational diabetes. Material and Methods: The study was conducted among 211 women with gestational diabetes mellitus, the patients of 1st Clinic of Obstetrics and Gynecology at the Medical University in Warsaw. We have reviewed the results of fasting plasma glucose, 50-g glucose screening test (GCT) and 2 hour 75-g glucose tolerance test in GDM patients with LGA and eutrophic newborns. The t-student or the Mann-Whitney test was used in order to compare both groups. P<0,05 was deemed statistically significant. Results: LGA was diagnosed in 10,4% of patients. We did not find any significant differences in gestational age when GDM was diagnosed, results of fasting glucose, GCT and OGTT among LGA (M) and control (K) group. However, when we compared the percentage of LGA in groups of women with different time of GDM diagnosis, the highest prevalence was noted in the group of first trimester diagnosis and between 28 and 32 weeks of pregnancy, which we found interesting. We compared the women and the results of the diagnostic tests with the group of standard time of GDM diagnosis (24-28 week of pregnancy) and the only difference was the late diagnosis. If 75-g glucose tolerance test had not included 1-hour after load glucose assignment, GDM would not have been diagnosed at all in 18,2% of female patients with LGA. We have not found any correlations between the results of the diagnostic tests, the time of the diagnosis or the mode of treatment GDM (diet alone or with insulin) and the birth weight. Conclusions: 1. Results of fasting glucose and glucose tolerance tests are not useful in the prediction of LGA in GDM pregnancies. 2. Diagnosis of GDM during the recommended period (between 24 and 28 weeks of pregnancy) may decrease the prevalence of LGA (comparing to later diagnostics). 3. 75-g glucose tolerance test should provide fasting, 1 and 2-hour after load glucose assignment.

/pdf/diagnostic-of-gestational-diabetes-mellitus-and-the-rs7zpfmyd6.pdf

Diagnostic of gestational diabetes mellitus and the prevalence of LGA (Large for Gestational Age)

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