Regulation of Interleukin-6 Expression in Human Decidual Cells and Its Potential Role in Chorioamnionitis

Chorioamnionitis frequently precedes both genital tract and placental inflammation and is both a primary cause of maternal morbidity and a major antecedent of preterm premature rupture of the membranes (PPROM) as well as preterm delivery (PTD). In most cases of chorioamnionitis, neutrophils dominate the decidua. In a subset of these cases, a predominance of monocytes is uniquely associated with both neonatal intraventricular hemorrhage and death. The multifunctional cytokine, interleukin-6, promotes local monocyte dominance via several mechanisms. In this study, immunostaining of placental sections revealed significantly higher interleukin-6 HSCOREs in decidual cells (DCs) but not in interstitial trophoblasts, in chorioamnionitis versus gestational age-matched control placentas (P < 0.05). In confluent leukocyte-free term DCs, secreted interleukin-6 levels in incubations with estradiol-17β were increased 2500-fold by IL-1β (P < 0.05). This up-regulation was inhibited by more than 50% in parallel incubations that included medroxyprogesterone acetate (n = 12, P < 0.05). Western blotting data confirmed these enzyme-linked immunosorbent assay results; quantitative RT-PCR findings demonstrated corresponding changes in interleukin-6 mRNA levels. Specific inhibitors of signaling for both nuclear factor-κB activation and p38-mitogen-activated protein kinase, but not for protein kinase C, significantly decreased IL-1β-enhanced interleukin-6 expression levels in cultured DCs. In conclusion, in situ and in vitro results indicate that significantly enhanced interleukin-6 expression levels in DCs during chorioamnionitis could be pivotal in skewing decidual monocyte differentiation to macrophages.During human pregnancy, chorioamnionitis (CAM) frequently precedes genital tract and placental inflammation. It is a primary cause of maternal morbidity and a major antecedent of preterm premature rupture of the membranes (PPROM) and preterm delivery (PTD).¹ The latter complicates about 13% of live births in the United States and is the leading cause of perinatal morbidity and mortality.^2,3 During CAM, microbial species usually first ascend from the vagina and cervix to the uterus where they induce deciduitis. Despite the presence of intact membranes, microorganisms can ultimately invade the chorion, amnion, amniotic fluid, and fetus. Although positive microbial cultures are found in the amniotic fluid in 23% of women with PPROM,^4,5 19% displayed no signs of overt amniotic fluid infection.⁵ However, these samples contained elevated levels of neutrophil collagenase and elastase^5,6 suggesting that pathogens remained localized to the decidua or the inciting microorganisms escaped detection.⁵Recently, the large prospective Alabama Preterm Birth Study confirmed the strong association between CAM and the detection of bacterial infections. This study also observed a positive correlation between neutrophil infiltration of the fetal membranes, chorionic plate, and umbilical cord, which serve as markers of CAM, with positive intrauterine bacterial cultures and the occurrence of both the neonatal inflammatory response syndrome and necrotizing enterocolitis.⁷ However, in a significant subset of cases, a mononuclear infiltrate in the fetal membranes and decidua basalis proved to be a harbinger of neonatal intraventricular hemorrhage. Specifically, mononuclear cell infiltration was present in 10% of the placental free membranes and decidua basalis, and within this group there was a 24% increase in neonatal intraventricular hemorrhage. This study also found a positive correlation between PTD and significantly elevated umbilical cord blood levels of interleukin (IL)-6.⁷ The latter is a multifunctional cytokine that regulates hematopoiesis, the acute phase reaction, and both pro- and anti-inflammatory events.⁸ Our laboratory recently demonstrated that decidual cells (DCs), the predominant endometrial cell type throughout pregnancy⁹ are a major source of elevated maternal plasma IL-6 levels¹⁰ implicated in inducing systemic endothelial cell activation and vascular damage in preeclampsia.¹¹In the current study we hypothesized that DCs could also contribute to the well documented CAM-related increase in IL-6 levels in maternal plasma¹² as well as cervical and amniotic fluid IL-6 levels¹³ that reliably predict PPROM and PTD¹⁴ while acting as an autocrine/paracrine modulator of the local immune cell population. To shed light on these questions, immunohistochemistry was used to localize IL-6 at the maternal-placental interface in DCs of placental sections from patients with CAM and gestational age-matched control placentas. The effects of IL-1β, a classic proinflammatory cytokine present at elevated levels in the amniotic fluid of women with CAM,¹⁵ were evaluated on IL-6 mRNA and protein expression in cultured human term DCs. In these experiments, IL-1β was added with either estradiol (E₂) as the control incubation or with E₂ plus medroxyprogesterone acetate (MPA) to mimic the steroid milieu of pregnancy. To eliminate the confounding effects of resident immune cells, experimental incubations were performed with DCs that were passaged until fluorescence-activated cell sorter analysis indicated that they were essentially leukocyte-free. To elucidate a potential mechanism regulating IL-1β-enhanced IL-6 expression in human decidual cells, experimental incubations included specific inhibitors of the intracellular signal transduction pathways for p38 mitogen-activated protein kinase (MAPK), protein kinase C (PKC), and nuclear factor (NF)-κB activation, which are all known to mediate IL-1β-enhanced IL-6 expression.^16,17 A functional in vitro assay determined whether elevated IL-6 levels in the conditioned medium of IL-1β-treated term DCs could promote differentiation of monocytes away from dendritic cells and toward macrophages.     

Turkish adolescents' knowledge on and attitude toward emergency contraception

ObjectiveAdolescents tend to be at risk for unwanted pregnancies, so detecting their level of knowledge on emergency contraception and providing them information is important to prevent such pregnancies. Hence, in two faculties at Gazi University, this study aimed to detect freshman students' level and need of knowledge on emergency contraception and to evaluate their attitude towards emergency contraception.Materials and MethodsThe study was performed with freshman students of the Occupational Education Faculty and the Technical Education Faculty. A questionnaire including questions about demographic properties, obstetrical history, status of contraceptive use, level of knowledge and opinions on emergency contraception was administered to the students. Data was analyzed statistically with the computer program EPI Info 6.0.ResultsA total number of 385 adolescents were included in the studys; 157 of whom were males (40.8%) and 228 of whom were females (59.2%). To the question “is there any way to prevent a possible pregnancy after an unprotected sexual intercourse?”, 166 students replied “yes” (50.5%), 39 “no” (11.9%) and 124 “I do not know” (37.7%). Of 166 students replying “yes,” 114 (68.7%) listed a possible contraceptive method. The mostly cited method was “morning after pills” (n = 62; 54.4%), followed by curettage (n = 15; 13.2%). Among all students, 158 (49.8%) informed us that they were aware of the presence of “morning after pills” whereas 159 (50.2%) claimed they were not. Eighty-six male students (70.5%) and 115 female students (72%) emphasized that they would use emergency contraception upon necessity.ConclusionHalf of the participants were familiar with various options to prevent pregnancy after an unprotected sexual intercourse episode, but they lacked specific knowledge about possible methods and ways to use them. Thus, it is essential that information about emergency contraception be included in adolescents' educational programs and that adolescents be provided with easily accessible medical services.     

Use of enzyme-linked immunosorbent assay for detection of natural leishmania infections in phlebotomine sand flies from southeastern Turkey

In total, 320 phelebotomine sand flies from Alibozlu (Osmaniye), Kizyusuflu (Osmaniye), and Sanliurfa in southeastern Turkey were tested for the detection and identification of Leishmania in vector sand flies by enzyme-linked immunosorbent assay with species-specific monoclonal antibodies. We used monoclonal antibodies that recognize both Leishmania tropica and Leishmania major, and a monoclonal antibody specific only to L. tropica. Phosphate-buffered saline and monoclonal antibody M2 recognizing Leishmania amazonensis were used as controls. Infection rates of sand flies were 0.9% in Alibozlu, 0% Kizyusuflu, and 3.6% in Sanliurfa. Positive sand flies were identified as Phlebotomus sergenti Parrot and Phlebotomus major syriacus Adler & Theodor.     

Selective laser trabeculoplasty induced changes in the thickness of ciliary body and iris evaluated by ultrasound biomicroscopy

Background

Selective laser trabeculoplasty (SLT) is widely used for the treatment of glaucoma. The main target tissue of this treatment modality is trabecular meshwork. We aimed to detect the SLT-induced changes in the thickness of ciliary body (CBT) and iris (IT), quantitatively.

Methods

Thirty-one patients treated by SLT were examined by ultrasound biomicroscopy (UBM) at different locations of ciliary body and iris at four quadrants, before and after (3rd, 7th, and 30th days) SLT. The IT was measured at various locations; 500???m anterior to the scleral spur (IT₁), 2?mm from the iris root (IT₂) and near the pupillary edge where the iris thickness was maximum (IT₃) at four quadrants. The CBT at positions 1 and 2?mm posterior to the scleral spur were measured (CBT_1-2). Additionally, intraocular pressure (IOP) levels were measured in all visits and post-laser 1 h.

Results

There were statistically significant higher CBT values at 3rd and 7th-day measurements in the study compared to pre-treatment levels (p?<?0.0001, p?<?0.0001, respectively). CBT₂ values at day 30 were similar compared to pre-treatment values (overall p?=?0.140), but CBT₁ values at day 30 were not exactly similar compared to pre-treatment values in superior and nasal quadrants (overall p?=?0.027). IT values obtained in the 3rd and 7th days were significantly higher in all quadrants and regions when compared to the pre-treatment values (p?<?0.0001, p?<?0.0001, respectively). There were no statistically significant differences in any of the IT values at the 30th day in comparison to the pre-treatment values (p?=?0.45).

Conclusions

The results suggest that SLT induces prominent increases in CBT and IT returning to baseline thickness in a month, which may be caused by inflammation, vascular engorgement, or mechanical muscular contraction.     

Using oral polio vaccine beyond the cold chain: A feasibility study conducted during the national immunization campaign in Mali

We conducted the first systematic documentation of using oral polio vaccine (OPV) out of the cold chain during national immunization day (NID) campaigns in Mali. Using a crossover intervention design, vaccinators compared the transport of OPV in vaccine carriers with or without ice packs. Vaccine integrity was assured through monitoring vaccine vial monitor (VVM) status. Despite ambient temperatures up to 40 °C, none of the VVMs on any of the vials used (n = 956) reached their discard point. Over 90% of vaccinators and supervisors preferred conducting NIDs without ice packs. In addition, using OPV out of the cold chain reduced vaccine wastage resulting from melting ice packs causing labels to detach from the vial.     

Alanine aminotransferase normalization at week 8 predicts viral response during hepatitis C treatment

AIM:To investigate alanine aminotransferase(ALT)and sustained virological response(SVR)in chronic hepatitis C(CHC)during peginterferon-ribavirin treatment.METHODS:One hundred and fifty-one genotype 1CHC patients underwent treatment for 48 wk with peginterferon and ribavirin,and were retrospectively divided into two groups as having a rapid virological response(RVR)(Group 1,n=52)and not having an RVR(Group 2,n=99).We also subdivided each group into two according to the initial ALT level being high(Group1h and Group 2h)or normal(Group 1n and Group 2n).HCV RNA and ALT levels were measured at baseline;at 4,12,24 and 48 wk during the treatment period;and at 24 wk follow-up.ALT levels were also obtained at 8 wk.According to the results of ALT,patients were enrolled in either the follow-up abnormal or follow-up normalized ALT groups at each interval.Patients with high and normal ALT levels were compared for each interval in terms of SVR.RESULTS:The SVR rates were 83%vs 40%(P=0.000),82%vs 84%(P=0.830),and 37%vs 44%(P=0.466)when comparing Group 1 with 2,1h with1n,and 2h with 2n,respectively.In Group 2h,the SVR rates were 34%vs 40%(P=0.701),11%vs 52%(P=0.004),12%vs 50%(P=0.007),7%vs 50%(P=0.003),6%vs 53%(P=0.001),and 0%vs 64%(P=0.000)when comparing patients with high and normalized ALT levels at week 4,8,12,24,48 and 72,respectively.The multiple logistic regression analysis revealed that RVR(OR=7.05;95%CI:3.1-16.05,P=0.000),complete early virological response(cEVR)(OR=17.55;95%CI:6.32-48.76,P=0.000),normalization of ALT at8 wk(OR=3.04;95%CI:1.31-7.06,P=0.008),and at 12 wk(OR=4.21;95%CI:1.65-10.76,P=0.002)were identified as independent significant predictive factors for SVR.CONCLUSION:Normalization of ALT at 8 wk may predict viral response during peginterferon-ribavirin treatment in genotype-1 CHC patients especially without RVR.     

Management of endometrial polyps diagnosed before or during ICSI cycles

This retrospective study aimed to shed light on the management options of endometrial polyps diagnosed before or during intracytoplasmic sperm injection (ICSI) treatment. The study included all fresh ICSI cycles performed in the Anatolia IVF Center between July 2005 and January 2009. Group 1 consisted of 47 patients who were diagnosed with an endometrial polyp before their ICSI cycle. All patients diagnosed with an endometrial polyp by transvaginal ultrasonography before the ICSI cycle underwent hysteroscopic polyp resection. Group 1 was compared with 47 matched control patients without endometrial polyps who underwent standard ICSI cycles (group 2). Group 3 included 128 patients diagnosed with an endometrial polyp during stimulation in their ICSI cycles. Group 3 was compared with 128 matched control patients without endometrial polyps who underwent standard ICSI cycles (group 4). Patients diagnosed with an endometrial polyp before ICSI cycles were similar to their controls with regard to clinical pregnancy (29.8% versus 38.3%) and live-birth (25.5% versus 31.9%) rates per transfer, as were patients diagnosed with an endometrial polyp during ovarian stimulation (clinical pregnancy rates 45.3% versus 46.9%; live-birth rates 40.6% versus 39.8%). In conclusion, further studies are required to identify the most appropriate management of endometrial polyps.     

Effect of air bubble localization after transfer on embryo transfer outcomes

Objectives

Our study aimed to provide information about the effects of air bubble localization after transfer on embryo transfer outcomes.

Study design

Retrospective analysis of 7489 ultrasound-guided embryo transfers. Group 1 included 6631 embryo transfers in which no movement of the air bubbles was observed after transfer. Group 2 consisted of 407 embryo transfers in which the air bubbles moved towards the uterine fundus spontaneously, a little time after transfer. Group 3 included 370 embryo transfers in which the air bubbles moved towards the uterine fundus with ejection, immediately after transfer. Group 4 consisted of 81 embryo transfers in which the air bubbles moved towards the cervical canal.

Results

The four patient groups were different from one another with respect to positive pregnancy tests. Post hoc test revealed that this difference was between group 4 and other groups.

Conclusions

An initial finding of our study was significantly decreased positive pregnancy test rates and clinical pregnancy rates with air bubbles moving towards the cervical canal after transfer. Although air bubbles moving towards the uterine fundus with ejection were associated with higher pregnancy rates, higher miscarriage rates and similar live birth rates were observed compared to air bubbles remaining stable after transfer.     

Effect of blood and mucus on the success rates of embryo transfers

Objectives

To provide information about the effects of blood and mucus on the embryo transfer (ET) catheters after transfer on embryo transfer outcomes.

Study design

Retrospective analysis of 8311 ultrasound-guided embryo transfers performed in a single center. In 6897 cases (82.9%), there were no blood on the catheter after ET, 1168 transfers were associated with mild blood (14.1%), 33 transfers with moderate blood (0.4%) and 213 transfers with severe blood (2.6%). A total of 6162 transfer catheters were free of mucus (74.8%), whereas mucus was detected on 2081 catheters (25.2%).

Results

The implantation rate (IR) was highest in the group with no blood on the transfer catheter, and lowest in the group with severe blood on the catheter. The clinical pregnancy rate (CPR) was lowest in the group with severe blood on the catheter. The presence of mucus on the catheter was found to have no effect on IR, CPR, biochemical pregnancy rates, miscarriage rates and live birth rates.

Conclusions

This study showed decreased IR, CPR and live birth rates in ETs associated with blood on the catheter. Mucus on the catheter appeared to be a simple contamination in this study and pregnancy rates remained unaffected.     

Is there a role for kisspeptin in pathogenesis of polycystic ovary syndrome?

Aim: To investigate association of kisspeptin levels in infertile women with different ovarian reserve patterns.

Materials and methods: In this prospective cross-sectional study, 157 participants were recruited. The women were divided into three groups: (i) adequate ovarian reserve (AOR) (n?=?57), (ii) high ovarian reserve (PCOS) (n?=?60), (iii) diminished ovarian reserve (DOR) (n?=?40). Weight, height, waist circumference (WC), hip circumference (HC), body mass index (BMI), waist/hip ratio (WHR) were measured. The blood samples were analyzed for estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), total testosterone (TT), 17-hydroxy progesterone (17OHP), dehydroepiandrosterone sulfate (DHEAS), antimullerian hormone (AMH), kisspeptin measurements.

Results: FSH concentration was higher and AMH concentration was lower in DOR group (p?p?p?=?.001, p?p?=?.003, respectively). The 17OHP level did not differ among the groups (p?=?.15). Women with PCOS possessed the highest kisspeptin level (p?=?.01). The kisspeptin level was negatively correlated with FSH level (r?=??0.18, p?=?.02) and positively correlated with TT and DHEAS levels (r?=?0.17, p?=?.02 and r?=?0.23, p?=?.003, respectively).

Conclusions: Women with PCOS had increased serum kisspeptin levels. Kisspeptin concentrations were negatively correlated with serum FSH and positively correlated with serum TT and DHEAS levels.