LIF endometrial expression is impaired in women with unexplained infertility while LIF-R expression in all infertility sub-groups.

The main objective of our study was to study LIF and LIF-R endometrial expression during the implantation window in the various sub-groups of infertile women according to infertility cause. A prospective observational case-control study was performed from March 2013 to February 2016. Infertile women consisted of the patients' group (group 2) while fertile women were the control group (group 1). Infertile women were divided according to infertility cause in women with tubal factor (group 2a), poor ovarian reserve (group 2b), endometriosis (group 2c) and unexplained infertility (group 2d). Endometrial biopsy was performed on 7th-8th postovulatory menstrual day. Leukemia Inhibitory Factor (LIF) and LIF-Receptor (LIF-R) expression in epithelial and stromal cells were assessed with Immunohistochemistry (IHC). There were 20 infertile with poor ovarian reserve, 15 with tubal factor, 10 with endometriosis and 15 with unexplained infertility included in the analysis. LIF expression in patients with unexplained infertility was significantly compared with controls (P=0.006). No significant difference was observed between patients with tubal factor, poor ovarian reserve and endometriosis compared with control group (P=0.78, P=0.44 and P=0.56 respectively). Analysis of LIF-R expression in sub-categories of infertility indicated that expression was significantly decreased in all sub-groups of infertility. Our study indicated impaired LIF expression levels only in women with unexplained infertility, while LIF-R expression was impaired in all sub-groups of infertile women. Further multicenter prospective studies should be performed in order to assess the exact etiopathogenetic role of these cytokines in the molecular background of infertility.

Association of two synonymous splicing-associated CpG single nucleotide polymorphisms in calpain 10 and solute carrier family 2 member 2 with type 2 diabetes.

Coding synonymous single nucleotide polymorphisms (SNPs) have attracted little attention until recently. However, such SNPs located in epigenetic, CpG sites modifying exonic splicing enhancers (ESEs) can be informative with regards to the recently verified association of intragenic methylation and splicing. The present study describes the association of type 2 diabetes (T2D) with the exonic, synonymous, epigenetic SNPs, rs3749166 in calpain 10 (CAPN10) glucose transporter (GLUT4) translocator and rs5404 in solute carrier family 2, member 2 (SLC2A2), also termed GLUT2, which, according to prior bioinformatic analysis, strongly modify the splicing potential of glucose transport-associated genes. Previous association studies reveal that only rs5404 exhibits a strong negative T2D association, while data on the CAPN10 polymorphism are contradictory. In the present study DNA from blood samples of 99 Greek non-diabetic control subjects and 71 T2D patients was analyzed. In addition, relevant publicly available cases (40) resulting from examination of 110 Personal Genome Project data files were analyzed. The frequency of the rs3749166 A allele, was similar in the patients and non-diabetic control subjects. However, AG heterozygotes were more frequent among patients (73.24% for Greek patients and 54.55% for corresponding non-diabetic control subjects; P=0.0262; total cases, 52.99 and 75.00%, respectively; P=0.0039). The rs5404 T allele was only observed in CT heterozygotes (Greek non-diabetic control subjects, 39.39% and Greek patients, 22.54%; P=0.0205; total cases, 34.69 and 21.28%, respectively; P=0.0258). Notably, only one genotype, heterozygous AG/CC, was T2D-associated (Greek non-diabetic control subjects, 29.29% and Greek patients, 56.33%; P=0.004; total cases, 32.84 and 56.58%, respectively; P=0.0008). Furthermore, AG/CC was strongly associated with very high (≥8.5%) glycosylated plasma hemoglobin levels among patients (P=0.0002 for all cases). These results reveal the complex heterozygotic SNP association with T2D, and indicate possible synergies of these epigenetic, splicing-regulatory, synonymous SNPs, which modify the splicing potential of two alternative glucose transport-associated genes.

LIF and LIF­R expression in the endometrium of fertile and infertile women: A prospective observational case­control study.

The aim of the present study was to determine the expression of leukemia inhibitory factor (LIF) and LIF receptor (LIF­R) in the endometrium of fertile and infertile women during the implantation window. A prospective study was conducted between March 2013 and March 2015 at Iakentro, Infertility Treatment Center (Thessaloniki, Greece) and the 3rd Department of Obstetrics and Gynecology, Aristotle University of Thessaloniki (Thessaloniki, Greece). The patient group consisted of women diagnosed with infertility, whereas the control group consisted of women who had delivered at least one live newborn (fertile women). An endometrial biopsy was obtained using a Pipelle on day 7 or 8 post­ovulation, and the expression of LIF and LIF­R was assessed by immunohistochemistry in epithelial and stromal cells. Primary outcomes included positive cellular percentage, staining intensity and H­score. P<0.05 was considered to indicate a statistically significant difference. Overall, 45 women were included in the present analysis (15 fertile women and 30 infertile women). Mean age was 32.8±6.0 years for the fertile group, and 37.6±3.7 for the infertile group. LIF and LIF­R expression was significantly reduced in the epithelial cells of infertile women (P=0.05 and P=0.006, respectively). However, no significant differences were detected with regards to the expression of LIF in stromal cells (P=0.95). In addition, LIF­R expression was relatively higher in the stromal cells of the fertile group; however, the difference did not reach statistical significance (P=0.10). In conclusion, endometrial expression of LIF and LIF­R is significantly reduced in the epithelial cells of infertile women. Expression patterns of LIF­R in stromal cells require further research in order to achieve definitive results.

Expression of progesterone receptors is significantly impaired in the endometrium of infertile women during the implantation window: a prospective observational study.

OBJECTIVE: To compare the expression of progesterone receptors (A + B) and type-B progesterone receptors in the epithelial and stromal cells of fertile and infertile women. METHODS: Women were divided into two groups, the group of fertile controls (group 1) and the group of infertile women (group 2) and were set on regular ultrasound imaging in order to detect ovulation. An endometrial biopsy was obtained on the seventh or eighth post-ovulatory day. Immunohistochemistry was performed to measure percentage of positive nuclei, intensity of staining and h-score for progesterone receptors (PgR) (A + B) as well as type-B progesterone receptors in epithelial and stromal cells. Secondary outcomes included endometrial tissue dating, the rate of tissues being out-of-phase and endometrial thickness. RESULTS: Endometrial issue was obtained from 15 fertile and 30 infertile women. Expression of PgR (A + B) and PgR type-B was significantly lower in the epithelial cells of infertile women. PgR (A + B) h-score was 220.0 ± 18.5 for fertile versus 147.3 ± 18.0 for infertile women (p = 0.02). PgR type-B h-score in epithelial cells was 166.8 ± 30.7 for fertile versus 90.8 ± 20.6 for infertile (p = 0.04). No significant difference was observed in stromal cells. CONCLUSIONS: Expression levels of PgR (A + B) as well as type-B receptors are significantly lower in the epithelial cells of infertile women during implantation window.