Limited knowledge exists about the effects of commonly used diuretic medications on the human normal flora. Thus, we investigated potential stimulatory effects of diuretic drug furosemide on urogenital tract microbiota in women. Three strains of E. coli and C. albicans with different biofilm forming capacities were obtained from female patients diagnosed with urinary tract infections. All tested strains were treated with two different concentrations of furosemide drug, in comparison to non-treated strains as the negative control. At specific time intervals, samples were obtained from growing culture and analyzed for their proliferation rate, aspartyl proteinase excretion and biofilm formation ability. E. coli and C. albicans strains significantly increased their aspartyl proteinase excretion under furosemide treatment. This effect was frequently observed after 16 hours of incubation at 37oC. This drug has also increased the biofilm forming capacities of E. coli and C. albicans strains. Interestingly, both E. coli and C. albicans non-biofilm former strains, gained the capacity of biofilm formation when treated with furosemide at certain concentrations. E. coli control became a weak biofilm former after 48 hours of incubation, while non-biofilm former C. albicans strain became a weak biofilm former in dose-dependent fashion, after 48 hours incubation with furosemide in concentration of 0.1 mg/mL, and after 16 hours of incubation with furosemide in concentration of 0.5 mg/mL. Loop diuretic drug furosemide is able to increase the microbial virulence and turn commensal microbes into opportunistic pathogens. Additionally, the results suggest that enzyme aspartyl proteinase might act as a signal molecule for the biofilm formation, leading to the increased microbial pathogenicity.

Purpose: To evaluate the results of the Kurjak antenatal neurodevelopmental test (KANET) in multicentric studies. Materials and methods: In KANET, normal, borderline, and abnormal fetuses postnatal development was followed. The Chi-square was considered significant if p < 0.05 was used in the statistical analysis. Specificity, sensitivity, prevalence, positive and negative predictive value, false-positive, and false-negative results were calculated. Results: In 3,709 singleton fetuses, KANET was assessed. Postnatal follow-up was available for 1,556 of 1,573 (42.4%) infants. For 2,136, the data were not available. Interrater reliability (κ coefficient) for low-risk fetuses was between 0.64 and 0.65 (agreement 94.2–97.3%), for high-risk fetuses was between 0.44 and 0.53 (agreement 70.5–78.9%), respectively. Borderline KANET was found in 153 (9.7%), abnormal in 52 (3.3%), both more prevalent in high-risk pregnancies ( χ 2 = 457.36; df = 2; p < 0.01). Abnormal KANET was connected with severe postnatal developmental delay ( χ 2 = 315.28; df = 6; p < 0.01). In 47 out of 1,102 children aged 2 years and more, abnormal KANET was found, among them in one with CP. In 1 out of 1,055 children with normal KANET, severe developmental delay was found. The KANET has low sensitivity for detection of developmental delay and CP. The KANET specificity is high for all tested variables, positive predictive value and the false-positive rate are high, the negative predictive value is high (99.77–99.95%), and the false-negative rate is low (0.23–0.05%). Conclusion: With normal KANET, there is a high probability of normal infant development. If the KANET score is borderline or abnormal in high-risk pregnancies, postnatal development may appear abnormal.

Background: Premature ovarian failure (POF) can be found in 1% of women at the age of 35-40, mostly due to unknown causes. PI3K-Akt signaling is associated with both ovarian function and growth of primordial follicles. In this study, we examined the effects of autologous in vitro ovarian activation with stem cells and autologous growth factors on reproductive and endocrine function in patients with ovarian impairment. Materials and Methods: The longitudinal prospective observational study included 50 patients (between 30 and 50 years) with a diagnosis of POF and infertility. This multicenter study was performed at Jevremova Special Hospital in Belgrade, Saint James Hospital (Malta), and Remedica Skoplje Hospital, between 2015 and 2018. All patients went through numerous laboratory testings, including hormonal status. The autologous bone marrow mesenchymal stem cells (BMSCs) and growth factors were used in combination for activation of ovarian tissue before its re-transplantation. The software package SPSS 20.0 was used for statistical analysis of the results. Results: Differences in follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and progesterone (PG) hormone concentrations before and after 3, 6, and 12 months post-transplantation were tested in correlation with the volume of transplanted ovarian tissue. A significant correlation (P=0.029) was found between the change in E2 level after 3 months and the volume of re-transplanted tissues. Also after re-transplantation, 64% of the patients had follicles resulting in aspiration of oocytes in 25% of positive women with follicles. Conclusion: The SEGOVA method could potentially solve many human reproductive problems in the future due to the large number of patients diagnosed with POF, as well as the possibility of delaying menopause, thus improving the quality of life and general health (Registration number: NCT04009473).

Multiple pathologies of the placenta can be identified during the first trimester of pregnancy thanks to the ultrasound diagnostics, and many molecular trophoblasts defects associated with it are still poorly understood. In other words, the 3Dand 4D-ultrasound are still the golden tool in early detection of abnormal fetal development. Using placenta as a marker in diagnosis of chromosomal abnormalities has detection sensitivity of only 3%, and is not syndrome specific, as in the case of Wolf-Hirschhorn syndrome where only few cases of hypotrophic placenta were reported. We describe the case of first-trimester pregnancy with diagnosis of pathologic placental morphology and intrauterine growth restriction of the fetus, with prenatal genetic screening testing which confirmed existence of Wolf-Hirschhorn syndrome. Suspicion for chromosomal abnormalities was raised due to placenta ultrasound examination only and this case report outlines the clinical significance of the placenta as a novel marker for Wolf-Hirschhorn syndrome, since other clinical indications in this case were not present. Keywords— Placenta, Wolf-Hirschhorn Syndrome, 3D-

Abstract An extensive review of the literature on the diagnostic and clinical accuracy of Kurjak’s antenatal neurodevelopmental test (KANET) and the summarized results of the multi-center study involving 10 centers revealed that four-dimensional ultrasonography (4D US) has become a powerful tool and KANET a valuable test that empowers the clinicians worldwide to evaluate the fetal behavior in a systematic way and contribute to the detection of fetuses that might be at high risk for neurological impairments and in particular cerebral palsy (CP). After 10 years of clinical use, many published papers and multi-center studies, hundreds of trained physicians and numerous tests performed all over the world, KANET has proven its value and has been showing encouraging results so far. The aim of this paper is to show the results from the studies done so far and to reveal the clinical value of the KANET. We expect that data from the larger ongoing collaborative study, the short- and long-term postnatal follow-up will continue to improve our knowledge. Ultimately, we all strive to prevent the CP disorders, autism spectrum disorder (ASD) and other neurological impairments and we are convinced that KANET might be helpful in the realization of this important aim.

Production of free oxygen radicals in greater volume than it is possible for human organism to neutralize, can affect quality of sperm. Reactive oxygen species (ROS) have an effect on mobility of human spermatozoids by changing the structure of membrane and the function of mitochondria, where they are primarily generated. This can affect DNA integrity of spermatozoids and compromise their vitality. Seven male patients, in age between 27 and 44, were examined by standard semen analysis, nitroblue tetrazolium (NBT) assay and sperm vitality test based on eosin and nigrosin staining. From patients with positive NBT test, 40% of the patients had normal spermiogram results, while 60% of them had decreased sperm concentration, motility, or both of these parameters below the referent value. In 20% cases the output of eosin-nigrosin test was below 58%. The NBT test and eoisn-nigrosin test are simple, effective and available tests which can help in better diagnosis of male infertility and should be integrated in routine

Background: The importance of homocysteine (Hcy) is increasingly recognized in last few decades as an independent risk factor for atherosclerosis and thrombosis, but there is lack of data referring to influence of Hcy on plasma oxidative stress parameters as well as the role of gasotransmitters in these effects. Therefore, this study aim was to assess the role of gasotransmitter inhibitors in Hcy-induced effects on plasma oxidative stress in rats. Material and Methods: Study involved 96 male Wistar albino rats divided into 8 groups: 1) Control group – saline (1ml 0.9 % NaCl i.p.); 2) DL-Hcy (8 mmol/kg i.p. DL homocysteine (DL-Hcy); 3) L-NAME (10 mg/kg i.p. Nω-Nitro-L-arginine methyl ester (L-NAME), inhibitor of NO production); 4) ZnPPR IX (30 mol/kg i.p. protoporphyrin IX zinc (ZnPPR IX), inhibitor of CO production); 5) DL-PAG (50 mg//kg i.p. DL-propargylglycine (DL-PAG), inhibitor of H2S production); 6) DL-Hcy+L-NAME (8 mmol/kg i.p. DL-Hcy + 10 mg/kg i.p. L-NAME); 7) DL-Hcy+ZnPPR IX (8 mmol/kg i.p. DL-Hcy + 30 mol/kg i.p. Zn PPR IX), and 8) DL-Hcy+DL-PAG (8 mmol/kg i.p. DL-Hcy + 50 mg//kg i.p. DL-PAG). In all experimental groups, tested substances were administered in a single dose, intraperitoneally, 60 minutes before animals’ euthanasia. In the collected blood samples malondialdehyde concentration, catalase, glutathione peroxidase and superoxide dismutase activity were measured. Results: Applied substances induced rapid and strong increase of plasma antioxidant enzymatic activity probably as a compensatory response to its pro-oxidant influence. Conclusion: The effects of Hcy on the activity of plasma antioxidant enzymes are in part mediated via interaction with gasotransmitters.

both in the prenatal and postnatal period2. The syndrome is like a big puzzle whose parts need to be carefully assembled to get a whole picture.2 Good multidisciplinary approach, proper communication and collaboration with parents and all physician’s involved are required in the diagnostic process. All available resources and tools are needed to increase diagnostic precision in the prenatal diagnosis of fetal syndromes.4