Sex steroids and sex hormone-binding globulin in postmenopausal women with nonalcoholic fatty liver disease.

OBJECTIVE: The evaluation of serum sex steroids and sex hormone-binding globulin (SHBG) levels in postmenopausal women with nonalcoholic fatty liver disease (NAFLD) and their association to the disease severity. DESIGN: Twenty-two postmenopausal women with biopsy-proven NAFLD and 18 matched controls were recruited. Blood samples for serum SHBG, total testosterone, estradiol levels and standard biochemical tests were obtained after overnight fasting. Free androgen index (FAI), calculated free (cFT) and bioavailable testosterone were estimated by standard formulas. RESULTS: The NAFLD group had lower serum SHBG levels and higher values of cFT, bioavailable testosterone and FAI, despite exhibiting similar to controls levels of serum total testosterone and estradiol. Serum SHBG levels (adjusted odds ratio [aOR]=0.912; 95% CI 0.854-0.973), bioavailable testosterone (aOR=1.254; 95% CI 1.010-1.556) and FAI (aOR=2.567; 95% CI 1.153-5.716), but not cFT, were associated with NAFLD independently of age, body mass index (BMI) and waist circumference. Serum estradiol levels were associated with the presence of nonalcoholic steatohepatitis (NASH) independently of age, BMI and waist circumference (aOR=0.727; 95% CI 0.537-0.985). CONCLUSIONS: Low SHBG levels and high metabolically active testosterone fractions were independently associated with NAFLD. Among NAFLD patients, serum estradiol levels were independently associated with NASH. However, these results need further validation from large-scale studies.

The role of 3-dimensional power Doppler imaging in the assessment of ovarian teratoma in pregnancy: a case report.

Background. Conventional sonography is the primary imaging tool for these pregnant women who present with an ovarian teratoma. In some cases, however, sonography diagnosis is difficult. We report a case of ovarian teratoma during pregnancy diagnosed by three-dimensional Power Doppler. The cyst was removed via laparotomy without fetal or maternal complications. Three-dimensional ultrasound with multiplanar view can better discriminate a benign ovarian teratoma from complex ovarian lesions or malignant tumors. Its role is significant especially during pregnancy as it may assist in determining which patients are requiring surgery and which are not. The results of three-dimensional sonography and magnetic resonance (MR) were equal but the role of MR imaging is limited in early pregnancy. Conclusions. Three-dimensional technique is a reliable diagnostic modality for preoperative assessment of an ovarian teratoma as it can be performed during the first trimester of pregnancy.

Early hCG addition to rFSH for ovarian stimulation in IVF provides better results and the cDNA copies of the hCG receptor may be an indicator of successful stimulation.

A simple, safe and cost-effective treatment protocol in ovarian stimulation is of great importance in IVF practice, especially in the case of previous unsuccessful attempts. hCG has been used as a substitute of LH because of the degree of homology between the two hormones. The main aim of this prospective randomized study was to determine, for the first time, whether low dose hCG added to rFSH for ovarian stimulation could produce better results compared to the addition of rLH in women entering IVF-ET, especially in those women that had previous IVF failures. An additional aim was to find an indicator that would allow us to follow-up ovarian stimulation and, possibly, modify it in order to achieve a better IVF outcome; and that indicator may be the cDNA copies of the LH/hCG receptor. Group A patients (n = 58) were administered hCG and Group B rLH (n = 56) in addition to rFSH in the first days of ovarian stimulation. The number of follicles and oocytes and, most importantly, implantation and pregnancy rates were shown to be statistically significantly higher in the hCG group. This study has also determined, for the first time to our best knowledge, m-RNA for LH/hCG receptors in the lymphocytes of peripheral blood 40 h before ovum pick-up. cDNA levels of the hCG receptor after ovarian stimulation were significantly higher among women receiving hCG compared to those receiving LH. In addition, higher levels were encountered among women with pregnancy compared to those without, although this was not statistically significant due to the small number of pregnancies. It seems that hCG permits a highly effective and more stable occupancy of rLH/hCG receptors and gives more follicles and more oocytes. The determination of cDNA copies could be, in the future, a marker during ovulation induction protocols and of course a predictor for the outcome of ART in the special subgroup of patients with previous failures.

Fournier's gangrene in a patient after third-degree burns: a case report.

INTRODUCTION: Fournier's gangrene is characterized by tissue ischemia leading to rapidly progressing necrotizing fasciitis. CASE PRESENTATION: We present the case of a patient with Fournier's gangrene after third-degree burns. Clinical manifestations, laboratory results and treatment options are discussed. CONCLUSION: Fournier's gangrene is a surgical emergency. Although it can be lethal, it is still a challenging situation in the field of surgical infections.

Embryoid bodies from mouse stem cells express oxytocin receptor, Oct-4 and DAZL.

Oxytocin is a nine amino acid peptide involved in a wide spectrum of physiological functions; predominantly those concerning reproduction and differentiation are of interest. Oxytocin receptors are expressed at early developmental stages of mammals, suggesting that oxytocin might be involved in the determination of the germ stem cell line, at the very early stages of mammalian development. In this respect, the proximate aim of the present study was to confirm and further analyze the existence of oxytocin receptors at a very early level of cell commitment, that is, the determination of germ cells derived from embryoid bodies. To achieve our purpose we have cultured mouse embryonic stem cells under conditions inducing formation of embryoid bodies. In this work, ES cells were allowed to aggregate in a novel medium, "Stefanidis medium" from day 0 to day 14 until formed EBs. RNA was isolated from EBs and using RT-PCR we showed that EBs expressed Oct-4, OTR, OT, and DAZL. To demonstrate simultaneous expression immunocytochemistry was preformed, in which EBs showed strong immunoreactivity for both, OTR and DAZL molecular markers. We found that 35% of the cells displayed OTR, using flow cytometry. In addition, this novel medium showed to increase OTR mRNA. We propose, that at least in murine induced embryoid bodies there is simultaneous expression of oxytocin receptors and germ cell markers (DAZL) in many cells (expressing Oct-4). We thus conclude that, the oxytocin might indeed be a molecule playing a leading role in germ cell determination.

Bidirectional communication between neural and cardiac cells in human amniotic fluid.

OBJECTIVE: To evaluate whether amniotic fluid cells contain cardiomyocyte- and neuron-like cells. STUDY DESIGN: In this experimental study, cells from human amniotic fluid samples were analyzed for mRNA expression of microtubule-associated protein 2 (MAP-2), vimentin and oxytocin (OT) receptor via RT-PCR. Immunocytochemistry was also performed with MAP-2, OT receptor, vimentin and troponin I antibodies. RESULTS: In all samples, MAP-2, vimentin and mRNA expression were detectable. OT receptor was also detectable. The cells showed strong immunoreactivity for molecular markers of neurogenic cells including MAP-2 and vimentin. The cells also showed strong immunoreactivity for molecular markers of cardiac muscle such as troponin I and for OT receptors. This report also shows that atosiban (an OT antagonist) added to culture medium of amniotic fluid cells did not induce neurogenic and cardiomyogenic differentiation. CONCLUSIONS: The observed concurrent development of cardiomyocyte- and neuron-like cells suggests that amniotic fluid contains progenitor cells and there is bidirectional communication between both cell types.

Nevirapine induces growth arrest and premature senescence in human cervical carcinoma cells.

OBJECTIVE: The aim of this study was to assess the impact of nevirapine on a cervix carcinoma cell line. METHODS: HeLa cells were cultured in Dulbecco's modified Eagle medium supplemented with 20% fetal bovine serum at 37 degrees C and humidified 10% CO(2) in air. Nevirapine was purified from commercially available Viramune (Boehringer-Ingelheim), diluted in dimethyl sulfoxide (DMSO, Sigma-Aldrich) in 350 microMu final concentration and added to cell cultures 5 h after seeding. The same DMSO volume (0.2% final concentration) was added to controls. RESULTS: We found that nevirapine treatment induces reversible growth arrest and produces morphological changes in treated cells. In contrast with previous reports the observed effects of nevirapine did not correlate with promotion of differentiation, but with induction of premature senescence. Premature senescence as a response to anti-tumour treatment is a common effect of the most anti-cancer chemotherapeutics. Nevirapine treated cells strongly accumulated SA-b-Gal activity and also expressed increased levels of p53 and p21 when analyzed via RT-PCR. In order to further explore the potent mechanisms of premature senescence induction we performed pChk2-Thr68 immunofluorescence analysis and found that nevirapine treated cells exhibited increased number of nuclear foci, indicating activated DNA damage response. CONCLUSION: We propose that at least in HeLa cell line nevirapine treatment exerts an effect far from the differentiation process, by introducing the cells into premature senescence. Based on these data, the effects of RT inhibitors should be further investigated since they may represent an additional agent against human cancer.

Oxytocin receptor- and Oct-4-expressing cells in human amniotic fluid.

BACKGROUND/AIMS: The present clinical and molecular study aimed at investigating the presence of the genes encoding oxytocin receptor (OT-R) and Oct-4 in human amniotic fluid cells. METHODS: Amniotic fluid samples were obtained from amniocentesis. Cells from human amniotic fluid samples were analyzed for mRNA expression of OT-R and Oct-4 via reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemistry was also performed with OT-R and Oct-4 antibodies. RESULTS: RT-PCR from 10 independent amniocentesis samples demonstrated the expression of OT-R and Oct-4 mRNA. The cells also showed strong immunoreactivity for molecular markers of OT-R and Oct-4. CONCLUSION: OT-R and Oct-4 are expressed in human amniotic fluid cells. The role of oxytocin in the physiology and pathophysiology of amniotic fluid cells remains to be settled.

Deleted in Azoospermia-Like (DAZL) gene-expressing cells in human amniotic fluid: a new source for germ cells research?

OBJECTIVE: To evaluate whether amniotic fluid cells contain a germ-like cell subpopulation. DESIGN: Experimental study. SETTING: University hospital. PATIENT(S): None. INTERVENTION(S): Cells from human amniotic fluid samples were analyzed for messenger RNA expression of Deleted in Azoospermia-Like gene (DAZL) and Oct-4 by reverse transcriptase polymerase chain reaction. DAZL and C-kit protein expression was assessed by flow cytometry. Immunocytochemistry also was performed to determine DAZL-, stage-specific embryonic antigen 4 (SSEA-4)-, and Oct-4-positive cells. MAIN OUTCOME MEASURE(S): DAZL gene expression in amniotic fluid cells. RESULT(S): Reverse transcriptase polymerase chain reaction, flow cytometric, and immunocytochemical analyses revealed that human amniotic fluid consists of a distinct cell population that expresses DAZL, C-kit, SSEA-4, and Oct-4. CONCLUSION(S): Our results suggest that human amniotic fluid represents a new source for the isolation of human DAZL-, C-kit-, SSEA-4-, and Oct-4-positive stem cells without raising the ethical issues associated with human embryonic research.