Eliana Garalejic

Is Manganese (II) Pentaazamacrocyclic Superoxide Dismutase Mimic Beneficial for Human Sperm Mitochondria Function and Motility

Mitochondria play an important role in sperm cell maturation and function. Here, we examined whether (and how) changes in sperm redox milieu affect the functional status of sperm mitochondria, that is, sperm functionality. Compared with the control, incubation in Tyrodes medium for 3 h, under noncapacitating conditions, decreased sperm motility, the amount of nitric oxide ((•)NO), the number of MitoTracker(®) Green FM (MT-G) positive mitochondria, and the expression of complexes I and IV of the mitochondrial respiratory chain. In turn, superoxide dismutase (SOD) mimic (M40403) treatment restored/increased these parameters, as well as the expression of endothelial nitric oxide synthase, manganese SOD, and catalase. These data lead to the hypothesis that M40403 improves mitochondrial functional state and motility of spermatozoa, as well as (•)NO might be involved in the observed effects of the mimic.

Correlation between Sperm Parameters and Protein Expression of Antioxidative Defense Enzymes in Seminal Plasma: A Pilot Study

Background. Semen analysis is the cornerstone in the evaluation of male (in)fertility. However, there are men with normal semen tests but with impaired fertilizing ability, as well as fertile men with poor sperm characteristics. Thus, there is rising interest to find novel parameters that will help to predict and define the functional capacity of spermatozoa. Methods. We examined whether there is a correlation between semen parameters (count, progressive motility, and morphology) and protein expression/activity of antioxidative defense enzymes in seminal plasma from 10 normospermic subjects. Results. Sperm progressive motility was in positive correlation with seminal plasma protein expression of both superoxide dismutase (SOD) isoforms (MnSOD and CuZnSOD) and catalase. Also, positive correlation was observed between sperm count and MnSOD protein expression, as well as between sperm morphology and protein expression of catalase in seminal plasma. In contrast, protein expression of glutathione peroxidase was not in correlation with any sperm parameter, while its activity negatively correlated with sperm morphology and motility. Conclusions. These data suggest that evaluation of protein expression of antioxidative defense enzymes in seminal plasma might be of importance in the evaluation of male fertility status and that could be used as an additional biomarker along with classic semen analysis in assessment of semen quality.

Licheniocin 50.2 and Bacteriocins from Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 Inhibit Biofilms of Coagulase Negative Staphylococci and Listeria monocytogenes Clinical Isolates.

Background Coagulase negative staphylococci (CoNS) and Listeria monocytogenes have important roles in pathogenesis of various genital tract infections and fatal foetomaternal infections, respectively. The aim of our study was to investigate the inhibitory effects of two novel bacteriocins on biofilms of CoNS and L. monocytogenes genital isolates. Methods The effects of licheniocin 50.2 from Bacillus licheniformis VPS50.2 and crude extract of bacteriocins produced by Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 (BGBU1-4 crude extract) were evaluated on biofilm formation and formed biofilms of eight CoNS (four S. epidermidis, two S. hominis, one S. lugdunensis and one S. haemolyticus) and 12 L. monocytogenes genital isolates. Results Licheniocin 50.2 and BGBU1-4 crude extract inhibited the growth of both CoNS and L. monocytogenes isolates, with MIC values in the range between 200–400 AU/ml for licheniocin 50.2 and 400–3200 AU/ml for BGBU1-4 crude extract. Subinhibitory concentrations (1/2 × and 1/4 × MIC) of licheniocin 50.2 inhibited biofilm formation by all CoNS isolates (p < 0.05, respectively), while BGBU1-4 crude extract inhibited biofilm formation by all L. monocytogenes isolates (p < 0.01 and p < 0.05, respectively). Both bacteriocins in concentrations of 100 AU/mL and 200 AU/mL reduced the amount of 24 h old CoNS and L. monocytogenes biofilms (p < 0.05, p < 0.01, p < 0.001). Conclusions This study suggests that novel bacteriocins have potential to be used for genital application, to prevent biofilm formation and/or to eradicate formed biofilms, and consequently reduce genital and neonatal infections by CoNS and L. monocytogenes.

Sensitivity and specificity of ultrasonography as a screening tool for gestational diabetes mellitus

Objective: To evaluate diagnostic accuracy and efficiency of ultrasound markers of gestational diabetes (GDM) and propose an ultrasound based scoring system suitable for screening (UGDS). Methods: 110 women with singleton pregnancies and established maternal and/or pregnancy related risk factors for GDM were scanned at/or after 24 weeks gestation followed by administration of a 3 hour 100-gram oral glucose tolerance test (oGTT). A number of ultrasound markers were determined/measured, including fetal adipose subcutaneous tissue, asymmetrical macrosomy, cardiac circumference, cardiac width, and interventricular septum thickness, immature appearance of placenta, intensified breathing movements, polyhydramnios and placental thickness. Each ultrasound GDM marker was assigned one point to create the ultrasound gestational diabetes screening score (UGDS). Results: All ultrasound GDM markers were positively correlated to the disease P < 0.0001. The strongest independent predictor of GDM was an immature appearance of placenta (RR 40.1 95% CI 5.9–271.0, P < 0.0001). Receiver operator characteristics (ROC) showed an area under the curve of 95.7% confirming good ability of UGDS to discriminate between positive and negative oGTT. We propose a UGDS score of 4 providing sensitivity of 90.9%, and specificity 89.6%. Conclusions: This study suggests that UGDS is a good predictor of GDM.

Antimicrobial Activity of Human Follicular Fluids

The aim of this study was to explore the antimicrobial activity of human follicular fluid (HFF), to test the hypothesis that different strains of the same bacterial species could display different patterns of susceptibility to antimicrobial action of HFF, and to preliminarily investigate the possible mechanism of antimicrobial action of this fluid. Antimicrobial activity of 60 samples of HFF toward 30 Streptococcus agalactiae strains was determined by the agar diffusion method and broth dilution method. To explore the mechanism of antimicrobial activity, biochemical analyses were performed with selected fluid samples. The obtained results indicate that 38.3% fluid samples did not inhibit bacterial growth, 53.3% showed moderate and 8.3% high antimicrobial activity. The tested effect of HFF on S. agalactiae strains was bactericidal and was not strain dependent. Lysozyme activity was detected in HFF exhibiting antimicrobial activity. There were no statistically significant differences in concentrations of estradiol, progesterone, transferrin, iron, total protein and albumin levels among tested samples regardless of the different rate of antimicrobial activity. The obtained results indicate that lysozyme is most probably a crucial antibacterial agent in this fluid; however, some other still unidentified factors may contribute to it.

Diagnosis of bacterial vaginosis

Bacterial vaginosis is a common, complex clinical syndrome characterized by alterations in the normal vaginal flora. When symptomatic, it is associated with a malodorous vaginal discharge and on occasion vaginal burning or itching. Under normal conditions, lactobacilli constitute 95% of the bacteria in the vagina. Bacterial vaginosis is associated with severe reduction or absence of the normal H2O2-producing lactobacilli and overgrowth of anaerobic bacteria and Gardnerella vaginalis, Atopobium vaginae, Mycoplasma hominis and Mobiluncus species. Most types of infectious disease are diagnosed by culture, by isolating an antigen or RNA/DNA from the microbe, or by serodiagnosis to determine the presence of antibodies to the microbe. Therefore, demonstration of the presence of an infectious agent is often a necessary criterion for the diagnosis of the disease. This is not the case for bacterial vaginosis, since the ultimate cause of the disease is not yet known. There are a variety of methods for the diagnosis of bacterial vaginosis but no method can at present be regarded as the best. Diagnosing bacterial vaginosis has long been based on the clinical criteria of Amsel, whereby three of four defined criteria must be satisfied. Nugents scoring system has been further developed and includes validation of the categories of observable bacteria structures. Up-to-date molecular tests are introduced, and better understanding of vaginal microbiome, a clear definition for bacterial vaginosis, and short-term and long-term fluctuations in vaginal microflora will help to better define molecular tests within the broader clinical context.

Results of modern tuboperitoneal infertility treatment

BACKGROUND/AIM A modem approach to surgical treatment of tuboperitoneal infertility is based on laporascopic techniques. The aim of this study was to compare results of tuboperitoneal infertility treatment by the use of laparoscopy and classical laparotomy. METHODS A retrospective-prospective study on 66 women treated operatively for tuboperitoneal infertility was performed. Data from patients anemnesis and those related to the surgical treatment results, obtained by the use of an inquiry, were used in retrospective and prospective analysis, respectively. Chi-square test was used in statistical analysis. P value < 0.05 was considered significant. RESULTS Classical laparotomy was used on 34 women in a period from 1996 to 1997, while 32 women were operated laparoscopically in a period from 1999 to 2000. The results were as follows: a total number of conceived women was 16 (24%), seven in the group I (20.6%) and nine in the group II (28.1%); 13 women were with one pregnancy, six in the group I (17.6%) and seven in the group II (22%). Twice pregnant were three women, one in the group I (2.9%) and two in the group II (6.2%). The resulting pregnancies were: five women with abortion spontaneous, two in the group I (5.9%) and three in the group II (9.40/%); two women with extrauterine pregnancy in the group I (5.9%); three with pretemporal birth, one in the group I (2.9%) and two in the group II (6.2%), while six women were with the temporal birth, two in the group I (5.9%) and four in the group II (12.5%). Statistical analysis showed that there was no significant difference in the results between these two groups. CONCLUSION Surgical treatment of tubeperitoneal infertility, regardless of the used methods (classical laparotomy or laparoscopy) was successful in a great number of women. These methods have a great advantage over in vitro fertilization, and they should not be ignored.

Level of NO/nitrite and 3-nitrotyrosine in seminal plasma of infertile men: Correlation with sperm number, motility and morphology

Nitric oxide (NO) is a signaling molecule responsible for initiation of molecular events that influence sperm functionality in a concentration-dependent manner. It is still not fully understood how seminal plasma NO contributes to sperm pathologies. The aim of this study was to elucidate whether and how NO is implicated in etiology of different sperm abnormalities. To this end we determine NO, nitrite and 3-nitrotyrosine (3-NT) content in seminal plasma of infertile men with specific pathologies (terato-, oligoterato- and oligoasthenoteratospermia) and relate it to infertile normospermic samples. To gain further understanding of NO metabolism in seminal plasma we determine protein expression of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS). Here we show that NO, nitrite and 3-NT levels in seminal plasma of men with suboptimal semen parameters are significantly lower compared to normospermic men. An increase in protein expression of eNOS and no change in protein expression of iNOS is observed in men with sperm pathologies. Association of seminal plasma 3-NT level with functional sperm parameters is observed - positive correlation with sperm count, motility and morphology in normospermia, teratospermia and oligoteratospermia, as well as negative correlation with sperm morphology and motility in oligoasthenoteratospermia. Present study revealed that suboptimal seminal plasma NO content is found in all examined sperm pathologies. This result unequivocally shows the importance of NO for sperm function and involvement of suboptimal NO level in etiology of sperm abnormalities. Lower seminal plasma 3-NT level and its significant association with sperm parameters, found in pathologies, strongly indicates that protein nitration is important for spermatozoa function and that failure to establish this post-translational protein modification might be involved in etiology of sperm abnormalities. According to our results, NO measurement can discriminate infertile men with sperm pathologies from infertile normospermic men but is not indicative of a specific type of sperm pathology.

Relationship between mid-trimester ultrasound fetal liver length measurements and gestational diabetes mellitus 妊娠中期三个月超声测量的胎儿肝脏长度与妊娠期糖尿病之间的关系

The aim of the present study was to investigate the relationship between mid‐trimester ultrasound fetal liver length (FLL) and gestational diabetes mellitus (GDM) in a high‐risk population.

Relationship between mid-trimester ultrasound fetal liver length measurements and gestational diabetes mellitus 妊娠中期三个月超声测量的胎儿肝脏长度与妊娠期糖尿病之间的关系: Fetal liver length predicts GDM

The aim of the present study was to investigate the relationship between mid‐trimester ultrasound fetal liver length (FLL) and gestational diabetes mellitus (GDM) in a high‐risk population.