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1.
Reprod Biol Endocrinol ; 19(1): 156, 2021 Oct 09.
Article in English | MEDLINE | ID: mdl-34627284

ABSTRACT

BACKGROUND: Human papillomavirus (HPV) has been shown to adversely affect human reproduction. We aimed to evaluate the prevalence of human papillomavirus (HPV) infection in men and its correlation with semen parameters and reproductive outcomes. METHODS: Semen samples and penile swabs were collected from potential sperm donors (SD, n = 97) and male partners of infertile couples (IM, n = 328). The presence of HPV DNA in semen samples and penile swabs was analyzed. Associations between hrHPV positive status and fertility outcomes as well as socio-behavioral and health characteristics were evaluated using the R software package. RESULTS: High-risk HPV (hrHPV) genotypes were detected in 28.9% of SD and 35.1% of IM (P = 0.312). Penile swabs were more frequently positive for hrHPV genotypes than semen samples in both IM (32.3% vs. 11.9%, P < 0.001) and SD (26.8% vs. 6.2%, P = 0.006). Men with hrHPV positive semen samples had lower semen volume (median volume 2.5 ml vs. 3 ml, P = 0.009), sperm concentration (median concentration 16 × 106/ml vs. 31 × 106/ml, P = 0.009) and total sperm count (median count 46 × 106 vs. 82 × 106, P = 0.009) than men with hrHPV negative samples. No association was identified between penile hrHPV status and semen parameters. CONCLUSIONS: Our findings indicate that penile HPV infection is common in both potential sperm donors and men from infertile couples. Although HPV positivity is higher in penile swabs, only HPV infection in semen samples affects sperm parameters. However, there was no association between hrHPV positivity in semen and fertility outcomes including abortion rate.


Subject(s)
Infertility/complications , Infertility/diagnosis , Papillomavirus Infections/complications , Adult , Czech Republic/epidemiology , Family Characteristics , Female , Fertilization in Vitro/statistics & numerical data , Humans , Infertility/epidemiology , Male , Middle Aged , Papillomaviridae/physiology , Papillomavirus Infections/diagnosis , Papillomavirus Infections/epidemiology , Pregnancy , Pregnancy Outcome/epidemiology , Prognosis , Semen/physiology , Semen/virology , Semen Analysis , Tissue Donors/statistics & numerical data , Treatment Outcome , Young Adult
2.
Virol J ; 18(1): 80, 2021 04 15.
Article in English | MEDLINE | ID: mdl-33858457

ABSTRACT

BACKGROUND: Data about the genotype-specific human papillomavirus (HPV) prevalence in the Czech Republic is limited. We aimed to evaluate the prevalence and concordance of genotype-specific HPV infection detected in semen samples, penile swabs and cervical swabs from non-vaccinated heterosexual couples without HPV-associated disease. METHODS: Semen samples and penile swabs were collected from male partners and cervical swabs were collected from female partners of heterosexual couples treated for infertility (n = 195). Presence of HPV DNA in semen samples and cervical swabs was analyzed using the cobas® HPV Test and PapilloCheck®. Only the PapilloCheck® test was used to detect HPV in penile swabs. The genotype-specific prevalence and concordance of HPV infection not targeted by vaccine were evaluated using Fisher exact test. RESULTS: Both partners were infected with any HPV type in 13.8% (27/195) of couples and, of these couples, 55.6% (15/27) harbored at least one mutual genotype. High-risk HPV (hrHPV) genotypes were detected in 12.3% (24/195) of semen samples, 31.3% (61/195) of penile swabs, and 19.5% (38/195) of cervical swabs (P < 0.001). The most prevalent hrHPV genotype were HPV53 (2.56%; 5/195) in semen samples, HPV16 (6.67%, 13/195) in penile swabs and HPV39 (3.59%, 7/195) in cervical swabs. Low-risk (lrHPV) genotypes were detected in 5.13% (10/195) of semen samples, 15.9% (31/195) of penile swabs, and 4.10% (8/195) of cervical swabs (P < 0.001). Male sexual partners of HPV-positive women were more likely to be infected with at least one of the same HPV types than female sexual partners of HPV-positive men (34.9% vs. 17.9%, P = 0.055). CONCLUSIONS: This study showed that the detection of HPV infection differ by anatomic site and gender. Regardless the anatomic site, high prevalence of HPV genital infection was found in both Czech men and women.


Subject(s)
Alphapapillomavirus , Papillomavirus Infections , Alphapapillomavirus/genetics , Czech Republic/epidemiology , Female , Genotype , Heterosexuality , Humans , Male , Papillomavirus Infections/epidemiology , Prevalence
3.
Article in English | MEDLINE | ID: mdl-31517300

ABSTRACT

OBJECTIVE: The aims of this study were to determine the prevalence of human papillomavirus (HPV) infection in women treated for infertility and oocyte donors, and to investigate the possible influence of HPV infection on reproductive outcomes. STUDY DESIGN: In this observational laboratory-based study, cervical swabs were collected from oocyte donors (n = 207), and women treated for infertility (n = 945) and analysed for the presence of high-risk HPV (hrHPV) genotypes using the cobas® 4800 HPV Test and PapilloCheck® HPV-Screening. Associations between hrHPV positive status and fertility outcome or socio-behavioral and health characteristics were evaluated using R statistical software. RESULTS: HrHPV prevalence was significantly higher in oocyte donors than in women treated for infertility (28.0% vs. 16.1%, P < 0.001). Women who became pregnant spontaneously (19.6%) and women not treated with in vitro fertilization (IVF, 18.1%) were more frequently hrHPV positive than women treated with IVF (12.7%, P = 0.077). Despite the high prevalence of hrHPV in both oocyte donors and infertile women, no associations between hrHPV positive status and pregnancy or abortion rates were found in IVF treated women or in oocyte recipients. Moreover, no associations between hrHPV positive status and abortion rates were found in spontaneously pregnant women. CONCLUSION: Despite the high prevalence of hrHPV in both oocyte donors and infertile women, HPV infection did not influence the outcomes of assisted reproductive technology.

4.
J Mol Diagn ; 20(6): 849-858, 2018 11.
Article in English | MEDLINE | ID: mdl-30165205

ABSTRACT

High-risk human papillomavirus (hrHPV) infection is a cause of cervical cancer development. The addition of hrHPV testing to cervical cancer screening and monitoring of cervical intraepithelial neoplasia treatment improves the efficacy of screening and treatment, respectively. Self-sampling for hrHPV testing seems a promising tool for increasing patient participation in cervical cancer screening. In this project, 1198 cervical swabs obtained by physicians and 176 cervicovaginal swabs obtained by self-sampling (not collected in parallel) were analyzed for the presence of 14 hrHPV genotypes using three commercially available assays in comparison. HPV DNA was detected in 21.2% of all samples (21% of cervical swabs and 22.7% of cervicovaginal swabs). The cobas 4800 HPV Test was the most sensitive (0.983) and specific (0.992) for hrHPV detection overall. The PapilloCheck HPV-Screening and LMNX Genotyping Kit HPV GP had comparable specificity with that of the cobas (0.989 and 0.955, respectively), but lesser sensitivity (0.897 and 0.909, respectively). In physician-obtained cervical swabs, the cobas showed the highest sensitivity and specificity (0.980 and 0.994, respectively) for hrHPV detection, whereas in cervicovaginal swabs, the cobas had the highest sensitivity (1.00), but the PapilloCheck had the highest specificity (0.993). In conclusion, all of the detection methods evaluated were highly sensitive and specific for hrHPV detection from both clinician-collected cervical swabs and self-sampled cervicovaginal swabs.


Subject(s)
DNA, Viral/analysis , Genotyping Techniques/methods , Papillomaviridae/genetics , Papillomavirus Infections/diagnosis , Papillomavirus Infections/virology , Reagent Kits, Diagnostic , Uterine Cervical Neoplasms/virology , Vagina/virology , Adolescent , Adult , Aged , DNA, Viral/genetics , Female , Humans , Middle Aged , Sensitivity and Specificity , Young Adult
5.
Article in English | MEDLINE | ID: mdl-29213142

ABSTRACT

BACKGROUND: In recent decades gamete donation has received growing attention. Data from the Czech National Registry of Assisted Reproduction show that the number of cycles using donated oocytes has been increasing every year. According to Czech law, gamete donation is anonymous. Since 2011, some members of the Czech parliament have repeatedly made requests to revoke the anonymity but anonymity is one of the preconditions for such donation in this country. The aim of this study was to find out how the gamete recipients feel towards informing their child about the circumstances of their conception and their access to the identity of the donor. METHODS: A total of 195 recipients (122 women undergoing treatment - 43 Czechs, 79 foreigners (Western Europe and the USA) and 73 male partners - 28 Czechs, 45 foreigners) participated in this survey. The data were obtained by anonymous questionnaire. RESULTS: A significant difference between the attitude of the future Czech and foreign parents regarding disclosing the mode of conception was found (P = 0.003). The vast majority of Czechs were against disclosure. The foreign recipients were somewhat more divided. Regarding the donor's identity, there was no difference in atttitude between the groups. Recipients rarely consider that the knowledge of the donor's identity will be important for their child. The recipients overall, were convinced that the psychological aspects of parenting are far more important to the child than genetics, and see no reasons for disclosing the donor´s identity. CONCLUSION: While the the foreign recipients were less adamant about non-disclosure, the overall finding was in accord with the current Czech law on anonymity and not in agreement with the proposed abolition. The recipient's attitudes towards disclosing were also culturally determined. The fact that some countries have revised their rules towards open idendity is not a rationale for such change in the Czech Republic.


Subject(s)
Directed Tissue Donation/legislation & jurisprudence , Disclosure/legislation & jurisprudence , Fertilization in Vitro , Guideline Adherence/legislation & jurisprudence , Tissue Donors/psychology , Tissue and Organ Procurement/legislation & jurisprudence , Adult , Child , Czech Republic , Family Characteristics , Female , Fertilization in Vitro/legislation & jurisprudence , Fertilization in Vitro/psychology , Humans , Information Seeking Behavior , Male , Parent-Child Relations , Pedigree , Pregnancy , Tissue Donors/legislation & jurisprudence
6.
J Nat Prod ; 79(4): 812-20, 2016 Apr 22.
Article in English | MEDLINE | ID: mdl-27015547

ABSTRACT

The effects in vitro of 2,3-dehydrosilybin and several galloyl esters and methyl ethers on the viability, proliferation, and migration of human umbilical vein endothelial cells (HUVECs) were evaluated. The monogalloyl esters were synthesized by a chemoselective esterification method or by Steglich esterification of suitably protected 2,3-dehydrosilybin (1) with protected gallic acid. 2,3-Dehydrosilybin (1) displayed more potent cytotoxic, antiproliferative, and antimigratory activities (IC50 12.0, 5.4, and 12.2 µM, respectively) than silybin. The methylated derivatives were less active, with the least potent being 3,7-di-O-methyl-2,3-dehydrosilybin (6). On the other hand, galloylation at C-7 OH and C-23 OH markedly increased the cytotoxicity and the effects on the proliferation and migration of HUVECs. The most active derivative was 7-O-galloyl-2,3-dehydrosilybin (13; IC50 value of 3.4, 1.6, and 4.7 µM in the cytotoxicity, inhibition of proliferation, and antimigratory assays, respectively). Overall, this preliminary structure-activity relationship study demonstrated the importance of a 2,3-double bond, a C-7 OH group, and a galloyl moiety in enhancing the activity of flavonolignans toward HUVECs.


Subject(s)
Silymarin/pharmacology , Cell Survival/drug effects , Free Radical Scavengers/chemistry , Gallic Acid/pharmacology , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Methyl Ethers/pharmacology , Molecular Structure , Silybin , Silymarin/chemistry , Structure-Activity Relationship
7.
Article in English | MEDLINE | ID: mdl-23446211

ABSTRACT

AIMS: Male infertility has been associated with imbalance in the polyunsaturated fatty acids arachidonic acid/docosahexaenoic acid (AA/DHA) ratio. This study aimed to assess whether use of the potent antioxidant, lycopene, would affect this imbalance in seminal plasma and to test whether modulation of the AA/DHA ratio correlates with male fertility. METHODS: The study included 44 males from infertile couples who were treated with lycopene for three months before scheduled IVF treatment. The control group was 13 fertile males who were also treated with lycopene for three months. AA and DHA levels were measured in seminal plasma before and after treatment. The outcome of IVF treatment after lycopene use in of males from infertile couples was recorded. RESULTS: The subjects were subdivided into 21 normozoospermic males from couples with idiopathic infertility and 23 males with semen abnormalities. Prior to the treatment with lycopene, the AA/DHA ratios in both subgroups of patients were significantly higher than in fertile controls and improved following treatment with lycopene. Improvement was more marked in the normospermic males. Seven spontaneous pregnancies (16%) occurred before the scheduled IVF treatment and 15 couples (42%) achieved pregnancy after IVF. CONCLUSIONS: Three months of treatment with lycopene led to a significant improvement in the AA/DHA ratio in seminal plasma of males from infertile couples and facilitated the spontaneous as well as IVF conception.


Subject(s)
Arachidonic Acid/metabolism , Carotenoids/therapeutic use , Docosahexaenoic Acids/metabolism , Infertility, Male/drug therapy , Oxidative Stress/drug effects , Semen/metabolism , Adult , Female , Humans , Infertility, Male/metabolism , Lycopene , Male , Pregnancy , Protozoan Proteins/metabolism , Semen/drug effects , Transcription Factors/metabolism , Treatment Outcome
8.
BMC Med Educ ; 14: 256, 2014 Nov 28.
Article in English | MEDLINE | ID: mdl-25430069

ABSTRACT

BACKGROUND: Psychological distress among medical students is commonly observed during medical education and is generally related to poor academic self-perception. We evaluated the role of peer social support at medical schools in the association between psychological distress and academic self-perception. METHODS: An online survey was conducted in a medical degree program for 138 international students educated in English in the Czech Republic. The Medical Student Well-Being Index was used to define the students' psychological distress. Perceived peer social support was investigated with the Multidimensional Scale of Perceived Social Support. Poor academic self-perception was defined as the lowest 30% of a subscale score of the Dundee Ready Education Environment Measure. Analyses evaluated the presence of additive interactions between psychological distress and peer social support on poor academic self-perception, adjusted for possible confounders. RESULTS: Both psychological distress and low peer social support were negatively associated with poor academic self-perception, adjusted for local language proficiency and social support from family. Students with psychological distress and low peer social support had an odds ratio of 11.0 (95% confidence interval (CI): 2.1-56.6) for poor academic self-perception as compared with those without distress who had high peer social support. The presence of an additive interaction was confirmed in that the joint association was four times as large as what would have been expected to be on summing the individual risks of psychological distress and low peer social support (synergy index = 4.5, 95% CI: 1.3-14.9). CONCLUSIONS: Psychological distress and low peer social support may synergistically increase the probability of poor academic self-perception among international medical students. Promoting peer social relationships at medical school may interrupt the vicious cycle of psychological distress and poor academic performance.


Subject(s)
Clinical Competence , Peer Group , Self Concept , Social Support , Stress, Psychological/epidemiology , Students, Medical/psychology , Adult , Confidence Intervals , Cross-Sectional Studies , Czech Republic , Education, Medical, Undergraduate/methods , Female , Humans , Internationality , Male , Multilingualism , Odds Ratio , Risk Assessment , Schools, Medical , Stress, Psychological/diagnosis , Young Adult
9.
Article in English | MEDLINE | ID: mdl-23446209

ABSTRACT

AIMS: Caesarean section is the most common obstetric operation associated with short and long term risks, one of which is uterine scar dehiscence. In this case report we describe four cases of in vitro fertilization and embryo transfer (IVF+ET) treatment where the embryo was transferred into the uterus with known scar dehiscence in the lower uterine segment after a previous Caesarean section (SC). METHODS: All transfers of embryos were ultrasound guided directly into the middle of uterine cavity. All resulting pregnancies continued without problems related to the dehiscent scar and babies were delivered in the third trimester by elective/emergency SC. RESULTS: Our cases suggest that IVF+ET can be offered as an infertility treatment option despite a dehiscent scar in the lower uterine segment after previous SC.


Subject(s)
Cesarean Section/adverse effects , Cicatrix/complications , Embryo Transfer/adverse effects , Fertilization in Vitro/adverse effects , Infertility/therapy , Surgical Wound Dehiscence/etiology , Uterus/pathology , Adult , Female , Humans , Pregnancy , Ultrasonography , Uterus/diagnostic imaging
10.
Article in English | MEDLINE | ID: mdl-24108198

ABSTRACT

AIM: The aim of this prospective study was to find possible relationship between ROS production measured by chemiluminescence and flow cytometry in human semen and sperm DNA damage estimated by Sperm Chromatin Structure Assay. METHODS: Study included 39 men from infertile couples and 23 fertile volunteers who served as a control group. Aliquot of neat semen was used for ROS detection by chemiluminescence. Aliquot of sperm suspension in phosphate buffered saline was used for the detection of ROS by flow cytometry. Another aliquot of sperm suspension was used for SCSA to measure DNA fragmentation index and High DNA stainability. RESULTS: DNA fragmentation index correlated negatively with sperm morphology and motility. High DNA stainability correlated positively with ROS production and negatively with sperm morphology and concentration. Although there were similar trends of rising DNA fragmentation index and ROS production among the three groups of men, the relationship did not reach statistical significance. CONCLUSIONS: Higher values of DNA fragmentation index and high DNA stainability may also reflect developmental and/or environmental problems and not only oxidative stress.


Subject(s)
DNA Damage , Reactive Oxygen Species/metabolism , Semen/metabolism , Spermatozoa , Adult , Chromatin , Genetic Techniques , Humans , Male , Prospective Studies
11.
Article in English | MEDLINE | ID: mdl-22580863

ABSTRACT

INTRODUCTION: Functional 3D training originating in rehabilitation and physiotherapy routines, is modelled on the basis of a patient's functional impairment and muscle function test results. 3-D functional rehabilitation comprises two key components. The first is three-dimensional movements of the body and the second is restoration of function, The aim of 3D training is not only to improve physical fitness and muscular coordination, it is recommended for all age groups regardless of health status. The aim of this review is to highlight the main principles, roles and methods of 3D functional training from a clinically-anatomical perspective and its application in clinical practice. Second, we present a case study of 3d training in an obese boy. METHODS: Database search using the lemmatization method and data bases: Web of Knowledge, Web of Science, UpToDate, PubMed and Google Scholar for the years 2005-2011. RESULTS AND CONCLUSIONS: Functional 3D training is an effective method for, among others, strengthening the postural muscles of the human body, increasing articular stability, strengthening the ligaments and stability of muscle groups,especially the back muscles and for improving lung capacity. The method of functional training is of considerable significance in activating the pelvic floor muscles and the pelvic-trochanteric muscles for gynegymnastics and incontinence prevention. It has application in the prevention of falls in patients with neurodegenerative diseases and, as shown below, in the therapy of obesity.


Subject(s)
Exercise Therapy , Obesity/therapy , Adolescent , Humans , Male , Obesity/physiopathology , Resistance Training
12.
J Med Chem ; 54(20): 7397-407, 2011 Oct 27.
Article in English | MEDLINE | ID: mdl-21928794

ABSTRACT

The synthesis of various silybin monogalloyl esters was developed, and their antiangiogenic activities were evaluated in a variety of in vitro tests with human umbilical vein endothelial cells (HUVECs). A structure-activity relationship (SAR) study found the regioselectivity of the silybin galloylation to be highly significant. Silybin (as an equimolar mixture of two diastereomers A and B) exhibited quite poor antiangiogenic activities, whereas its B stereoisomer is more active than silybin A. The galloylation of phenolic OH groups of natural silybin (a mixture of both isomers) leads to increases in their antiangiogenic activities, which is more apparent with the 7-OH than the 20-OH. In contrast, gallates at aliphatic OH groups either had a comparable activity to the parent compound or are even worse than silybin, which was observed in the case of 3-O-galloylsilybin. The most effective compound from this series (7-O-galloylsilybin) has also been prepared from stereochemically pure silybins A and B to evaluate the effect of stereochemistry on the activity. As with silybin itself, the B isomer of 7-O-galloylsilybin was more active than the A isomer.


Subject(s)
Angiogenesis Inhibitors/chemical synthesis , Gallic Acid/analogs & derivatives , Gallic Acid/chemical synthesis , Silymarin/analogs & derivatives , Angiogenesis Inhibitors/chemistry , Angiogenesis Inhibitors/pharmacology , Cell Differentiation/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Collagen , Drug Combinations , Endothelial Cells/drug effects , Endothelial Cells/physiology , Esters , Gallic Acid/pharmacology , Human Umbilical Vein Endothelial Cells , Humans , Laminin , Proteoglycans , Silybin , Silymarin/chemical synthesis , Silymarin/pharmacology , Stereoisomerism , Structure-Activity Relationship
13.
Am J Reprod Immunol ; 66(3): 179-84, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21276122

ABSTRACT

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.


Subject(s)
Carotenoids/pharmacology , Receptors, Immunologic/analysis , Semen/drug effects , Biomarkers/analysis , Humans , Lycopene , Male , Prostate/chemistry , Prostate/drug effects , Receptor for Advanced Glycation End Products , Semen/chemistry
15.
Fertil Steril ; 94(7): 2604-8, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20413114

ABSTRACT

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.


Subject(s)
Flow Cytometry/methods , Luminescent Measurements/methods , Reactive Nitrogen Species/analysis , Reactive Oxygen Species/analysis , Semen/chemistry , Adult , Case-Control Studies , Humans , Hydrogen Peroxide/analysis , Hydrogen Peroxide/metabolism , Infertility, Male/diagnosis , Infertility, Male/metabolism , Leukocyte Count , Male , Peroxynitrous Acid/analysis , Peroxynitrous Acid/metabolism , Reactive Nitrogen Species/metabolism , Reactive Oxygen Species/metabolism , Semen/metabolism , Semen Analysis/methods , Young Adult
16.
Reprod Biol Endocrinol ; 7: 118, 2009 Oct 27.
Article in English | MEDLINE | ID: mdl-19860878

ABSTRACT

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.


Subject(s)
Excipients/pharmacology , Reactive Oxygen Species/analysis , Semen/chemistry , Semen/drug effects , Humans , Male , Reactive Oxygen Species/metabolism , Semen/metabolism , Semen Analysis , Semen Preservation/methods , Sodium Chloride/pharmacology , Sperm Retrieval
17.
J Agric Food Chem ; 57(15): 6584-9, 2009 Aug 12.
Article in English | MEDLINE | ID: mdl-19572653

ABSTRACT

The effect of Lonicera caerulea L. (blue honeysuckle) phenolic fraction (18.5% anthocyanins) on cell viability and against oxidative damage in low density lipoproteins (oxLDL), in rat microsomes and in primary cultures of rat hepatocytes and human umbilical vein endothelial cells (HUVEC), was tested. The phenolic fraction was nontoxic to rat hepatocytes and HUVEC at tested concentrations (1-1000 microg/mL) and time intervals up to 24 h inclusive. Phenolic fraction inhibited rat liver microsome peroxidation, induced by tert-butyl hydroperoxide (tBH), with IC(50) values of 160 +/- 20 microg/mL. The fraction at 0.5, 1.0, and 2.0 microg/mL delayed LDL oxidation, induced by Cu(2+), by 130 +/- 20%, 200 +/- 30%, and 400 +/- 10%, respectively. The treatment of HUVEC with oxidatively modified LDL induced an increase in lactate dehydrogenase (LDH) leakage and thiobarbituric acid reactive substances (TBARS) formation, and resulted in lower formazan formation from 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) uptake, most pronounced for 200 microg/mL (24 h oxidation) after 2 h of incubation. The protective effect of the phenolic fraction against cell damage caused by oxLDL was noted at 0.1 microg/mL for HUVEC and against tBH at 1000 microg/mL for both HUVEC and hepatocytes. The observed protective effects were probably due to the antioxidant properties of L. caerulea constituents, mainly anthocyanins. Microsome peroxidation and LDL oxidation inhibition results provide promising perspectives into the prevention of some oxidative stress-associated diseases. Other data are important in in vitro systems but seem to be accidental in vivo.


Subject(s)
Antioxidants/pharmacology , Caprifoliaceae/chemistry , Endothelium, Vascular/metabolism , Hepatocytes/metabolism , Oxidative Stress/drug effects , Plant Extracts/pharmacology , Adolescent , Adult , Animals , Cells, Cultured , Endothelium, Vascular/drug effects , Female , Hepatocytes/drug effects , Humans , Male , Pregnancy , Rats , Rats, Wistar , Young Adult
18.
Article in English | MEDLINE | ID: mdl-19365527

ABSTRACT

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.


Subject(s)
Infertility, Male/physiopathology , Neutrophils , Semen/chemistry , Semen/cytology , Humans , Male , Neutrophils/metabolism , Reactive Oxygen Species/analysis
19.
Reprod Biol Endocrinol ; 7: 9, 2009 Feb 03.
Article in English | MEDLINE | ID: mdl-19192281

ABSTRACT

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".


Subject(s)
Embryo Disposition , Embryo, Mammalian/cytology , Fertilization in Vitro , Adult , Cell Nucleus/ultrastructure , Cleavage Stage, Ovum/cytology , Cleavage Stage, Ovum/ultrastructure , Embryo Implantation , Embryo Transfer , Embryo, Mammalian/ultrastructure , Female , Humans , Maternal Age , Pregnancy , Pregnancy Rate , Retrospective Studies , Treatment Outcome
20.
Klin Mikrobiol Infekc Lek ; 15(6): 192-5, 2009 Dec.
Article in Czech | MEDLINE | ID: mdl-20077395

ABSTRACT

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.


Subject(s)
Bacteria/isolation & purification , Infertility/microbiology , Reactive Oxygen Species/metabolism , Semen/metabolism , Semen/microbiology , Adult , Humans , Male
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