The Hydrogen Storage Nanomaterial MgH2 Improves Irradiation-induced Male Fertility Impairment by Suppressing Oxidative Stress

ObjectiveThis study aimed to reveal the protective effect of hydrogen storage nanomaterial MgH2 on radiation-induced male fertility impairment.MethodsThe characterization of MgH2 were analyzed by scanning electron microscopy (SEM) and particle size analyzer. The safety of MgH2 were evaluated in vivo and in vitro. The radioprotective effect of MgH2 on the reproductive system were analyzed in mice, including sperm quality, genetic effect, spermatogenesis, and hormone secretion. ESR, flow cytometry and western blotting assay were used to reveal the underlying mechanisms.ResultsMgH2 had an irregular spherical morphology and a particle size of approximately 463.2 nm, and the content of Mg reached 71.46%. MgH2 was safe and nontoxic in mice and cells. After irradiation, MgH2 treatment significantly protected testicular structure, increased sperm density, improved sperm motility, reduced deformity rates, and reduced the genetic toxicity. Particularly, the sperm motility were consistent with those in MH mice and human semen samples. Furthermore, MgH2 treatment could maintain hormone secretion and testicular spermatogenesis, especially the generation of Sertoli cells, spermatogonia and round sperm cells. In vitro, MgH2 eliminated the [·OH], suppressed the irradiation-induced increase in ROS production, and effectively alleviated the increase in MDA contents. Moreover, MgH2 significantly ameliorated apoptosis in testes and cells and reversed the G2/M phase cell cycle arrest induced by irradiation. In addition, MgH2 inhibited the activation of radiation-induced inflammation and pyroptosis.ConclusionMgH2 improved irradiation-induced male fertility impairment by eliminating hydroxyl free radicals.

Estimation of haemodynamic changes in varicocele testis and results after microsurgical sub-inguinal varicocelectomy

Introduction: Varicocele is among the most frequently encountered, surgically correctable causes of male infertility. Since varicocele is a primary abnormality of testicular vessels, a thorough understanding of haemodynamic changes in the microcirculation of varicocele testis is needed. Objectives: To estimate changes in the microcirculation of varicocele testis and correlate them with symptomatic and semen parameters changes before and after varicocelectomy. Material and methods: This prospective, observational study includes 47 patients of age group 18–40 years who underwent microsurgical varicocelectomy during June 2016 to April 2018 at a tertiary medical institute. Testicular haemodynamic was evaluated using a colour spectral doppler study (PSV-Peak systolic velocity/ESV-End diastolic velocity/RI-Resistive index/PI-Pulsatility index). Semen analysis parameters and testicular blood flow were compared with pre-operative values. Results: RI in the capsular artery of varicocele affected testis, which was 0.61 ± 0.11 (mean ± SD), significantly reduced to 0.56 ± 0.09 (mean ± SD) in post-operative follow-up, indicating improved perfusion. Pre-operative sperm density was noted to be 15.94 ± 7.88 (mean ± SD), which improved to 28.41 ± 10.99, Progressive sperm motility from 33.81 ± 10.88 to 38.32 ± 9.21 and morphology from 36.13 ± 10.68 to 41.43 ± 9.29 on 6-month follow-ups ( p < 0.05). Conclusion: Varicocele testis is associated with altered testicular haemodynamic, which insults spermatogenesis. Microsurgical varicocelectomy with testicular artery and lymphatic preservation leads to correction of this altered haemodynamic state and improves the testicular microcirculation.

The Impact of the Time Interval Between Cycles on Pregnancy Outcome of Ovulation Induction Cycle Intrauterine Artificial Insemination

ObjectiveThe objective of this study was to estimate whether the time interval between two intrauterine inseminations (IUI) treatments needs to be extended by one menstrual cycle or more in patients undergoing successive cycles of ovulation stimulation, and whether this will have an impact on the clinical pregnancy rate (CPR).Study DesignRetrospective cohort study.Study siteThe study site was the reproductive medicine center of a teaching hospital.Patient(s)The subjects were women and their husbands who received two or more intrauterine insemination in our reproductive medicine center due to mild infertility in the period from January 2017 to December 2019. Patients were divided into 2 groups according to the number of days between the last menstrual day(LMD)and the previous IUI operation day(POD), continuous group (the time from the LMD to POD ≤ 34 days) and delayed group (the time from the LMD to POD ≥ 35 days). We excluded cycles with intervals of more than 180 days.In order to avoid the inclusion of multiple repeat cycles for the same couple, only the first two cycles of IUI treatment in the same couple were allowed to be included in this study. That is, when they failed the first IUI cycle, they were given a second IUI treatment.Intervention(s)No intervention.Main Outcome Measure(s)A total of 550 cycles met the inclusion criteria, and 374 (68.0%) cycles met the inclusion criteria for the continuous group,the remaining 176 (32.0%) cycles with at least one or more menstruations between two IUI cycles were included in the delayed group.The primary outcome measure was clinical pregnancy rate (CPR), with secondary outcomes including abortion rate. Differences in clinical pregnancy rate (CPR)、abortion rate were compared between the two groups.ResultThere was no significant difference between the continuous group and the delayed group in female age, male age, infertility duration, infertility type, female BMI, endometrial classification, endometrial thickness, semen volume before treatment, sperm density before treatment, percentage of forward motile sperm before treatment, sperm density after treatment, and percentage of forward motile sperm after treatment. There were no statistical differences between the delayed group vs continuous group regarding the clinical pregnancy rate (20.5 % vs 21.9 %) and abortion rate (27.8% vs 22.0%)（P>0.05）. The above factors were included for binary logistic regression analysis. It was found that the increase of endometrial thickness promoted the clinical pregnancy rate, which was statistically significant (OR=1.205, 95% CI 1.05-1.384,P=0.008). Compared with primary infertility, secondary infertility can promote the improvement of clinical pregnancy rate, which is statistically significant (OR=2.637,95%CI 1.313-5.298,P=0.006). The effect of time interval between IUI on clinical pregnancy was not statistically significant (OR=1.007,95% CI 0.513-1.974,P=0.985).ConclusionsOverall, prolonging the interval between two IUI did not significantly improve pregnancy outcomes. Unless there are clear clinical indications, it is not necessary to deliberately prolong the interval between two treatments.

Dracaena arborea (Dracaenaceae) Increases Sexual Hormones and Sperm Parameters, Lowers Oxidative Stress, and Ameliorates Testicular Architecture in Rats with 3 Weeks of Experimental Varicocele

Varicocele is a disease characterized by an abnormal dilation of the pampiniform plexus that drains the testis. The main objective of this work was to evaluate the curative effects of aqueous and ethanolic extracts of Dracaena arborea on some reproductive and antioxidant markers in rats with experimental varicocele. Following varicocele induction, rats (5 per group) were randomly partitioned into untreated varicocele, vitamin E-treated (150 mg/kg), aqueous extract-treated (500 mg/kg), and ethanolic extract-treated (100 mg/kg) animals. Two other groups served as normal and sham-operated. After 2 or 4 weeks of treatments, body and sex organ weights, spermatozoa characteristics, antioxidant status, NO level, sex hormones, and testis histology were measured. Animals with 3 weeks of varicocele showed a significant ( p < 0.05–0.001) decrease in body and sex organ weights, total proteins, sperm characteristics, testosterone concentration, SOD, catalase, and total peroxidase activities. An increase in the plasmatic FSH, LH, and testicular MDA and NO concentrations was also recorded. Moreover, marked disorganization of the testicular architecture was observed. Treatment with D. arborea significantly reversed these impairments due to varicocele. For instance, after 4 weeks, treatment with aqueous extract of D. arborea significantly ( p < 0.05–0.001) increased testes and epididymis weights, sperm viability (89.12 ± 1.09 vs 68.22 ± 1.42), sperm density (148.50 ± 2.59 vs 110.25 ± 2.51), and sperm motility (68.16 ± 2.39 vs 55.88 ± 3.20) in the left side, compared with varicocele-untreated rats. The extract also significantly ( p < 0.05–0.001) decreased malondialdehyde level (2.19 ± 0.04 vs 3.50 ± 0.13) but elevated catalase (0.97 ± 0.03 vs 0.55 ± 0.03), SOD (0.5 ± 0.03 vs 0.15 ± 0.03), and peroxidase (65.80 ± 2.9 vs 40.95 ± 2.44) activities. Present results showed that D. arborea extracts possess antioxidant effects and improve sperm quality in male rats with an existing varicocele.

Effects of the Aqueous Extract of Cnestis ferruginea Leaves on the Male Reproductive System in Alloxan-induced Diabetic Mice

Diabetes mellitus is the most common endocrine disease and one of the most common chronic disorders. Diabetes mellitus has been associated with impaired reproductive health, mainly in men. The present study aimed to evaluate effects of the aqueous extract of Cnestis ferruginea leaves on the male reproductive system in alloxan-induced diabetic mice. The determination of phenolic compounds content in the aqueous extract was performed by conventional methods. Diabetes was induced in adult male mice by intraperitoneal injection with a single dose of 220 mg/Kg body weight of alloxan. Animal’s treatment with 100 and 200 mg/Kg of body weight of the aqueous extract was started 34 days after induction of diabetes. Sperm density, morphology and motility were assessed by standard methods. Serum levels of testosterone, FSH, and LH were measured. In addition, the testes were removed for histological study. Proportions of total polyphenols, flavonoids and tannins in the aqueous extract of Cnestis ferruginea were 5260.32 +/- 26 mg EAG /100g, 1384.43 +/- 4 mg EQ /100g and 7380.95 +/- 121 mg EC /100g respectively. 100 and 200 mg/Kg of body weight doses induced highly significant (P ˂ .001) reductions of 68% and 71% in blood glucose levels respectively, significant (P ˂ .05) increases in testes weight of 9% and 10% respectively, highly significant (P ˂ .001) increases in testosterone, pituitary gonadotropins and sperm parameter levels. In addition, regeneration of seminiferous tubules and interstitial cells was observed. Cnestis ferruginea leaves are rich in phenolic compounds. These compounds have anti-diabetic and fertilizing activities on the male reproductive system of diabetic mice.

Comparison of Success Rate and Complications of Open Inguinal and Subinguinal Varicocelectomy: A Randomized Controlled Trial

Background: Varicocele is the most common surgically correctable cause of infertility in men. The aim of this study was comparing the success rate and complications of open inguinal and subinguinal varicocelectomy.Methods: This randomized clinical trial was conducted from March 2017 to 2018. Sixty six patients that met inclusion criteria for varicocelectomy (persistent pain or impaired spermogram) were sequentially randomly allocated to inguinal and subinguinal varicocelectomy. Sperm analysis was performed in all patients before surgery. One week, one and three months after the operation patients were visited and evaluated for pain intensity (using visual analog scale) and complications( recurrence, hematoma, scrotal swelling, infection, hydrocele formation and numbness). Follow- up spermogram was done three months after varicocelectomy to evaluate the success of operation. Results: Mean time of operation was17.33±2.18 and 22.48± 4.45 minutes in the subinguinal and inguinal varicocelectomy method, respectively (p=0.001, CI95%=3.41-6.89). Semen analysis three months after surgery showed that there was no significant differences between semen volume (p=0.85), sperm count (p=0.09), sperm density (p=0.13), normal morphology (p=0.07) and motility (p=0.47) in both groups. There was no significant difference in pain density one week (p=0.51), one (p=0.29) and three months (p=0.67) after surgery between two groups. There wasn't any significant difference between two groups regarding recurrence of varicocele (p=0.67). Conclusions: In this study subinguinal varicocelectomy was superior to inguinal method due to less time of operation. However, there was no significant difference in postoperative complications and improvement of spermogram. Further studies with longer period of follow-up are recommended to compare fertility rate in inguinal and subinguinal varicocelectomy.Trial registration: IRCT, IRCT201710131323N12, Registered 20 October 2017, Retrospectively registered, https://www.irct.ir/trial/581

Letrozole Versus Clomiphene Citrate for Superovulation in Intrauterine Insemination Cycles - Retrospective Comparative Study Conducted in the Department of Reproductive Medicine, Mahatma Gandhi Medical College & Hospital, Jaipur

BACKGROUND Superovulation with intrauterine insemination (IUI) increases the probability of pregnancy by increasing number of oocytes and sperm density. The main oral agents used for superovulation are clomiphene citrate and letrozole. Clomiphene citrate a selective estrogen receptor modulator with predominant antiestrogenic action has adverse effects on endometrium and cervical mucous. Letrozole an aromatase inhibitor acts by reducing estrogen production by blocking androgen conversion to estrogen in ovary and peripheral tissues without affecting endometrium and cervical mucous. We wanted to compare the efficacy of letrozole v/s clomiphene citrate as first line ovulation induction drug in infertile patients undergoing IUI. METHODS Based on the inclusion and exclusion criteria, we included 160 patients in our study. They were divided into two groups of 80 each based on the drug given for ovulation induction. The drug was given for 5 days from D3 to D7 of menstrual cycle. IUI was done 38 - 40 hours after trigger and after confirmation of ovulation by sonography. RESULTS The mean age, body mass index (BMI), duration of infertility, type and cause of infertility in both groups were similar. Ovulation rate was 85 % in letrozole group and 71.25 % in clomiphene citrate group, which was statistically significant (P0.035). There was no statistically significant difference between endometrial thickness and total days till ovulation between two groups. Monofolliculogenesis and clinical pregnancy rate were statistically significantly higher in letrozole group. CONCLUSIONS Our study shows that letrozole has better pregnancy rate in comparison to clomiphene citrate (C. C.) in IUI cycles with less risk of anovulation, thin endometrium and multi follicular growth. KEY WORDS Clomiphene Citrate (C.C.), Letrozole (LTZ), Ovulation Induction (OI), Intrauterine Insemination (IUI)

P–126 A comparison of the efficacy of sperm freezing using high security tubes versus high security straws

Study question Are there any advantages in using High security tubes rather than High Security straws for conventional slow sperm freezing? Summary answer Freezing sperm in High Security tubes (HST) improved post-thaw recovery rate and motility, and also reduced processing and handling compared to High Security straws (HSS). What is known already The use of High Security freezing consumables (HSFC) in an IVF setting is a safe and effective way of eliminating concerns related to viral cross-contamination during storage. The lower diameter of HSS does make them susceptible to warming during handling. The HSFC used in this study is the only CE marked products that are made of resin, leak-proof and shatter-proof in all cryogenic temperatures even in LN2. No previous studies have compared the use of HST with HSS for conventional human sperm freezing. This study sets out to investigate the performance of HST compared to HSS. Study design, size, duration The study was designed as a controlled split-sample study with blind post-thaw analysis. Following the routine WHO analysis of 20 semen samples, the remainder of each of the samples was evenly divided and cryopreserved by conventional slow freezing in each of the two different HSFC. The freeze was conducted simultaneously by the same practitioner, employing the same freezing protocol and cryoprotectant. The pre-freeze and post-thaw concentration, total and progressive sperm motility were recorded. Participants/materials, setting, methods At one IVF clinic, semen samples with sperm density ≥15million/ml, ≥40% motility, ≥1.5ml were included. Cryoprotectant (SpermFreeze, Fertipro) was added dropwise to unprepared semen and kept at room temperature for 10 minutes before loading into HSFC (0.5ml CBS™HSS; CBS™HST). HSFC were heat-sealed (SYMS; SYMSIII sealers) and placed in vapour for 30 minutes before plunging into LN2. Samples were thawed by immersion in a 37Cº water bath for 5 minutes and analysed using WHO methods. Main results and the role of chance Paired-t test was used to compare the percentage motility between the different HSFC. All analysis was considered statistically significant when p &lt; 0.01. We demonstrated that the sperm recovery rate (Percentage total motility post-thaw/ Percentage total motility pre-freeze) in HST was 66.63 ± 14.94 (mean ± standard deviation) compared to 40.80 ± 14.69 in HSS. In the HSS, the percentage post-thaw total motility was 19.99 ± 7.21 and the percentage post-thaw progressive motility was 12.26 ± 2.59. In the HST, the percentage post-thaw total motility was 32.57 ± 8.33 and the percentage post-thaw progressive motility was 23.08 ± 5.53. The overall improvement when using HST against HSS was 12.53 ± 5.69, 10.44 ± 5.29 for the total motility and the progressive motility respectively. Comments were recorded regarding the handling and the condition of the HSS and HST for each freeze event. Neither device displayed any leakage of LN2 or any explosion during the warming. The freezing process was easier and faster using HST rather than HSS. It was also noted that the entire sample can be recovered from the HST, unlike the HSS. Limitations, reasons for caution The study looked at sperm recovery in terms of motility only. DNA damage was not considered as a parameter of sperm quality. Also, fertilization, pregnancy rates, live birth rates and the use of poorer quality sperm samples have not been investigated. Wider implications of the findings: For conventional sperm freezing, the use of HST resulted in improved sperm motility and progression post-thaw, when compared to HSS. This finding supports the use of HST to improve the post thaw quality of sperm, benefitting patients with own frozen samples, recipients of donor sperm and donor sperm banks. Trial registration number Not applicable.

Laparoscopic varicocelectomy in male infertility

Summary Background The suitability of laparoscopic varicocelectomy for assisted reproductive technology depends on the improvement of semen parameters. The present study analyzed the improvement of semen parameters following laparoscopic varicocele ligation. Material and methods A retrospective study of the laparoscopic varicocele clippings at the Department of Urology of University Hospital of Kiel between the years 2007 and 2019 was conducted. The semen analyses according to WHO standards (sperm count, density, motility and morphology) were conducted before and 12 months after surgery. Screening for surgical complications took place at the time of the follow-up seminal analysis. Included were patients with oligozoospermia, asthenozoospermia and/or teratozoospermia (group 1, OAT) or with nonobstructive azoospermia (group 2, NOA). Results This study included data of 27 patients and 22 patients presented preoperative OAT (81%, group 1). Another 5 patients showed NOA (19%, group 2). Data of group 1 showed that semen parameters normalized in 32% of the patients after surgery. Significant improvement in total sperm count (p < 0.005), sperm density (p < 0.005) and total motile sperm count (p < 0.005) was observed. No deterioration of semen parameters was observed. In group 2 we detected spermatozoa in 1 case in the postoperative ejaculate. None of the patients showed complications according to the Clavien-Dindo classification, postoperative hydrocele formation or recurrence of varicocele at the time of control spermiogram. Conclusion Laparoscopic varicocelectomy is a valid therapeutic approach to improve semen parameters for further assisted reproductive techniques. Spermatogenesis may be induced for patients with NOA. Normalization of semen parameters can be achieved for patients with OAT.

Can Unlikely Neanderthal Chloride Channel CLC-2 Gene Variants Provide Insights in Modern Human Infertility?

Background/Aims: Neanderthals, although well adapted to local environments, were rapidly replaced by anatomically modern humans (AMH) for unknown reasons. Genetic information on Neanderthals is limited restricting applicability of standard population genetics. Methods: Here, we apply a novel combination of restricted genetic analyses on preselected physiological key players (ion channels), electrophysiological analyses of gene variants of unclear significance expressed in Xenopus laevis oocytes using two electrode voltage clamp and transfer of results to AMH genetics. Using genetic screening in infertile men identified a loss of CLC-2 associated with sperm deficiency. Results: Increased genetic variation caused functionally impaired Neanderthals CLC-2 channels. Conclusion: Increased genetic variation could reflect an adaptation to different local salt supplies at the cost of reduced sperm density. Interestingly and consistent with this hypothesis, lack of CLC-2 protein in a patient associates with high blood K+ concentration and azoospermia.