DNA Fragmentation In Patients Research Articles

P-057 Do the reduction of sexual abstinence influence sperm DNA fragmentation? Exploring the relationship between the two parameters

Abstract Study question Is the reduction of sexual abstinence (SA) really beneficial on sperm DNA fragmentation? Summary answer Despite recent studies support short SA’s positive impact on sperm parameters, apparently a reduced SA has not a clear impact on sperm DNA fragmentation. What is known already Sperm DNA fragmentation (sDF) has been linked to male infertility, affecting the success of fertility treatments. It’s strictly associated with failure and/or no longer time to conceive, impaired embryo development or higher miscarriege rates. The exact mechanism for age-dependent patterns of sperm decline is still not fully understood. Several factors, such as the increase in free radicals in conjuction with the rising in SA, have been discussed in literature. Recent studies support short SA’s positive impact on sperm parameters, despite controversial results; hence, we decided to analyze the trend in our facility: Humanitas Fertility Center. Study design, size, duration The present retrospective analysis, performed between 2021 and 2023, aims to investigate whether or not a reduced SA is beneficial on sperm DNA fragmentation in patients undergoing IVF. In our study 828 patients (from 21 to 59 years old) underwent both sperm analysis and sDF evaluation through Sperm Chromatin Structure Assay (SCSA). A DNA fragmentation index (DFI) lower than 30% indicates a sperm population with a good chromatin integrity, according to current knowledge. Participants/materials, setting, methods Patients were split in 4 groups, based on their age (≤38 and >38 years) and days of SA (0-3 and 4-7 days). Group 1 (n = 111/828): ≤38 years, ≤3 days; Group 2 (n = 359/828): ≤38 years, >3 days; Group 3 (n = 83/828): >38 years, ≤3 days; Group 4 (n = 275/828): >38 years, >3 days. Statistics was performed to analyze the variance among DFI values belonging to the four groups analyzed. Main results and the role of chance The results are expressed as mean(±SD). In Group 1, DFI %: 22,5(±14,7); in Group 2, DFI %: 21,5(±14,3); in Group 3, DFI %: 22,3(±14,9); in Group 4, DFI %: 26(±16,3). Surprisingly we observed the same behavior between most of the groups analyzed, except for the one belonging to the older patients. As a matter of fact, we found no statistical difference by comparing the DFI values belonging to samples of the same age stratification (≤38 or > 38 years), indeed Group 1 vs. Group 2: P=0,546; Group 3 vs. Group 4: P=0,1152. Different results were observed by comparing groups with the same SA days (≤3 or > 3), but belonging to a different age stratification, as it follows: Group 1 vs. Group 3: P=0,6384; Group 2 vs. Group 4: P=0,0001. This leads us to think that only the mixture of advanced paternal age (>38 years) and the increase of SA defines a worsening of DFI, according to our patients’ stratification. Limitations, reasons for caution There’s no homogeneity in the number of patients recruited with ≤3 days of SA compared to those with >3 days, as expected. Men undergoing fertility treatments follow strict SA guidelines; moreover, there’s no consensus about to request a second semen sample in clinical practice, in order to eliminate SA. Wider implications of the findings Exploring the inclusion of a broader spectrum of patients with low SA, despite their age, or instructing them to undergo semen analysis with a 0-day SA, could yield interesting insights. This could lead to targeted interventions and treatments, offering further hope for individuals with sDF issues. Trial registration number Not applicable

Exploring Gut Microbiome Composition and Circulating Microbial DNA Fragments in Patients with Stage II/III Colorectal Cancer: A Comprehensive Analysis.

Colorectal cancer (CRC) significantly contributes to cancer-related mortality, necessitating the exploration of prognostic factors beyond TNM staging. This study investigates the composition of the gut microbiome and microbial DNA fragments in stage II/III CRC. A cohort of 142 patients with stage II/III CRC and 91 healthy controls underwent comprehensive microbiome analysis. Fecal samples were collected for 16S rRNA sequencing, and blood samples were tested for the presence of microbial DNA fragments. De novo clustering analysis categorized individuals based on their microbial profiles. Alpha and beta diversity metrics were calculated, and taxonomic profiling was conducted. Patients with CRC exhibited distinct microbial composition compared to controls. Beta diversity analysis confirmed CRC-specific microbial profiles. Taxonomic profiling revealed unique taxonomies in the patient cohort. De novo clustering separated individuals into distinct groups, with specific microbial DNA fragment detection associated with certain patient clusters. The gut microbiota can differentiate patients with CRC from healthy individuals. Detecting microbial DNA fragments in the bloodstream may be linked to CRC prognosis. These findings suggest that the gut microbiome could serve as a prognostic factor in stage II/III CRC. Identifying specific microbial markers associated with CRC prognosis has potential clinical implications, including personalized treatment strategies and reduced healthcare costs. Further research is needed to validate these findings and uncover underlying mechanisms.

Dynamics of sperm DNA fragmentation in patients with clinically significant varicocele after Marmar operation and laparoscopic varicocelectomy

BACKGROUND: Infertility is detected in 15% of sexually active couples who engage in unprotected sexual intercourse. At the same time, the male factor of infertility accounts from 30 to 50%. It is known that in 15–25% the cause of frozen pregnancy and miscarriage is the male factor of infertility associated with violations of the integrity of sexual chromatin – pathological fragmentation of sperm DNA (FDNAS). One of the main pathophysiological mechanisms of the occurrence of pathozoospermia and FDNAS in secretory male infertility is oxidative stress, which occurs in patients with varicocele. It has been proven that varicocelectomy increases the frequency of spontaneous pregnancy and improves the results of IVF-procedures. However, there are currently insufficient studies devoted to the comparative evaluation of the effectiveness of various methods of varicocelectomy in patients with pathological FDNAS.
 AIM: To conduct a comparative assessment of the dynamics of sperm DNA fragmentation after subinguinal and laparoscopic varicocelectomy in male infertility.
 MATERIALS AND METHODS: The study included the results of examination and treatment of 87 men who complained of infertility in marriage for 1–9 years (on average, 3.8 ± 2.7 years). The age of the patients was 24–42 years (the average, 28.6 ± 11.3 years). All patients were diagnosed with grade II left-sided varicocele. Other pathological conditions potentially affecting ejaculate parameters were excluded. All patients underwent ejaculate analysis — sperm analysis, determined the degree of FDNAS by TUNEL method and the level of IgG class antisperm antibodies in the ejaculate by direct Mar-test. The patients were divided into 2 groups by randomization. The group 1 included 41 patients who underwent subinguinal varicocelectomy (Marmar operation), the group 2 included 46 patients who underwent laparoscopic varicocelectomy. Studies of the ejaculate — a sperm-analysis (according to WHO criteria, 2010), a MAR test and FDNAS — were carried out before, 3 and 6 months after surgery.
 RESULTS: In all patients of both groups in six months after surgical correction of varicocele the concentration of spermatozoa, the number of progressively mobile and normal forms of spermatozoa (according to the strict Kruger criteria) were significantly increased. The MAR test value in all patients of both groups before surgery did not exceed 10%. There were no statistically significant changes in this indicator in the postoperative period. FDNAS values in patients of group 1 before and 3 months after surgical treatment no statistically significant differences were found (27.6 ± 6.7% and 22.4 ± 8.3%, respectively, p 0.1). In patients in group 2, 3 months after surgical treatment a statistically significant decrease in the FDNAS value was noted compared with that before treatment (from 26.1 ± 8.9% to 13.3 ± 7.7%, p 0.001). 6 months after surgery, patients in both groups showed a significant (p 0.001) decrease in FDNAS values compared to the value before treatment: up to 14.6 ± 7.8% in patients of group 1 and up to 12.1 ± 8.0% — in patients of group 2.
 CONCLUSIONS: 1. Performing subinguinal and laparoscopic varicocelectomy contributes to the significant improvement of the main parameters of sperm in six months after the surgical operation. 2. The DNA fragmentation level significantly decrease already in 3 months after laparoscopic varicocelectomy, and just in 6 months after Marmar operation.

Long-term effect of cytotoxic treatments on sperm DNA fragmentation in patients affected by testicular germ cell tumor.

Testicular germ cell tumor (TGCT) is the most frequent neoplasia in men of reproductive age, with a 5-year survival rate of 95%. Antineoplastic treatments induce sperm DNA fragmentation (SDF), especially within the first year post-therapy. Data in the literature are heterogeneous concerning longer follow-up periods and the large majority is limited to 2 years. To define the timing for the recovery of sperm DNA damage and the proportion of patients with severe DNA damage (SDD) at 2 and 3 years from the end of therapy. SDF was evaluated in 115 TGCT patients using TUNEL assay coupled with flow cytometry before (T0 ) and 2 (T2 ) and 3 (T3 ) years post-treatment. Patients were divided based on the type of treatment: carboplatin (CP), bleomycin-etoposide-cisplatin (BEP), radiotherapy (RT). For 24 patients paired SDF data were available at all time-points (T0 -T2 -T3 ). 79 cancer-free, fertile normozoospermic men served as controls. SDD was defined as the 95th percentile in controls (SDF = 50%). Comparing patients versus controls we observed: i) no differences at T0 and T3 ; ii) significantly higher SDF levels (p<0.05) at T2 in all treatment groups. Comparing pre and post-therapy in the 115 patients, the median SDF values were higher in all groups at T2 , reaching significance (p<0.05) only in the CP group. While the median SDF values were higher also in the strictly paired cohort at T2 , about 50% of patients returned to baseline. The proportion of SDD in the entire cohort was 23.4% and 4.8% of patients at T2 and T3 , respectively. Currently, TGCT patients are advised to wait two years post-therapy before seeking natural pregnancy. Our results suggest that this period may not be sufficient for all patients. The analysis of SDF may represent a useful biomarker for pre-conception counselling following cancer treatment. This article is protected by copyright. All rights reserved.

Investigation of the effect of COVID-19 infection on sperm DNA fragmentation

Aim. To compare sperm parameters and the DNA fragmentation in patients after the COVID-19 infection and in men without a history of infection.Materials and methods. Sperm parameters and DNA fragmentation index were evaluated in 172 patients examined in 2020–2022. Based on the data on COVID-19 infection in the anamnesis, the patient’s cohort was divided into three groups: group 1 (n = 16) and group 2 (n = 65) – patients who had an infectious disease less than 75 days and more than 75 days before the delivery of the ejaculate, respectively, and group 3 (control, n = 91) – men have no coronavirus infection. A standard semen examination was performed according by the WHO guidelines (2010) recommendation. Sperm DNA fragmentation was evaluated by fluorescent labeling of single- and double-strand DNA breaks (TUNEL method).Results. The percentage of spermatozoa with fragmented DNA in group 1 varies from 2 to 48 %, in group 2 – from 0.9 to 39 %, in group 3 (control group) – from 1.3 to 52.9 %. The average number of gametes with DNA breaks in group 1 (11.7 ± 3.3 %) is greater than in group 2 (10.6 ± 0.8 %) and in group 3 (10.7 ± 0.8%), but without a statistically significant differences. The proportions of patients whose DNA fragmentation index exceeds the reference value (≤15 %) in groups 1, 2 and 3 are 18.8; 13.8 and 18.7 %, respectively. The count of progressively motile (PR) and morphologically normal spermatozoa in group 2 is statistically significantly greater than in group 3 (22.5 ± 1.6 % vs 18.0 ± 1.2 % and 5.52 ± 0.47 % vs 4.03 ± 0.31 %, respectively).Conclusion. The content of sperm with fragmented DNA is higher in men examined less than 75 days after COVID-19 disease, than in patients, examined more than 75 days after COVID-19 and in men who have no history of this infection. In group 1 an increased DNA fragmentation index was noted mainly in patients who had suffered from the disease in a moderate form.

The impact of autoimmune systemic inflammation and associated medications on male reproductive health in patients with chronic rheumatological, dermatological, and gastroenterological diseases: A systematic review.

Autoimmune disorders currently affect 5%-8% of the global population, characterized by an aberrant chronic inflammatory response to self-antigens. The aim of this study was to systematically review the current available evidence investigating the impact of systemic autoimmune diseases and associated immunosuppressive treatment on fertility parameters of adult men. Clinical trials, observational studies, and case reports written in English and reporting semen analysis, evaluation of seminal oxidative stress, and/or sperm DNA fragmentation in patients affected by psoriasis and psoriatic arthritis, celiac disease, inflammatory bowel diseases, systemic lupus erythematosus, ankylosing spondylitis, hidradenitis suppurativa, uveitis, dermatomyositis, and rheumatoid arthritis were collected by searching on PubMed, EMBASE, OVID, Scopus, and Cochrane Library databases, with no limit of time. The study quality and the extent of bias in design, methods, and outcome assessment were evaluated by applying the Joanna Briggs Institute Critical Appraisal tools. Evidence suggested that various autoimmune diseases or relevant medications can adversely affect male fertility parameters and that patients may benefit of counseling and sperm cryopreservation. Clinical trials further investigating any adverse effect of autoimmunity and related thereby on male infertility are warranted, to develop appropriate guidelines for males diagnosed and treated for autoimmune disorders.

Detection of microbial DNA in the blood of colorectal cancer patients for the diagnosis of microbial translocation.

171 Background: Dysbiosis has been associated with diseases and is of a major public health importance. Can lead to the passage of viable bacteria, their products or their fragments from the intestinal lumen through the mesenteric lymph nodes and other sites, known as bacterial translocation. The aim of the study was to determine whether microbial translocation occurs in stage II/III-IV colorectal cancer (CRC) patients and evaluate the usefulness of blood PCR for diagnosis of such translocation. Also to correlate the presence of Toll-Like Receptor and Vitamin D Receptor polymorphisms with the detection of microbial DNA fragments in the blood of CRC patients. Methods: Peripheral blood was obtained from 397 CRC patients (adjuvant n = 202 and metastatic n = 195) and 32 healthy individuals. DNA from all subjects was analyzed using PCR for amplification of genomic DNA encoding 16S rRNA, β-galactosidase gene of E. coli, Glutamine synthase gene of B. fragilis and 5.8S rRNA found in C. albicans. Results: Significantly higher rates of 16S rRNA, β-galactosidase, Glutamine synthase and 5.8S rRNA detection was observed in the pool of CRC patients than the controls ( p&lt; 0.001). All microbial DNA fragments detected were also significantly associated with the metastatic disease ( p&lt; 0.001) leading to shorter survival rates ( p&lt; 0.001). Moreover, individuals with the homozygous mutant alleles of either TLR or VDR gene polymorphisms had significantly higher detection rates of microbial DNA fragments. Conclusions: The detection of microbial DNA fragments in patients with CRC highlights the role of these microbes in cancer development, progression and therefore in patients’ survival.

Evaluation of sperm DNA fragmentation and chromatin structure in infertile men with immotile short‐tail sperm defect

Teratozoospermia is characterised by the presence of spermatozoa with abnormal morphology. One of the morphological disorders that lead to male infertility is immotile short-tail sperm (ISTS) defect. In this study, we evaluated the levels of chromatin packing and DNA fragmentation in patients with immotile short-tail sperm defect. Semen samples were obtained from 31 infertile men with ISTS as case group and 31 normozoospermic men as a control group. Protamine status was evaluated using chromomycin A3 (CMA3) staining and sperm DNA fragmentation assessed by sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labelling (TUNEL). The percentage of positive CMA3 spermatozoa was significantly higher in patients' samples (22.6±6.9) compared with controls (16.3±4.2) (p<.05) and also mean (±SD) of sperm DNA fragmentation was significantly higher in patients compared with controls, as measured by TUNEL assay (10.45±4.60 vs. 7.03±2.86, p<.05) and SCSA (24.80±13.1 vs. 15.2±7.2, p<.05). According to our study, sperm DNA fragmentation and chromatin packing abnormality are significantly higher in the ISTS samples compared with normal samples. A possible explanation for this relationship is that sperm chromatin condensation and sperm flagellum formation occur during the same phase of spermatogenesis.

Genome-wide cell-free DNA fragmentation in patients with cancer.

Cell-free DNA (cfDNA) in the blood provides a noninvasive diagnostic avenue for patients with cancer1. However, characteristics of the origins and molecular features of cfDNA are poorly understood. We developed an approach to evaluate fragmentation patterns of cfDNA across the genome and found that cfDNA profiles of healthy individuals reflected nucleosomal patterns of white blood cells, while patients with cancer had altered fragmentation profiles. We applied this method to analyze fragmentation profiles of 236 patients with breast, colorectal, lung, ovarian, pancreatic, gastric, or bile duct cancer and 245 healthy individuals. A machine learning model incorporating genome-wide fragmentation features had sensitivities of detection ranging from 57% to >99% among the seven cancer types at 98% specificity, with an overall AUC of 0.94. Fragmentation profiles could be used to identify the tissue of origin of the cancers to a limited number of sites in 75% of cases. Combining our approach with mutation based cfDNA analyses detected 91% of cancer patients. The results of these analyses highlight important properties of cfDNA and provide a proof of principle approach for screening, early detection, and monitoring of human cancer.

Evaluation of sperm DNA fragmentation index, Zinc concentration and seminal parameters from infertile men with varicocele.

The aim of this study was to investigate the effect of varicocele on DNA fragmentation index (DFI), zinc concentration and seminal parameters in infertile patients. In this prospective study, 179 men with at least 1-year history of infertility and varicocele were examined for semen quality at Hanoi Medical University Hospital (HMUH), Hanoi, Vietnam. In addition, an inverse correlation between zinc concentration and the degree of sperm DNA fragmentation in patients with clinical varicocele was found. The difference in mean values of sperm DNA fragmentation index in patients with various grades of varicoceles can be neglected, whereas most patients with varicocele of grades II and III had DFI >30%. Varicocele is associated with high levels of DNA damage in spermatozoa and reduced zinc levels that correlate with different grades of disease. Therefore, DNA fragmentation index and zinc concentration can be used as essential additional diagnostic test for patients with clinical varicocele. A study should be conducted to evaluate the benefits of zinc supplementation to improve seminal parameters in patients with varicocele.

LONG-TERM RESULTS OF TREATMENT OF PATIENTS WITH CHRONIC NONBACTERIAL PROSTATITIS AND INCREASED LEVEL OF SPERM DNA FRAGMENTATION WITH THE PROSTATILEN® AC DRUG

The study objectiveis to evaluate the results of treatment of patients with chronic nonbacterial prostatitis, increased level of sperm DNA fragmentation, and concomitant reproductive function disorders with the Prostatilen® AC drug 1, 2, and 3 months after the end of therapy.Materials and methods. Thirty five men aged 21–46 years (mean age 31.3 ± 4.3 years) with a verified diagnosis of “chronic nonbacterial prostatitis, associated reproductive function disorders, history of early-stage missed miscarriage in a wife/sex partner” were examined. All patients received Prostatilen® AC as rectal suppositories daily for 10 days with a repeat course 20 days later. Before and after the treatment, as well as after 1, 2, and 3 months after its end, the fraction of spermatozoa with fragmented DNA was measured in all patients using sperm chromatin structure assay with flow cytofluorometry. The risk of decreased fertility was considered low if the number of spermatozoa with fragmented DNA was 15 % or lower (this value is considered the norm). If DNA fragmentation was 16 % or higher, it was considered increased.Results. The fraction of spermatozoa with fragmented DNA significantly decreased during the treatment, and 1 month after the end of the therapy it was 7.5 ± 4.2 % (prior to treatment it was 17.8 ± 6.7 %) (p &lt;0.005). Prior to Prostatilen® AC treatment, enhanced DNA fragmentation was observed in 18 (51 %) patients, 1 month after treatment – only in 6 (17 %); the difference is statistically significant (p &lt;0.001). Two months after the end of treatment, increased level of DNA fragmentation was detected in 7 (20 %) patients, 3 months after – in 15 (42 %) patients. Aside from that, if the fraction of spermatozoa with fragmented DNA was more than 30 % before treatment, then a decrease in this value during treatment wasn’t statistically significant.Conclusion. Prostatilen® AC promotes a reduction in the level of sperm DNA fragmentation in patients with chronic nonbacterial prostatitis. A steady positive effect persists for 2 months with a tendency to weaken at the end of month 3 which indicates advisability of repeated courses of Prostatilen® AC treatment 2 months after the end of therapy.

Cryopreservation triggers DNA fragmentation and ultrastructural damage in spermatozoa of oligoasthenoteratozoospermic men

Objective: Oligoasthenoteratozoospermia (OAT) is one of the causes of male infertility. Cryopreservation is valuable for the management of infertility. This study aimed to reveal the effects of cryopreservation on sperm motility, morphology, ultrastructural details and DNA fragmentation in patients with OAT. Materials and Methods: In this observational study, ejaculates were collected from 40 male volunteers of whom 20 were OAT and 20 were normospermic. The ejaculates were stored in liquid nitrogen at -196oC and analysed following thawing after 1 or 3 months. Ejaculates were evaluated in terms of motility, morphology and DNA fragmentation before and after thawing. Results: Sperm motility and morphology were both affected by cryopreservation in samples from both groups. After thawing, spermatozoa with morphological abnormalities were increased significantly in both groups. Ultrastructural investigations showed alteration in integrity of the membranes and increased acrosomal defects. Post-thaw investigations revealed prominent increases in the number of DNA fragmented cells in both groups when compared to the results before freezing. OAT groups revealed a significantly higher number of DNA fragmented spermatozoa compared to the normospermic group for both time periods. Conclusion: Cryopreservation produces ultrastructural damage and induces DNA fragmentation in both normospermic and OAT groups. The damage is more severe in the OAT group.

Varicocele and DNA Fragmentation in Senegalese Infertile Males—A Study of 95 Cases

Varicocele is the first etiology of infertility in men worldwide and affects almost half of the men dealing with infertility in Senegal. Varicocele is associated with a high rate of sperm DNA fragmentation so this test is being more and more recommended especially when patients have to benefit from an ART (Assisted Reproductive Technology) procedure. The aim of our study was to assess if our patients with varicocele are concerned by higher DNA fragmentation rate in spermatozoon. Sperm samples were collected (95 cases of varicocele and 30 controls) and analyzed between January 2013 and June 2016 in a prospective study. Sperm DNA damage, characterized by the rate of DNA fragmentation was evaluated by TUNEL assay using fluorescence microscopy. There was a significant higher percentage of DNA fragmentation in patients with varicocele (p < 0.001), compared to controls. We plan to study a bigger cohort to assess this tendency and study the benefits of surgery in varicocele by evaluating DNA fragmentation after the procedure.

EFFECT OF PROSTATILEN® AC ON SPERM DNA FRAGMENTATION DURING TREATMENT OF PATIENTS WITH CHRONIC NONBACTERIAL PROSTATITIS AND CONCOMITANT DISORDERS OF THE REPRODUCTIVE FUNCTION

The study objective is to analyze the effect of Prostatilen® AC on sperm DNA fragmentation during treatment of patients with chronic nonbacterial prostatitis and concomitant disorders of the reproductive function. Materials and methods . The study is based on the results of treatment of 25 men aged 24 to 45 years (mean age 35.3 ± 4.4 years) with a verified diagnosis of chronic nonbacterial prostatitis and complaints of early-stage missed miscarriage in a spouse/sexual partner. All patients received Prostatilen® AC daily in rectal suppositories formulation. The duration of treatment was 10 days with retreatment after 20 days. In all patients before treatment and 20 days after it, spermiogram parameters (5th ed., WHO, 2010) and sperm DNA fragmentation level using SCSA (sperm chromatin structure assay) by FACSCantoll with monoclonal antibodies (Roche, Germany) were determined, and all patients underwent the MAR (mixed antiglobulin reaction) test with normal value considered to be 10 % or less. The normal value of sperm DNA fragmentation was considered to be 15 % or less (low risk of fertility impairment). The analysis of the obtained data was carried out using the IBM SPSS Statistics program 22. Results . Before the treatment, pathologic level of sperm DNA fragmentation was observed in 6 (43 %) of 14 patients with normozoospermia and in 7 (63 %) of 11 patients with pathozoospermia (χ² = 1.06; p <0.3). Thus, there weren’t any significant difference between the rates of occurrence of increased sperm DNA fragmentation in patients with normo- and pathozoospermia. A correlation was found between the level of sperm DNA fragmentation and the results of MAR test before treatment (r = 0.8, p <0.05), which varied between 0 and 99 % (mean 16.48 ± 31.64 %). Meanwhile, increased sperm DNA fragmentation was observed in 7 (53 %) of 13 patients with pathological MAR test results, and in 2 (40 %) of 5 patients with normal MAR test results (χ² = 0.67; p <0.01). The level of sperm DNA fragmentation prior to treatment varied between 1 and 55 % (mean 18.99 ± 6.87 %), and after treatment it varied between 1 and 26 % (mean 9.76 ± 4.32 %). Increased level of sperm DNA fragmentation was observed in 13 (52 %) of 25 patients before treatment with Prostatilen® AC and only in 3 (12 %) patients after the treatment (χ² = 0.86; p < 0.001). Conclusion . Prostatilen® AC as treatment of chronic nonbacterial prostatitis promotes a decrease in the level of sperm DNA fragmentation.

A Phytotherapic Approach to Reduce Sperm DNA Fragmentation in Patients with Male Infertility

Infertility affects 50 to 80 million (between 8 and 12% of couples). Male factor is a cause of infertility in almost half of the cases, mainly due to oligoasthenoteratozoospermia. DNA fragmentation is now considered an important factor in the aetiology of male infertility. We studied the effects on semen analysis and on DNA fragmentation of in vivo admnistration of Myo-Inositol and Tribulus Terrestris plus Alga Ecklonia plus Biovis (Tradafertil; Tradapharma Sagl, Swizerland) in men with previously diagnosed male infertility. Sixty patients were enrolled in the present study and were randomized into two subgroups: the group A who received Myo-inositol 1000 mg, Tribulus Terrestris 300 mg, Alga Ecklonia Bicyclis 200 mg and Biovis one tablet a day for 90 days, and the group B (placebo group) who received one placebo tablet a day for 90 days. The primary efficacy outcome was the improvement of semen characteristics after 3 months' therapy and the secondary outcome was the reduction of the DNA fragmentation after treatment. The groups were homogenous for age, hormonal levels, sperm concentration and all parameters of sperm analysis. Sperm concentration and progressive motility improved after treatment with Tradafertil (3.82 Mil/ml vs. 1.71 Mil/ml; p<0.05; 4.86% vs. 1.00%; p<0.05) as well as the DNA fragmentation (-1.64% vs -0.39%, p<0.001). No side effects were revealed. In conclusion, we can affirm that Tradafertil is safe and tolerable. It is a new phytotherapic approach to Oligoasthenoteratospermia (OAT) syndrome that could lead to good results without interacting with hypothalamic-pituitary-gonadal axis.

Association between MTHFR C677T polymorphism and folate, vitamin B12, homocysteine, and DNA fragmentation in patients with ovarian cancer

Abstract Objective Methylenetetrahydrofolate reductase (MTHFR) is an important enzyme that regulates the metabolism of methionine and folate. MTHFR C677T polymorphism was reported to be associated with breast and ovarian cancer. The aim of this study was to evaluate the association between the MTHFR C677T (rs1801133) polymorphism and homocysteine, vitamin B12, and folate levels, and DNA fragmentation in patients with ovarian cancer and healthy controls. Materials and methods This case-control study was conducted in Istanbul University Cerrahpasa Medical Faculty. We studied 50 ovarian cancer patients and 54 healthy controls. The MTHFR C677T polymorphism was determined by PCR followed by restriction fragment length polymorphism (RFLP) and agarose gel electrophoresis. DNA fragmentation was assessed by the comet assay. Homocysteine levels were measured by ELISA, whereas vitamin B12 and folate levels were measured by chemiluminescence methods. Results We found no correlation between the MTHFR C677T polymorphism and ovarian cancer. No significant difference was found in homocysteine, folate, and vitamin B12 levels between patient and control groups. Increased DNA fragmentation was detected in patients with ovarian cancer. Conclusion Our findings suggest that MTHFR C677T polymorphism, as well as homocysteine, folic acid, and vitamin B12 levels, are not associated with an increased risk for ovarian cancer.

Estimation of changes in spermogram and index of sperm DNA fragmentation in patients with premature ejaculation receiving Neurodoz biocomplex

Background. An incidence of premature ejaculation (PE) in men of reproductive age of 24–45 years is 23 %. The treatment of PE is based on antidepressants from the group of selective serotonin re-uptake inhibitors. However, drugs of this group have gametotoxic action. Objective . Evaluate the effectiveness of Neurodoz biocomplex, its effect on the parameters of spermogram and sperm DNA integrity in patients with PE. Materials and methods. The study involved 16 patients with PE and normozoospermia aged from 26 to 35 years. Ejaculate and sperm DNA fragmentation were assessed by Halosperm (Halotech®) method before and after the therapy. Duration of treatment was 2 months. Evaluation of efficacy was based on the duration of sexual intercourse, as well as on the visual analogue scale and the questionnaire data of patients with premature ejaculation. Results. In 8 weeks there was a decrease in ejaculate volume from to 3.6 ± 1.1 down to 3.2 ± 1.1 mL (p = 0.312) and an increase in the concentration of sperm (34.2 ± 9.8) × 106 up to (35.6 ± 9.7) × 106 (p = 0.688). Sperm motility (category a + b) increased from 36.6 ± 4.3 to 37.2 ± 4.5 % (p = 0.703), the percentage of morphologically normal forms increased from 14.6 ± 1.4 up to 14.8 ± 1.1 (p = 0.656). DNA fragmentation of sperm has decreased from 15.6 ± 2.7 down to 15.4 ± 2.7 % (p = 0.635). Duration of intravaginal latency increased from 101.8 ± 34.4 (45–150) up to 217.3 ± 28.9 (190–280) sec (p < 0.001). An overall score according to the premature ejaculation questionnaire decreased from 13.9 ± 3.0 (9–20) down to 4.4 ± 1.9 (0–7) (p < 0.001). Visual analogue scale symptoms improved from 7.3 ± 1.7 (5–10) to 2.6 ± 1.2 (0–4) (p < 0.001). Conclusion. Neurodoz is an effective and safe agent for PE correction. Giving an absence of negative impact of biocomplex on spermatogenesis in patients with normozoospermia, it is possible to administer it to men who are trying to conceive. Further study is necessary in order to assess an impact of the drug on semen parameters in subfertile patients

Relationship Between Sperm DNA Fragmentation and Nuclear Vacuoles.

The aim of the present study is to assess the correlation between the presence, quantity and size of nuclear vacuoles and DNA damage and chromatin status in sperm samples of men who underwent to assisted reproduction technology. Forty six males who underwent to assisted reproductive technology (ART) were considered. According to their latest semen analysis (<3 months), were grouped into: (A) strict morphology index ≤4% (26) and (B) strict morphology index ≥14% (20). Motile sperm were selected by density gradient, and MSOME study was conducted to assess the number and size of nuclear vacuoles. DNA fragmentation (TUNEL) and DNA strand status (acridine orange) were assessed over the selected spermatozoa accordingly to their vacuole pattern. In group A, sperm without vacuoles (1°) have similar levels of DNA fragmentation (TUNEL) in compare to the rest of observed patterns (2°- 6°). Regarding to AO, spermatozoa with large or several vacuoles that cover more than 30-50% of the nuclear surface are AO+, but not necessarily TUNEL positive. The first three patterns of vacuoles patterns had lower levels of AO in compare to grades 4° and 6°. In group B, those sperm with one or more vacuoles greater than 30%-50% (4° and 6°), had a significant increase in TUNEL values, in relation to group 1°- 3°. Considering AO, it was found that the 4° and 6° pattern had a significantly elevated level of this marker, as same of group A (P <0.05). There is no relationship between the greater number and size of sperm vacuoles with high levels of DNA fragmentation in patients with severe teratozoospermia (Kruger <4%). Conversely, this relationship is evident in normal semen samples (normal morphology. Sperm selection by IMSI technique, to select non-fragmented sperm in patients with Kruger <4%, is not necessarily secured when non-vacuolated sperm is selected.

Increased DNA Fragmentation in Patients with Infertility in Dakar (Senegal)

The presence of DNA fragmentation in semen is associated with poor results in ART (Assisted Re-productive Technology). This has led to the introduction of sperm DNA integrity testing in the clinical assessment of male infertility. The TUNEL assay is one of the most commonly used tests to measure DNA fragmentation in research studies, but it is now being optimized for routine clinical use. So, the objectives of our study were to install the TUNEL assay in our laboratory and to use it to assess the degree of DNA fragmentation in our patients dealing with couple infertility. Forty seven (47) and 25 samples from patients followed up for infertility and known fertile men were analyzed between August 2013 and July 2015 respectively. The sperm samples were evaluated according to the World Health Organization Guidelines. Sperm DNA damage was assessed by the TUNEL assay, with a standardized protocol that we adapted for reading using fluorescence microscopy instead of flow cytometry. The Sperm DNA Fragmentation Index was significantly higher in patients with infertility compared to the controls (22.2% ± 5.6% vs. 16.7% ± 0.7%) with a p < 0.05. This Index was also high in patients with varicocele compared to the control group (22.8% ± 9.2 vs. 16.7% ± 0.7%) (p < 0.01). After establishing the ROC curve (Receiver Operating Characteristics curve), we found a 15% threshold of DNA fragmentation, separating the patients with infertility from the control group. In our study, a 15% threshold positivity of DNA fragmentation using the TUNEL assay was highly associated with infertility. The results indicate that high level DNA fragmentation is associated with infertility.

The study of spermatic DNA fragmentation and sperm motility in infertile subjects

Although the pathophysiology of the testicular damage associated with varicocele remains unclear, sperm DNA damage has been identified as a potential explanation for this cause of male infertility. The current study was designed to determine the extent of sperm nuclear DNA damage in patients with varicocele, and to examine its relationship with parameters of seminal motility. Semen samples from 60 patients with clinical varicocele and 90 infertile men without varicocele were examined. Varicocele sperm samples were classified as normal or pathological according to the 1999 World Health Organizzation guidelines. Sperm DNA damage was evalutated using the Halosperm kit, an improved Sperm Chromatin Dispersion (SCD) test. The DNA fragmentation index (DFI: percentage of sperm with denatured nuclei) values was significantly higher in patients with varicocele, either with normal or abnormal (DFI 25.8 ± 3.2 vs 17.4 ± 2.8 - P < 0,01) semen profiles. In addition, an inverse correlation was found between spermatic motility and the degree of spermatic DNA fragmentation in patients with clinical varicocele. Varicocele is associated with high levels of DNA-damage in spermatozoa. In addition, in subjects with varicocele, abnormal spermatozoa motility is associated with higher levels of sperm DNA fragmentation. DNA fragmentation may therefore be an essential additional diagnostic test that should be recommended for patients with clinical varicocele.