Krunoslav Kuna

Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa

We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower “linearity” and “straightness”, and as showed by this research “elongation”, were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20), asthenozoospermia (n = 54) and oligoasthenozoospermia (n = 23). The findings indicate that sperm quality of patients with normal spermiogram diagnosed according to WHO criteria, may be compromised by showing premature spontaneous hyperactivation which can decrease the chances of natural conception. We assessed synergistic effects of multiple chemical elements in ejaculated semen to find if premature spontaneous hyperactivation of spermatozoa can be a sign of imbalanced semen composition especially of elements K, Ca, Cu and Zn. Human semen samples showing low or high baseline status of chemical elements concentrations were found in samples from all three diagnostic groups. However, correlation of K/Ca and Cu/Zn ratios, taking into account samples from all three groups of men, were negative at statistical significance level p = 0.01. We tested if the negative correlation between K/Ca and Cu/Zn ratio works for greater number of semen samples. We found the negative correlation to be valid for 175 semen samples at statistical significance of p = 0.00002. The ratio of K/Ca and Cu/Zn, i.e. increased concentrations of K and Zn in comparison to concentrations of Ca and Cu, were associated with a decrease of “straightness” in the group of men with normal spermiogram and pronounced spontaneous hyperactivation of spermatozoa, implying that these elements act in synergy and that the balance of elements and not their absolute concentrations plays the major role in premature spermatozoa hyperactivation in ejaculated semen.

Paraoxonase 1 activity and phenotype distribution in premenopausal and postmenopausal women

Introduction Postmenopausal women have higher risk of cardiovascular disease. One of the contributing factors could be reduced activity of anti-atherogenic enzyme paraoxonase 1 (PON1). The aim of this study was to examine differences in the lipid status, paraoxonase and arylesterase PON1 activities and PON1 phenotype in women with regular menstrual cycle and in postmenopausal women. Materials and methods: The study included 51 women in reproductive age (25 in follicular and 26 in luteal phase of the menstrual cycle) and 23 women in postmenopause. Lipid parameters in sera were determined using original reagents and according to manufacturer protocol. PON1 activity in serum was assessed by spectrophotometric method with substrates: paraoxon and phenylacetate. PON1 phenotype was determined by double substrate method. Results: Compared to the women in follicular and luteal phase, postmenopausal women have significantly higher concentration of triglyceride [0.9 (0.7–1.3), 0.7 (0.6–1.0) vs. 1.5 (0.9–1.7) mmol/L; P = 0.002], cholesterol [5.10 (4.78–6.10), 5.05 (4.70–5.40) vs. 6.30 (5.73–7.23) mmol/L; P < 0.001], LDL [3.00 (2.56–3.63), 3.00 (2.70–3.70) vs. 3.90 (3.23–4.50) mmol/L; P < 0.001], and apolipoprotein B [0.88 (0.75–1.00), 0.79 (0.68–1.00) vs. 1.07 (0.90–1.24) mmol/L; P = 0.002]. PON1 basal [104 (66–260), 106 (63–250) vs. 93 (71–165) U/L; P = 0.847] and salt-stimulated paraoxonase activity [210 (131–462), 211 (120–442) vs. 180 (139–296) U/L; P = 0.857] as well as arylesterase activity [74 (63–82), 70 (54–91) vs. 70 (60–81) kU/L; P = 0.906] and PON1 phenotype (P = 0.810) were not different in the study groups. Conclusion: There are no differences in PON1 activity and PON1 phenotype between women with regular menstrual cycle and postmenopausal women.

Endometrioza trbušne stijenke jedanaest godina nakon carskog reza: prikaz slučaja

Endometriosis is a common chronic disease characterized by growth of the endometrial gland and stroma outside the uterus. Symptoms affect physical, mental and social well-being. Extrapelvic location of endometriosis is very rare. Abdominal wall endometriosis occurs in 0.03%-2% of women with a previous cesarean section or other abdominopelvic operation. The leading symptoms are abdominal nodular mass, pain and cyclic symptomatology. The number of cesarean sections is increasing and so is the incidence of abdominal wall endometriosis as a potential complication of the procedure. There are cases of malignant transformation of abdominal wall endometriosis. Therefore, it is important to recognize this condition and treat it surgically. We report a case of a 37-year-old woman with abdominal wall endometriosis 11 years after cesarean section. She had low abdominal pain related to menstrual cycle, which intensified at the end of menstrual bleeding. A nodule painful to palpation was found in the medial part of previous Pfannenstiel incision. Ultrasound guided biopsy was performed and the diagnosis of endometriosis confirmed. Surgery is the treatment of choice for abdominal wall endometriosis. Excision with histologically proven free surgical margins of 1 cm is mandatory to prevent recurrence. A wide spectrum of mimicking conditions is the main reason for late diagnosis and treatment of abdominal wall endometriosis. In our case, the symptoms lasted for eight years and had intensified in the last six months prior to surgery.

PP-016 - Croatian IVF legislation and the effect of restrictions on IVF outcome

The in vitro fertilization (IVF) legislation can directly affect the success of IVF treatment. The illustration and support for these statements can be found in the Italian restrictive law (Law 40/2004) and the effect of its implementation. The aim of this study was to compare data on assisted reproduction treatment and outcome before and after the introduction of this new legislation and give answer to a question on the possible negative effects of restrictive IVF regulation using data from Croatia. Data were analysed retrospectively: the first group consisted of data from patients which underwent IVF/ICSI procedure during period of restrictions (group I) and the second one of data from patients which underwent IVF/ICSI procedure after the new law was enforced (group II).In both groups there were more antagonist cycles (71.9%) than agonist cycles (28.1%) ; but after the new law was enforced there were more antagonist cycles (81.4% for group II compared to 59.5% for group I). Regarding ICSI and IVF procedure ; for ICSI 68.7% in group I vs. 67.4% in group II, and for IVF cycles 31.3% in group I vs. 32.6% in group II. Ratio of ICSI and IVF was similar (χ²=0.598, df=1, p >.01) although we expected more ICSI cycles in group I. During restrictions (in group I, 49.2%) usually three embryos were implemented but after the enforcement of the new law (group II, 63.4%) mostly two embryos were implemented (χ²=77.70, df=2, p .01) leaving us with a conclusion that the pregnancy rate around 30% was achieved during enforcement of both laws. These results show that IVF regulation affects the treatment and outcomes in ART, and that for some issues in ART restrictive regulations has negative effects. Our findings represent clinical contribution for countries still debating the enforcement of restrictions in ART legislation.

Prevention and Treatment of Ovarian Hyperstimulation Syndrome

Ovarian hyperstimulation syndrome (OHSS) is a clinical symptom complex associated with ovarian enlargement resulting from exogenous gonadotropin therapy. In severe cases, a critical condition develops with massive ascites, marked ovarian enlargement, pleural effusion, electrolyte imbalance, hypovolemia with hypotension, oliguria, hemoconcentration, and thromboembolism (Madill JJ et al., 2008). Moderate to severe ovarian hyperstimulation syndrome (OHSS) has been calculated to occur in 0.2% to 2% of all ovarian stimulation cycles (Binder H et al., 2007).

Sperm Cell in ART

Infertility today represents a global problem. Male factor contributes in approximately 50% of infertile couples. In the last decade we are witnesses of the decreased quality of semen and the increased frequency of testicular cancer and cryptorchidism. Currently, the assessment of semen quality is based on the routine semen analysis including sperm count, morphology and motility. Although variation and combination among these three main factors articulate few diagnosis, nowadays developed assisted reproduction techniques (ART), especially intracytoplasmatic sperm injection (ICSI) may be used to treat most of the male infertility problems. In general we can say that traditional semen parameters provide a limited degree of diagnostic information, thus we are aware that these indexes of diagnosis should be revisited, which includes more specific test of sperm assessments, such as DNA tests and sperm proteome. Spermatogenesis is a process that includes physiological, morphological and biochemical changes. After a complex process from the round diploid spermatogonia to haploid spermatozoa, a mature sperm just has an ability to fertilize a mature oocyte. If any errors occur during spermatogenesis process, appropriate sperm will not be produced. Thus, in our ART practice, it is important to understand normal physiology of spermatogenesis and find the reason of abnormal situation.