Determination of maternal serum pro-inflammatory cytokine changes in intrauterine growth restriction.

OBJECTIVE: To evaluate maternal serum inflammatory marker changes in intrauterine growth restriction (IUGR) pregnancies. PATIENTS AND METHODS: 50 healthy pregnant women and 50 patients diagnosed with IUGR were enrolled. Maternal serum high sensitivity C-reactive protein (hsCRP), erythrocyte sedimentation rate (ESR), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-10 levels were measured before delivery and neonatal outcomes were evaluated. RESULTS: Birth weight, Apgar scores, and cord blood gas pH were lower in the IUGR group (p<0.001, p<0.001, p<0.001 and p=0.006, respectively). While the levels of ESR, hsCRP, IL-6, and TNF-α were higher, the IL-10 level was found to be lower in the IUGR group (p<0.001, p=0.033, p<0.001, p=0.004 and p<0.001, respectively). As ESR, hsCRP, and IL-6 levels increased, birth weight, Apgar scores, and cord blood gas pH decreased (p<0.001, p<0.001, p<0.001, p<0.001, p=0.02, p=0.002, p=0.001, p=0.03, p<0.001, p<0.001, p<0.001 and p=0.02, respectively). As TNF-α level increased, only birth weight and Apgar score at the 1st minute decreased (p=0.006 and p=0.048, respectively). As IL-10 level decreased, birth weight, Apgar scores, and cord blood gas pH decreased (p<0.001 for all). IL-6 (>3.2 pg/ml) had a sensitivity of 100%, specificity of 100%, PPV of 100% and NPV of 100%. CONCLUSIONS: While birth weight, Apgar score and cord blood pH decreased in IUGR cases, ESR, hsCRP, IL-6 and TNF-α levels increased. Combined measurement of these markers can be used for the diagnosis of IUGR.

Role of vitamin D on gestational hypertension, diabetes mellitus, timing and mode of delivery.

OBJECTIVE: To determine the efficacy of VD in preventing the development of gestational diabetes mellitus (GDM) and pregnancy-induced hypertension (PIH). The secondary purpose is to investigate the effect of VD on the mode and time of delivery. PATIENTS AND METHODS: A vitamin D value of <20 ng/mL during pregnancy is considered a deficiency according to the Endocrine Society, and 400-600 IU/day VD replacement is recommended. Forty patients whose serum VD levels were below 20 ng/mL during routine pregnancy follow-up and who were planned for VD replacement therapy were included in the study. They were divided into two equal groups with 20 patients in each group. Twenty pregnant women with serum VD levels greater than 20 ng/mL were considered as the control group. While 400 IU/day VD replacement was applied to the patients in Group 1, 600 IU/day VD was given to Group 2. Group 3 consisted of control patients who did not undergo VD replacement. VD replacement was continued from the 14th week of pregnancy until delivery. Each group of participants was screened with a 50-g GCT at 24-28 weeks of gestation. Following 50-g GCT if serum glucose level was found >140 mg/dL, patients underwent 100-g OGTT. GDM was diagnosed in the presence of at least two of the following results: fasting serum glucose ≥92 mg/dL and/or 1-hour glycemia ≥180 mg/dL, and/or 2-hour glycemia ≥153 mg/dL. PIH was defined as systolic blood pressure >140 mmHg and diastolic blood pressure >90 mmHg. Patients in each group delivered by cesarean section or normal vaginal route. In addition to the incidence of PIH and GDM, the time and mode of delivery were recorded. RESULTS: PIH was detected in two patients in each of the 400 IU/day and 600 IU/day vitamin D replacement groups (10%). In the control group, PIH developed in 3 patients (15%). Although PIH was detected in an extra case in the control group, no significant difference was found between the replacement group and the control group in terms of PIH (p<0.44). While GDM was not detected in the 400 IU/day vitamin D group, GDM was detected in one patient (5%) in the 600 IU/day vitamin D group. No case of GDM was found in the control group either. There was no significant difference between the VD replacement and the control groups in terms of GDM rates. No significant difference was found between the VD replacement and the control groups in terms of mode of delivery. While the C/S ratio was 65% in the 400 IU/day vitamin D group, this ratio was 75% in the 600 IU/day vitamin D group. There was an insignificant trend of increase in C/S ratios in the group given 600 IU/day of vitamin D. The C/S ratio of the control group, which could not be given VD replacement, was found to be 70%. CONCLUSIONS: VD replacement therapy during pregnancy does not prevent the development of PIH and GDM, and does not significantly contribute to the time and mode of delivery.

Impact of spontaneous preterm birth on amniotic fluid NF-κB, IL-6, TNF-α and IL-1ß levels in singleton pregnancies conceived after IVF/ICSI treatment or natural conception.

OBJECTIVE: The aim of our study was to compare the amniotic fluid NF-κB, TNF-α, IL-1ß and IL-6 levels of patients who developed spontaneous preterm birth (sPTB) after IVF/ICSI or natural pregnancy, among themselves and with the pregnant women who gave term birth. PATIENTS AND METHODS: A total of 43 patients who had spontaneous preterm birth before 37 weeks were included in the study. While 23 out of 43 patients conceived after IVF/ICSI, the remaining 20 patients conceived spontaneously. Women in both participant groups delivered by cesarean section or vaginally. Ten patients who did not have a history of preterm labor were accepted as the control group. Amniotic fluid was taken with the aid of a 10 cc injector following spontaneous or artificial rupture of membranes from patients who presented with spontaneous preterm labor with intact membranes and started normal labor. Samples of amniotic fluid accumulated in the speculum were collected from patients with ruptured membranes at the first admission. Amniotic fluid was collected with the help of an injector just before the amniotic membrane was cut in patients who decided to have a cesarean section. NF-κB, IL-6, TNF-α and IL-1ß concentrations in amniotic fluid samples were measured quantitatively by enzyme-linked immunosorbent assay (ELISA) using human NF-κB, IL-6, TNF-α, and IL-1ß ELISA kits. RESULTS: The maternal age, parity and gestational age at the time of delivery, fetal birth weight were similar in the IVF/ICSI and natural conception groups. The amniotic fluid NF-κB, TNF-α, IL-1ß and IL-6 levels of sPTB patients in the IVF/ICSI group and those in the natural conception group were found to be similar. The tendency to increase in cytokine levels in term pregnant women compared to sPTB groups did not reach significance. Amniotic fluid proinflammatory cytokine levels of sPTB patients in both natural conception and IVF/ICSI groups were found to be similar to healthy controls with term delivery. Amniotic fluid proinflammatory cytokine levels of sPTB patients in both natural conception and IVF/ICSI groups were found to be similar to healthy controls with term delivery. There was no difference between the amniotic fluid proinflammatory cytokine levels of the patients who delivered vaginally or by cesarean section. CONCLUSIONS: Whether sPTB develops after ICSI or after natural conception, the mechanism is the same and largely overlaps with the term birth mechanism.

The effect of Nateglinide and Octreotide on follicular morphology and free radical scavenging system in letrazole-induced rat model of PCOS.

OBJECTIVE: To investigate the effects of octreotide and nateglinide on ovarian follicle count, ovarian tissue damage, biochemical parameters and free radical scavenging system in letrazole-induced rat model of PCOS. MATERIALS AND METHODS: Forty-two female Sprague-Dawley rats were divided into six groups. Group 1 (Control Group): after localizing the ovaries and the uterine horns, the abdominal wall was closed without any surgical procedure. Group 2 (PCOS Group): PCOS was induced by administrating Letrozole orally for 21 successive days. At the end of 21 days, rats underwent ovarian biopsies. The experimental PCOS model was considered successful in the presence of atretic follicles without granulosa cell stratification. Group 3 (PCOS + Nateglinide Group): Nateglinide was administered by oral dropper for 30 days to the rats in which PCOS model was created. Group 4 (Nateglinid only Group): 30 days of NG was applied to the rats without PCOS. Group 5 (PCOS+Octreotide Group): 0.1 mg/kg/day Octreotide was given intraperitoneally for 4 weeks to the rats in which PCOS model was created. Group 6 (Octreotide only Group): animals without PCOS given 0.1 mg/kg/day Octreotide at the end of the treatment, bilateral oophorectomy was performed and blood samples were collected from all groups. Ovarian tissue was stained immunohistochemically with TLR-4 in addition to conventional staining. In addition to follicle classification, ovarian damage was graded. Serum insulin, FSH and LH, TNF-α, IL-6, SHBG, SOD, IGF-1, MDA and GSH levels were also measured. RESULTS: The cystic and degenerated follicle density of PCOS group was high compared with the other groups. Both cystic and degenerated follicles were significantly reduced in PCOS+NG and PCOS+OC groups compared to PCOS group. There was no difference between the groups in terms of serum LH, FSH and insulin levels (p>0.05). Serum testosterone level was significantly higher in the PCOS group compared to the other groups (p<0.01). Adding OC or NG to PCOS groups did not cause significant changes in testosterone levels. TNF-α and IL-6 levels were high in PCOS group (p<0.03). IGF-1 and MDA levels were higher in PCOS than in other groups (p<0.03, p<0.01 respectively). Adding OC or NG to the treatment normalized IGF-1 and MDA levels. Serum GSH levels were significantly lower in the PCOS group (p<0.05). Adding NG to the treatment increased GSH levels. CONCLUSIONS: Both NG and OCT reverses atretic and degenerate follicle damage due to PCOS through TLR-4, antioxidant and anti-inflammatory pathways.

Comparison of immunohistochemical characteristics of endometriomas with non-endometriotic benign ovarian cysts.

OBJECTIVE: The aim of the study was to investigate the existence of neuroendocrine cells and to compare the density of those in normal ovarian tissue, endometriotic and non-endometriotic benign ovarian cysts. PATIENTS AND METHODS: Twenty patients with the diagnosis of endometrioma and 30 control subjects consisting of ovarian serous cystadenoma (n=10), ovarian mucinous cystadenoma (n=10) and normal ovarian tissue (n=10) were included. The tissues were prepared and assessed according to staining density by using the H-score method. RESULTS: Tissues with mucinous cystadenoma were significantly more stained with PAS and VanGieson, when compared to women with endometrioma. Macrophage deposition was higher in cyst samples with endometrioma and in normal ovarian tissue when compared to serous cystadenoma and mucinous cystadenoma. Normal ovarian tissue was significantly more stained with PGP9.5, NSE and SYN when compared to endometrioma and non-endometriotic benign ovarian cyst. PGP9.5 staining was higher in normal ovarian tissue when compared with endometriotic lesions (p<.001). Endometrioma samples were significantly more stained with p53 when compared to non-endometriotic cysts and normal ovarian tissue. c-Kit staining was mild and not statistically significant among all groups. CONCLUSIONS: During endometrioma transformation, expression intensity of neuroendocrine markers decreases compared to normal ovarian tissue and other benign ovarian cysts.

DNA repair gene (XRCC1 and XPD) polymorphism and risk of primary ovarian failure.

OBJECTIVE: DNA repair genes may be related with the onset of primary ovarian failure (POF). The study was planned to investigate whether the polymorphisms in the DNA repair genes modulate the risk of POF. PATIENTS AND METHODS: This prospective study included 25 women diagnosed with POF and 25 healthy controls. The genotyping and allele of XRCC1 and XPD genes were determined by using Polymerase Chain Reaction and fluorescence melting curve analysis. RESULTS: The genotype and allele distribution of the Arg194Trp and Arg399Lys polymorphism of the XRCC1 gene did not differ statistically between those of the POF and control groups. The frequency of the C (Gln) allele was significantly lower in patients diagnosed with POF when compared to that in healthy controls [48% vs. 76%, p=0.040, OR: 3.43 (CI: 1.03-11.48)]. The Lys/Lys genotype for XPD-Lys751Gln polymorphism in patients diagnosed with POF was more common than in healthy controls (p=0.028, 52% vs. 24%). CONCLUSIONS: The genotype distribution and allele frequency of XPD-Lys751Gln, XRCC1-Arg194Trp and XRCC1-Arg399 Gln did not regulate the risk of developing POF. Gln/Gln+Lys/Gln and XPD-Lys751Gln polymorphism may have a possible protective role against the development of POF.

Surgical removal of intramural fibroids improves the TNF-α induced inflammatory events in endometrium.

OBJECTIVE: To investigate whether surgical removal of intramural fibroids makes any changes in the expression of endometrial tumor necrosis factor alpha (TNF-α) and nuclear factor kappa B (NF-κB). PATIENTS AND METHODS: 20 patients who had no pathology other than intramural fibroid as a cause of subfertility were included in the study. 6 patients who planned tubal ligation were considered as the control group. Type 3 or 4 fibroid diagnosis was made according to the revised International Federation of Gynecology and Obstetrics (FIGO). At the end of the diagnostic and confirmatory tests, 8 patients were diagnosed with type 3 and 12 patients with type 4 fibroids. While the patients in the fibroid group went to myomectomy, the patients in the control group went to tubal ligation. Before the myomectomy, endometrial sampling was performed with a pipelle cannula under anesthesia. The same procedure was repeated three months after the treatment. The same endometrial sampling procedure was also performed on the patients in the control group. TNF-alpha and NF-κB levels were measured with enzyme-linked immunosorbent assay (ELISA) in endometrial samples taken before and three months after myomectomy. RESULTS: Pre-myomectomy TNF-α levels of the patients in the type 3 fibroid group were significantly higher than the control group (5.10±1.30 vs. 2.50±0.11, p<0.01). Similarly, the pre-myomectomy TNF-α levels of the patients in the type 4 fibroid group were significantly higher than the control group (4.73±1.76 vs. 2.50±0.11, p<0.01). There was no significant difference between pre-myomectomy endometrial TNF-α levels of patients in type 3 and 4 fibroid groups (5.10±1.30 vs. 4.73±1.76, p>0.05). Removal of type 3 fibroids by myomectomy significantly decreased TNF-α levels (5.10±1.30 vs. 2.20±0.44, p<0.03). Similarly, removal of type 4 fibroids by myomectomy significantly decreased TNF-α levels (4.73±1.76 vs. 2.60±0.30, p<0.04). Pre-myomectomy NF-κB levels of the patients in the type 3 fibroid group were significantly higher than the control group (2.42±0.39 vs. 1.09±0.60, p<0.02). Similarly, the pre-myomectomy NF-κB levels of the patients in the type 4 fibroid group were significantly higher than the control group (2.04±0.50 vs. 1.09±0.60, p<0.01). There was no significant difference between the pre-myomectomy endometrial NF-κB levels of the patients in the type 3 and 4 fibroid groups (2.42±0.39 vs. 2.04±0.50 p>0.05). Removal of type 3 fibroids by myomectomy significantly decreased NF-κB levels (2.42±0.39 vs. 1.02±0.33, p<0.01). Similarly, removal of type 4 fibroids by myomectomy significantly decreased NF-κB levels (2.04±0.50 vs. 0.97±0.02, p<0.02). CONCLUSIONS: Surgical removal of type 3 or type 4 fibroids contributes positively to receptivity by down-regulating endometrial TNF-α and NF-ƙB.