Identification of serum miR-1246 and miR-150-5p as novel diagnostic biomarkers for high-grade serous ovarian cancer.

Epithelial ovarian cancer (EOC) is one of the leading cancers in women, with high-grade serous ovarian cancer (HGSOC) being the most common and lethal subtype of this disease. A vast majority of HGSOC are diagnosed at the late stage of the disease when the treatment and total recovery chances are low. Thus, there is an urgent need for novel, more sensitive and specific methods for early and routine HGSOC clinical diagnosis. In this study, we performed miRNA expression profiling using the NanoString miRNA assay in 34 serum samples from patients with HGSOC and 36 healthy women. We identified 13 miRNAs that were differentially expressed (DE). For additional exploration of expression patterns correlated with HGSOC, we performed weighted gene co-expression network analysis (WGCNA). As a result, we showed that the module most correlated with tumour size, nodule and metastasis contained 8 DE miRNAs. The panel including miR-1246 and miR-150-5p was identified as a signature that could discriminate HGSOC patients with AUCs of 0.98 and 1 for the training and test sets, respectively. Furthermore, the above two-miRNA panel had an AUC = 0.946 in the verification cohorts of RT-qPCR data and an AUC = 0.895 using external data from the GEO public database. Thus, the model we developed has the potential to markedly improve the diagnosis of ovarian cancer.

Implantation of embryos. The way for the improvement of the cumulative life birth rate (CLBR) in the assisted reproductive technology (ART).

Despite the willingness, humans belong to the species with the limited procreation possibilities. Infertility affects about 15% of population is the very important subject for the reproductive medicine. An assisted reproductive technology (ART) offers the significant chance for the infertile couples, but it does not give the guarantee for the pregnancy and for the birth of a healthy child. The implantation of embryo, despite numerous trials and attempts, remain the last barrier in the assisted reproduction technologies; thus, the endometrial receptivity becomes the subject of permanent interest. In this review we have tried to present various methods of improvement of an endometrial receptivity with the conclusion that we still wait for the valuable prognostic factor in the treatment of infertility by ART which could predict the chance for the birth of a healthy child.

Recent advances in prophylactics and treatment of osteoporosis.

Recently, a dramatic increase in the average life expectancy has been noted, regarded as the one of the greatest achievements of the last decades, but in consequence osteoporosis affects millions of people all over the world. Currently osteoporosis is defined as a skeletal disorder characterized by compromised bone strength which leads to an increased risk of fractures. The most commonly used tool to evaluate the 10-year risk of fractures is the fracture risk assessment tool validated in many independent cohorts. Osteoporosis itself has no symptoms, but fractures are common symptoms of osteoporosis which can result in disability and mortality. Hence, osteoporosis is called a silent epidemic as well as a silent killer. The best way to assess patients with osteoporosis is by using the most widely employed techniques - dual-energy X-ray absorptiometry or quantitative computed tomography. There are a lot of precisely documented risk factors of osteoporosis - e.g. cigarette smoking, alcohol use, getting little or no exercise, being small-framed or thin, a diet low in foods containing calcium and vitamin D - and their limitation or elimination is the best way for prophylactics of this dangerous disease. Some other risk factors such as age and sex of patients should not be omitted in the decision making process. In the literature there are numerous therapeutic proposals and different guidelines. In this review we present the recent advances in the prophylactics and treatment of osteoporosis.

miRNAs as Predictive Factors in Early Diagnosis of Gestational Diabetes Mellitus.

Introduction: Circulating miRNAs are important mediators in epigenetic changes. These non-coding molecules regulate post-transcriptional gene expression by binding to mRNA. As a result, they influence the development of many diseases, such as gestational diabetes mellitus (GDM). Therefore, this study investigates the changes in the miRNA profile in GDM patients before hyperglycemia appears. Materials and Methods: The study group consisted of 24 patients with GDM, and the control group was 24 normoglycemic pregnant women who were matched for body mass index (BMI), age, and gestational age. GDM was diagnosed with an oral glucose tolerance test between the 24th and 26th weeks of pregnancy. The study had a prospective design, and serum for analysis was obtained in the first trimester of pregnancy. Circulating miRNAs were measured using the NanoString quantitative assay platform. Validation with real time-polymerase chain reaction (RT-PCR) was performed on the same group of patients. Mann-Whitney U-test and Spearman correlation were done to assess the significance of the results. Results: Among the 800 miRNAs, 221 miRNAs were not detected, and 439 were close to background noise. The remaining miRNAs were carefully investigated for their average counts, fold changes, p-values, and false discovery rate (FDR) scores. We selected four miRNAs for further validation: miR-16-5p, miR-142-3p, miR-144-3p, and miR-320e, which showed the most prominent changes between the studied groups. The validation showed up-regulation of miR-16-5p (p<0.0001), miR-142-3p (p=0.001), and miR-144-3p (p=0.003). Conclusion: We present changes in miRNA profile in the serum of GDM women, which may indicate significance in the pathophysiology of GDM. These findings emphasize the role of miRNAs as a predictive factor that could potentially be useful in early diagnosis.

Serum C18:1-Cer as a Potential Biomarker for Early Detection of Gestational Diabetes.

We hypothesized that sphingolipids may be early biomarkers of gestational diabetes mellitus (GDM). Here, 520 women with normal fasting plasma glucose levels were recruited in the first trimester and tested with a 75 g oral glucose tolerance test in the 24th-28th week of pregnancy. Serum sphingolipids concentrations were measured in the first and the second trimester by ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC/MS/MS) in 53 patients who were diagnosed with GDM, as well as 82 pregnant women with normal glucose tolerance (NGT) and 32 non-pregnant women. In the first trimester, pregnant women showed higher concentrations of C16:0, C18:1, C22:0, C24:1, and C24:0-Cer and lower levels of sphinganine (SPA) and sphingosine-1-phosphate (S1P) compared to non-pregnant women. During pregnancy, we observed significant changes in C16:0, C18:0, C18:1, and C24:1-Cer levels in the GDM group and C18:1 and C24:0-Cer in NGT. The GDM (pre-conversion) and NGT groups in the first trimester differed solely in the levels of C18:1-Cer (AUC = 0.702 p = 0.008), also considering glycemia. Thus, C18:1-Cer revealed its potential as a GDM biomarker. Sphingolipids are known to be a modulator of insulin resistance, and our results indicate that ceramide measurements in early pregnancy may help with GDM screening.

Ceramides and Sphingosino-1-Phosphate in Obesity.

Obesity is a growing worldwide problem, especially in developed countries. This disease adversely affects the quality of life and notably contributes to the development of type 2 diabetes, metabolic syndrome, and cardiovascular disorders. It is characterised by excessive lipids accumulation in the subcutaneous and visceral adipose tissue. Considering the secretory function of adipose tissue, this leads to impaired adipokines and cytokines release. Changes in adipose tissue metabolism result in chronic inflammation, pancreatic islets dysfunction and peripheral insulin resistance. In addition to saturating various adipocytes, excess lipids are deposited into non-adipose peripheral tissues, which disturbs cell metabolism and causes a harmful effect known as lipotoxicity. Fatty acids are metabolised into bioactive lipids such as ceramides, from which sphingolipids are formed. Ceramides and sphingosine-1-phosphate (S1P) are involved in intracellular signalling, cell proliferation, migration, and apoptosis. Studies demonstrate that bioactive lipids have a crucial role in regulating insulin signalling pathways, glucose homeostasis and ß cell death. Data suggests that ceramides may have an opposite cellular effect than S1P; however, the role of S1P remains controversial. This review summarises the available data on ceramide and sphingolipid metabolism and their role in obesity.

Proven and unproven methods for diagnosis and treatment of infertility.

Infertility and subfertility affect a significant part of the population. Among various definitions of this pathology, the most common one is provided by the World Health Organisation; it says that infertility is 'a disease of the reproductive system defined by the failure to achieve a clinical pregnancy after 12 months or more of regular unprotected sexual intercourses'. The global prevalence of infertility is about 50-70 million couples. Since the process of human reproduction involves a large number of factors, a broad spectrum of infertility tests might be applied. Some of the tests focus directly on the pregnancy, some are only partially effective, whereas others should not be recommended for infertile couples. The aim of this review is to present a critical evaluation of the infertility diagnostic process and to discuss the recommendations for its most effective treatment.

Myoinositol in the Prevention of Gestational Diabetes Mellitus: Is It Sensible?

Gestational diabetes mellitus (GDM) is a complication that increasingly affects pregnant women. Due to the risk of adverse outcomes in the mother as well as in the fetus which is caused by GDM, appropriate diagnosis and treatment is very essential. Nevertheless, it is important to find new, effective ways of prevention of GDM to avoid side effects. A promising example of such an action may be supplementation of myoinositol. As shown in studies, myoinositol may reduce the risk of developing gestational diabetes mellitus by improving insulin sensitivity.

Molecular mechanisms underlying mifepristone's agonistic action on ovarian cancer progression.

BACKGROUND: Recent clinical trials on ovarian cancer with mifepristone (MF) have failed, despite in vitro findings on its strong progesterone (P4) antagonist function. METHODS: Ovarian cancer human and murine cell lines, cultured high-grade human primary epithelial ovarian cancer (HG-hOEC) cells and their explants; as well as in vivo transgenic mice possessing ovarian cancer were used to assess the molecular mechanism underlying mifepristone (MF) agonistic actions in ovarian cancer progression. FINDINGS: Herein, we show that ovarian cancer cells express traceable/no nuclear P4 receptor (PGR), but abundantly P4 receptor membrane component 1 (PGRMC1). MF significantly stimulated ovarian cancer cell migration, proliferation and growth in vivo, and the translocation of PGRMC1 into the nucleus of cancer cells; the effects inhibited by PGRMC1 inhibitor. The beneficial antitumor effect of high-doses MF could not be achieved in human cancer tissue, and the low tissue concentrations achieved with the therapeutic doses only promoted the growth of ovarian cancers. INTERPRETATION: Our results indicate that treatment of ovarian cancer with MF and P4 may induce similar adverse agonistic effects in the absence of classical nuclear PGRs in ovarian cancer. The blockage of PGRMC1 activity may provide a novel treatment strategy for ovarian cancer. FUND: This work was supported by grants from the National Science Centre, Poland (2013/09/N/NZ5/01831 to DP-T; 2012/05/B/NZ5/01867 to MC), Academy of Finland (254366 to NAR), Moikoinen Cancer Research Foundation (to NAR) and EU PARP Cluster grant (UDA-POIG.05.01.00-005/12-00/NCREMFP to SW).

A comparison of the International Association of Diabetes and Pregnancy Study Groups Recommendations with Former Criteria for Diagnosing Gestational Diabetes Mellitus: A Retrospective Cohort Study.

AIM: The aim of the study was to compare the frequency of gestational diabetes mellitus (GDM) and pregnancy outcomes in women diagnosed by WHO 1999 and IADPSG criteria. METHODS: This was a retrospective cohort study comprising 1508 women who underwent a 75-g OGTT after the 24th week of gestation at the University Hospital of Bialystok between 2004 and 2012. RESULTS: GDM was diagnosed by WHO 1999 criteria in 486 (32.2%) patients and by IADPSG criteria in 397 (26.3%) women. Three hundred fifty five (23.5%) patients fulfilled both criteria, whereas 111 (7.4%) and 39 (2.6%) subjects met only WHO 1999 or IADPSG criteria, respectively. Isolated fasting hyperglycemia was found in 3.4% of patients fulfilling WHO 1999 criteria and in 17.6% of women who met IADPSG criteria. In total, fasting glycemic value was diagnostic in 42.8% of the participants fulfilling the new criteria. The main risk factor for GDM was family history of diabetes (OR 2.285 [95%CI: 1.772-2.945], p=0.00001). The rates of cesarean section and macrosomia were higher in the group with GDM than in the healthy women (54.7% vs 41.9% and 18.9% vs 13.9%, respectively), but the differences were not significant. Three months postpartum the disturbances of glucose tolerance were found in 21% of the patients with GDM. CONCLUSIONS: The introduction of the IADPS criteria did not increase the prevalence of GDM, but increased the number of patients with fasting hyperglycemia. Twelve weeks postpartum the patients with prior GDM had significantly higher post-load glucose levels than the healthy women.

Systematic biobanking, novel imaging techniques, and advanced molecular analysis for precise tumor diagnosis and therapy: The Polish MOBIT project.

Personalized and precision medicine is gaining recognition due to the limitations by standard diagnosis and treatment; many areas of medicine, from cancer to psychiatry, are moving towards tailored and individualized treatment for patients based on their clinical characteristics and genetic signatures as well as novel imaging techniques. Advances in whole genome sequencing have led to identification of genes involved in a variety of diseases. Moreover, biomarkers indicating severity of disease or susceptibility to treatment are increasingly being characterized. The continued identification of new genes and biomarkers specific to disease subtypes and individual patients is essential and inevitable for translation into personalized medicine, in estimating both, disease risk and response to therapy. Taking into consideration the mostly unsolved necessity of tailored therapy in oncology the innovative project MOBIT (molecular biomarkers for individualized therapy) was designed. The aims of the project are: (i) establishing integrative management of precise tumor diagnosis and therapy including systematic biobanking, novel imaging techniques, and advanced molecular analysis by collecting comprehensive tumor tissues, liquid biopsies (whole blood, serum, plasma), and urine specimens (supernatant; sediment) as well as (ii) developing personalized lung cancer diagnostics based on tumor heterogeneity and integrated genomics, transcriptomics, metabolomics, and radiomics PET/MRI analysis. It will consist of 5 work packages. In this paper the rationale of the Polish MOBIT project as well as its design is presented. (iii) The project is to draw interest in and to invite national and international, private and public, preclinical and clinical initiatives to establish individualized and precise procedures for integrating novel targeted therapies and advanced imaging techniques.

Seroprevalence of parvovirus B19 antibodies among young pregnant women or planning pregnancy, tested for toxoplasmosis.

INTRODUCTION AND OBJECTIVE: Acute parvovirus B19 (B19V) infection is a proven risk for pregnant women and fetus. The aim of this study was to determine the prevalence of B19V antibodies among pregnant women or planning pregnancy, who were referred for preventive toxoplasmosis screening. MATERIAL AND METHODS: Between 2007-2010, 55 women in the age between 21 and 40 years were tested for both B19V IgG and IgM antibodies and sociodemographic information was collected. RESULTS: Among the study group, the mean age was 30 years, 43.6% of women were positive only for B19V IgG antibodies, 9% were positive for both B19V IgG and IgM antibodies and 11% were positive only for B19V IgM antibodies. Women negative for B19 IgG antibodies (47.3%) were considered as a high-risk group of B19V viremia. The serological profile indicating infection with Toxoplasma gondii was considered as a risk factor for fetal distress. The T. gondii IgG antibodies were detected in 51% cases, in 32.7% antibodies were positive for both IgG and IgM, while in 16.3% cases both IgG and IgM were negative. CONCLUSIONS: B19V infection and overlapping of other independent risk factors during pregnancy pose a significant hazard to fetus during development. Therefore, we recommend further broadening the epidemiological database of B19V infection prevalence among women. B19V infection should be taken into account during differential diagnosis as a cause of miscarriage.

Increased Maternal and Cord Blood Betatrophin in Gestational Diabetes.

AIM: The aim of the study was to compare maternal and cord blood levels of betatrophin--a new peptide potentially controlling beta cell growth--as well as in its mRNA expression in subcutaneous adipose tissue, visceral adipose tissue and placental tissue obtained from pregnant women with normal glucose tolerance (NGT) and gestational diabetes (GDM). METHODS: Serum betatrophin and irisin concentrations were measured by ELISA in 93 patients with GDM and 97 women with NGT between 24 and 28 week of gestation. Additionally, maternal and cord blood betatrophin and irisin, as well as their genes (C19orf80 and Fndc5) expression were evaluated in 20 patients with GDM and 20 women with NGT at term. RESULTS: In both groups, serum betatrophin concentrations were significantly higher in the patients with GDM than in the controls (1.91 [1.40-2.60] ng/ml vs 1.63 [1.21-2.22] ng/ml, p=0.03 and 3.45 [2.77-6.53] ng/ml vs 2.78 [2.16-3.65] ng/ml, p=0.03, respectively). Cord blood betatrophin levels were also higher in the GDM than in the NGT group (20.43 [12.97-28.80] ng/ml vs 15.06 [10.11-21.36] ng/ml, p=0.03). In both groups betatrophin concentrations in arterial cord blood were significantly higher than in maternal serum (p=0.0001). Serum irisin levels were significantly lower in the patients with GDM (1679 [1308-2171] ng/ml) than in the healthy women between 24 and 28 week of pregnancy (1880 [1519-2312] ng/ml, p=0.03). Both C19orf80 and Fndc5 mRNA expression in fat and placental tissue did not differ significantly between the groups studied. CONCLUSIONS: Our results suggest that an increase in maternal and cord blood betatrophin might be a compensatory mechanism for enhanced insulin demand in GDM.

The association of bone turnover markers with pro- and anti-inflammatory adipokines in patients with gestational diabetes.

OBJECTIVE: The aim of the study was to assess differences in circulating osteocalcin (OC) and osteoprotegerin (OPG), as well as in their expression in subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and placental tissue obtained from patients with gestational diabetes mellitus (GDM) and normal glucose tolerance (NGT). MATERIALS AND METHOD: Serum levels of OC, OPG and soluble nuclear factor-kB ligand (sRANKL) were measured in 49 women with GDM and 30 subjects with NGT between weeks 24-32 of gestation, and three months after childbirth. OC and OPG mRNA expression was measured in 23 patients with GDM and 23 women with NGT at term, using quantitative real-time RT-PCR. RESULTS: The patients with GDM had decreased OC mRNA expression in SAT (p=0.015), lower adiponectin mRNA expression in VAT (p=0.039), and a lower circulating adiponectin level (p=0.04). Multiple regression analysis revealed that serum adiponectin was significantly associated with OC mRNA expression in SAT (b=0.49, p=0.03). Three months postpartum, the OPG/sRANKL ratio was markedly higher in the subjects with prior GDM (p=0.03) and correlated positively with HbA1c (R=0.33; p=0.04), fasting insulin (R=0.35; p=0.03) and HOMA-IR (R=0.34; p=0.04). CONCLUSIONS: In the patients with GDM decreased OC mRNA expression in SAT might be associated with a reduced stimulatory effect on adiponectin expression in adipose tissue. On the other hand, higher OPG/sRANKL ratio suggests a better protection against bone loss in the subjects with prior GDM.

Disturbances of modulating molecules (FOXP3, CTLA-4/CD28/B7, and CD40/CD40L) mRNA expressions in the orbital tissue from patients with severe graves' ophthalmopathy.

Purpose. To evaluate the relationship between the expression of orbital tissue mRNA for FOXP3, CTLA-4/CD28/CD80/CD86, and CD40/CD40 and the severity of Graves' orbitopathy (GO). Material and Methods. Orbital tissue was obtained from 26 patients with GO, with mild (n = 6) or severe GO (n = 20), and 7 healthy controls. The expression of mRNA of FOXP3, CTLA-4/CD28/CD80/CD86, CD40/CD40L was measured by RT-PCR. TCR and CD3 were evaluated by immunohistochemistry. Results. Higher mRNA for FoxP3 (relative expression: 1.4) and CD40 (1.27) and lower expression of CTLA-4 (0.61) were found in the GO tissues versus controls. In severe GO as compared to mild GO higher mRNA expression for FoxP3 (1.35) and CD40 (1.4) and lower expression for CTLA-4 (0.78), CD28 (0.62), and CD40L (0.56) were found. A positive correlation was found between FOXP3 mRNA and CD3 infiltration (R = 0.796, P = 0.0000001). Conclusions. The enhanced FOXP3 mRNA expression in GO samples may suggest the dysfunction of FOXP3 cells in the severe GO. The diminished mRNA expression of CTLA-4 in severe GO may indicate inadequate T regulatory function. The enhanced mRNA expression of CD40 in severe GO and negative correlation to CRP mRNA may suggest their role in the active and inactive GO.

The IL-6/IL-6R/sgp130 system and Th17 associated cytokines in patients with gestational diabetes.

INTRODUCTION: Interleukin-6 (IL-6) is a pleiotropic cytokine which signals through a cell surface receptor complex consisting of a cognatereceptor subunit (IL-6R) and glycoprotein 130 (gp130), which is considered an antagonist to the IL-6R/IL-6 pathway. The aim of the present study was to assess IL-6/IL-6R/gp130 system and Th17 associated cytokines in different time points during and after pregnancy in women with gestational diabetes mellitus (GDM) and normal glucose tolerance (NGT). MATERIAL AND METHODS: Serum levels of IL-6, sIL6R, sgp130, IL-17 and IL-23 were measured in 91 women divided into three groups: GDMin the 24th-28th week of gestation (visit 1), NGT at the 1st visit and GDM in the 29th-32nd week, and NGT at both visits. RESULTS: The patients with GDM recognised at the 1st visit had significantly higher IL-6 (p = 0.02) and sgp130 (p = 0.03) concentrations than had the women with NGT, whereas the women with GDM diagnosed at the 2nd visit had elevated sIL-6R concentrations (p = 0.03). The patients with low sIL-6R but high sgp130 concentration had significantly higher glucose levels (p = 0.04) and lower IL-6 values (p = 0.04) than had the patients with low sIL-6R and sgp130 concentrations. IL-17 and IL-23 were detected in approximately one-third of the population studied. A trend towards higher IL-17 levels was observed in the subjects with GDM, but the differences were not significant. CONCLUSIONS: Our results suggest that an increased serum sgp130 concentration in the patients with GDM might represent a compensatory mechanism, controlling intracellular IL-6 signalling and preventing the activation of the IL-6/IL-6R pathway.

Serum irisin concentration in women with gestational diabetes.

Irisin is a novel myokine and adipokine which induces an increase in total body energy expenditure, improving insulin sensitivity and glucose tolerance in experimental animals. In the present study, serum irisin concentration was measured by an enzyme immunoassay in 130 women with gestational diabetes mellitus (GDM) and 140 BMI-matched patients with normal glucose tolerance (NGT). Median irisin level was significantly lower in the patients with GDM than in the NGT subjects (1703.3 [1354.8-2097.9 ng/ml] versus 1873.8 [1519.8-2294.8 ng/ml], p = 0.01); however, 3 months after childbirth its concentrations did not differ markedly between the two groups (1165.9 [872.1-1497.5] ng/ml versus 1139.0 [984.0-1376.7] ng/ml). In the whole group, irisin concentration correlated negatively with 2 h glucose level (R = -0.14, p = 0.03). In the women with NGT, irisin concentration correlated positively with IS(OGTT) (R = 0.22, p = 0.04) and the disposition index (DI(120)) (R = 0.24, p = 0.03), as well as negatively with 2 h insulin level (R = -0.23, p = 0.03) and HOMA-IR (R = -0.24, p = 0.02). Multiple regression analysis revealed that 2 h glucose and DI(120) were the only variables significantly influencing serum irisin (ß = 0.158, p = 0.03 and ß = 0.159, p = 0.02, respectively). Our results suggest that serum irisin concentration increases markedly in pregnant women, but this increase seems to be significantly lower in patients with GDM.

[The role of inositol deficiency in the etiology of polycystic ovary syndrome disorders]. / Rola niedoboru inozytolu w patofizjologii zaburzen wystepujacych w zespole policystycznych jajników.

Inositol acts as a second messenger in insulin signaling pathway Literature data suggest inositol deficiency in insulin-resistant women with the polycystic ovary syndrome. Supplementation of myo-inisitol decreases insulin resistance as it works as an insulin sensitizing agent. The positive role of myo-inositol in the treatment of polycystic ovary syndrome has been of increased evidence recently The present review presents the effects of myo-inositol on the ovarian, hormonal and metabolic parameters in women with PCOS.