Ramadan A Saleh

Role of reactive oxygen species in the pathophysiology of human reproduction

OBJECTIVE To summarize the role of reactive oxygen species (ROS) in the pathophysiology of human reproduction. DESIGN Review of literature. SETTING Fertility research center and obstetrics and gynecology department in a tertiary care facility. RESULT(S) ROS plays an essential role in the pathogenesis of many reproductive processes. In male-factor infertility. oxidative stress attacks the fluidity of the sperm plasma membrane and the integrity of DNA in the sperm nucleus. Reactive oxygen species induced DNA damage may accelerate the process of germ cell apoptosis, leading to the decline in sperm counts associated with male infertility. ROS mediated female fertility disorders share many pathogenic similarities with the ones on the male side. These similarities include a potential role in the pathophysiology of endometriosis and unexplained infertility. High follicular fluid ROS levels are associated with negative IVF outcomes, particularly in smokers. Moreover, oxidative stress may be responsible in hydrosalpingeal fluid mediated embryotoxicity as well as poor in vitro embryonic development. CONCLUSION(S) High levels of ROS are detrimental to the fertility potential both in natural and assisted conception states.

Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: a prospective study

OBJECTIVE To investigate levels of seminal oxidative stress (OS) and sperm quality in a group of infertile men with a history of cigarette smoking. DESIGN A prospective clinical study. SETTING Male infertility clinic, Urological Institute, the Cleveland Clinic Foundation, Cleveland, Ohio. PATIENT(S) Infertile men who smoked cigarettes (n = 20), infertile men who were nonsmokers (n = 32), and healthy nonsmoking donors (n = 13). INTERVENTION(S) Genital examination, standard semen analysis, sperm DNA damage. MAIN OUTCOME MEASURE(S) Levels of seminal reactive oxygen species (ROS) and total antioxidant capacity (TAC) measured by a chemiluminescence assay and seminal OS assessed by calculating a ROS-TAC score. Sperm DNA damage was measured by sperm chromatin structure assay. RESULT(S) Smoking was associated with a 48% increase in seminal leukocyte concentrations (P<.0001), a 107% increase in ROS levels (P=.001), and a 10-point decrease in ROS-TAC scores (P=.003). Differences in standard sperm variables and DNA damage indices between the infertile smokers and infertile nonsmokers were not statistically significant. CONCLUSION(S) Infertile men who smoke cigarettes have higher levels of seminal OS than infertile nonsmokers. Given the potential adverse effects of seminal OS on fertility, physicians should advise infertile men who smoke cigarettes to quit.

Evaluation of nuclear DNA damage in spermatozoa from infertile men with varicocele

OBJECTIVE To examine levels of sperm DNA damage and oxidative stress (OS) in infertile men with varicocele. DESIGN Prospective controlled study. SETTING Male infertility clinic, Glickman Urological Institute, Cleveland Clinic Foundation, Cleveland, Ohio. PATIENT(S) Thirty-one infertility patients and 16 fertile controls. INTERVENTION(S) Sperm DNA fragmentation index (DFI), levels of seminal reactive oxygen species (ROS), and total antioxidant capacity (TAC) were assessed using the sperm chromatin structure assay, chemiluminescence assay, and enhanced chemiluminescence assay, respectively. ROS-TAC score was calculated as a measure of OS. MAIN OUTCOME MEASURE(S) Median (interquartile range) DFI and ROS-TAC scores. RESULT(S) Sixteen of the 31 patients had left varicocele [grade I (n = 3), grade II (n = 10), and grade III(n = 3)], and the remaining 15 had normal genital examination. Patients with varicoceles had significantly higher percent DFI than controls (25%, range: 20%-35%; vs. 15%, range: 10%-22%). Patients with varicoceles had significantly lower ROS-TAC scores (21, range: 9.5-31) than the infertile patients with normal genital examination (34, range: 28-42) or the controls (40.3, range: 38-44). CONCLUSION(S) Infertile men with varicoceles showed significantly increased spermatozoal DNA damage that appears to be related to high levels of OS in semen.

Increased sperm nuclear DNA damage in normozoospermic infertile men: a prospective study

OBJECTIVE To evaluate levels of sperm nuclear DNA damage in infertile men with normal and abnormal standard semen parameters. DESIGN Prospective study. SETTING Male infertility clinic. PATIENT(S) Ninety-two men seeking infertility treatment and 16 fertile volunteers. INTERVENTION(S) Standard semen analysis was performed according to the World Health Organization guidelines. MAIN OUTCOME MEASURE(S) Sperm DNA damage was assessed by sperm chromatin structure assay and the results expressed as %DFI. RESULT(S) Of the 92 patients, 21 (23%) had normal standard sperm parameters (concentration, motility, and normal sperm forms), while 71 (77%) had an abnormality in one or more of these parameters. The %DFI [median (25th and 75th percentiles)] in infertile men with normal sperm parameters [23 (15, 32)] was significantly higher than fertile donors [15 (11, 20)] (P=.02), but not significantly different from infertile men with abnormal sperm parameters [28 (18, 41)] (P=.27). CONCLUSION(S) The results of this study indicate that a significant increase in SCSA-defined DNA damage can be found in sperm from infertile men with normal standard sperm parameters. Therefore, sperm DNA damage analysis may reveal a hidden abnormality of sperm DNA in infertile men classified as idiopathic based on apparently normal standard sperm parameters.

Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa

To investigate the role of increased seminal leukocytes in enhancing reactive oxygen species (ROS) production by human spermatozoa.A prospective study. Male infertility clinic.Forty-eight infertile men. Standard semen analysis. Assessment of sperm nuclear DNA damage by sperm chromatin structure assay. Incubation of spermatozoa from nonleukocytospermic samples with blood neutrophils. Spontaneous and phorbol 12-myristate 13-acetate (PMA)-induced ROS production in pure-sperm suspensions (after removal of leukocytes) as measured by a chemiluminescence assay. Levels of spontaneous and PMA-induced ROS production in pure-sperm suspensions from the infertile men with a diagnosis of leukocytospermia (n = 16) were significantly higher compared with the case of infertile men without leukocytospermia (n = 32) and with the case of a control group of healthy volunteers (n = 13). A similar pattern of increased ROS was observed when spermatozoa were incubated with blood neutrophils. Leukocytospermia was associated with a significant decrease in sperm motility and increase in DNA damage. Increased seminal leukocytes may play a role in stimulating ROS production by human spermatozoa. Such stimulation may be mediated via direct cell-cell contact or by soluble products released by leukocytes. Poor sperm quality in leukocytospermic samples may be due to leukocyte-mediated oxidative stress.

Increased DNA damage in sperm from leukocytospermic semen samples as determined by the sperm chromatin structure assay

OBJECTIVE To determine DNA damage as measured by the sperm chromatin structure assay (SCSA) in subsets of human spermatozoa at different stages of maturation in patients who are undergoing infertility evaluation. DESIGN Prospective study. SETTING Andrology laboratory at a tertiary care hospital. PATIENT(S) Fifty-six patients undergoing infertility evaluation. Patients with normal semen parameters (n = 17), abnormal semen parameters (n = 29), leukocytospermia (n = 10), and a group of healthy fertile men (n = 18) were included in the study. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) The shift of green (native DNA) to red (denatured, single-stranded DNA) fluorescence was measured and quantified using the expression alpha(t) (red fluorescence/[red + green fluorescence] per cell). Sperm DNA damage was examined in subsets of spermatozoa isolated by a three-step density gradient. The DNA damage was correlated with classic semen characteristics. RESULT(S) Leukocyte concentration in semen was directly correlated with chromatin alterations in immature and mature sperm. Leukocyte concentration in semen was also directly correlated with immature germ cell concentration and the percentage of abnormal forms in semen. CONCLUSION(S) The increase in chromatin alterations and DNA damage in sperm, as defined by the sperm chromatin structure assay from leukocytospermic samples may be related to alterations in the regulation of spermatogenesis.

Sexual dysfunction in men undergoing infertility evaluation: a cohort observational study

OBJECTIVE To study psychosexual problems in men undergoing infertility evaluation. DESIGN A cohort observational study. SETTING Male infertility diagnostic center. PATIENT(S) Four hundred twelve men undergoing infertility evaluation between 1999 and 2001. INTERVENTION(S) Baseline and follow-up data on sexual functions. Semen analysis for samples collected by masturbation. A second analysis was requested in 2 weeks upon finding an abnormality of semen parameters. Penile vibratory stimulation to help men who failed to collect semen on their second or subsequent appointments. MAIN OUTCOME MEASURE(S) Sexual functions (erection and orgasm) during the time of infertility evaluation. RESULT(S) Seven of 412 men were excluded from the analysis due to a past history of sexual dysfunction. Of the remaining 405 men, 46 (11%) failed to collect semen by masturbation for a second semen analysis after repeated (2-4 times) attempts at 2- to 3-day intervals. Nine of these men (20%) were able to collect semen using vibratory stimulation. All 46 men experienced problems with erection or orgasm and had severe anxiety during attempts to masturbate and during sexual contact with their partners. CONCLUSION(S) Our study indicates that some men may experience sexual dysfunction of a psychogenic nature in response to the diagnosis of infertility.

Increased levels of oxidants and reduced antioxidants in semen of infertile men with varicocele.

Significantly higher levels of oxidants (malonaldehyde and nitric oxide) and reduced levels of antioxidants (superoxide dismutase, glutathione peroxidase, catalase, and ascorbic acid) are seen in semen of infertile men with varicocele. Seminal oxidative stress (OS) seen in men with varicocele is associated with sperm motility and grade of varicocele.

Prevalence of female genital cutting in Upper Egypt: 6 years after enforcement of prohibition law

The objective of this study was to evaluate the prevalence of female genital cutting (FGC) in Upper Egypt, after 6 years of putting prohibition law into action. A total number of 3730 girls between the ages of 10-14 years were recruited to participate in this study. They were mainly preparatory school students (three urban and three rural areas). Social workers interviewed them as to whether they had undergone circumcision within the last 6 years or not. Subsequently, a questionnaire was sent to parents of girls who were positive for circumcision as to the circumstances surrounding the procedure. The prohibition law of FGC seems not to have altered the prevalence of this procedure. The majority of girls (84.9%) had had circumcision within the last 6 years with high prevalence in rural areas (92.5%). Circumcision was done for a combination of reasons, according to parents, with high rates of non-medical personnel participation (64.15%). This studys results indicate that the practice of FGC in Upper Egypt remains high despite enforcement of law. Extensive efforts are needed both to revise public awareness and to change attitudes regarding FGC.

Histopathologic patterns of testicular biopsies in infertile azoospermic men with varicocele

In azoospermic infertile men with varicocele, testicular biopsy revealed histopathologic patterns that varied from disorganized spermatogenesis with low or moderate sperm scores to early (primary spermatocytes stage) or late (spermatid stage) arrested spermatogenesis or germ cell aplasia and Sertoli cells only. Diagnostic testicular biopsy can be helpful for accurate management of azoospermic infertile men with varicoceles before surgical repair.