Comparison of DNA fragmentation levels in spermatozoa with different sex chromosome complements.

RESEARCH QUESTION: Do spermatozoa with different sex chromosome complements (X and Y; aneuploidy and monosomy) exhibit different degrees of DNA damage? DESIGN: A prospective, observational study to measure the DNA fragmentation level and sex chromosome complement simultaneously using combined sperm chromosome dispersion (SCD) and fluorescence in-situ hybridization tests. Two methods were used to evaluate SCD images: a traditional semi-quantitative method to categorize halo size and a newly developed quantitative method based on the Matlab image analysis programme to more precisely measure the halo area and calculate the halo size index (HSI). RESULTS: The HSI (which was inversely proportional to DNA fragmentation level) of Y chromosome-bearing spermatozoa was significantly (P < 0.05) lower than that of X chromosome-bearing spermatozoa in both normozoospermic and pathozoospermic groups. The HSI of sex chromosome-aneuploid spermatozoa was also significantly (P < 0.05) lower than that of monosomic spermatozoa. CONCLUSIONS: Our results indicated that Y chromosome-bearing spermatozoa are more susceptible to DNA damage than X chromosome-bearing spermatozoa, and the segregation errors during the meiotic division of spermatogenesis (resulting in aneuploidy) constitute an important contributory cause of DNA damage.

The changing pattern of uterine contractions before and after fresh embryo transfer and its relation to clinical outcome.

In this prospective cohort study of 286 women undergoing fresh embryo transfer after IVF, uterine contraction frequency and direction were measured before (-5 min), 5 min after (+5 min) and 60 min after (+60 min) embryo transfer. Mean ± SD uterine contraction frequency at -5 min was 1.8 ± 1.1 contractions per min, increasing significantly (P < 0.05) to 2.0 ± 1.1 at +5 min, and returning back to baseline 1.8 ± 1.1 at +60 min. At -5 min, the proportion of women the with retrograde, antegrade, indeterminate direction and absent contractions were 33%, 44%, 17% and 6%; at +5 min, 40%, 42%, 13% and 5%, and at +60 min, 42%, 38%, 14% and 6%. No significant change was observed in the proportion of direction at these three time points. Logistic regression analysis showed live birth rate was significantly reduced in older women (P = 0.035) and in those with higher uterine contraction frequency at +5 min (P = 0.006). Frequency of uterine contraction immediately after embryo transfer (+5 min) seemed to be a significant predictor of IVF outcome and may help to identify women who could benefit from the use of muscle relaxant therapy to improve outcome.

Assessment of the embryo flash position and migration with 3D ultrasound within 60 min of embryo transfer.

STUDY QUESTION: Does the air bubble (embryo flash) position and migration as visualized with 3D ultrasound (US) within 60 min of embryo transfer correlate with clinical outcome following fresh ART transfer cycles? SUMMARY ANSWER: The location of the embryo flash and the direction of its movement at 60 min, but not at 1 or 5 min after transfer, are associated with clinical pregnancy. WHAT IS KNOWN ALREADY: Studies assessing the relation between the pregnancy rate and the position of the catheter tip and/or the position of the air bubbles following embryo transfer show conflicting results to date. STUDY DESIGN, SIZE AND DURATION: This was a prospective cohort study including 277 infertile women undergoing ART between July 2011 and August 2013. PARTICIPANTS/MATERIALS, SETTING AND METHODS: Good prognosis patients undergoing fresh ART cycles within a single tertiary University unit were assessed by 3D US at 1, 5 and 60 min after embryo transfer. The distance of the embryo flash from the fundus was measured at these time points, along with the direction of the embryo flash movement within 60 min of transfer. MAIN RESULTS AND THE ROLE OF CHANCE: Within 60 min of embryo transfer, 76.4% (198/259) of the embryo flashes migrated towards the fundus, 12.4% (32/259) migrated towards the cervix and 11.2% (29/259) remained static. There was no significant association between the embryo position or movement and the pregnancy rate at 1 and 5 min. At 60 min, however, the pregnancy and implantation rates among subjects with embryo flashes located <15 mm from the fundus was significantly higher than those with embryo flashes located >15 mm from the fundus (46.5 and 32.8% versus 25.8 and 18.2%, respectively; P < 0.05). The pregnancy and implantation rates when the embryo flash was seen moving towards the cervix (25.0 and 15.0%) was significantly lower (P < 0.05 and P < 0.01, respectively) compared with those remaining static (55.2 and 37.7%) or moving towards the fundus (45.5 and 32.8%). LIMITATIONS AND REASONS FOR CAUTION: Although the air bubbles seen at the time of embryo transfer are thought to represent the position of the embryo, they are in fact a surrogate marker of the embryo itself, as this cannot be directly visualized by US. WIDER IMPLICATIONS OF THE FINDINGS: Following embryo transfer the majority, but not all, embryo flashes undergo significant migration towards the fundus. The location of the embryo flash and the direction of its movement at 60 min, but not at 1 or 5 min after transfer, is associated with clinical outcome. These findings may challenge the traditional notion that the exact position of the embryo flash immediately following embryo transfer is related to clinical outcome. STUDY FUNDING/COMPETING INTERESTS: The authors have no relevant funding or conflicts of interest to disclose.

Blood metal/metalloid concentration of male subjects undergoing IVF/ICSI treatment outcomes: A prospective cohort study.

BACKGROUND: Previous epidemiology studies reported that heavy metal/metalloid exposure is associated with the impairment of semen quality. However, it is still not clear whether the in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment outcome will be affected after the heavy metal/metalloid exposure of the male partners. METHODS: A prospective cohort study with a 2-year followed-up was conducted in a tertiary IVF center. A total of 111 couples undergoing IVF/ICSI treatment were initially recruited from November 2015 to November 2016. Male blood concentrations of heavy metal/metalloid including Ca, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Mo, Cd, Hg, and Pb were measured by inductively coupled plasma mass spectrometry, and the lab and pregnancy outcome data were followed up. The associations between male blood heavy metal/metalloid concentration and the clinical outcomes were analyzed by Poisson regression analysis. RESULTS: Our results showed that none of the heavy metal/metalloid of male partners we investigated are significantly associated with the oocyte fertilization and good embryo (P ≥ 0.05); however, antral follicle count (AFC) was a protective factor for the oocyte fertilization (RR: 1.07, 95 % CI: 1.04-1.10). The blood Fe concentration of the male partner was positively associated (P < 0.05) with pregnancy in the first fresh cycle (RR:170.93, 95 % CI: 4.13-7082.04), cumulative pregnancy (RR: 23.61, 95 % CI: 3.25-171.64) and cumulative live birth (RR: 36.42, 95 % CI: 1.21-1092.54). In the first frozen embryo cycles, pregnancy was significantly associated (P < 0.05) with the blood Mn (RR: 0.01, 95 % CI:0.00-0.11) and Se concentration (RR: 0.01, 95 % CI:8.25 E-5-0.47) and female age (RR: 0.86, 95 % CI:0.75-0.99); live birth was significantly associated (P < 0.05) with the blood Mn concentration (RR: 0.00, 95 % CI: 1.14E-7-0.51). CONCLUSIONS: Our results suggested that the higher male blood Fe concentration was positively associated with pregnancy in the fresh embryo transfer cycle, cumulative pregnancy, and cumulative live birth, whereas the higher male blood Mn and Se concentration were associated with lower chance of pregnancy and live birth in the frozen embryo transfer cycle. However, the underline mechanism of this finding still needs further investigation.

In vitro rescue immature oocytes - a literature review.

Immature oocytes retrieved from in vitro fertilization (IVF) and clinical in vitro maturation (IVM) is a common problem, especially in patients with advanced age, poor ovarian response (POR), or polycystic ovary syndrome (PCOS). Considering there is no common name to describe this group of oocytes, we suggest naming all of immature oocytes retrieved from IVF and clinical IVM cycles as 'Medical Unusable Oocytes' (MUO) as none of them will be used for subsequent treatment and will eventually be discarded. Scientists attempt to improve the clinical utilization rate of MUO instead of discarding them. Rescue IVM and mitochondria supplementation may be available approaches to mature MUO. We propose a specific definition of rescue IVM, namely the cultivation and maturation of immature oocytes in vitro collected from IVF cycles with human chorionic gonadotropin (hCG) trigger. Rescue IVM is usually mixed up with clinical IVM. Clarification of the differences between rescue IVM and clinical IVM is necessary. This manuscript aims to clarify the rather confusing IVM procedures and review existing methods of improving rescue IVM, currently available information on the success rate, and explore the future possibility of rescue IVM serving as a promising tool in reproductive medicine.

Associations between blood metal/ metalloid concentration and human semen quality and sperm function: A cross-sectional study in Hong Kong.

BACKGROUND: The association between metal/metalloid exposure on human sperm quality is still inconclusive. There is a lack of data on the effect of metal/metalloid on sperm function. METHODS: The aim of this study was to clarify the association between blood metal/metalloid concentration and traditional and functional sperm parameters, the blood concentration of Pb, Hg, Cd, As, Ni, Mo, Zn, Cu, Se, Fe, Mg, Cr and Ca of 288 men in Hong Kong were assessed by inductively coupled plasma-mass spectrometry, and sperm parameters including sperm concentration, motility, morphology, vitality, total sperm count, total motile sperm count, sperm DNA fragmentation and sperm acrosome reaction were measured. Demographic and lifestyle questionnaires were also provided for all participants. Multivariable linear regression analysis was performed to test the association between blood metal/ metalloid concentration and semen parameters after adjusting for relevant confounding variables. RESULTS: The results showed that moderate to high level of blood Pb concentration (>27.19 µg/L) appeared to be negatively associated with sperm morphology (P < 0.05); high level of blood Cd concentration (>1.44 µg/L) was negatively associated with sperm acrosome reaction (P < 0.05); Mo was positively associated with semen volume (P < 0.05), however, high level of blood Mo concentration (>13.52 µg/L) was negatively associated with sperm vitality (P < 0.05); high level of blood Zn concentration (>6.20 mg/L) was positively associated with sperm vitality (P < 0.05); moderate level of blood Fe concentration (526.89-566.63 mg/L) was positively associated with sperm acrosome reaction (P < 0.05); moderate level of blood Ca concentration (55.92-66.10 mg/L) was positively associated with semen volume (P < 0.05); however, lower level of blood Ca concentration (45.90-55.92 mg/L) was negatively associated with sperm morphology (P < 0.05). CONCLUSIONS: Our results suggested that the sperm function could be affected by blood Cd and Fe concentration and traditional sperm parameters could be affected by blood concentration of Mo, Zn, Pb and Ca.

Influence of the Period of Abstinence on Semen Quality in a Patient with Systemic Lupus Erythematosus: A Case Report and Review of the Literature.

Systemic lupus erythematosus (SLE) is a chronic inflammatory disease that can affect fertility. There is currently little information regarding the semen profile of males with SLE. Moreover, there is no consensus on an appropriate period of sexual abstinence for semen analysis and on the use of DNA fragmentation assay, together with multiple semen analyses to document the semen profile in this clinical population. In this case report, multiple semen analyses, with DNA fragmentation assays, were performed for a male with SLE undergoing fertility treatment at our andrology unit. A 4-day period of abstinence improved the semen concentration, total sperm count, total progressive motile sperm, and sperm morphology, with minimal DNA fragmentation. In conclusion, multiple semen analyses obtained after different periods of sexual abstinence, together with DNA fragmentation assays, may be useful to develop a semen profile for patients with SLE, providing information on the optimal abstinence period to yield the best semen quality for subsequent fertility treatment. For patients with fluctuating semen results, concomitant semen cryopreservation should be considered to preserve the better quality semen before starting assisted reproductive technologies if pregnancy is planned in the future.

A double-blinded, randomized placebo-controlled trial on the effect of traditional Chinese medicine formula Wuzi Yanzong pill on improving semen qualities in men with suboptimal parameters.

BACKGROUND: In Hong Kong, one of six couples is affected by subfertility problems. Male infertility contributes to half of the infertility cases. In male infertility, there is no effective treatment for patients with idiopathic infertility/poor semen parameters. Recent meta-analysis results suggest that a traditional Chinese medicine (TCM) formula - Wuzi Yanzong pill - showed a curative effect on male fertility. However, the heterogeneity of the studies could not draw a definitive conclusion on the therapeutic effect of this formula. The aim of this study is to conduct a well-designed randomized controlled trial to investigate the effect of TCM formula Wuzi Yanzong pill on improving semen qualities in men with suboptimal parameters. METHODS: This study is a double-blinded, randomized placebo-controlled trial conducted in a public hospital in Hong Kong. Participants will be randomized, using computer-generated random numbers, with a 1:1 ratio to either the Wuzi Yanzong pill formula group or the placebo group. Both groups will be administered the drugs for 12 weeks. Participants will have a total of four visits for their semen and blood assessments for a 6-month period, and we will follow up for another 6 months to record their conception outcome. The primary outcome is to compare the total motile sperm count, natural conception rate, and pregnancy outcome to those under placebo treatment. Secondary objectives are sperm functions and assisted reproductive technology outcome. DISCUSSION: To date, there are no studies using the disclosed Wuzi Yanzong formula or double-blinded, randomized trials. The Wuzi Yanzong TCM formula may provide a good clinical solution for subfertile males for which contemporary western medicine has no cure. Therefore, a well-designed randomized trial for evaluating the effect of Wuzi Yanzong TCM formula is urgently needed. TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR), ChiCTR-INR-17010790 . Registered on 27 February 2017. Centre for Clinical Research and Biostatistics - Clinical Trials Registry, CUHK_CCRB00548 . Registered on 27 February 2017.

Lifestyle and demographic factors associated with human semen quality and sperm function.

In this study, we sought to investigate the effect of lifestyle and demographic factors on classic and functional semen parameters. Three hundred and twenty-eight subjects who underwent semen analysis were recruited. Routine SA, sperm vitality, acrosome reaction (AR) assay and sperm DNA fragmentation index (DFI) were analyzed. Demographic and lifestyle information, including (1) BMI, (2) current smoking and alcohol drinking frequency, (3) sleep habits, (4) daily fluid intake, (5) weekly meat intake, (6) sports frequency, (7) trouser cell phone use, (8) age, and (9) abstinence time, were collected. Generalized additive models were used to analyze the possible non-linear association. The results showed that total sperm count (TSC) was significantly associated with age (P = 0.001), abstinence time (P = 0.001) and daily coffee intake (P = 0.044). Semen volume was significantly associated with age (P < 0.001) and daily coffee intake (P < 0.001). Sperm concentration was significantly associated with abstinence time (P = 0.011) and average sleep duration (P = 0.010). Sperm motility was significantly associated with age (P = 0.002) and daily juice intake (P = 0.001). Total motile sperm count was significantly associated with age (P = 0.003) and abstinence time (P = 0.009). DFI was significantly associated with age (P = 0.002), irregular sleeping habit (P = 0.008) and abstinence time (P = 0.032). The percentage of AR sperm was significantly associated with daily juice intake (P = 0.013). In conclusion, DFI and TSC were the most sensitive semen parameters for demographic and lifestyle features, whereas age had more influence on semen parameters than other demographic and lifestyle features. ABBREVIATIONS: BMI: body mass index; SA: semen analysis; AR: acrosome reaction; DFI: DNA fragmentation index; GAM: generalized additive model; TSC: total sperm count; TMC: total motile sperm count; IUI: intrauterine insemination; SCSA: sperm chromatin structure assay; SD: standard deviation; IQR: interquartile range; CBAVD: congenital bilateral absence of vas deferens; NEQAS: national external quality assessment service; HTF: human tubal fluid; HSA: human serum albumin.

Effects of mechanical stresses on sperm function and fertilization rate in mice.

In this study, we investigated whether any of the observed changes in mouse sperm function tests secondary to mechanical stresses (centrifugation and pipetting) correlate with sperm fertilization ability. Chinese Kunming mice were used as sperm and oocyte donors. Sperm samples were allocated evenly into centrifugation, pipette, and control groups. Sperm plasma membrane integrity (PMI), mitochondrial membrane permeability (MMP), baseline and stimulated intracellular ROS, and sperm fertilization ability were measured by hypo-osmotic swelling, flow cytometry, and fertilization tests. Parallel studies were conducted and all tests were repeated six times. Our results showed that after centrifugation, the progressive motility, average path velocity, and overall sperm motility and PMI decreased significantly (p < 0.05). In addition, the MMP level decreased significantly in viable sperm when the centrifugation condition reached 1,400 g × 15 minutes (p < 0.05). When pipetting was performed two or more times, progressive motility, average path velocity, and overall sperm motility decreased significantly (p < 0.05); when it was performed four or more times, sperm membrane integrity and intracellular basal ROS level of viable sperm was also significantly decreased (p < 0.05). In conclusion, various mechanical stresses seem to affect sperm function, however this does not appear to alter fertilization rate. Laboratory handling steps should be minimized to avoid unnecessary mechanical stresses being applied to sperm samples.