A randomized controlled trial of lycopene treatment on soluble receptor for advanced glycation end products in seminal and blood plasma of normospermic men.

PROBLEM: The aim of present study was to investigate the effects of antioxidant lycopene on soluble receptor for advanced glycation end products (sRAGE) levels in blood and seminal plasma in normospermic males. METHODS: Study included 15 fertile volunteers and 13 normospermic male partners from infertile relationships. The treatment was 12-week administration of 20 mg of lycopene or placebo followed by crossover and treatment for a further 12 weeks. The ELISA kit Quantikine(®) was used to determine sRAGE levels. RESULTS: Lycopene administration decreased sRAGE levels in seminal plasma in fertile volunteers (controls) as well as in male partners in the infertile relationships group (P=0.008 and P=0.012, respectively). No significant effect of lycopene on sRAGE in blood plasma was found in either group, but seminal plasma sRAGE was significantly suppressed. CONCLUSION: Lycopene decreased sRAGE in seminal, but not in blood plasma. This may be because of selective local uptake of lycopene in the male reproductive tract, namely in prostate. Decreased sRAGE may be caused by lycopene suppression of oxidative stressors and explain in part the putative improvement in fertility reported after lycopene treatment.

Comparison of chemiluminescence and flow cytometry in the estimation of reactive oxygen and nitrogen species in human semen.

OBJECTIVE: To determine the relative contribution of different cell types in washed sperm to the overall intracellular production of H(2)O(2) and peroxynitrite. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Thirty-one fertile volunteers and 166 men undergoing fertility assessment were included. INTERVENTION(S): Aliquots of sperm suspension in phosphate-buffered saline solution were used for the reactive oxygen species (ROS) detection by chemiluminescence and for the detection of H(2)O(2) and peroxynitrite by flow cytometry, with use of specific fluorescent probes, carboxy-2',7'-dichlorodihydrofluorescein diacetate dye for H(2)O(2) and dihydrorhodamine 123 for peroxynitrite. Gated analysis determined the relative contribution of spermatozoa, leukocytes, and "other round cells." MAIN OUTCOME MEASURE(S): Simultaneous estimates of global ROS production assessed by chemiluminescence assay compared with flow cytometric measurements. RESULT(S): The estimates of ROS with use of chemiluminescence positively correlated with the estimates of H(2)O(2) (r = 0.53) and peroxynitrite (r = 0.62) as assessed with flow cytometry. H(2)O(2) and peroxynitrite were measurable also in samples in which chemiluminescence did not detect measurable values. Increased production of H(2)O(2) by one cell type was associated with a relative increase in its peroxynitrite production. CONCLUSION(S): The levels of ROS production measured by chemiluminescence and flow cytometry were related. Each cell type in semen contributed differently to the global intracellular levels of H(2)O(2) and peroxynitrite.

The measurement of reactive oxygen species in human neat semen and in suspended spermatozoa: a comparison.

BACKGROUND: It is generally accepted that oxidative stress is an important factor in male infertility because it may impair the physiological function of spermatozoa at the molecular level. Nevertheless, although several approaches have been reported, the imbalance between production of reactive oxygen species (ROS) and activity of the antioxidant defense system in semen is difficult to investigate and remains poorly understood. METHODS: This study compares measurement of ROS production in neat semen and in washed spermatozoa obtained from the same ejaculate, and suspended in phosphate buffered saline using exactly the same luminol-mediated chemiluminescence method. Ninety one samples were obtained from males of infertile couples and 34 from volunteers with proven fertility. RESULTS: As expected, ROS levels were markedly lower in neat semen than in washed spermatozoa suspensions where seminal plasma with its potent antioxidant capacity was removed. In the cases of both neat semen and washed spermatozoa, ROS production was lowest in samples from normozoospermic males and highest in samples containing more than half million peroxidase-positive leukocytes per milliliter. For all samples, there was a significant positive correlation between ROS production by neat semen and that by washed spermatozoa suspension. CONCLUSION: Measurement of ROS production in neat semen better reflects actual oxidative status because it detects only the overproduction of ROS which are not effectively scavenged by antioxidant capacity of seminal fluid. The results of our study show a good commutability of both measurements for identification of semen samples with high ROS production. The measurement in neat semen is even less time consuming and therefore easier to implement into laboratory routine.

Reactive oxygen species in human semen in relation to leukocyte contamination.

AIMS: Excessive production of reactive oxygen species (ROS) in semen has been linked to male infertility. Main sources of ROS in male genital tract are immature and/or damaged spermatozoa and a subpopulation of leukocytes known as polymorphonuclear neutrophils (PMN). METHODS: Study group included male partners of infertile couples, 67 normospermic males (group B) and 98 males with sperm abnormalities in one or more parameters (group C), 36 fertile volunteers (group A) served as controls. Sperm parameters were determined according to WHO guidelines. The ROS production was measured by chemiluminiscence in sperm suspension in phosphate buffered saline. RESULTS: All fertile volunteers in the control group had seminal PMN concentrations below 0.5x10(6)/ml. Therefore study subjects, 67 normospermic and 98 men with sperm abnormalities, were further subdivided into two subgroups of PMN concentrations: (1) < 0.5x10(6)/ml and (2) 0.5 to 1.0x10(6)/ml. The ROS production in individuals varied greatly from 1.0x10(2) to 1.7 x10(7) RLU/min per 20x10(6) spermatozoa. The ROS production in both subgroups of normospermic men and the subgroup (1) of men with sperm abnormalities was not different from the ROS production in fertile controls. The ROS production in the subgroup (2) with sperm abnormalities was significantly higher than in controls (P = 0.00004). CONCLUSIONS: Our findings suggest that the contribution of PMN to the ROS production in semen is negligible only up to a concentration of 0.5x10(6)/ml. This suggests that the current WHO Guidelines threshold of 1.0x10(6) PMN per ml of semen is too high and might be re-evaluated.

Evaluation of day one embryo quality and IVF outcome--a comparison of two scoring systems.

BACKGROUND: The aim of our retrospective study was to compare the clinical usefulness of two non-invasive embryo scoring systems based either on a simplified pronuclear morphology of the zygote or on early cleavage rate, as well as their combination, for the selection of embryos with the best implantation potential in embryo transfer (ET). METHODS: Over a period of five years, the quality of 2708 embryos from 364 IVF cycles in women under the age of 39 years was assessed using these scoring systems in a university assisted reproduction centre. ET was always performed on day 3 of cultivation. The outcome of ETs of 702 embryos scored in the respective systems or their combination was retrospectively analyzed in terms of biochemical (bPR) and clinical pregnancy rates (cPR) and implantation rate (IR). Mann-Whitney U test and t-test for differences between relative values were used, p < 0.05 was considered statistically significant. RESULTS: There was no difference in outcome parameters in 109 cycles where only Pattern "0" zygotes, according to our simplified pronuclear morphology classification, were transferred and 140 cycles where only "other" pattern zygotes were transferred, regardless of their cleavage rate. On the contrary, significantly greater cPR and IR (p = 0.003 and p = 0.006, respectively) were achieved in 120 cycles where only early cleavage (EC) embryos were transferred compared with 152 cycles where only non early cleavage (NEC) embryos were transferred regardless of their pronuclear morphology. The best outcome in terms of cPR (56%) and IR (43%) was found in 50 cycles when Pattern "0" and EC embryos only were used for transfer. CONCLUSION: The results indicate that early cleavage is a better independent marker of implantation potential than zygote morphology. The best outcome can be achieved if both embryo scoring systems are used jointly and the embryo is classified as EC and Pattern "0".

[Bacteriospermia and the production of reactive oxygen species in the semen of males from infertile couples]. / Bacteriospermie a produkce of reaktivních kyslíkových cástic v ejakulátu muzu z neplodných páru.

OBJECTIVE: To estimate the incidence of bacteriospermia, the representation of specific micro-organisms and the generation of reactive oxygen species (ROS) in the semen of males from infertile couples in comparison with the semen of fertile volunteers. MATERIAL AND METHODS: Males from infertile couples were divided according to WHO criteria of their spermiogram results into those with normospermia (Group A, n = 65) and those with semen abnormalities (Group B, n = 116). The control group consisted of 44 fertile volunteers (Group C, n = 44). Aerobic culture was performed to ascertain the genus and species of the present microorganisms. ROS production was estimated by the chemiluminescence method. Statistical analysis was performed using the Mann-Whitney test. RESULTS: The incidence of positive cultures in the semen between Groups A, B and C (69 %, 74 % and 66 %, respectively) did not differ significantly, with Staphylococcus and Streptococcus species being most frequently identified. A significant difference was found only between the occurrence of microorganism Escherichia coli in the groups A, B and C (11.1 %, 9.3 % and 3.2 %). There were no statistically significant differences between ROS production in semen with positive or negative culture in any of the studied groups. CONCLUSIONS: In all the studied groups, the incidence of bacteriospermia as well as ROS production in the semen were similar. Staphylococcus and Streptococcus species were found in all groups. A significantly higher frequency of Escherichia coli in the semen samples of males from infertile couples in comparison with those from fertile volunteers was observed. No differences in ROS production in semen samples with positive and negative culture results were found in any of the studied groups.

Antioxidant capacity of seminal plasma measured by TAS Randox.

BACKGROUND: An imbalance between reactive oxygen species (ROS) generation in sperm and antioxidant capacity of seminal plasma has been linked to male infertility. The antioxidant power of biological fluids can be evaluated either by measurement of individual antioxidants or total antioxidant capacity (TAC). The aim of this study was to assess whether TAS Randox can also be used for seminal plasma antioxidant capacity estimation. MATERIAL AND METHODS: Freshly thawed aliquots of seminal plasma and blood serum of 38 males from infertile couples and 24 healthy normospermic controls were simultaneously assayed using TAS Randox reagents on the Hitachi Modular P800 Analyzer. Semen analysis was performed according to WHO guidelines. ROS in fresh sperm suspension in phosphate buffered saline was measured by chemiluminescence immediately after separation of seminal plasma. RESULTS: Semen analysis showed that in our study group only 14 males were normospermic and 24 males had mostly combined pathologies. The medians for ROS production were similar in both the study and the control groups (4850 and 5450 RLU/min, resp). Seminal plasma TAS levels were significantly lower (p<0.02) in the study group while blood serum TAS levels were similar in both groups. A significant positive correlation (p<0.05) between TAS in seminal plasma and serum was found, seminal plasma levels being on average 1.4 times higher. CONCLUSIONS: The TAS Randox kit may be used for clinical studies intended to identify decreased antioxidant power in the seminal fluid of infertile men.

Uterine rupture during pregnancy and delivery among women attending the Al-Tthawra Hospital in Sana'a City Yemen Republic.

BACKGROUND: About 20 percent of the population in developing countries is composed of women of reproductive age. These women face one of the catastrophic risks of pregnancy "uterine rupture". Studies conducted in the developing world give strong evidence that uterine rupture is a major health problem in these countries with the rate being high in rural areas. AIM: The purpose of the study was to estimate the incidence and determine the risk factors and outcome of uterine rupture among women using the referral hospital Al-thawra in Sana'a City, Yemen republic and to extrapolate the data to the whole of Yemen. METHODS: The data was collected retrospectively; by interviewing, examining and following up all the cases of uterine rupture coming to the hospital during a period of 9 months between September 1996 and May 1997. A descriptive analysis and distribution frequency of the commonest causes of uterine rupture in 37 cases are presented taking into account medical, reproductive, health services provided and sociodemographic factors. RESULTS: Incidence of uterine rupture in Yemen was found to be (0.63), obstructed labor 83 %, contracted pelvis 19 %, previous surgery in 48 %, Oxytocine infusion in 42 %. Grand-multiparty was in 65 % and maternal age over 35 years in 50 %. Antenatal care was only in 34 %. CONCLUSION: The high percentage of malpresentation, cephalopelvic disproportion, previous uterine surgery accompanied by the high percentage of use of Oxytocin in this study highlights very clearly the role of this medication in increasing the risk of uterine rupture in Yemen.

The occurrence of reactive oxygen species in the semen of males from infertile couples.

The aim of this pilot study was to establish a reactive oxygen species (ROS) evaluation method as a step in the routine diagnosis of men from infertile couples, which attend the Centre of Assisted Reproduction at the Teaching Hospital in Olomouc. Standard semen analyses were performed manually according to WHO guidelines. The number of peroxidase-positive leukocytes in the semen was determined using the Endtz test. The levels of ROS were estimated by chemiluminescence assay using luminol (5-amino-2,3 dihydro-1,4 phthalazinedione) as a probe. The semen samples were collected from 68 patients. Normospermia was found in 15 patients (22.1 %). The semen samples of 3 normospermic patients were classified as ROS-positive. Elevated ROS production was recorded in all subgroups of patients irrespective of any pathology found. We confirmed that spermatozoa might be the source of ROS as well as the seminal leukocytes. Apart from the leukocytes, sperm cells with residual cytoplasm and immature spermatozoa are considered to be a major source of ROS. Thus it is suggested that sperm morphology abnormalities should be evaluated more carefully.