Analysis of the Functional Aspects of Sperm and Testicular Oxidative Stress in Individuals Undergoing Metabolic Surgery.

PURPOSE: Metabolic surgery is a recommended treatment for obese patients that results in BMI reduction; however, the observed impact of this therapy on male fertility is inconsistent. This research aimed to study the effects of BMI changes after metabolic surgery on seminal analysis, sex hormonal profile, sperm functional integrity, and the seminal plasma lipid peroxidation levels. MATERIALS AND METHODS: A prospective study was performed in 15 patients for whom metabolic surgery was recommended. The patients were evaluated by the techniques proposed in this study before and after the surgical procedure for 12 months. In each analysis, the male sex hormonal profile, semen analysis, sperm functional integrity, and seminal lipid peroxidation levels were assessed. RESULTS: The surgery resulted in BMI reduction and improvement in seminal characteristics and male sex hormone profile. The semen analysis showed increases in volume, sperm progressive motility, and in sperm morphology and a decrease in immotile sperms. Sperm mitochondrial activity and sperm DNA integrity were improved, and the levels of seminal lipid peroxidation were decreased. The hormonal profile showed lower levels of estradiol and highest levels of luteinizing hormone (LH), sex hormone-binding globulin (SHBG), and testosterone. CONCLUSION: BMI changes resulting from this treatment and its metabolic consequences can be associated with changes in the male fertile potential, leading to an improvement in the seminal quality, male sex hormone profile, sperm functional aspects, and levels of seminal lipid peroxidation, thus decreasing the testicular oxidative stress.

Effect of smoking on the functional aspects of sperm and seminal plasma protein profiles in patients with varicocele.

STUDY QUESTION: What are the effects of smoking on the functional aspects of the sperm, the levels of lipid peroxidation and the protein profile of seminal plasma in patients with varicocele? SUMMARY ANSWER: In men with varicocele, smoking is associated with altered semen quality, decreased sperm functional integrity and seminal oxidative stress. Alterations in seminal plasma protein profiles are also present and may explain the altered semen phenotype. WHAT IS KNOWN ALREADY: Varicocele is a major cause of male infertility. It reduces testicular blood renewal with a consequent accumulation of toxic substances. Thus, it can potentiate the toxic effects of environmental exposure to genotoxic substances such as those found in cigarette smoke. STUDY DESIGN, SIZE AND DURATION: A cross-sectional study was performed in 110 patients presenting with variococele to the Human Reproduction Section of the Sao Paulo Federal University (2006-2010). The patients were divided into a control group of non-smokers, a moderate smokers group and a heavy smokers group. PARTICIPANTS/MATERIALS, SETTING AND METHODS: Semen parameters were analysed by standard methods. Sperm DNA integrity and mitochondrial activity were assessed by Comet assays and by 3,3'-diaminobenzidine deposition, respectively. The level of lipid peroxidation in semen was determined by malondialdehyde quantification. Proteomic studies were performed by 2D-electrophoresis and mass spectrometry. MAIN RESULTS AND THE ROLE OF CHANCE: Both groups of smokers showed reduced semen quality in comparison with the control group. In the groups of smokers, sperm DNA integrity and mitochondrial activity were also decreased and lipid peroxidation levels were increased. Proteomic analyses revealed 20 proteins differentially expressed between the study groups. LIMITATIONS AND REASONS FOR CAUTION: A study including smokers without varicocele is still warranted as these results apply only to smokers who present varicocele. WIDER IMPLICATIONS OF THE FINDINGS: Patients with varicocele who are exposed to tobacco smoking present more important alterations to semen quality and sperm functional integrity and show changes in the seminal plasma proteome. This suggests testicular, and possibly systemic, adverse effects of smoking. STUDY FUNDING/COMPETING INTEREST(S): Funding for the study was provided by Fundação de Amparo à Pesquisa do Estado de São Paulo (Fapesp) (2007/59423-7) and by the Division of Urology, Human Reproduction Section at the São Paulo Federal University.

Adolescent varicocele: improved sperm function after varicocelectomy.

OBJECTIVE: To assess the effect of varicocelectomy on sperm function (DNA integrity and mitochondrial activity) and levels of lipid peroxidation in seminal plasma of adolescents. DESIGN: Prospective study. SETTING: Patients recruited from a local public school. PATIENT(S): Adolescents (14-19 years old), Tanner stages IV or V with varicocele grades II or III, attending a local public school. INTERVENTION(S): Two semen collections with a one week interval between collections before bilateral varicocele repair using subinguinal microsurgical varicocelectomy, and two semen collections with a one week interval between collections three months after the surgery. MAIN OUTCOME MEASURE(S): Rate of sperm DNA fragmentation as assessed by the Comet assay and categorized as classes I (no DNA fragmentation) to IV (high DNA fragmentation). Rate of mitochondrial activity as assessed by the diaminobenzidine assay and categorized as grades I (all mitochondria active) to IV (all mitochondria inactive). Levels of lipid peroxidation in seminal plasma by a colorimetric method that quantifies a lipid peroxidation subproduct (malondialdehyde). RESULT(S): Concerning DNA integrity, the samples after varicocelectomy showed more spermatozoa with intact nuclear DNA (grade I) and less spermatozoa with Comet grades II, III, and IV. Regarding mitochondrial activity, the samples after varicocelectomy showed less cells with inactive mitochondria (class III). No differences were observed in classes I, II, and IV. Concerning lipid peroxidation, no significant differences were observed. CONCLUSION(S): This study was able to demonstrate that varicocelectomy in adolescents is associated with increased sperm DNA integrity and mitochondrial activity. However, levels of seminal products of lipid degradation (malondialdehyde) are not different.

Is sperm cryopreservation an option for fertility preservation in patients with spinal cord injury-induced anejaculation?

OBJECTIVE: To evaluate the effects of cryopreservation on sperm mitochondrial activity and nuclear DNA integrity in men with spinal cord injury. DESIGN: Prospective controlled study. SETTING: Patients in an academic research environment. PATIENT(S): Men with and without spinal cord injury-induced anejaculation. INTERVENTION(S): Electroejaculation or penile vibrating stimulation semen cryopreservation using a commercial TEST-yolk-buffer technique. MAIN OUTCOME MEASURE(S): Rate of sperm DNA fragmentation as assessed by the comet assay, graded in Classes I (high DNA integrity) to IV (high DNA fragmentation). Mitochondrial activity as assessed by a method in which active mitochondria precipitate 3,3'-diaminobenzidine. Cells were classified as I (all active) to IV (all inactive). Semen was cryopreserved in a Test-yolk buffer, and motility, DNA fragmentation, and mitochondrial activity were analyzed precryopreservation and postthaw. RESULT(S): Before cryopreservation, when the study (SCI) and control groups were compared, no statistically significant differences were found with respect to concentration or total sperm count; however, the SCI group presented significantly lower ejaculate volume, decreased sperm morphology, and an increase in the round cell and neutrophils counts. In both groups, cryopreservation was associated with an increase in DNA fragmentation, a decrease in mitochondrial activity, and a decrease in motility, of which the latter was of greater importance in the control group. CONCLUSION(S): Cryopreservation causes a decrease in conventional seminal variables as well as in mitochondrial activity and DNA fragmentation. However, these were no more detrimental to sperm from men with SCI than to sperm from the control group.

Effect of leukocytospermia and processing by discontinuous density gradient on sperm nuclear DNA fragmentation and mitochondrial activity.

PURPOSE: To assess the effect of leukocytospermia and semen processing on sperm DNA and mitochondria. METHODS: Twenty-two patients with and 41 without leukocytospermia were included. Sperm DNA fragmentation was assessed by the Comet assay, and mitochondrial activity by a colorimetric method for active mitochondria. Semen was processed using Percoll, and motility, DNA fragmentation, and mitochondrial activity were analyzed pre- and post-processing. RESULTS: No differences were observed in age, abstinence, volume, sperm morphology, progressive motility, concentration, and vitality (p>0.10). Variables were grouped according to time (pre- vs post-processing) and group (leukocytospermia vs non-leukocytospermia) because no interactions could be observed. Leukocytospermia was associated to increased DNA fragmentation, while semen processing led to a decrease in DNA fragmentation and to increased mitochondrial activity. CONCLUSION: While semen processing selects sperm with higher rates of DNA integrity independent of the presence or absence of leukocytes in semen, samples without leukocytospermia present more sperm without DNA fragmentation. Semen processing also selects sperm with higher mitochondrial activity.

Sperm nuclear DNA fragmentation and mitochondrial activity in men with varicocele.

OBJECTIVE: To verify the impact of varicocele on semen quality and sperm function (DNA integrity and mitochondrial activity). DESIGN: Prospective study. SETTING: Patients in an academic research environment. PATIENT(S): Seventeen patients with a clinical diagnosed varicocele of grade II or III and 20 men without a varicocele. MAIN OUTCOME MEASURE(S): Rate of sperm DNA fragmentation as assessed by the Comet assay and categorized as classes I (no DNA fragmentation), II (little DNA fragmentation), III (meaningful DNA fragmentation), and IV (high DNA fragmentation). Rate of mitochondrial activity as assessed by the diaminobenzidine (DAB) assay and categorized as grades I (all mitochondria active), II (most mitochondria active), III (most mitochondria inactive), and IV (all mitochondria inactive). RESULT(S): No statistically significant differences were found between the study and control groups with respect to age, ejaculatory abstinence, and round cell count. Men with varicocele had significantly higher ejaculate volume, concentration of immotile sperm, and neutrophil count and lower mean percentage of sperm concentration, progressive motility, and morphology than men in the control group. The study group presented a lower percentage of sperm with little DNA fragmentation (class II) and a higher percentage of sperm with DNA fragmentation (class IV). In addition, the study group presented a greater percentage of sperm with inactive mitochondria (class III). CONCLUSION(S): Compared with men without varicocele, men with varicocele had a higher percentage of cells with DNA fragmentation and sperm with inactive mitochondria. Indeed, varicocele causes a decrease in motility, concentration, and morphology and an increase in volume and concentration of immotile sperm and neutrophils. The sperm functional evaluation (DNA fragmentation and mitochondrial activity) could be important factors in deciding treatment options for men with varicocele.