BioMed Research International: Obstetrics and Gynecology The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. Maternal Body Mass Index and Risk of Obstetric Anal Sphincter Injury Tue, 15 Apr 2014 14:04:29 +0000 Objective. To estimate the association between maternal obesity and risk of three different degrees of severity of obstetric anal sphincter injury. Methods. The study population consisted of 436,482 primiparous women with singleton term vaginal cephalic births between 1998 and 2011 identified in the Swedish Medical Birth Registry. Women were grouped into six categories of BMI. BMI 18.5–24.9 was set as reference. Primary outcome was third-degree perineal laceration, partial or total, and fourth-degree perineal laceration. Adjustments were made for year of delivery, maternal age, fetal head position at delivery, infant birth weight and instrumental delivery. Results. The overall prevalence of third- or four-degree anal sphincter injury was 6.6% (partial anal sphincter injury 4.6%, total anal sphincter injury 1.2%, unclassified as either partial and total 0.2%, or fourth degree lacerations 0.6%). The risk for a partial, total, or a fourth-degree anal sphincter injury decreased with increasing maternal BMI most pronounced for total anal sphincter injury where the risk among morbidly obese women was half that of normal weight women, OR 0.47 95% CI 0.28–0.78. Conclusion. Obese women had a favourable outcome compared to normal weight women concerning serious pelvic floor damages at birth. Marie Blomberg Copyright © 2014 Marie Blomberg. All rights reserved. Application of MicroRNA in Diagnosis and Treatment of Ovarian Cancer Tue, 15 Apr 2014 12:54:20 +0000 Ovarian cancer has a poor prognosis because early detection is difficult and recurrent ovarian cancer is usually drug-resistant. The morbidity and mortality of ovarian cancer are high worldwide and new methods of diagnosis and therapy are needed. MicroRNAs (miRNAs) are posttranscriptional regulators of gene expression that are involved in carcinogenesis, metastasis, and invasion. Thus, miRNAs are likely to be useful as diagnostic and prognostic biomarkers and for cancer therapy. Many miRNAs have altered expression in ovarian cancer compared to normal ovarian tissues and these changes may be useful for diagnosis and treatment. For example, deficiencies of enzymes including Dicer and Drosha that are required for miRNA biogenesis may be adverse prognostic factors; miRNAs such as miR-214 and miR-31, which are involved in drug resistance, and the miR-200 family, which is implicated in metastasis, may serve as biomarkers; and transfection of downregulated miRNAs and inhibition of upregulated miRNAs may be effective for treatment of ovarian cancer. Chemotherapy targeting epigenetic mechanisms associated with miRNAs may also be effective to reverse gene silencing. Kouji Banno, Megumi Yanokura, Miho Iida, Masataka Adachi, Kanako Nakamura, Yuya Nogami, Kiyoko Umene, Kenta Masuda, Iori Kisu, Hiroyuki Nomura, Fumio Kataoka, Eiichiro Tominaga, and Daisuke Aoki Copyright © 2014 Kouji Banno et al. All rights reserved. Stem Cells as New Agents for the Treatment of Infertility: Current and Future Perspectives and Challenges Mon, 14 Apr 2014 09:55:32 +0000 Stem cells are undifferentiated cells that are present in the embryonic, fetal, and adult stages of life and give rise to differentiated cells that make up the building blocks of tissue and organs. Due to their unlimited source and high differentiation potential, stem cells are considered as potentially new therapeutic agents for the treatment of infertility. Stem cells could be stimulated in vitro to develop various numbers of specialized cells including male and female gametes suggesting their potential use in reproductive medicine. During past few years a considerable progress in the derivation of male germ cells from pluripotent stem cells has been made. In addition, stem cell-based strategies for ovarian regeneration and oocyte production have been proposed as future clinical therapies for treating infertility in women. In this review, we summarized current knowledge and present future perspectives and challenges regarding the use of stem cells in reproductive medicine. Vladislav Volarevic, Sanja Bojic, Jasmin Nurkovic, Ana Volarevic, Biljana Ljujic, Nebojsa Arsenijevic, Majlinda Lako, and Miodrag Stojkovic Copyright © 2014 Vladislav Volarevic et al. All rights reserved. Singleton Pregnancy Outcomes after In Vitro Fertilization with Fresh or Frozen-Thawed Embryo Transfer and Incidence of Placenta Praevia Sun, 13 Apr 2014 00:00:00 +0000 The aim of the study was to compare the single pregnancy and neonate outcome after fresh and frozen-thawed embryo transfer in the in vitro fertilization programme (IVF). The study focused on clinical and laboratory factors affecting the abnormal placentation, especially placenta praevia, in patients conceiving in the IVF programme. The results confirm that neonates born after frozen-thawed embryo transfer had significantly higher mean birth weight than after fresh embryo transfer (ET). Moreover, the birth weight distribution in singletons was found to shift towards “large for gestation” (LGA) after frozen-thawed ET. On the other hand, the pregnancies after fresh ET were characterized by a higher incidence of placenta praevia and 3rd trimester bleeding. Placenta praevia was more common in IVF patients with fresh ET in a stimulated cycle than in patients with ET in a spontaneous cycle. It occurred more frequently in patients with transfer of 2 embryos. From this point of view, single ET and ET in a spontaneous cycle should be encouraged in good prognosis patients in the future with more than two good quality embryos developed. An important issue arose of how the ovarian hormonal stimulation relates to abnormal placentation and if the serum hormone levels interfere with in the IVF treatment results. Sara Korosec, Helena Ban Frangez, Ivan Verdenik, Urska Kladnik, Vanja Kotar, Irma Virant-Klun, and Eda Vrtacnik Bokal Copyright © 2014 Sara Korosec et al. All rights reserved. Evaluation of the Usefulness of the MRI Jelly Method for Diagnosing Complete Cul-de-Sac Obliteration Thu, 10 Apr 2014 14:09:53 +0000 Objective. We conducted a single-center study to evaluate the usefulness of the magnetic resonance (MR) imaging jelly method for diagnosing endometriosis-associated adhesions in the Pouch of Douglas. Methods. Thirty women with menstrual pain, dyspareunia, and chronic pelvic pain were enrolled in the study. All had been scheduled for laparoscopic surgery on the basis of pelvic and/or ultrasonographic (US) evaluation. All underwent MR imaging both with and without application of US jelly to the vagina and rectum. The images were compared and analyzed postsurgically in a random and blinded fashion by a radiology specialist and a radiology fellow. The radiologists’ interpretations of the images were compared to the surgical findings recorded on DVDs. Results. Adhesions in the Pouch of Douglas were found in 21 patients. The sensitivity and specificity of MR imaging without jelly administration were 85.7% and 55.6%, respectively, for the specialist and 81.0% and 55.6%, respectively, for the fellow; with jelly administration, values were 95.2% and 88.9% for the specialist and 90.5% and 66.7% for the fellow. Opacity produced by the jelly increased the sensitivity and specificity for both radiologists. Conclusion. The MRI jelly method is a potentially useful, beneficial, and simple approach for diagnosing Pouch of Douglas adhesions. Iwaho Kikuchi, Ryohei Kuwatsuru, Kana Yamazaki, Jun Kumakiri, Yoichi Aoki, and Satoru Takeda Copyright © 2014 Iwaho Kikuchi et al. All rights reserved. Clinical and Structural Features of Sperm Head Vacuoles in Men Included in the In Vitro Fertilization Programme Wed, 09 Apr 2014 14:13:36 +0000 The human sperm head vacuoles and their role in male infertility are still poorly understood. The aim of this study was to identify the clinical and ultrastructural features of human sperm head vacuoles in men included in the in vitro fertilization programme: men with normal (normozoospermia) and impaired sperm morphology (teratozoospermia). The sperm samples were observed under 6000-time magnification using motile sperm organelle morphology examination (MSOME). The proportion of sperm with head vacuoles was evaluated and related to the outcome of in vitro fertilization. The sperm of men with impaired sperm morphology was characterized by a higher proportion of sperm head vacuoles. The sperm head vacuoles were related to impaired semen quality (sperm concentration, motility, and morphology) but were not influenced by male factors (semen volume, height, age, weight, or body mass index). Moreover, sperm head vacuoles were related to impaired fertilization rate merely after classical in vitro fertilization (IVF), while there was no relation to pregnancy. In a subgroup of men, the sperm was fixed and observed by transmission electron microscopy (TEM). The ultrastructural study revealed that sperm head vacuoles are large nuclear indentations of various sizes and positions, packed with membranous material organized in membrane whorls (MW). Nina Fekonja, Jasna Štrus, Magda Tušek Žnidarič, Katja Knez, Eda Vrtacnik Bokal, Ivan Verdenik, and Irma Virant-Klun Copyright © 2014 Nina Fekonja et al. All rights reserved. Estimating Rate of Insulin Resistance in Patients with Preeclampsia Using HOMA-IR Index and Comparison with Nonpreeclampsia Pregnant Women Wed, 09 Apr 2014 13:27:37 +0000 Women with preeclampsia, independent of obesity and glucose intolerance, exhibit insulin resistance during pregnancy. The purpose of the present study is to determine whether early diagnosis of insulin resistance during pregnancy can predict preeclampsia. Through a case-control study, 675 pregnant women were selected and their first trimester blood was taken. Their fasting blood glucose and insulin were also measured after diagnosis of preeclampsia by 20 weeks of pregnancy. Based on the experiments conducted on 675 women who were 20 weeks past their pregnancy, 375 cases with preeclampsia were selected and assigned to the case group. 35 other pregnant women were put in the control group. Diagnosis criteria for the participants included blood pressure above 140/90 and proteinuria above 300 mg or above +1. Both groups were matched according to age, parity, gestational age, and BMI. Homa-Irand rate of insulin resistance was calculated by HOMA-IR and patients were followed up. Homeostatic model assessments (HOMA-IR) revealed that the average insulin resistance increased during pregnancy among both the case and control groups. There was a significant difference between insulin resistance of these two groups in both first trimester and third trimester and after developing preeclampsia ( < 0.001, = 0.021). Insulin-resistance of the group with preeclampsia was higher in first trimester prior to diagnosis as well as the third trimester after diagnosis compared to natural pregnancy under similar conditions. Measurement of insulin resistance in first trimester may be useful in predicting the risk of preeclampsia. Farideh Rezaei Abhari, Maryam Ghanbari Andarieh, Asadollah Farokhfar, and Soleiman Ahmady Copyright © 2014 Farideh Rezaei Abhari et al. All rights reserved. Evaluation of Two Different Analytical Methods for Circulating Tumor Cell Detection in Peripheral Blood of Patients with Primary Breast Cancer Tue, 08 Apr 2014 09:25:58 +0000 Background. Evidence is accumulating that circulating tumor cells (CTC) out of peripheral blood can serve as prognostic marker not only in metastatic but also in early breast cancer (BC). Various methods are available to detect CTC. Comparisons between the different techniques, however, are rare. Material and Methods. We evaluate two different methods for CTC enrichment and detection in primary BC patients: the FDA-approved CellSearch System (CSS; Veridex, Warren, USA) and a manual immunocytochemistry (MICC). The cut-off value for positivity was ≥1 CTC. Results. The two different nonoverlapping patient cohorts evaluated with one or the other method were well balanced regarding common clinical parameters. Before adjuvant CHT 21.1% (416 out of 1972) and 20.6% (247 out of 1198) of the patients were CTC-positive, while after CHT 22.5% (359 out of 1598) and 16.6% (177 out of 1066) of the patients were CTC-positive using CSS or MICC, respectively. CTC positivity rate before CHT was thus similar and not significantly different (), while CTC positivity rate immediately after CHT was significantly lower using MICC compared to CSS (). Conclusion. Using CSS or MICC for CTC detection, we found comparable prevalence of CTC before but not after adjuvant CHT. B. A. S. Jaeger, J. Jueckstock, U. Andergassen, J. Salmen, F. Schochter, V. Fink, M. Alunni-Fabbroni, M. Rezai, Th. Beck, M. W. Beckmann, K. Friese, T. W. P. Friedl, W. Janni, and B. Rack Copyright © 2014 B. A. S. Jaeger et al. All rights reserved. Standardised Models for Inducing Experimental Peritoneal Adhesions in Female Rats Tue, 08 Apr 2014 00:00:00 +0000 Animal models for adhesion induction are heterogeneous and often poorly described. We compare and discuss different models to induce peritoneal adhesions in a randomized, experimental in vivo animal study with 72 female Wistar rats. Six different standardized techniques for peritoneal trauma were used: brushing of peritoneal sidewall and uterine horns (group 1), brushing of parietal peritoneum only (group 2), sharp excision of parietal peritoneum closed with interrupted sutures (group 3), ischemic buttons by grasping the parietal peritoneum and ligating the base with Vicryl suture (group 4), bipolar electrocoagulation of the peritoneum (group 5), and traumatisation by electrocoagulation followed by closure of the resulting peritoneal defect using Vicryl sutures (group 6). Upon second look, there were significant differences in the adhesion incidence between the groups (). Analysis of the fraction of adhesions showed that groups 2 (0%) and 5 (4%) were significantly less than the other groups (). Furthermore, group 6 (69%) was significantly higher than group 1 (48%) () and group 4 (47%) (). There was no difference between group 3 (60%) and group 6 (). From a clinical viewpoint, comparison of different electrocoagulation modes and pharmaceutical adhesion barriers is possible with standardised models. Bernhard Kraemer, Christian Wallwiener, Taufiek K. Rajab, Christoph Brochhausen, Markus Wallwiener, and Ralf Rothmund Copyright © 2014 Bernhard Kraemer et al. All rights reserved. High HPV Infection Prevalence in Men from Infertile Couples and Lack of Relationship between Seminal HPV Infection and Sperm Quality Sun, 06 Apr 2014 11:11:53 +0000 Human papillomaviruses (HPV) are the most frequently sexually transmitted viruses and etiological agents of several human cancers. Controversial results of the role of HPV in infertile population on sperm parameters have been published. The aim of this study was to estimate the type-specific prevalence of HPV DNA infection of the external genitalia and semen in 340 Slovenian men from infertile couples and to establish the relationship between seminal HPV DNA infection and abnormal sperm parameters. Self-taken swabs of the entire penile surface and semen samples were collected, and HPV detection and genotyping were performed. HPV DNA was detected in 37.12% of external genitalia and in 13.61% of semen samples with high HPV type concordance of both sampling sites. The most prevalent HPV types in the male external genitalia were HPV-CP6108 and HPV-84. The most prevalent HPV types in semen were HPV-53 and HPV-CP6108. The prevalence of HPV infection between normozoospermic men and men with abnormal sperm parameters did not differ significantly. Sperm quality did not differ significantly between men with seminal HPV infection and uninfected men. In conclusion, the men from infertile couples are equally susceptible to HPV infection regardless of their fertile potential; seminal HPV infection does not impair sperm quality. Barbara Golob, Mario Poljak, Ivan Verdenik, Mojca Kolbezen Simoniti, Eda Vrtačnik Bokal, and Branko Zorn Copyright © 2014 Barbara Golob et al. All rights reserved. Proteomes of Animal Oocytes: What Can We Learn for Human Oocytes in the In Vitro Fertilization Programme? Thu, 03 Apr 2014 11:25:25 +0000 Oocytes are crucial cells for mammalian reproduction, yet the molecular principles underlying oocyte development are only partially understood. Therefore, contemporary proteomic approaches have been used increasingly to provide new insights into oocyte quality and maturation in various species such as mouse, pig, and cow. Especially, animal studies have helped in elucidating the molecular status of oocytes during in vitro maturation and other procedures of assisted reproduction. The aim of this review is to summarize the literature on mammalian oocyte proteome and secretome research in the light of natural and assisted reproduction and on lessons to be learned for human oocytes, which have so far remained inaccessible for proteome analysis. Irma Virant-Klun and Jeroen Krijgsveld Copyright © 2014 Irma Virant-Klun and Jeroen Krijgsveld. All rights reserved. Good Preservation of Stromal Cells and No Apoptosis in Human Ovarian Tissue after Vitrification Thu, 03 Apr 2014 07:47:01 +0000 The aim of this study was to develop a vitrification procedure for human ovarian tissue cryopreservation in order to better preserve the ovarian tissue. Large size samples of ovarian tissue retrieved from 15 female-to-male transgender subjects (18–38 years) were vitrified using two solutions (containing propylene glycol, ethylene glycol, and sucrose at different concentrations) in an open system. Light microscopy, transmission electron microscopy, and TUNEL assay were applied to evaluate the efficiency of the vitrification protocol. After vitrification/warming, light microscopy showed oocyte nucleus with slightly thickened chromatin and irregular shape, while granulosa and stromal cells appeared well preserved. Transmission electron microscopy showed oocytes with slightly irregular nuclear shape and finely dispersed chromatin. Clear vacuoles and alterations in cellular organelles were seen in the oocyte cytoplasm. Stromal cells had a moderately dispersed chromatin and homogeneous cytoplasm with slight vacuolization. TUNEL assay revealed the lack of apoptosis induction by vitrification in all ovarian cell types. In conclusion after vitrification/warming the stromal compartment maintained morphological and ultrastructural features similar to fresh tissue, while the oocyte cytoplasm was slightly damaged. Although these data are encouraging, further studies are necessary and essential to optimize vitrification procedure. Raffaella Fabbri, Rossella Vicenti, Maria Macciocca, Gianandrea Pasquinelli, Roberto Paradisi, Cesare Battaglia, Nicola Antonio Martino, and Stefano Venturoli Copyright © 2014 Raffaella Fabbri et al. All rights reserved. Macrophage Capping Protein CapG Is a Putative Oncogene Involved in Migration and Invasiveness in Ovarian Carcinoma Wed, 02 Apr 2014 11:07:32 +0000 The actin binding protein CapG modulates cell motility by interacting with the cytoskeleton. CapG is associated with tumor progression in different nongynecologic tumor entities and overexpression in breast cancer cell lines correlates with a more invasive phenotype in vitro. Here, we report a significant CapG overexpression in 18/47 (38%) of ovarian carcinomas (OC) analyzed by qRealTime-PCR analyses. Functional analyses in OC cell lines through siRNA mediated CapG knockdown and CapG overexpression showed CapG-dependent cell migration and invasiveness. A single nucleotide polymorphism rs6886 inside the CapG gene was identified, affecting a CapG phosphorylation site and thus potentially modifying CapG function. The minor allele frequency (MAF) of SNP rs6886 (c.1004A/G) was higher and the homozygous (A/A, His335) genotype was significantly more prevalent in patients with fallopian tube carcinomas (50%) as in controls (10%). With OC being one of the most lethal cancer diseases, the detection of novel biomarkers such as CapG could reveal new diagnostic and therapeutic targets. Moreover, in-depth analyses of SNP rs6886 related to FTC and OC will contribute to a better understanding of carcinogenesis and progression of OC. J. Glaser, M. H. D. Neumann, Qi Mei, B. Betz, N. Seier, I. Beyer, T. Fehm, H. Neubauer, D. Niederacher, and M. C. Fleisch Copyright © 2014 J. Glaser et al. All rights reserved. Expression Profiles of Epithelial-Mesenchymal Transition-Associated Proteins in Epithelial Ovarian Carcinoma Tue, 01 Apr 2014 09:04:33 +0000 Epithelial-mesenchymal transition (EMT) has been suggested to contribute to tumor progression and acquisition of therapeutic resistance. To assess the clinical significance of EMT-associated proteins, we evaluated the expression of Snail and Slug, the key regulators of EMT, in the primary ovarian cancer samples () by immunohistochemistry. Snail was differentially expressed according to the histologic subtype () and was predominantly expressed in serous and endometrioid types. In the serous and endometrioid adenocarcinomas, the expression of Snail remained high across the stage and grade, suggesting its role in the early phase of carcinogenesis. However, the expression of Snail and Slug was not related to chemoresistance and poor prognosis and did not serve as independent predictive or prognostic marker. Mi-Kyung Kim, Min A Kim, Haeryoung Kim, Yong-Beom Kim, and Yong-Sang Song Copyright © 2014 Mi-Kyung Kim et al. All rights reserved. Review of Clinical Trials on Effects of Oral Antioxidants on Basic Semen and Other Parameters in Idiopathic Oligoasthenoteratozoospermia Mon, 31 Mar 2014 16:37:53 +0000 Infertility affects 50 to 80 million people worldwide. Male factor is a cause of infertility in almost half of cases, mainly due to oligoasthenoteratozoospermia (OAT). With common diagnostic methods no cause can be found in approximately 30% of cases of male infertility due to OAT and these are considered idiopathic. Reactive oxygen species (ROS) play an important role in male infertility and are proved to be higher in infertile men; antioxidants could oppose their effect. The aim of this paper was to review the literature on clinical trials in the period from year 2000 to year 2013 studying the effects of various types of antioxidant supplements on basic and other sperm parameters and pregnancy rates in subfertile males with idiopathic OAT. The majority of studies were randomized and placebo controlled and confirmed beneficial effect of antioxidants on at least one of the semen parameters; the biggest effect was determined on sperm motility. In many of these trials combinations of more antioxidants were assessed. The optimal dosages of one or more antioxidants were not defined. We concluded that antioxidants play an important role in protecting semen from ROS and can improve basic sperm parameters in case of idiopathic OAT. Senka Imamovic Kumalic and Bojana Pinter Copyright © 2014 Senka Imamovic Kumalic and Bojana Pinter. All rights reserved. Looking for Celiac Disease in Italian Women with Endometriosis: A Case Control Study Mon, 31 Mar 2014 12:13:13 +0000 In the last years, a potential link between endometriosis and celiac disease has been hypothesized since these disorders share some similarities, specifically concerning a potential role of oxidative stress, inflammation, and immunological dysfunctions. We investigated the prevalence of celiac disease among Italian women with endometriosis with respect to general population. Consecutive women with a laparoscopic and histological confirmed diagnosis of endometriosis were enrolled; female nurses of our institution, without a known history of endometriosis, were enrolled as controls. IgA endomysial and tissue transglutaminase antibodies measurement and serum total IgA dosage were performed in both groups. An upper digestive endoscopy with an intestinal biopsy was performed in case of antibodies positivity. Presence of infertility, miscarriage, coexistence of other autoimmune diseases, and family history of autoimmune diseases was also investigated in all subjects. Celiac disease was diagnosed in 5 of 223 women with endometriosis and in 2 of 246 controls (2.2% versus 0.8%; ). Patients with endometriosis showed a largely higher rate of infertility compared to control group (27.4% versus 2.4%; ). Our results confirm that also in Italian population an increased prevalence of celiac disease among patients with endometriosis is found, although this trend does not reach the statistical significance. Luca Santoro, Sebastiano Campo, Ferruccio D’Onofrio, Antonella Gallo, Marcello Covino, Vincenzo Campo, Guglielmo Palombini, Angelo Santoliquido, Giovanni Gasbarrini, and Massimo Montalto Copyright © 2014 Luca Santoro et al. All rights reserved. First Trimester Biomarkers in the Prediction of Later Pregnancy Complications Sun, 30 Mar 2014 09:45:12 +0000 Adverse obstetric outcomes, such as preeclampsia, preterm birth, gestational diabetes, and fetal growth restriction, are poorly predicted by maternal history and risk factors alone, especially in nulliparae. The ability to predict these outcomes from the first trimester would allow for the early initiation of prophylactic therapies, institution of an appropriate model and location of care, and recruitment of a truly “high risk” population to clinical trials of interventions to prevent or ameliorate these conditions. To this end, development of adequately sensitive and specific predictive tests for these outcomes has become a significant focus of perinatal research. This paper reviews the biomarkers involved in these multiparametric tests and also outlines the performance of these tests and issues regarding their introduction into clinical practice. Stefan C. Kane, Fabricio da Silva Costa, and Shaun Brennecke Copyright © 2014 Stefan C. Kane et al. All rights reserved. Meconium Indicators of Maternal Alcohol Abuse during Pregnancy and Association with Patient Characteristics Sun, 30 Mar 2014 08:09:54 +0000 Aim. Identification of women with moderate alcohol abuse during pregnancy is difficult. We correlated self-reported alcohol consumption during pregnancy and patient characteristics with objective alcohol indicators measured in fetal meconium. Methods. A total of 557 women singleton births and available psychological tests, obstetric data and meconium samples were included in statistical analysis. Alcohol metabolites (fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG)), were determined from meconium and correlated with patient characteristics. Results. We found that 21.2% of the 557 participants admitted low-to-moderate alcohol consumption during pregnancy. Of the parameters analyzed from meconium, only EtG showed an association with alcohol history (). This association was inverse in cases with EtG value above 120 ng/g. These values indicate women with most severe alcohol consumption, who obviously denied having consumed alcohol during pregnancy. No other associations between socioeconomic or psychological characteristics and the drinking status (via meconium alcohol metabolites) could be found. Conclusion. Women who drink higher doses of ethanol during pregnancy, according to metabolite measures in meconium, might be less likely to admit alcohol consumption. No profile of socioeconomic or psychological characteristics of those women positively tested via meconium could be established. Tamme W. Goecke, Pascal Burger, Peter A. Fasching, Abdulsallam Bakdash, Anne Engel, Lothar Häberle, Franziska Voigt, Florian Faschingbauer, Eva Raabe, Nicolai Maass, Michael Rothe, Matthias W. Beckmann, Fritz Pragst, and Johannes Kornhuber Copyright © 2014 Tamme W. Goecke et al. All rights reserved. Time-Lapse Dynamics of the Mouse Oocyte Chromatin Organisation during Meiotic Resumption Sun, 30 Mar 2014 07:30:33 +0000 In the mammalian oocyte, distinct patterns of centromeres and pericentromeric heterochromatin localisation correlate with the gamete’s developmental competence. Mouse antral oocytes display two main types of chromatin organisation: SN oocytes, with a ring of Hoechst-positive chromatin surrounding the nucleolus, and NSN oocytes lacking this ring. When matured to MII and fertilised, only SN oocytes develop beyond the 2-cell, and reach full term. To give detailed information on the dynamics of the SN or NSN chromatin during meiosis resumption, we performed a 9 hr time-lapse observation. The main significant differences recorded are: (1) reduction of the nuclear area only in SN oocytes; (2) ~17 min delay of GVBD in NSN oocytes; (3) chromatin condensation, after GVBD, in SN oocytes; (4) formation of 4-5 CHCs in SN oocytes; (5) increase of the perivitelline space, ~57 min later in NSN oocytes; (6) formation of a rosette-like disposition of CHCs, ~84 min later in SN oocytes; (7) appearance of the MI plate ~40 min later in NSN oocytes. Overall, we described a pathway of transition from the GV to the MII stage that is punctuated of discrete recordable events showing their specificity and occurring with different time kinetics in the two types of oocytes. Martina Belli, Giulia Vigone, Valeria Merico, Carlo Alberto Redi, Silvia Garagna, and Maurizio Zuccotti Copyright © 2014 Martina Belli et al. All rights reserved. Oxidatively Modified Proteins in the Serous Subtype of Ovarian Carcinoma Thu, 27 Mar 2014 13:45:46 +0000 Serous subtype of ovarian cancer is considered to originate from fallopian epithelium mucosa that has been exposed to physiological changes resulting from ovulation. Ovulation influences an increased in inflammation of epithelial ovarian cells as results of constant exposure of cells to ROS. The imbalance between ROS and antioxidant capacities, as well as a disruption of redox signaling, causes a wide range of damage to DNA, proteins, and lipids. This study applied spectrophotometric, dinitrophenylhydrazone (DNPH) assay, two-dimensional gel electrophoresis, and Western blot analyses to assess the levels of oxidatively modified proteins in 100 primary serous epithelial ovarian carcinoma and normal/surrounding tissues. These samples were obtained from 56 Caucasian and 44 African-American patients within the age range of years. Analyses showed that the levels of reactive protein carbonyl groups increased as stages progressed to malignancy. Additionally, the levels of protein carbonyls in serous ovarian carcinoma among African Americans are 40% () higher relative to Caucasian at similar advanced stages. Results suggest that oxidative stress is involved in the modification of carbonyl protein groups, leading to increased aggressiveness of epithelial ovarian tumors and may contribute to the disease's invasiveness among African Americans. Sharifeh Mehrabi, Edward E. Partridge, William Seffens, Xuebiao Yao, and Felix O. Aikhionbare Copyright © 2014 Sharifeh Mehrabi et al. All rights reserved. BACs-on-Beads Technology: A Reliable Test for Rapid Detection of Aneuploidies and Microdeletions in Prenatal Diagnosis Thu, 27 Mar 2014 11:39:01 +0000 The risk of fetal aneuploidies is usually estimated based on high resolution ultrasound combined with biochemical determination of criterion in maternal blood, with invasive procedures offered to the population at risk. The purpose of this study was to investigate the effectiveness of a new rapid aneuploidy screening test on amniotic fluid (AF) or chorionic villus (CV) samples based on BACs-on-Beads (BoBs) technology and to compare the results with classical karyotyping by Giemsa banding (G-banding) of cultured cells in metaphase as the gold standard technique. The prenatal-BoBs kit was used to study aneuploidies involving chromosomes 13, 18, 21, X, and Y as well as nine microdeletion syndromes in 321 AF and 43 CV samples. G-banding of metaphase cultured cells was performed concomitantly for all prenatal samples. A microarray-based comparative genomic hybridization (aCGH) was also carried out in a subset of samples. Prenatal-BoBs results were widely confirmed by classical karyotyping. Only six karyotype findings were not identified by Prenatal-BoBs, all of them due to the known limitations of the technique. In summary, the BACs-on-Beads technology was an accurate, robust, and efficient method for the rapid diagnosis of common aneuploidies and microdeletion syndromes in prenatal samples. Sandra García-Herrero, Inmaculada Campos-Galindo, José Antonio Martínez-Conejero, Vicente Serra, Inés Olmo, Coral Lara, Carlos Simón, and Carmen Rubio Copyright © 2014 Sandra García-Herrero et al. All rights reserved. Can Ki-67 Play a Role in Prediction of Breast Cancer Patients' Response to Neoadjuvant Chemotherapy? Tue, 25 Mar 2014 11:11:32 +0000 Background. Currently the choice of breast cancer therapy is based on prognostic factors. The proliferation marker Ki-67 is used increasingly to determine the method of therapy. The current study analyses the predictive value of Ki-67 in foreseeing breast cancer patients’ responses to neoadjuvant chemotherapy. Methods. This study includes patients with invasive breast cancer treated between 2008 and 2013. The clinical response was assessed by correlating Ki-67 to histological examination, mammography, and ultrasonography findings. Results. The average Ki-67 value in our patients collectively () is 34.9 ± 24.6%. The average Ki-67 value is the highest with 37.4 ± 24.0% in patients with a pCR. The Ki-67 values do not differ significantly among the 3 groups: pCR versus partial pathological response versus stable disease/progress (). However, Ki-67 values of patients with luminal, Her2 enriched, and basal-like cancers differed significantly from each other. Furthermore, within the group of luminal tumors Ki-67 values of patients with versus without pCR also differed significantly. Conclusion. Our data shows that the Ki-67 value predicts the response to neoadjuvant chemotherapy as a function of the molecular subtype, reflecting the daily routine concerning Ki-67 and its impressing potential and limitation as a predictive marker for neoadjuvant chemotherapy response. Juhasz-Böss Ingolf, Mavrova Russalina, Moga Simona, Radosa Julia, Schmidt Gilda, Rainer M. Bohle, Hasenfus Andrea, Solomayer Erich, and Herr Daniel Copyright © 2014 Juhasz-Böss Ingolf et al. All rights reserved. Male and Couple Fertility Impairment due to HPV-DNA Sperm Infection: Update on Molecular Mechanism and Clinical Impact—Systematic Review Tue, 25 Mar 2014 08:56:54 +0000 Recent evidences identify Human Papillomavirus (HPV) sperm infection as a possible cause of male and couple infertility. It acts through different mechanisms at various steps of human conception and early gestational development. We performed a systematic review to assess the role of HPV semen infection on male and couple infertility. Analysis of available and eligible data does not permit us to fund clear evidences about clinical impact of HPV infection on fertility, although sperm parameters impairment is the most widely recognized effect. Regarding biomolecular implications, the available data are often conflicting. More studies are required to define the role of HPV sperm infection in clinical practice. The great majority of evidences are obtained by in vitro studies and this fact represents a limitation for the clinical management of HPVDNA sperm infection. Understanding the biological significance of HPV-DNA semen infection could permit us to explain most of the idiopathic male and couple infertility, leading to a better management of infertile men and a better timing for sperm banking storage before ART cycles. Salvatore Gizzo, Bruno Ferrari, Marco Noventa, Emanuele Ferrari, Tito Silvio Patrelli, Michele Gangemi, and Giovanni Battista Nardelli Copyright © 2014 Salvatore Gizzo et al. All rights reserved. New Advances of Preimplantation and Prenatal Genetic Screening and Noninvasive Testing as a Potential Predictor of Health Status of Babies Mon, 24 Mar 2014 07:57:24 +0000 The current morphologically based selection of human embryos for transfer cannot detect chromosome aneuploidies. So far, only biopsy techniques have been able to screen for chromosomal aneuploidies in the in vitro fertilization (IVF) embryos. Preimplantation genetic diagnosis (PGD) or screening (PGS) involves the biopsy of oocyte polar bodies or embryonic cells and has become a routine clinical procedure in many IVF clinics worldwide, including recent development of comprehensive chromosome screening of all 23 pairs of chromosomes by microarrays for aneuploidy screening. The routine preimplantation and prenatal genetic diagnosis (PND) require testing in an aggressive manner. These procedures may be invasive to the growing embryo and fetus and potentially could compromise the clinical outcome. Therefore the aim of this review is to summarize not only the new knowledge on preimplantation and prenatal genetic diagnosis in humans, but also on the development of potential noninvasive embryo and fetal testing that might play an important role in the future. Tanya Milachich Copyright © 2014 Tanya Milachich. All rights reserved. Comparative Genomic Hybridization Selection of Blastocysts for Repeated Implantation Failure Treatment: A Pilot Study Sun, 23 Mar 2014 13:04:19 +0000 The aim of this study is to determine if the use of preimplantation genetic screening (PGS) by array comparative genomic hybridization (array CGH) and transfer of a single euploid blastocyst in patients with repeated implantation failure (RIF) can improve clinical results. Three patient groups are compared: 43 couples with RIF for whom embryos were selected by array CGH (group RIF-PGS), 33 couples with the same history for whom array CGH was not performed (group RIF NO PGS), and 45 good prognosis infertile couples with array CGH selected embryos (group NO RIF PGS). A single euploid blastocyst was transferred in groups RIF-PGS and NO RIF PGS. Array CGH was not performed in group RIF NO PGS in which 1-2 blastocysts were transferred. One monoembryonic sac with heartbeat was found in 28 patients of group RIF PGS and 31 patients of group NO RIF PGS showing similar clinical pregnancy and implantation rates (68.3% and 70.5%, resp.). In contrast, an embryonic sac with heartbeat was only detected in 7 (21.2%) patients of group RIF NO PGS. In conclusion, PGS by array CGH with single euploid blastocyst transfer appears to be a successful strategy for patients with multiple failed IVF attempts. Ermanno Greco, Sara Bono, Alessandra Ruberti, Anna Maria Lobascio, Pierfrancesco Greco, Anil Biricik, Letizia Spizzichino, Alessia Greco, Jan Tesarik, Maria Giulia Minasi, and Francesco Fiorentino Copyright © 2014 Ermanno Greco et al. All rights reserved. Cryopreservation of Embryos and Oocytes in Human Assisted Reproduction Sun, 23 Mar 2014 11:47:41 +0000 Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification) of human embryos and oocytes are summarized. János Konc, Katalin Kanyó, Rita Kriston, Bence Somoskői, and Sándor Cseh Copyright © 2014 János Konc et al. All rights reserved. Guideline Concordant Therapy Prolongs Survival in HER2-Positive Breast Cancer Patients: Results from a Large Population-Based Cohort of a Cancer Registry Thu, 20 Mar 2014 14:16:35 +0000 Even though randomized controlled clinical trials demonstrated improved survival by adjuvant trastuzumab treatment of HER2-positive breast cancer patients, data on its effect in clinical routine are scarce. This study evaluated the use and efficacy of trastuzumab in routine treatment of HER2-positive breast cancer patients. Data from the clinical cancer registry Regensburg (Germany) were analyzed. The present study investigated 6,991 female patients with primary invasive breast cancer. In premenopausal HER2-positive patients a considerable increase of trastuzumab therapy was observed from 58.1% in 2006 to 90.9% in 2011, whereas in postmenopausal patients trastuzumab was rather used on a constant rate of 49.1%. Best overall survival (OS) was found in HER2/steroid hormone receptor-positive patients receiving guideline concordant treatment with trastuzumab plus chemotherapy (CHT) plus antihormone therapy (AHT) with a 7-year OS rate of 96% compared to the non-trastuzumab group with a 7-year OS rate of 92%. In multivariable analysis, HER2-positive patients treated with CHT or AHT who did not get trastuzumab, had a worse 7-year OS (65%, versus 79%, ) than the control groups. This population-based study demonstrated that guideline concordant use of adjuvant trastuzumab improves OS for HER2-positive breast cancer patients treated in routine clinical care. E. C. Inwald, O. Ortmann, F. Zeman, M. Koller, F. Hofstädter, M. Gerstenhauer, and M. Klinkhammer-Schalke Copyright © 2014 E. C. Inwald et al. All rights reserved. Awareness about a Life-Threatening Condition: Ectopic Pregnancy in a Network for Surveillance of Severe Maternal Morbidity in Brazil Wed, 19 Mar 2014 09:28:02 +0000 Objective. To assess occurrence of severe maternal complications associated with ectopic pregnancy (EP). Method. A multicenter cross-sectional study was conducted, with prospective surveillance of potentially life-threatening conditions (PLTC), maternal near miss (MNM), and maternal death (MD). EP complications, patient sociodemographic/obstetric characteristics, and conditions of severity management were assessed, estimating prevalence ratios with respective 95% CI. Factors independently associated with greater severity were identified using multiple regression analysis. Results. Of the 9.555 severe maternal morbidity patients, 312 women (3.3%) had complications after EP: 286 (91.7%) PLTC, 25 (8.0%) MNM, and 1 (0.3%) MD. Severe maternal outcome ratio (SMOR) was 0.3/1000 LB among EP cases and 10.8/1000 LB among other causes. Complicated EP patients faced a higher risk of blood transfusion, laparotomy, and lower risk of ICU admission and prolonged hospitalization than women developing complications resulting from other causes. Substandard care was the most common in more severe maternal morbidity and EP cases (22.7% MNM and MD versus 15% PLTC), although not significant. Conclusion. Increased maternal morbidity due to EP raised awareness about the condition and its impact on female reproductive life. No important risk factors for greater severity were identified. Care providers should develop specific guidelines and interventions to prevent severe maternal morbidity. Edilberto Alves Rocha Filho, Danielly Scaranello Santana, Jose Guilherme Cecatti, Maria Laura Costa, Samira Maerrawe Haddad, Mary Angela Parpinelli, Maria Helena Sousa, Rodrigo Soares Camargo, Rodolfo Carvalho Pacagnella, Fernanda Garanhani Surita, and Joao Luiz Pinto e Silva Copyright © 2014 Edilberto Alves Rocha Filho et al. All rights reserved. Time-to-Delivery after Maternal Transfer to a Tertiary Perinatal Centre Tue, 18 Mar 2014 12:15:14 +0000 Objectives. To determine, in women transferred antenatally for acute admission with high risk pregnancies, the numbers who deliver, the average time from transfer to delivery, and whether the reason for transfer influences the time-to-delivery. Methods. A retrospective analysis of time-to-delivery was performed in a population of women transferred to the Royal Brisbane and Women’s Hospital, QLD. Data were obtained from the hospital obstetric, neonatal, and admission databases. Results. A total of 941 women were transferred antenatally with high risk pregnancies where delivery was deemed potentially imminent. Of these 821 (87%) delivered at RBWH. The remaining 120 women (13%) were discharged prior to delivery and then delivered elsewhere. Of the 821 maternal transfers that delivered, the median time to delivery was 24.4 hrs. There were 43% who delivered within 24 hours of admission and 29% who either delivered after 7 days or delivered elsewhere. Most transfers for fetal abnormality delivered in the first 24 hours while most transfers for antepartum haemorrhage and preterm prelabour membrane rupture delivered beyond 24 hours. Conclusion. There are significant differences in time-to-delivery following transfer depending on the reason for transfer and many infants transferred in utero will not deliver imminently. Fiona H. Hutchinson and Mark W. Davies Copyright © 2014 Fiona H. Hutchinson and Mark W. Davies. All rights reserved. Recent Progress in Cryopreservation of Bovine Oocytes Sun, 16 Mar 2014 06:47:21 +0000 Principle of oocyte cryoinjury is first overviewed and then research history of cryopreservation using bovine oocytes is summarized for the last two decades with a few special references to recent progresses. Various types of cryodevices have been developed to accelerate the cooling rate and applied to the oocytes from large domestic species enriched with cytoplasmic lipid droplets. Two recent approaches include the qualitative improvement of IVM oocytes prior to the vitrification and the short-term recovery culture of vitrified-warmed oocytes prior to the subsequent IVF. Supplementation of L-carnitine to IVM medium of bovine oocytes has been reported to reduce the amount of cytoplasmic lipid droplets and improve the cryotolerance of the oocytes, but it is still controversial whether the positive effect of L-carnitine is reproducible. Incidence of multiple aster formation, a possible cause for low developmental potential of vitrified-warmed bovine oocytes, was inhibited by a short-term culture of the postwarm oocytes in the presence of Rho-associated coiled-coil kinase (ROCK) inhibitor. Use of an antioxidant -tocopherol, instead of the ROCK inhibitor, also supported the revivability of the postwarm bovine oocytes. Further improvements of the vitrification procedure, combined with pre- and postvitrification chemical treatment, would overcome the high sensitivity of bovine oocytes to cryopreservation. In-Sul Hwang and Shinichi Hochi Copyright © 2014 In-Sul Hwang and Shinichi Hochi. All rights reserved. Quantitative Proteomics Analysis of Altered Protein Expression in the Placental Villous Tissue of Early Pregnancy Loss Using Isobaric Tandem Mass Tags Thu, 13 Mar 2014 10:00:08 +0000 Many pregnant women suffer miscarriages during early gestation, but the description of these early pregnancy losses (EPL) can be somewhat confusing because of the complexities of early development. Thus, the identification of proteins with different expression profiles related to early pregnancy loss is essential for understanding the comprehensive pathophysiological mechanism. In this study, we report a gel-free tandem mass tags- (TMT-) labeling based proteomic analysis of five placental villous tissues from patients with early pregnancy loss and five from normal pregnant women. The application of this method resulted in the identification of 3423 proteins and 19647 peptides among the patient group and the matched normal control group. Qualitative and quantitative proteomic analysis revealed 51 proteins to be differentially abundant between the two groups (≥1.2-fold, Student's -test, ). To obtain an overview of the biological functions of the proteins whose expression levels altered significantly in EPL group, gene ontology analysis was performed. We also investigated the twelve proteins with a difference over 1.5-fold using pathways analysis. Our results demonstrate that the gel-free TMT-based proteomic approach allows the quantification of differences in protein expression levels, which is useful for obtaining molecular insights into early pregnancy loss. Xiaobei Ni, Xin Li, Yueshuai Guo, Tao Zhou, Xuejiang Guo, Chun Zhao, Min Lin, Zuomin Zhou, Rong Shen, Xirong Guo, Xiufeng Ling, and Ran Huo Copyright © 2014 Xiaobei Ni et al. All rights reserved. Differential Gene Expression Profiling of Enriched Human Spermatogonia after Short- and Long-Term Culture Wed, 12 Mar 2014 12:12:54 +0000 This study aimed to provide a molecular signature for enriched adult human stem/progenitor spermatogonia during short-term (<2 weeks) and long-term culture (up to more than 14 months) in comparison to human testicular fibroblasts and human embryonic stem cells. Human spermatogonia were isolated by CD49f magnetic activated cell sorting and collagen−/laminin+ matrix binding from primary testis cultures obtained from ten adult men. For transcriptomic analysis, single spermatogonia-like cells were collected based on their morphology and dimensions using a micromanipulation system from the enriched germ cell cultures. Immunocytochemical, RT-PCR and microarray analyses revealed that the analyzed populations of cells were distinct at the molecular level. The germ- and pluripotency-associated genes and genes of differentiation/spermatogenesis pathway were highly expressed in enriched short-term cultured spermatogonia. After long-term culture, a proportion of cells retained and aggravated the “spermatogonial” gene expression profile with the expression of germ and pluripotency-associated genes, while in the majority of long-term cultured cells this molecular profile, typical for the differentiation pathway, was reduced and more genes related to the extracellular matrix production and attachment were expressed. The approach we provide here to study the molecular status of in vitro cultured spermatogonia may be important to optimize the culture conditions and to evaluate the germ cell plasticity in the future. Sabine Conrad, Hossein Azizi, Maryam Hatami, Mikael Kubista, Michael Bonin, Jörg Hennenlotter, Markus Renninger, and Thomas Skutella Copyright © 2014 Sabine Conrad et al. All rights reserved. Genetic Variants in the Genes of the Stress Hormone Signalling Pathway and Depressive Symptoms during and after Pregnancy Wed, 12 Mar 2014 11:42:31 +0000 Purpose. The aim of this study was to investigate whether single nucleotide polymorphisms (SNPs) in genes of the stress hormone signaling pathway, specifically FKBP5, NR3C1, and CRHR1, are associated with depressive symptoms during and after pregnancy. Methods. The Franconian Maternal Health Evaluation Study (FRAMES) recruited healthy pregnant women prospectively for the assessment of maternal and fetal health including the assessment of depressiveness. The German version of the 10-item Edinburgh Postnatal Depression Scale (EPDS) was completed at three time points in this prospective cohort study. Visit 1 was at study entry in the third trimester of the pregnancy, visit 2 was shortly after birth, and visit 3 was 6–8 months after birth. Germline DNA was collected from 361 pregnant women. Nine SNPs in the above mentioned genes were genotyped. After construction of haplotypes for each gene, a multifactorial linear mixed model was performed to analyse the depression values over time. Results. EPDS values were within expected ranges and comparable to previously published studies. Neither did the depression scores differ for comparisons among haplotypes at fixed time points nor did the change over time differ among haplotypes for the examined genes. No haplotype showed significant associations with depressive symptoms severity during pregnancy or the postpartum period. Conclusion. The analysed candidate haplotypes in FKBP5, NR3C1, and CRHR1 did not show an association with depression scores as assessed by EPDS in this cohort of healthy unselected pregnant women. Michael Schneider, Anne Engel, Peter A. Fasching, Lothar Häberle, Elisabeth B. Binder, Franziska Voigt, Jennifer Grimm, Florian Faschingbauer, Anna Eichler, Ulf Dammer, Dirk Rebhan, Manuela Amann, Eva Raabe, Tamme W. Goecke, Carina Quast, Matthias W. Beckmann, Johannes Kornhuber, Anna Seifert, and Stefanie Burghaus Copyright © 2014 Michael Schneider et al. All rights reserved. Decreasing Quality of the New Generations of Anti-Müllerian Hormone Assays Tue, 11 Mar 2014 08:02:14 +0000 Anti-Müllerian hormone (AMH) measurements are widely used to optimize the stimulation protocols. First generation AMH kits correlated well with ovarian reserve and response to stimulation. In the present study we aimed to asses if the new generation kits share the same accurate correlations. Retrospective data were collected from 8323 blood samples. For comparison we used Immunotech I generation kit (ImI 4035 samples), Beckman Coulter II generation kit RUO (BCII RUO 3449, samples) and Beckman Coulter II generation kit with IVD certificate (BCII IVD 839 samples). We compared average AMH concentrations measured with different kits, as well as correlation between kits. We also compared average AMH concentrations in sera collected on different cycle days and samples of different quality of preservation. AMH serum concentrations differed for each kit, ranging 4.4 ± 4.12 (mean ± SD) for the ImI, 2.68 ± 3.15 for the BCII RUO, and 1.64 ± 2.85 for BCII IVD. The mean differences from an adjusted regression model were −48.7%, −40%, and −69.2%, respectively. In conclusion, the changes of the BC AMH kits are unpredictable; however, the improvement of them is still possible. It would be very dangerous to use elaborated stimulation protocol (based on the Ist generation AMH results) with the results from the IInd generation assays. Krzysztof Lukaszuk, Beata Ludwikowska, Joanna Liss, Michal Kunicki, Miroslaw Sawczak, Aron Lukaszuk, Lukasz Plociennik, Grzegorz Jakiel, Tomasz Wasniewski, Izabela Woclawek-Potocka, and Dorota Bialobrzeska Copyright © 2014 Krzysztof Lukaszuk et al. All rights reserved. High-Throughput Analysis of Ovarian Granulosa Cell Transcriptome Mon, 10 Mar 2014 11:47:55 +0000 The quality of follicular oocytes depends on interactions with surrounding granulosa cells. Development of molecular techniques and methods enables better understanding of processes underlying mammalian reproduction on cellular level. The success in reproductive biology and medicine in different species depends on reliable assessment of oocyte and embryo viability which presently mainly bases on embryo morphology. Although successful pregnancies have been achieved using this approach, its precision still should be improved and completed with other, more objective, and accurate assessment strategies. Global profiling of gene expression in follicular cumulus cells using microarrays is continuously leading to the establishment of new biomarkers which can be used to select oocytes with highest developmental potential. Even more potential applications and greater precision could be achieved using next generation sequencing (NGS) of granulosa and cumulus cell RNA (RNA-seq). However, due to the high cost, this method is not used as frequently as microarrays at the moment. In any case, high-throughput technologies offer the possibilities and advantages in ovarian somatic cell analysis on scale that has not been noted so far. The aim of this work is to present current directions and examples of global molecular profiling of granulosa cells and underline its impact on reproductive biology and medicine. Ewa Chronowska Copyright © 2014 Ewa Chronowska. All rights reserved. Cumulative Delivery Rate after Providing Full Reimbursement In Vitro Fertilization Programme: A 6-Years Survey Sun, 09 Mar 2014 06:57:29 +0000 Since 1983, Slovenia has been offering well-established, successful, and fully reimbursed IVF programme to infertile couples. On the grounds of data gathered at the Slovenian IVF units we aimed to determine whether the fully accessible IVF treatment system can provide notable success considering cumulative delivery rate (cDR). Longitudinal analysis of getting cDR was performed in 810 IVF cycles of 395 couples who for the first time attended the IVF programme in year 2006 and were followed until year 2012. We calculated the actual and the optimistic cDR. In women aged <38 years the actual cDR was 54% and optimistic DR was 83%, respectively. In women aged ≥38 years the actual cDR was 24 % and optimistic cDR was 27%. These results enable us to report that prospects of the treatment for the women aged <38 years, if they undergo all 6 available IVF cycles, are very positive and quite comparable to the chances of spontaneous conception. Even in older patients it is beneficial to repeat the IVF procedures. Therefore we consider the existing infertility treatment system in Slovenia as an example of good medical practice with high level of beneficence offered to the patients. Urban Vrtacnik, Eda Vrtacnik Bokal, and Rok Devjak Copyright © 2014 Urban Vrtacnik et al. All rights reserved. Quantitative Elastography for Cervical Stiffness Assessment during Pregnancy Wed, 05 Mar 2014 16:42:40 +0000 Aim. Feasibility and reliability of tissue Doppler imaging-(TDI-) based elastography for cervical quantitative stiffness assessment during all three trimesters of pregnancy were evaluated. Materials and Methods. Prospective case-control study including seventy-four patients collected between the 12th and 42nd weeks of gestation. The tissue strain (TS) was measured by two independent operators as natural strain. Intra- and interoperator intraclass correlation coefficient (ICC) agreements were evaluated. Results. TS measurement was always feasible and exhibited a high performance in terms of reliability (intraoperator ICC-agreement = 0.93; interoperator ICC agreement = 0.89 and 0.93 for a single measurement and for the average of two measurements, resp.). Cervical TS showed also a significant correlation with gestational age, cervical length, and parity. Conclusions. TS measurement during pregnancy demonstrated high feasibility and reliability. Furthermore, TS significantly correlated with gestational age, cervical length, and parity. A. Fruscalzo, A. P. Londero, C. Fröhlich, U. Möllmann, and R. Schmitz Copyright © 2014 A. Fruscalzo et al. All rights reserved. Polymorphisms in the RANK/RANKL Genes and Their Effect on Bone Specific Prognosis in Breast Cancer Patients Wed, 05 Mar 2014 13:20:42 +0000 The receptor activator of NF-B (RANK) pathway is involved in bone health as well as breast cancer (BC) pathogenesis and progression. Whereas the therapeutic implication of this pathway is established for the treatment of osteoporosis and bone metastases, the application in adjuvant BC is currently investigated. As genetic variants in this pathway have been described to influence bone health, aim of this study was the prognostic relevance of genetic variants in RANK and RANKL. Single nucleotide polymorphisms in RANK(L) (rs1054016/rs1805034/rs35211496) were genotyped and analyzed with regard to bone metastasis-free survival (BMFS), disease-free survival, and overall survival for a retrospective cohort of 1251 patients. Cox proportional hazard models were built to examine the prognostic influence in addition to commonly established prognostic factors. The SNP rs1054016 seems to influence BMFS. Patients with two minor alleles had a more favorable prognosis than patients with at least one common allele (HR 0.37 (95% CI: 0.17, 0.84)), whereas other outcome parameters remained unaffected. rs1805034 and rs35211496 had no prognostic relevance. The effect of rs1054016(RANKL) adds to the evidence that the RANK pathway plays a role in BC pathogenesis and progression with respect to BMFS, emphasizing the connection between BC and bone health. Alexander Hein, Christian M. Bayer, Michael G. Schrauder, Lothar Häberle, Katharina Heusinger, Reiner Strick, Matthias Ruebner, Michael P. Lux, Stefan P. Renner, Rüdiger Schulz-Wendtland, Arif B. Ekici, Arndt Hartmann, Matthias W. Beckmann, and Peter A. Fasching Copyright © 2014 Alexander Hein et al. All rights reserved. Superoxide Dismutase: A Predicting Factor for Boar Semen Characteristics for Short-Term Preservation Wed, 05 Mar 2014 13:08:23 +0000 Superoxide dismutase (SOD), total antioxidant capacity (TAC), and thiobarbituric acid reactive substances (TBARS) in seminal plasma were evaluated on the basis of receiver operating characteristics (ROC) analysis as predictors for distinguishing satisfactory from unsatisfactory boar semen samples after storage. SOD on day 0 correlated significantly with progressive motility (; ) and viability (; ) after storage; TBARS correlated only with motility (; ). Semen samples that, after 3 days of storage, fulfilled all criteria for semen characteristics (viability > 85%, motility > 70%, progressive motility > 25%, and normal morphology > 50%) had significantly lower SOD levels on the day 0 than those with at least one criterion not fulfilled () following storage. SOD levels of less than 1.05 U/mL predicted with 87.5% accuracy that fresh semen will suit the requirements for satisfactory semen characteristics after storage, while semen with SOD levels higher than 1.05 U/mL will not fulfill with 100% accuracy at least one semen characteristic after storage. These results support the proposal that SOD in fresh boar semen can be used as a predictor of semen quality after storage. Maja Zakošek Pipan, Janko Mrkun, Marjan Kosec, Alenka Nemec Svete, and Petra Zrimšek Copyright © 2014 Maja Zakošek Pipan et al. All rights reserved. Expression of Mesenchymal Stem Cells-Related Genes and Plasticity of Aspirated Follicular Cells Obtained from Infertile Women Mon, 03 Mar 2014 09:32:27 +0000 After removal of oocytes for in vitro fertilization, follicular aspirates which are rich in somatic follicular cells are discarded in daily medical practice. However, there is some evidence that less differentiated cells with stem cell characteristics are present among aspirated follicular cells (AFCs). The aim of this study was to culture AFCs in vitro and to analyze their gene expression profile. Using the RT2 Profiler PCR array, we investigated the expression profile of 84 genes related to stemness, mesenchymal stem cells (MCSs), and cell differentiation in AFCs enriched by hypoosmotic protocol from follicular aspirates of infertile women involved in assisted reproduction programme in comparison with bone marrow-derived mesenchymal stem cells (BM-MSCs) and fibroblasts. Altogether the expression of 57 genes was detected in AFCs: 16 genes (OCT4, CD49f, CD106, CD146, CD45, CD54, IL10, IL1B, TNF, VEGF, VWF, HDAC1, MITF, RUNX2, PPARG, and PCAF) were upregulated and 20 genes (FGF2, CASP3, CD105, CD13, CD340, CD73, CD90, KDR, PDGFRB, BDNF, COL1A1, IL6, MMP2, NES, NUDT6, BMP6, SMURF2, BMP4, GDF5, and JAG1) were downregulated in AFCs when compared with BM-MSCs. The genes which were upregulated in AFCs were mostly related to MSCs and connected with ovarian function, and differed from those in fibroblasts. The cultured AFCs with predominating granulosa cells were successfully in vitro differentiated into adipogenic-, osteogenic-, and pancreatic-like cells. The upregulation of some MSC-specific genes and in vitro differentiation into other types of cells indicated a subpopulation of AFCs with specific stemness, which was not similar to those of BM-MSCs or fibroblasts. Edo Dzafic, Martin Stimpfel, Srdjan Novakovic, Petra Cerkovnik, and Irma Virant-Klun Copyright © 2014 Edo Dzafic et al. All rights reserved. Fetus Sound Stimulation: Cilia Memristor Effect of Signal Transduction Wed, 26 Feb 2014 07:14:54 +0000 Background. This experimental study evaluates fetal middle cerebral artery (MCA) circulation after the defined prenatal acoustical stimulation (PAS) and the role of cilia in hearing and memory and could explain signal transduction and memory according to cilia optical-acoustical properties. Methods. PAS was performed twice on 119 no-risk term pregnancies. We analyzed fetal MCA circulation before, after first and second PAS. Results. Analysis of the Pulsatility index basic (PIB) and before PAS and Pulsatility index reactive after the first PAS (PIR 1) shows high statistical difference, representing high influence on the brain circulation. Analysis of PIB and Pulsatility index reactive after the second PAS (PIR 2) shows no statistical difference. Cilia as nanoscale structure possess magnetic flux linkage that depends on the amount of charge that has passed between two-terminal variable resistors of cilia. Microtubule resistance, as a function of the current through and voltage across the structure, leads to appearance of cilia memory with the “memristor” property. Conclusion. Acoustical and optical cilia properties play crucial role in hearing and memory processes. We suggest that fetuses are getting used to sound, developing a kind of memory patterns, considering acoustical and electromagnetically waves and involving cilia and microtubules and try to explain signal transduction. Svetlana Jankovic-Raznatovic, Svetlana Dragojevic-Dikic, Snezana Rakic, Branka Nikolic, Snezana Plesinac, Lidija Tasic, Zivko Perisic, Mirjana Sovilj, Tatjana Adamovic, and Djuro Koruga Copyright © 2014 Svetlana Jankovic-Raznatovic et al. All rights reserved. Does Loop Electrosurgical Excision Procedure of the Uterine Cervix Affect Anti-Müllerian Hormone Levels? Sun, 23 Feb 2014 10:00:16 +0000 Background. A delayed time to pregnancy was recently reported for women who had a loop electrosurgical excision procedure (LEEP) to remove cervical intraepithelial neoplasia (CIN) grade 2 or 3. The objective of the current study was to determine if treatment of CIN with LEEP is associated with decreased levels of anti-Müllerian hormone (AMH), a marker of ovarian reserve. Methods. AMH levels were measured in 18 women treated with LEEP and 18 age-matched controls, who had colposcopy only and did not require LEEP. Cases and controls had their blood drawn at study entry time zero and again 6 months later. Results. The mean AMH level decreased significantly from baseline to follow-up; however, no significant differences were observed when stratifying by LEEP status, suggesting that both groups experienced a similar decrease in AMH levels during the follow-up period. Although women treated with LEEP had lower overall AMH levels than controls at both baseline and follow-up, these differences were not statistically significant. Conclusion. Overall, the delayed time to pregnancy observed in women treated with LEEP is likely not due to a LEEP-associated decrease in ovarian reserve as measured by AMH; thus, other mechanism are responsible for the delayed time to pregnancy associated with LEEP. Martha M. Sklavos, Cassandra N. Spracklen, Audrey F. Saftlas, and Ligia A. Pinto Copyright © 2014 Martha M. Sklavos et al. All rights reserved. Expression of Neuroendocrine Markers in Different Molecular Subtypes of Breast Carcinoma Wed, 19 Feb 2014 13:30:30 +0000 Background. Carcinomas of the breast with neuroendocrine features are incorporated in the World Health Organization classification since 2003 and include well-differentiated neuroendocrine tumors, poorly differentiated neuroendocrine carcinomas/small cell carcinomas, and invasive breast carcinomas with neuroendocrine differentiation. Neuroendocrine differentiation is known to be more common in certain low-grade histologic special types and has been shown to mainly cluster to the molecular (intrinsic) luminal A subtype. Methods. We analyzed the frequency of neuroendocrine differentiation in different molecular subtypes of breast carcinomas of no histologic special type using immunohistochemical stains with specific neuroendocrine markers (chromogranin A and synaptophysin). Results. We found neuroendocrine differentiation in 20% of luminal B-like carcinomas using current WHO criteria (at least 50% of tumor cells positive for synaptophysin or chromogranin A). In contrast, no neuroendocrine differentiation was seen in luminal A-like, HER2 amplified and triple-negative carcinomas. Breast carcinomas with neuroendocrine differentiation presented with advanced stage disease and showed aggressive behavior. Conclusions. We conclude that neuroendocrine differentiation is more common than assumed in poorly differentiated luminal B-like carcinomas. Use of specific neuroendocrine markers is thus encouraged in this subtype to enhance detection of neuroendocrine differentiation and hence characterize the biological and therapeutic relevance of this finding in future studies. David L. Wachter, Arndt Hartmann, Matthias W. Beckmann, Peter A. Fasching, Alexander Hein, Christian M. Bayer, and Abbas Agaimy Copyright © 2014 David L. Wachter et al. All rights reserved. Whole Ovine Ovaries as a Model for Human: Perfusion with Cryoprotectants In Vivo and In Vitro Wed, 19 Feb 2014 13:08:52 +0000 These experiments were performed to test the perfusion of ovine as a model for human ovaries by cryoprotectants in vivo at high temperature when the permeability of capillaries is high and when blood is insensibly replaced by the solution of cryoprotectants. By our hypothetical supposition, ovaries could be saturated by cryoprotectants before their surgical removal. The objective was to examine the effectiveness of perfusion of ovine ovaries with vascular pedicle in vivo and in vitro. Arteria ovarica was cannuled and ovaries were perfused by Leibovitz L-15 medium + 100 IU/mL heparin + 5% bovine calf serum + 6% dimethyl sulfoxide + 6% ethylene glycol + 0.15 M sucrose + Indian ink in vivo and in vitro. In the first and second cycle of experiments, ovaries ( and ) were perfused in vivo and in vitro, respectively, during 60 min with the rate of perfusion 50 mL/h (0.8 mL/min). It was established with in vivo perfusion that only about 10% of ovarian tissues were perfused due to an appearance of multiple anastomoses when the perfusion medium goes from arteria ovarica to arteria uterina without inflow into the ovaries. It was concluded that in vitro perfusion of ovine intact ovaries with vascular pedicle by freezing medium is more effective than this manipulation performed in vivo. Vladimir Isachenko, Gohar Rahimi, Maria Dattena, Peter Mallmann, Saltanat Baikoshkarova, Elisabeth Kellerwessel, Marat Otarbaev, Tamara Shalakhmetova, and Evgenia Isachenko Copyright © 2014 Vladimir Isachenko et al. All rights reserved. Maternal OGTT Glucose Levels at 26–30 Gestational Weeks with Offspring Growth and Development in Early Infancy Thu, 13 Feb 2014 12:24:35 +0000 Aims. We aim to evaluate the association of maternal gestational oral glucose tolerance test (OGTT) glucose concentrations with anthropometry in the offspring from birth to 12 months in Tianjin, China. Methods. A total of 27,157 pregnant women underwent OGTT during 26–30 weeks gestation, and their children had body weight/length measured from birth to 12 months old. Results. Maternal OGTT glucose concentrations at 26–30 gestational weeks were positively associated with Z-scores for birth length-for-gestational age and birth weight-for-length. Compared with infants born to mothers with normal glucose tolerance, infants born to mothers with gestational diabetes mellitus (impaired glucose tolerance/new diabetes) had higher mean values of Z-scores for birth length-for-gestational age (0.07/0.23; normal group −0.08) and birth weight-for-length (0.27/0.57; normal group −0.001), smaller changes in mean values of Z-scores for length-for-age (0.75/0.62; normal group 0.94) and weight-for-length (0.18/−0.17; normal group 0.37) from birth to month 3, and bigger changes in mean values in Z-scores for weight-for-length (0.07/0.12; normal group 0.02) from month 9 to 12. Conclusions. Abnormal maternal glucose tolerance during pregnancy was associated with higher birth weight and birth length, less weight and length gain in the first 3 months of life, and more weight gain in the months 9–12 of life. Gongshu Liu, Nan Li, Shurong Sun, Jing Wen, Fengjun Lyu, Wen Gao, Lili Li, Fang Chen, Andrea A. Baccarelli, Lifang Hou, and Gang Hu Copyright © 2014 Gongshu Liu et al. All rights reserved. Effect of Hyaluronan on Developmental Competence and Quality of Oocytes and Obtained Blastocysts from In Vitro Maturation of Bovine Oocytes Thu, 13 Feb 2014 00:00:00 +0000 The objective of the present study was to evaluate the effect of hyaluronan (HA) during IVM on meiotic maturation, embryonic development, and the quality of oocytes, granulosa cells (GC), and obtained blastocysts. COCs were matured in vitro in control medium and medium with additional 0.035% or 0.07% of exogenous HA. The meiotic maturity did not differ between the analysed groups. The best rate and the highest quality of obtained blastocysts were observed when 0.07% HA was used. A highly significant difference () was noted in the mean number of apoptotic nuclei per blastocyst and in the DCI between the 0.07% HA and the control blastocysts (). Our results suggest that addition of 0.035% HA and 0.07% HA to oocyte maturation media does not affect oocyte nuclear maturation and DNA fragmentation. However, the addition of 0.07% HA during IVM decreases the level of blastocysts DNA fragmentation. Finally, our results suggest that it may be risky to increase the HA concentration during IVM above 0.07% as we found significantly higher Bax mRNA expression levels in GC cultured with 0.07% HA. The final concentration of HA being supplemented to oocyte maturation media is critical for the success of the IVP procedure. Jolanta Opiela, Joanna Romanek, Daniel Lipiński, and Zdzisław Smorąg Copyright © 2014 Jolanta Opiela et al. All rights reserved. Evaluation of Granulocyte Colony-Stimulating Factor Effects on Treatment-Resistant Thin Endometrium in Women Undergoing In Vitro Fertilization Wed, 12 Feb 2014 10:02:06 +0000 The aim of the study was to assess the granulocyte colony-stimulating factor (G-CSF) effects on unresponsive thin (<7 mm) endometrium in women undergoing in vitro fertilization (IVF). We included thirty-seven subjects who had thin unresponsive endometrium on the day of triggering ovulation. These patients also failed to achieve an adequate endometrial thickness in at least one of their previous IVF cycles. In all the subjects at the time of infusion of G-CSF, endometrial thickness was 6,74 ± 1,75 mm, and, after infusion, it increased significantly to 8,42 ± 1,73 mm. When we divided the group into two subgroups according to whether the examined women conceived, we showed that the endometrium expanded significantly from 6,86 ± 1,65 to 8,80 ± 1,14 mm in the first group (who conceived) and from 6,71 ± 1,80 to 8,33 ± 1,85 mm in the second, respectively. There were no significant differences between the two subgroups in respect to the endometrial thickness both before and after G-CSF infusion. The clinical pregnancy rate was 18,9%. We concluded that the infusion of G-CSF leads to the improvement of endometrium thickness after 72 hours. Michał Kunicki, Krzysztof Łukaszuk, Izabela Woclawek-Potocka, Joanna Liss, Patrycja Kulwikowska, and Joanna Szczyptańska Copyright © 2014 Michał Kunicki et al. All rights reserved. Fluid Consumption by Mexican Women during Pregnancy and First Semester of Lactation Sun, 02 Feb 2014 09:49:40 +0000 The objective of this study was to describe daily fluid consumption in a sample of pregnant or lactating adult women. Women between 18 and 45 years of age, residents of Mexico City, stratified by socioeconomic status were asked to register their total fluid intake during 7 consecutive days. A total of 153 pregnant and 155 lactating women were recruited. On average, they drank 2.62 L/day and 2.75 L/day, respectively. Forty-one percent of pregnant women drank less than the recommended 2.3 L fluids/day, and 54% of women drank less than the recommended intake of 2.7 L/day during the first semester of lactation. Plain water contributed to 33% of total fluid intake, and sugar-sweetened beverages (SSB) contributed to 38% of total fluid intake. Up to 50% of pregnant and lactating women drank more than 1 L/day of SSB, which contributed to 632 kcal/day (27.5% of recommended dietary intake) and to 700 kcal/day (28% of recommended dietary intake), respectively. The high rates of overweight and obesity found in Mexican population, particularly among women, should alert us to the consumption of SSB during pregnancy and lactation, as excessive intake of these beverages may increase the risks of obesity, diabetes mellitus, and other chronic disorders. Homero Martinez Copyright © 2014 Homero Martinez. All rights reserved. A Comparison of Capillary and Venous Blood Haematocrits of Pregnant Women in Nigeria: The Impact on Diagnosis and Prevalence of Anaemia in Pregnancy Tue, 28 Jan 2014 09:22:38 +0000 Background. Volume of red cells in capillary blood varies from that of venous blood. The magnitude of this variation as well as its impact on the diagnosis of anaemia in pregnancy needs to be studied. This study demonstrates the disparity between capillary and venous PCV in pregnancy. Objectives. To determine whether capillary blood PCV (cPCV) differed from venous blood PCV (vPCV) of normal pregnant women in Enugu, Nigeria, and its effect on diagnosis and prevalence of anaemia. Methods. PCV was estimated using pairs of venous and capillary blood samples from 200 consecutive pregnant women at the Antenatal Clinic of University of Nigeria Teaching Hospital, Enugu, Nigeria. Results. Participants’ cPCV %, was significantly lower than their vPCV %, (, ). However, women’s cPCV had strong positive correlation with their vPCV (, ). The prevalence of anaemia among participants using capillary and venous blood was 33.5% (67/200) and 28.0% (56/200), respectively ( (CI 95%: 0.85, 1.98),  ). Conclusions. Capillary blood PCV was lower than vPCV among pregnant women in Enugu, Nigeria. Nevertheless, the prevalence of anaemia derived from cPCV did not differ significantly from that of vPCV. Cyril Chukwudi Dim, Emmanuel Onyebuchi Ugwu, Ugochukwu Bond Anyaehie, and Kingsley Chukwu Obioha Copyright © 2014 Cyril Chukwudi Dim et al. All rights reserved. Hormonal Contraceptive Use and the Prevalence of Endometriotic Lesions at Different Regions within the Peritoneal Cavity Mon, 20 Jan 2014 11:57:16 +0000 Endometriosis is an estrogen-dependent disease that can lead to chronic pain and subfertility. Endometriotic lesions found in different locations are heterogeneous and may represent a collection of related but distinct conditions. Whether there is a relationship between hormonal contraceptive (HC) use and endometriosis is still controversial. The purpose of this study was to determine whether HC use affected the prevalence of endometriotic lesions differently based on lesion location. Data was retrospectively collected from 161 patients presenting to the Berne University Women’s Hospital between 2008 and 2012 for laparoscopic investigation. Women with histologically proven endometriosis were included in the study and patients were grouped according to lesion location and HC use. The results of the study indicate that HC users are significantly less likely to have endometriotic lesions on the ovaries, although in contrast, no difference was observed in the incidence of lesions in the rectovaginal septum (RVS) or peritoneal region. In addition, women using HC who were diagnosed with endometriotic lesions on the peritoneum were significantly younger than women with lesions in other locations. In conclusion, women with endometriosis who are currently using HC are less likely to have ovarian endometriotic lesions than in alternate locations. Brett D. McKinnon, Dominic Bertschi, Juilette Wanner, Nick A. Bersinger, and Michael D. Mueller Copyright © 2014 Brett D. McKinnon et al. All rights reserved. Biochemical Markers of Spontaneous Preterm Birth in Asymptomatic Women Sun, 19 Jan 2014 00:00:00 +0000 Preterm birth is a delivery that occurs at less than 37 completed weeks of gestation and it is associated with perinatal morbidity and mortality. Spontaneous preterm birth accounts for up to 75% of all preterm births. A number of maternal or fetal characteristics have been associated with preterm birth, but the use of individual or group biochemical markers have advanced some of the understanding on the mechanisms leading to spontaneous preterm birth. This paper provides a summary on the current literature on the use of biochemical markers in predicting spontaneous preterm birth in asymptomatic women. Evidence from the literature suggests fetal fibronectin, cervical interleukin-6, and -fetoprotein as promising biochemical markers in predicting spontaneous preterm birth in asymptomatic women. The role of gene-gene and gene-environment interactions, as well as epigenetics, has the potential to further elucidate and improve understanding of the underlying mechanisms or pathways of spontaneous preterm birth. Refinement in study design and methodology is needed in future research for the development and validation of individual or group biochemical marker(s) for use independently or in conjunction with other potential risk factors such as genetic variants and environmental and behavioral factors in predicting spontaneous preterm birth across diverse populations. Ronna L. Chan Copyright © 2014 Ronna L. Chan. All rights reserved. The Effect of Multiple Single Nucleotide Polymorphisms in the Folic Acid Pathway Genes on Homocysteine Metabolism Sun, 12 Jan 2014 13:22:23 +0000 Objective. To investigate the joint effects of the single nucleotide polymorphisms (SNPs) of genes in the folic acid pathway on homocysteine (Hcy) metabolism. Methods. Four hundred women with normal pregnancies were enrolled in this study. SNPs were identified by MassARRAY. Serum folic acid and Hcy concentration were measured. Analysis of variance (ANOVA) and support vector machine (SVM) regressions were used to analyze the joint effects of SNPs on the Hcy level. Results. SNPs of MTHFR (rs1801133 and rs3733965) were significantly associated with maternal serum Hcy level. In the different genotypes of MTHFR (rs1801133), SNPs of RFC1 (rs1051266), TCN2 (rs9606756), BHMT (rs3733890), and CBS (rs234713 and rs2851391) were linked with the Hcy level adjusted for folic acid concentration. The integrated SNPs scores were significantly associated with the residual Hcy concentration (RHC) (). The Hcy level was significantly higher in the group with high SNP scores than that in other groups with SNP scores of less than 0.2 (). Moreover, this difference was even more significant in moderate and high levels of folic acid. Conclusion. SNPs of genes in the folic acid pathway possibly affect the Hcy metabolism in the presence of moderate and high levels of folic acid. Shuang Liang, Yuanpeng Zhou, Huijun Wang, Yanyan Qian, Duan Ma, Weidong Tian, Vishwani Persaud-Sharma, Chen Yu, Yunyun Ren, Shufeng Zhou, and Xiaotian Li Copyright © 2014 Shuang Liang et al. All rights reserved. Mannose-Binding Lectin Codon 54 Gene Polymorphism and Vulvovaginal Candidiasis: A Systematic Review and Meta-Analysis Mon, 06 Jan 2014 09:03:07 +0000 Mannose-binding lectin (MBL) plays a key role in the human innate immune response. It has been shown that polymorphisms in the MBL2 gene, particularly at codon 54 (variant allele B; wild-type allele designated as A), impact upon host susceptibility to Candida infection. This systematic review and meta-analysis were performed to assess the association between MBL2 codon 54 genotype and vulvovaginal candidiasis (VVC) or recurrent VVC (RVVC). Studies were searched in MEDLINE, SCOPUS, and ISI Web of Science until April 2013. Five studies including 704 women (386 cases and 318 controls) were part of the meta-analysis, and pooled ORs were calculated using the random effects model. For subjects with RVVC, ORs of AB versus AA and of BB versus AA were 4.84 (95% CI 2.10–11.15; for heterogeneity ; %) and 12.68 (95% CI 3.74–42.92; for heterogeneity , %), respectively. For subjects with VVC, OR of AB versus AA was 2.57 (95% CI 1.29–5.12; for heterogeneity ; %). This analysis indicates that heterozygosity for the MBL2 allele B increases significantly the risk for both diseases, suggesting that MBL may influence the women’s innate immunity in response to Candida. Bojan Nedovic, Brunella Posteraro, Emanuele Leoncini, Alberto Ruggeri, Rosarita Amore, Maurizio Sanguinetti, Walter Ricciardi, and Stefania Boccia Copyright © 2014 Bojan Nedovic et al. All rights reserved. Comparative Assessment of Lymph Node Micrometastasis in Cervical, Endometrial and Vulvar Cancer: Insights on the Real Time qRT-PCR Approach versus Immunohistochemistry, Employing Dual Molecular Markers Thu, 02 Jan 2014 11:19:06 +0000 To address the value of qRT-PCR and IHC in accurately detecting lymph node micrometastasis in gynecological cancer, we performed a systematic approach, using a set of dual molecular tumor-specific markers such as cytokeratin 19 (CK19) and carbonic anhydrase 9 (CA9), in a series of 46 patients (19 with cervical cancer, 18 with endometrial cancer, and 9 with vulvar cancer). A total of 1281 lymph nodes were analyzed and 28 were found positive by histopathology. Following this documentation, 82 lymph nodes, 11 positive and 71 negative, were randomly selected and further analyzed both by IHC and qRT-PCR for CK19 and CA9 expression. All 11 (100%) expressed CK19 by IHC, while only 6 (54.5%) expressed CA9. On the contrary, all the histologically negative for micrometastases lymph nodes were also negative by IHC analysis for both markers. The comparative diagnostic efficacy of the two markers using qRT-PCR, however, disclosed that the analysis of the same aliquots of the 82 lymph nodes led to 100% specificity for the CK19 biomarker, while, in contrast, CA9 failed to recapitulate a similar pattern. These data suggest that qRT-PCR exhibits a better diagnostic accuracy compared to IHC, while CK19 displays a consistent pattern of detection compared to CA9. Kalliopi I. Pappa, Alexandros Rodolakis, Ioanna Christodoulou, Maria Gazouli, Sofia Markaki, Aris Antsaklis, and Nicholas P. Anagnou Copyright © 2014 Kalliopi I. Pappa et al. All rights reserved. Effect of Paracetamol, Dexketoprofen Trometamol, Lidocaine Spray, and Paracervical Block Application for Pain Relief during Suction Termination of First-Trimester Pregnancy Tue, 24 Dec 2013 09:12:35 +0000 The aim of the study was to investigate the analgesic efficacy of preoperative oral dexketoprofen trometamol, intravenous paracetamol, lidocaine spray, and paracervical block with ultracaine on curettage procedure. A total of 111 subjects with the request of pregnancy termination between 5 and 7 weeks of gestation were included in the study. The first group (control group) consisted of 20 participants without medication. The second group consisted of 25 participants receiving 2 puffs of lidocaine sprays on cervical mucosa. The third group consisted of 20 participants receiving oral 25 mg dexketoprofen trometamol. The fourth group consisted of 23 participants receiving 1000 mg intravenous paracetamol and the fifth group consisted of 23 participants receiving paracervical block with ultracaine. Paracervical block reduced pain score significantly in both intraoperative and postoperative periods. All analgesic procedures were significantly effective for reducing pain in postoperative period. Paracervical block may be the best method for reducing pain scores in intraoperative and postoperative periods during curettage procedure. All analgesic procedures such as lidocaine, paracetamol, ultracaine, and paracervical block with ultracaine can be used for reducing pain score in postoperative period. This trial is registered with NCT01947205. Gökhan Açmaz, Hüseyin Aksoy, Nil Özoğlu, Ülkü Aksoy, and Evrim Albayrak Copyright © 2013 Gökhan Açmaz et al. All rights reserved. Prognostic Significance of Immunohistochemical Phenotypes in Patients Treated for High-Grade Cervical Intraepithelial Neoplasia Wed, 18 Dec 2013 14:49:28 +0000 Strong evidence exists that the host’s immune system plays a crucial role for the development of human papillomavirus-related cervical premalignant and malignant lesions. In particular, effective cell-mediated immunity (CMI) promotes spontaneous infection clearance and cancer precursors regression in healthy subjects, while immunosuppressed individuals are more likely to experience infection persistence, cervical intraepithelial neoplasia (CIN) lesions, and cervical cancer. In this study, the prognostic significance of immunohistochemical profiling of CD4+ T-cells, CD8+ T-cells, dendritic cells (CD11c+), T-bet+, and GATA-3+ transcription factors has been studied in surgical specimens of 34 consecutive women affected by high-grade cervical intraepithelial neoplasia (CIN2-3) submitted to cervical conization. Results have been correlated with the clinical outcomes at 24 months after treatment and statistically analyzed. Higher rates of CD4+ T-cells, CD11c+ dendritic cells, and T-bet+ transcription factor positivity showed a strong statistically significative correlation with favourable clinical outcomes (). These data reinforce the evidence of the relevance of the host’s immune status in the natural history of HPV-related cervical disease and add a prognostic significance of the cervical immunological profile in terms of predicting significant lower recurrence rates. Massimo Origoni, Marta Parma, Giacomo Dell'Antonio, Chiara Gelardi, Chiara Stefani, Stefano Salvatore, and Massimo Candiani Copyright © 2013 Massimo Origoni et al. All rights reserved. Survey of the Situation of Infertile Women Seeking In Vitro Fertilization Treatment in China Tue, 03 Dec 2013 08:43:50 +0000 Background. In previous studies, people’s knowledge of reproductive health and infertile women’s psychological states was surveyed in several countries. However, there has been limited information concerning the psychological states of infertile women seeking treatment and the outcomes of in vitro fertilization (IVF) in China. Methods. Infertile women were asked to complete short questionnaires on the day that their oocytes were retrieved; these questionnaires covered the durations of their infertility, levels of education, sources of pressure, and psychological states. Data concerning IVF outcomes were provided by embryologists and clinicians. The correlations between the duration of infertility and educational level, psychological state and education level, and psychological state and outcome of IVF were analyzed in the cohort study. Results. The duration of infertility in more than half of the females was longer than 5 years. Compared with less-educated women, women with higher levels of education sought treatment earlier and their rates of depressive symptoms were lower. There is an association between negative emotions and outcome of IVF. Conclusions. The survey of the situations of infertile women seeking IVF treatment in China indicates the importance of popularizing knowledge concerning reproductive health. Improving medical conditions, reducing the costs of treatment, and developing social culture will aid in relieving the stress of infertile women and improving assisted reproductive treatment. Xuan Jin, Gongxian Wang, Sisun Liu, Jing Zhang, Fang Zeng, Yun Qiu, and Xiaojin Huang Copyright © 2013 Xuan Jin et al. All rights reserved. Risks for Gestational Diabetes Mellitus and Pregnancy-Induced Hypertension Are Increased in Polycystic Ovary Syndrome Mon, 25 Nov 2013 09:08:06 +0000 Objectives. To evaluate pregnancy outcomes and its determinants in women with polycystic ovary syndrome (PCOS). Methods. Two-hundred and twenty pregnant PCOS and 594 healthy women were followed from early pregnancy. Incidences of gestational diabetes mellitus (GDM), pregnancy-induced hypertension (PIH), preterm birth, twinning, and fetal growth restriction (FGR) were determined. Results. The incidence of GDM was notably higher among all PCOS combined (54.9%; OR: 2.9, 95% CI: 2.0–4.1) and PCOS subgroups, whether they conceived spontaneously (51.5%; OR: 3.3, 95% CI: 2.0–5.4), or via IVF-ET or ovarian stimulation, compared with controls (14.3%; ). The incidence of PIH was also higher among all PCOS (10.4%; OR: 2.2, 95% CI: 1.1–4.4) and the subgroup conceiving spontaneously (11.8%; OR: 2.6, 95% CI: 1.1–6.2; ) but not for those conceiving with IVF-ET (9.1%) or ovarian stimulation (9.4%). Lean women with PCOS (BMI kg/m2) had higher incidences of GDM (51.1% versus 14.5%; OR: 5.6, 95% CI: 3.4–9.0) and PIH (8.9% versus 3.2%; OR: 3.0, 95% CI: 1.3–7.1) than lean controls. PCOS woemn with normal glucose tolerance had higher risk for PIH than their comparable control group (OR: 4.0, 95% CI: 1.3–11.7). Conclusion. This study suggested that PCOS is an independent risk factor for the development of GDM and PIH. This trial is registered with ChiCTR-RCC-11001824. Yunhui Wang, Xiaomiao Zhao, Huidan Zhao, Hong Ding, Jianping Tan, Jingte Chen, Rui Zhang, Ricardo Azziz, and Dongzi Yang Copyright © 2013 Yunhui Wang et al. All rights reserved. Changes in Maternal Serum Transforming Growth Factor Beta-1 during Pregnancy: A Cross-Sectional Study Mon, 18 Nov 2013 17:29:37 +0000 Changes in circulating levels of maternal serum transforming growth factor beta-1 (TGF-β1), collected from 98 women (AGA) at different gestational ages (10–38 weeks) were measured and comparisons were made between levels in pregnant and nonpregnant controls and also between 10 women with small-for-gestational age (SGA) and 7 with appropriate-for-gestational age (AGA) fetuses. Maternal serum TGF-β1 levels at all stages of pregnancy were higher than those in normal healthy nonpregnant adults. The mean TGF-β1 levels in SGA pregnancies at 34-week gestation (32.5 + 3.2 ng/mL) were significantly less than those in AGA pregnancies (39.2 + 9.8 ng/mL) while at 38-week gestation, the levels were similar in the two groups (36.04 + 4.3 versus 36.7 + 7.0 ng/mL). This differential change in TGF-β1 levels is probably an important modulating factor in the aetiopathogenesis of abnormal intrauterine fetal growth. Mandeep Singh, Ngozi C. Orazulike, Jill Ashmore, and Justin C. Konje Copyright © 2013 Mandeep Singh et al. All rights reserved. An Influence of Immunomodulation on Th1 and Th2 Immune Response in Endometriosis in an Animal Model Tue, 05 Nov 2013 15:59:49 +0000 Aim. To assess the role of the Th1 and Th2 cellular response in the etiology of endometriosis observed in a rat model, with the use of the RESAN immunomodulator. Materials and Methods. A comparative analysis of cytokines in blood serum typical of Th1 (TNF-α and INF-γ) and Th2 (IL-4, IL-6, IL-10) cell response in groups of rats, in which RESAN preparation was used as prophylaxis (Gr. I) or treatment (Gr. II) of endometriosis. Results. The results indicated an increase in the level of cytokines in blood serum typical of Th2 cell response by comparing the second and third stages of the experiment in the second group of rats and a decrease in IL-4 and IL-10 between III and IV stages. There was a significant difference in cytokine levels during the third stage of the experiment by comparing I and II groups of rats. In the III group of rats, levels of IL-10 significantly increased between the II and III stages of the experiment. Conclusion. RESAN preparation shows Th2 cell response, inhibiting the development of endometriosis in a rat model. Due to successful prophylactic action, one may speculate that RESAN vaccine may be effective as a complementary treatment after surgical excision. K. Szymanowski, J. Niepsuj-Biniaś, A. Dera-Szymanowska, M. Wołuń-Cholewa, A. Yantczenko, E. Florek, T. Opala, M. Murawski, and K. Wiktorowicz Copyright © 2013 K. Szymanowski et al. All rights reserved. Frontomaxillary Facial Angle Measurement in Screening for Trisomy 18 at 11 + 0 to 13 + 6 Weeks of Pregnancy: A Double-Centre Study Tue, 01 Oct 2013 10:04:48 +0000 Objective. The aim of this study was to evaluate the effectiveness of prenatal screening for trisomy 18 with the use of the frontomaxillary facial angle (FMF angle) measurement. Material and Methods. The study involved 1751 singleton pregnancies at 11–13 + 6 weeks, examined between 2007 and 2011. Serum PAPP-A and free beta-hCG levels were assessed, and crown-rump length, nuchal translucency, and FMF angle were measured in all patients. 1350 fetuses with known follow-up were included in the final analysis. Results. Highly significant () negative correlation between the CRL and the FMF angle was found. There were 30 fetuses with trisomy 18. FMF angle was highly significantly larger () in fetuses with trisomy 18 as compared to chromosomally normal fetuses. Two models of first trimester screening were compared: Model 1 based on maternal age, NT, and first trimester biochemistry test (DR 80–85% and FPR 0.3–0.6%), and Model 2 = Model 1 + FMF angle measurement (DR 87.3–93.3% and FPR 0.8–1.3%). Conclusions. The use of FMF angle measurement increases the effectiveness of the screening for trisomy 18. Introduction of the FMF angle as an independent marker for fetal trisomy 18 risk requires further prospective research in large populations. Bartosz Czuba, Wojciech Cnota, Agata Wloch, Piotr Wegrzyn, Krzysztof Sodowski, Miroslaw Wielgos, and Dariusz Borowski Copyright © 2013 Bartosz Czuba et al. All rights reserved. 2-Hour Postload Serum Glucose Levels and Maternal Blood Pressure as Independent Predictors of Birth Weight in “Appropriate for Gestational Age” Neonates in Healthy Nondiabetic Pregnancies Wed, 18 Sep 2013 08:49:22 +0000 Introduction. Increased neonatal birth weight (NBW), often associated with diabetic pregnancies, is a recognized indicator of childhood obesity and future metabolic risk. Predictors of NBW in healthy non-diabetic pregnancies are not yet established. Here, we investigated the association of maternal parameters of healthy non-diabetic mothers with NBW of their “appropriate-for-gestational age” neonates. Methods. The study involved 36 healthy mother/infant pairs. Examined parameters included NBW, maternal age, first and last trimester (BMI), weight gain, fasting serum lipids and glucose, 2-hour postload glucose levels and blood pressure. Results. Postload-glucose levels were significantly higher in mothers of heavier neonates. ANOVA results indicated that 15% increase in postload-glucose levels corresponded to more than 0.5 Kg increase in NBW in the third tertile. NBW correlated positively with postload glucose levels, and negatively with systolic blood pressure. Regression analysis showed that the main predictors of NBW were postload-glucose levels (, ), followed by systolic blood pressure (, ), together predicting 31.7% NBW variation. Conclusion. This study highlights that increased maternal postload sugar levels and blood pressure, within the normal range, highly predicts NBW of healthy mothers. These findings may provide focus for early dietary intervention measures to avoid future risks to the mother and baby. Jumana Saleh, Lovina Machado, and Zahra Razvi Copyright © 2013 Jumana Saleh et al. All rights reserved. Effect of Supracervical Apposition and Spontaneous Labour on Apoptosis and Matrix Metalloproteinases in Human Fetal Membranes Wed, 11 Sep 2013 11:42:23 +0000 Background. Apoptosis and matrix metalloproteinase (MMP-9) are capable of hydrolysing components of the extracellular matrix and weakening the fetal membranes which leads to eventual rupture, a key process of human parturition. The aim of this study was to determine the effect of supracervical apposition and spontaneous labour on apoptosis and MMP-9 in human fetal membranes at term. Methods. Fetal membranes were obtained from term non-labouring supracervical site (SCS) and compared to (i) a paired distal site (DS) or (ii) site of rupture (SOR) after spontaneous labour onset. Results. The expression of the proapoptotic markers Bax, Smac, Fas, FasL, caspase-3, and PARP, was significantly higher in the non-labouring SCS chorion compared to paired DS. Bax, Smac, FasL, caspase-3, and PARP staining was higher in the non-labouring SCS fetal membranes than that in the post-labour SOR. MMP-9 expression and activity were higher in the post-labour SOR fetal membranes compared to non-labouring SCS fetal membranes. Conclusion. Components of the apoptotic signalling pathways and MMP-9 may play a role in rupture and labour. Non-labouring SCS fetal membranes display altered morphology and altered apoptotic biochemical characteristics in preparation for labour, while the laboured SOR displays unique MMP characteristics. Mahalia Chai, Susan P. Walker, Clyde Riley, Gregory E. Rice, Michael Permezel, and Martha Lappas Copyright © 2013 Mahalia Chai et al. All rights reserved. Maternal Plasma miRNAs Expression in Preeclamptic Pregnancies Mon, 09 Sep 2013 12:15:46 +0000 Objective. Preeclampsia (PE) is a pregnancy-specific syndrome and one of the leading causes of maternal and fetal morbidity and mortality. The pathophysiological mechanisms of PE remain poorly known. Recently, circulating miRNAs are considered as potential useful noninvasive biomarkers. The aim of this study was to identify differentially expressed plasma miRNAs in preeclamptic pregnancies compared with normal pregnancies. Methods. Maternal plasma miRNA expression profiles were detected by SOLiD sequencing. Differential expressions between mPE/sPE and control group were found. Next, four differentially expressed plasma miRNAs were chosen to validate their expression in other large scale samples by real-time PCR. Results. In terms of sequencing results, we identified that 51 miRNAs were differentially expressed. Four differentially expressed plasma miRNAs (miR-141, miR-144, miR-221, and miR-29a) were selected to validate the sequencing results. RT-PCR data confirmed the reliability of sequencing results. The further statistical analysis showed that maternal plasma miR-141 and miR-29a are significantly overexpressed in mPE (). Maternal plasma miR-144 is significantly underexpressed in mPE and sPE (). Conclusions. Results showed that there were differentially expressed maternal plasma miRNAs in patients with preeclampsia. These plasma miRNAs might be used as notable biomarkers for diagnosis of preeclampsia. Hailing Li, Qinyu Ge, Li Guo, and Zuhong Lu Copyright © 2013 Hailing Li et al. All rights reserved. Clinical Prediction of Deeply Infiltrating Endometriosis before Surgery: Is It Feasible? A Review of the Literature Thu, 05 Sep 2013 13:41:35 +0000 Background. Endometriosis is a chronic benign gynecologic disease that can cause pelvic pain and infertility affecting almost 10% of reproductive-age women. Deeply infiltrating endometriosis (DIE) is a specific entity responsible for painful symptoms which are related to the anatomic location of the lesions. Definitive diagnosis requires surgery, and histological confirmation is advisable. The aim of this paper is to review the current literature regarding the possibility of diagnosing DIE accurately before surgery. Despite its low sensitivity and specificity, vaginal examination and evaluation of specific symptoms should not be completely omitted as a basic diagnostic tool in detecting endometriosis and planning further therapeutic interventions. Recently, transvaginal ultrasound (TVUS) has been reported as an excellent tool to diagnose DIE lesions in different locations (rectovaginal septum, retrocervical and paracervical areas, rectum and sigmoid, and vesical wall) with good accuracy. Conclusion. There are neither sufficiently sensitive and specific signs and symptoms nor diagnostic tests for the clinical diagnosis of DIE, resulting in a great delay between onset of symptoms and diagnosis. Digital examination, in addition to TVS, may help to gain better understanding of the anatomical extent and dimension of DIE which is of crucial importance in defining the best surgical approach. Márcia Mendonça Carneiro, Ivone Dirk de Sousa Filogônio, Luciana Maria Pyramo Costa, Ivete de Ávila, and Márcia Cristina França Ferreira Copyright © 2013 Márcia Mendonça Carneiro et al. All rights reserved. Comparison between Amnisure Placental Alpha Microglobulin-1 Rapid Immunoassay and Standard Diagnostic Methods for Detection of Rupture of Membranes Sun, 01 Sep 2013 09:00:53 +0000 Objective. To determine the diagnostic accuracy of placental alpha microglobulin-1 assay and standard diagnostic methods for detecting rupture of membrane. Study Design. Prospective diagnostic study, between June 2011 to November 2011 at a tertiary centre. Initial evaluation included both the standard diagnostic methods for rupture of membranes and placental alpha microglobulin-1 immunoassay. The actual rupture of membranes was diagnosed on review of the medical records after delivery (absence of membrane or a positive pad chart). Main Outcome Measures. Placental alpha microglobulin-1 immunoassay and standard diagnostic methods for diagnosis of rupture of membrane. Results. A total of 211 patients were recruited. At initial presentation, 187 patients (88.6%) had ruptured membranes, while 24 patients (11.4%) had intact membranes. Placental alpha microglobulin-1 immunoassay confirmed rupture of membranes at initial presentation with a sensitivity of 95.7% (179 of 187), specificity of 100% (24 of 24), positive predictive value of 100% (179 of 179), and negative predictive value of 75.0% (24 of 32). By comparison, the conventional standard diagnostic methods had a sensitivity of 78.1% (146 of 187), specificity of 100% (24 of 24), positive predictive value of 100% (146 of 146), and negative predictive value of 36.9% (24 of 65) in diagnosing rupture of membrane. Conclusion. Placental alpha-microglobulin-1 immunoassay is a rapid and accurate method for confirming the diagnosis of rupture of membrane. It was superior to conventional standard diagnostic methods (pooling, nitrazine, and ferning), the nitrazine test alone or fern test alone. Beng Kwang Ng, Pei Shan Lim, Mohamad Nasir Shafiee, Nur Azurah Abdul Ghani, Nor Azlin Mohamed Ismail, Mohd Hashim Omar, and Muhammad Abdul Jamil Muhammad Yassin Copyright © 2013 Beng Kwang Ng et al. All rights reserved. Endometrial Polyps in Women Affected by Levothyroxine-Treated Hypothyroidism—Histological Features, Immunohistochemical Findings, and Possible Explanation of Etiopathogenic Mechanism: A Pilot Study Thu, 22 Aug 2013 11:01:48 +0000 The aim of the study was to investigate the possible overexpression of estrogen (ERs) and progesterone (PRs) receptors both in EPs glandular and stromal cells in postmenopausal women with levothyroxine-treated hypothyroidism in comparison to EPs detected in women with physiological thyroid hormone levels. During the study period (January-February 2013) 22 patients were eligible (12 treated, 10 controls). The two groups were homogenous for general, EPs sonographic and hysteroscopic features. None of the cases of atypia was found. Immunohistochemistry showed that the two groups were similar for ERs and PRs intensity rates in EPs glandular cells despite a trend of ERs percentage expression more than 60% in 2/3 of treated patients versus 1/3 of controls. In stromal EPs components, ERs intensity was high positive in 10 (83,3%) treated cases while it was high positive in 1 control (10%). Percentage of ERs stromal expression showed a different trend between the two groups despite a borderline statistical significance. Our hypothesis is based on a possible double action of hypothyroidism and thyroxine intake: the subclinical TSH increased levels and its possible circadian oscillation could stimulate the endometrial TSHRs (increasing type 2 DIO activity); the circulating levels of exogenous thyroxine could be locally metabolized in active form by type 2 DIO stimulating ERs. Carlo Saccardi, Salvatore Gizzo, Kathrin Ludwig, Maria Guido, Mara Scarton, Michele Gangemi, Raffaele Tinelli, and Pietro Salvatore Litta Copyright © 2013 Carlo Saccardi et al. All rights reserved. Levels of Key Enzymes of Methionine-Homocysteine Metabolism in Preeclampsia Tue, 20 Aug 2013 11:57:17 +0000 Objective. To evaluate the role of key enzymes in the methionine-homocysteine metabolism (MHM) in the physiopathology of preeclampsia (PE). Methods. Plasma and placenta from pregnant women (32 controls and 16 PE patients) were analyzed after informed consent. Protein was quantified by western blot. RNA was obtained with RNA purification kit and was quantified by reverse transcritase followed by real-time PCR (RT-qPCR). Identification of the C677T and A1298C methylenetetrahydrofolate reductase (MTHFR) single-nucleotide polymorphisms (SNPs) and A2756G methionine synthase (MTR) SNP was performed using PCR followed by a high-resolution melting (HRM) analysis. S-adenosyl methionine (SAM) and S-adenosyl homocysteine (SAH) were measured in plasma using high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS). The SNP association analysis was carried out using Fisher’s exact test. Statistical analysis was performed using a Mann-Whitney test. Results. RNA expression of MTHFR and MTR was significantly higher in patients with PE as compared with controls. Protein, SAM, and SAH levels showed no significant difference between preeclamptic patients and controls. No statistical differences between controls and PE patients were observed with the different SNPs studied. Conclusion. The RNA expression of MTHFR and MTR is elevated in placentas of PE patients, highlighting a potential compensation mechanism of the methionine-homocysteine metabolism in the physiopathology of this disease. Alejandra Pérez-Sepúlveda, Pedro P. España-Perrot, Ximena Fernández B, Verónica Ahumada, Vicente Bustos, José Antonio Arraztoa, Aneta Dobierzewska, Horacio Figueroa-Diesel, Gregory E. Rice, and Sebastián E. Illanes Copyright © 2013 Alejandra Pérez-Sepúlveda et al. All rights reserved. Maternal Deaths in NSW (2000–2006) from Nonmedical Causes (Suicide and Trauma) in the First Year following Birth Mon, 19 Aug 2013 10:10:01 +0000 Introduction. Trauma, including suicide, accidental injury, motor traffic accidents, and homicides, accounts for 73% of all maternal deaths (early and late) in NSW annually. Late maternal deaths are underreported and are not as well documented or acknowledged as early deaths. Methods. Linked population datasets from births, hospital admissions, and death registrations were analysed for the period from 1 July 2000 to 31 December 2007. Results. There were 552 901 births and a total of 129 maternal deaths. Of these deaths, 37 were early deaths (early MMR of 6.7/100 000) and 92 occurred late (late MMR of 16.6/100 000). Sixty-seven percent of deceased women had a mental health diagnosis and/or a mental health issue related to substance abuse noted. A notable peak in deaths appeared to occur from 9 to 12 months following birth with the odds ratio of a woman dying of nonmedical causes within 9–12 months of birth being 3.8 (95% CI 1.55–9.01) when compared to dying within the first 3 months following birth. Conclusion. Perinatal services are often constructed to provide short-term support. Long-term identification and support of women at particular risk of maternal death due to suicide and trauma in the first year following birth may help lower the incidence of late maternal deaths. Charlene Thornton, Virginia Schmied, Cindy-Lee Dennis, Bryanne Barnett, and Hannah Grace Dahlen Copyright © 2013 Charlene Thornton et al. All rights reserved. Hemoglobin Concentration and Pregnancy Outcomes: A Systematic Review and Meta-Analysis Thu, 25 Jul 2013 14:46:06 +0000 Objective. To conduct a systematic review and meta-analysis of hemoglobin effect on the pregnancy outcomes. Methods. We searched MEDLINE and SCOPUS from January 1, 1990 to April 10, 2011. Observational studies addressing association between hemoglobin and adverse pregnancy outcomes were selected. Two reviewers independently extracted data. A mixed logistic regression was applied to assess the effects of hemoglobin on preterm birth, low birth weight, and small for gestational age. Results. Seventeen studies were included in poolings. Hemoglobin below 11 g/dL was, respectively, 1.10 (95% CI: 1.02–1.19), 1.17 (95% CI: 1.03–1.32), and 1.14 (95% CI: 1.05–1.24) times higher risk of preterm birth, low birth weight, and small for gestational age than normal hemoglobin in the first trimester. In the third trimester, hemoglobin below 11 g/dL was 1.30 (95% CI: 1.08–1.58) times higher risk of low birth weight. Hemoglobin above 14 g/dL in third trimester decreased the risk of preterm term with ORs of 0.50 (95% CI: 0.26–0.97), but it might be affected by publication bias. Conclusions. Our review suggests that hemoglobin below 11 g/dl increases the risk of preterm birth, low birth weight, and small gestational age in the first trimester and the risk of low birth weight in the third trimester. Bunyarit Sukrat, Chumpon Wilasrusmee, Boonying Siribumrungwong, Mark McEvoy, Chusak Okascharoen, John Attia, and Ammarin Thakkinstian Copyright © 2013 Bunyarit Sukrat et al. All rights reserved. Does Cryopreservation of Ovarian Tissue Affect the Distribution and Function of Germinal Vesicle Oocytes Mitochondria? Wed, 17 Jul 2013 09:56:17 +0000 The aim of this study was to evaluate mitochondrial alteration and ATP content of germinal vesicle (GV) oocytes isolated from fresh and vitrified ovaries. After superovulation, the ovaries from adult mice were collected and divided into control and vitrified groups. GV oocytes were isolated mechanically from each group. Half were cultured for 24 hours and their maturation was assessed. Metaphase II oocytes were collected and submitted to in vitro fertilization and their fertilization rates and development to the blastocyst stage were evaluated. In the remaining GV oocytes, ATP levels were quantified, and mitochondrial distribution, mitochondrial membrane potential, and intracellular free calcium were detected with rhodamine 123, JC-1 and Flou-4 AM staining, using laser-scanning confocal microscopy. Maturation and fertilization rates of GV oocytes and the developmental rates of subsequent embryos were significantly lower in vitrified samples (). The ATP content and Ca2+ levels differed significantly in fresh and vitrified GV oocytes (). Most mitochondria were seen as large and homogenous aggregates (66.6%) in fresh GV oocytes compared to vitrified oocytes (50%). No significant differences in mitochondrial membrane potential were found between the groups. The lower maturation and fertilization rates of GV oocytes from vitrified ovaries may be due to changes in their mitochondrial function and distribution. Mojdeh Salehnia, Virpi Töhönen, Saeed Zavareh, and Jose Inzunza Copyright © 2013 Mojdeh Salehnia et al. All rights reserved. Autophagy and Human Parturition: Evaluation of LC3 Expression in Placenta from Spontaneous or Medically Induced Onset of Labor Tue, 16 Jul 2013 12:52:12 +0000 Induction of labor is one of the most used procedures in obstetrics, performed to achieve vaginal delivery through cervical ripening and stimulation of uterine contractions. We investigated the impact of induction of labor upon placental autophagy, a catabolic pathway activated in response to alteration of the physiological intracellular conditions. We collected 28 singleton placentas at the time of uncomplicated term vaginal delivery (7 spontaneous onset of labor, 21 induced labor). Autophagy was evaluated by immunohistochemistry, immunofluorescence, and immunoblotting. No significant difference in the autophagy expression was found between spontaneous or induced onset of labor. We found an inverse relationship between autophagy expression and the maternal prepregnancy body mass index, irrespective of the mode of labor onset. This result could be related to the nutritional maternal habits before and throughout pregnancy rather than rapid metabolic changes during labor. Laura Avagliano, Eleonora Virgili, Chiara Garò, Federica Quadrelli, Patrizia Doi, Michele Samaja, Gaetano Pietro Bulfamante, and Anna Maria Marconi Copyright © 2013 Laura Avagliano et al. All rights reserved. Interinstitutional Variation of Caesarean Delivery Rates According to Indications in Selected Obstetric Populations: A Prospective Multicenter Study Tue, 25 Jun 2013 17:34:23 +0000 The aim of the study was to identify which groups of women contribute to interinstitutional variation of caesarean delivery (CD) rates and which are the reasons for this variation. In this regard, 15,726 deliveries from 11 regional centers were evaluated using the 10-group classification system. Standardized indications for CD in each group were used. Spearman’s correlation coefficient was used to calculate (1) relationship between institutional CD rates and relative sizes/CD rates in each of the ten groups/centers; (2) correlation between institutional CD rates and indications for CD in each of the ten groups/centers. Overall CD rates correlated with both CD rates in spontaneous and induced labouring nulliparous women with a single cephalic pregnancy at term (). Variation of CD rates was also dependent on relative size and CD rates in multiparous women with previous CD, single cephalic pregnancy at term (). As for the indications, “cardiotocographic anomalies” and “failure to progress” in the group of nulliparous women in spontaneous labour and “one previous CD” in multiparous women previous CD correlated significantly with institutional CD rates (, , and , resp.). These results supported the conclusion that only selected indications in specific obstetric groups accounted for interinstitutional variation of CD rates. Gianpaolo Maso, Monica Piccoli, Marcella Montico, Lorenzo Monasta, Luca Ronfani, Sara Parolin, Carmine Gigli, Daniele Domini, Claudio Fiscella, Sara Casarsa, Carlo Zompicchiatti, Michela De Agostini, Attilio D’Atri, Raffaela Mugittu, Santo La Valle, Cristina Di Leonardo, Valter Adamo, Mara Fracas, Giovanni Del Frate, Monica Olivuzzi, Silvio Giove, Maria Parente, Daniele Bassini, Simona Melazzini, Secondo Guaschino, Caterina Businelli, Franco G. Toffoletti, Diego Marchesoni, Alberto Rossi, Sergio Demarini, Laura Travan, Giorgio Simon, Sandro Zicari, Giorgio Tamburlini, and Salvatore Alberico Copyright © 2013 Gianpaolo Maso et al. All rights reserved. Effects of Processing and Storage on Pediococcus pentosaceus SB83 in Vaginal Formulations: Lyophilized Powder and Tablets Mon, 17 Jun 2013 18:43:51 +0000 Vaginal probiotics have an important role in preventing the colonization of the vagina by pathogens. This study aimed to investigate different formulations with Pediococcus pentosaceus SB83 (lyophilized powder and tablets with and without retarding polymer) in order to verify its stability and antilisterial activity after manufacture and during storage. The bacteriocinogenic activity of P. pentosaceus SB83 against Listeria monocytogenes was evaluated in simulated vaginal fluid. Suspension of Pediococcus pentosaceus SB83 reduced the pathogen only after 2 h and the lyophilized bacteria after 24 h of contact, and, in the tablets, P. pentosaceus SB83 lost the antimicrobial activity. The pH of simulated vaginal fluid decreased for all the tested conditions. As lyophilized powder demonstrated better results concerning antimicrobial activity, this formulation was selected to evaluate the antilisterial activity during the 12 months of storage. During storage at room temperature, lyophilized bacteria totally inhibited the pathogen only until one month of storage. At 4°C, P. pentosaceus SB83 showed antimicrobial activity during all the time of storage investigated. Therefore, the better formulation of P. pentosaceus SB83 is the lyophilized powder stored at 4°C, which may be administered intravaginally as a washing solution. Sandra Borges, Paulo Costa, Joana Silva, and Paula Teixeira Copyright © 2013 Sandra Borges et al. All rights reserved. Comparison of Hay’s Criteria with Nugent’s Scoring System for Diagnosis of Bacterial Vaginosis Thu, 13 Jun 2013 08:55:37 +0000 Although Nugent’s criterion is considered as the gold standard for the diagnosis of bacterial vaginosis (BV), the method requires an experienced slide reader and considerable time and skill. In this study, we compared the method of Hay and Ison with Nugent’s scoring criteria. Vaginal specimens were collected from a total of 213 women, presenting with or without the symptoms of vaginitis. Diagnosis of BV was done using Nugent’ and Hay’s method. Sensitivity, specificity, and predictive values for positive and negative test were calculated for Hay’s method using Nugent’s method as the gold standard. We diagnosed 70 cases (32.86%) of BV by Nugent’s method and 87 (40.85%) cases by the Hay’s method. Sensitivity, specificity, predictive value of positive result, predictive value of negative result, and Kappa value when evaluating Hay’s criteria using Nugent’s criteria as the gold standard were ≥97.2%, ≥88.1%, ≥80.4%, ≥97.1%, and ≥0.830, respectively, when Hay’s grade II and/or Nugent’s intermediate score were considered either as negative or positive or excluded. Using Nugent score for the intermediate group is the most difficult. Hay’s method shows good agreement with the gold standard method of Nugent et al. and can be used as an alternative to Nugent’s criteria in busy tertiary care hospitals. Rohit Chawla, Preena Bhalla, Sanjim Chadha, Sujatha Grover, and Suneela Garg Copyright © 2013 Rohit Chawla et al. All rights reserved. Metabolomics Application in Maternal-Fetal Medicine Sun, 09 Jun 2013 11:51:57 +0000 Metabolomics in maternal-fetal medicine is still an “embryonic” science. However, there is already an increasing interest in metabolome of normal and complicated pregnancies, and neonatal outcomes. Tissues used for metabolomics interrogations of pregnant women, fetuses and newborns are amniotic fluid, blood, plasma, cord blood, placenta, urine, and vaginal secretions. All published papers highlight the strong correlation between biomarkers found in these tissues and fetal malformations, preterm delivery, premature rupture of membranes, gestational diabetes mellitus, preeclampsia, neonatal asphyxia, and hypoxic-ischemic encephalopathy. The aim of this review is to summarize and comment on original data available in relevant published works in order to emphasize the clinical potential of metabolomics in obstetrics in the immediate future. Vassilios Fanos, Luigi Atzori, Karina Makarenko, Gian Benedetto Melis, and Enrico Ferrazzi Copyright © 2013 Vassilios Fanos et al. All rights reserved. Identification of Women for Referral to Colposcopy by Neural Networks: A Preliminary Study Based on LBC and Molecular Biomarkers Wed, 03 Oct 2012 10:31:54 +0000 Objective of this study is to investigate the potential of the learning vector quantizer neural network (LVQ-NN) classifier on various diagnostic variables used in the modern cytopathology laboratory and to build an algorithm that may facilitate the classification of individual cases. From all women included in the study, a liquid-based cytology sample was obtained; this was tested via HPV DNA test, E6/E7 HPV mRNA test, and p16 immunostaining. The data were classified by the LVQ-NN into two groups: CIN-2 or worse and CIN-1 or less. Half of the cases were used to train the LVQ-NN; the remaining cases (test set) were used for validation. Out of the 1258 cases, cytology identified correctly 72.90% of the CIN-2 or worst cases and 97.37% of the CIN-1 or less cases, with overall accuracy 94.36%. The application of the LVQ-NN on the test set allowed correct classification for 84.62% of the cases with CIN-2 or worse and 97.64% of the cases with CIN-1 or less, with overall accuracy of 96.03%. The use of the LVQ-NN with cytology and the proposed biomarkers improves significantly the correct classification of cervical precancerous lesions and/or cancer and may facilitate diagnosis and patient management. Petros Karakitsos, Charalampos Chrelias, Abraham Pouliakis, George Koliopoulos, Aris Spathis, Maria Kyrgiou, Christos Meristoudis, Aikaterini Chranioti, Christine Kottaridi, George Valasoulis, Ioannis Panayiotides, and Evangelos Paraskevaidis Copyright © 2012 Petros Karakitsos et al. All rights reserved. Quantitative Proteomic (iTRAQ) Analysis of 1st Trimester Maternal Plasma Samples in Pregnancies at Risk for Preeclampsia Tue, 10 Apr 2012 13:58:58 +0000 A current major obstacle is that no reliable screening markers exist to detect pregnancies at risk for preeclampsia. Quantitative proteomic analysis employing isobaric labelling (iTRAQ) has been suggested to be suitable for the detection of potential plasma biomarkers, a feature we recently verified in analysis of pregnancies with Down syndrome foetuses. We have now examined whether this approach could yield biomarkers to screen pregnancies at risk for preeclampsia. In our study, we used maternal plasma samples obtained at 12 weeks of gestation, six from women who subsequently developed preeclampsia and six with uncomplicated deliveries. In our analysis, we observed elevations in 10 proteins out of 64 proteins in the preeclampsia study group when compared to the healthy control group. These proteins included clusterin, fibrinogen, fibronectin, and angiotensinogen, increased levels of which are known to be associated with preeclampsia. An elevation in the immune-modulatory molecule, galectin 3 binding protein, was also noted. Our pilot study, therefore, indicates that quantitative proteomic iTRAQ analysis could be a useful tool for the detection of new preeclampsia screening markers. Varaprasad Kolla, Paul Jenö, Suzette Moes, Olav Lapaire, Irene Hoesli, and Sinuhe Hahn Copyright © 2012 Varaprasad Kolla et al. All rights reserved. Ovarian Surface Epithelium in Patients with Severe Ovarian Infertility: A Potential Source of Cells Expressing Markers of Pluripotent/Multipotent Stem Cells Tue, 06 Dec 2011 13:50:19 +0000 The aim of this study was to confirm the presence of stem cells in the ovarian surface epithelium of patients with premature ovarian failure and no mature follicles and oocytes. In these patients, small round cells of unknown origin expressing SOX-2 marker of pluripotency were observed among the epithelial cells just after the ovarian surface epithelium scraping. These cells were an integral part of the ovarian surface epithelium. When the scraped cells were cultured in a medium with added follicular fluid to provide some ovarian niche, primitive oocyte-like cells and typical round-shaped cell clusters positively stained on alkaline phosphatase, and markers of pluripotency, such as SOX-2 and SSEA-4, were developed. These markers were expressed early and also later in the culture. Single oocyte-like cells expressed genes OCT4A, SOX-2, NANOG, NANOS, STELLA, CD9, LIN28, KLF4, GDF3, and MYC, characteristic for pluripotent stem cells. The results of this study confirmed the presence of putative stem cells in the ovarian surface epithelium of these patients and provided some basis to create a stem cell line in the future. Irma Virant-Klun, Thomas Skutella, Martin Stimpfel, and Jasna Sinkovec Copyright © 2011 Irma Virant-Klun et al. All rights reserved. Association of Angiotensin-Converting Enzyme Intron 16 Insertion/Deletion and Angiotensin II Type 1 Receptor A1166C Gene Polymorphisms with Preeclampsia in South East of Iran Tue, 26 Jul 2011 19:05:26 +0000 Some evidence suggests that a variety of genetic factors contributed in pathogenesis of the preeclampsia. The aim of this study was to assess the association between the angiotensin-converting enzyme (ACE) I/D and angiotensin II type1 receptor A1166C polymorphisms with preeclampsia. This study was performed in 125 preeclamptic pregnant women and 132 controls. The I/D Polymorphism of the ACE gene was assessed by polymerase chain reaction and the A1166C Polymorphism of the AT1R gene was determined by restriction fragment length polymorphism. The genotype and allele frequencies of I/D polymorphism differed between two groups. The risk of preeclampsia was 3.2-fold in pregnant women with D allele (OR, 3.2 [95% CI, 1.1 to 3.8]; 𝑃=0.01). The distribution of the AT1R gene A1166C polymorphism was similar in affected and control groups. Our results supported that presence of the I/D polymorphism of ACE gene is a marker for the increased risk of preeclampsia. Saeedeh Salimi, Mojgan Mokhtari, Minoo Yaghmaei, Mohammad Jamshidi, and Anoosh Naghavi Copyright © 2011 Saeedeh Salimi et al. All rights reserved. Clinical Therapeutics in Pregnancy Wed, 06 Jul 2011 15:53:15 +0000 Most drugs are not tested for use during pregnancy, consequently, labeling, which may include information about fetal safety, includes nothing about dosing, efficacy, or maternal safety. Yet these are concerns of health care providers considering treatment of disease during pregnancy. Therefore, the practitioner treats the pregnant woman with the same dose recommended for use in adults (typically men) or may decide not to treat the disease at all. However, is the choice of not treating a woman during pregnancy better than dealing with the challenges which accompany treatment? This paper, which summarizes metabolic and physiologic changes induced by pregnancy, illustrates that standard adult dosing is likely to be incorrect during pregnancy. Maisa N. Feghali and Donald R. Mattison Copyright © 2011 Maisa N. Feghali and Donald R. Mattison. All rights reserved. Differential Expression of MicroRNAs between Eutopic and Ectopic Endometrium in Ovarian Endometriosis Wed, 10 Mar 2010 10:52:09 +0000 Endometriosis, defined as the presence of endometrial tissue outside the uterus, is a common gynecological disease with poorly understood pathogenesis. MicroRNAs are members of a class of small noncoding RNA molecules that have a critical role in posttranscriptional regulation of gene expression by repression of target mRNAs translation. We assessed differentially expressed microRNAs in ectopic endometrium compared with eutopic endometrium in 3 patients through microarray analysis. We identified 50 microRNAs differentially expressed and the differential expression of five microRNAs was validated by real-time RT-PCR in other 13 patients. We identified in silico their predicted targets, several of which match the genes that have been identified to be differentially expressed in ectopic versus eutopic endometrium in studies of gene expression. A functional analysis of the predicted targets indicates that several of these are involved in molecular pathways implicated in endometriosis, thus strengthening the hypothesis of the role of microRNAs in this pathology. Nicoletta Filigheddu, Ilaria Gregnanin, Paolo E. Porporato, Daniela Surico, Beatrice Perego, Licia Galli, Claudia Patrignani, Andrea Graziani, and Nicola Surico Copyright © 2010 Nicoletta Filigheddu et al. All rights reserved.