Effect of the technical variability of counting chambers upon the interpretation of sperm concentration results.

RESEARCH QUESTION: Does the choice of sperm-counting chamber affect the proportion of samples generating results with an erroneous interpretation? DESIGN: Laboratories in an external quality assurance programme were sent 141 semen samples over a 12-year period and asked to return the sperm concentration and whether or not the result was abnormal. Only those using 5th edition of the World Health Organization manual (WHO5) interpretation criteria were included. Submissions from specialist fertility laboratories were used to calculate assigned values for each sample. Laboratory50 values determined the sperm concentration at which the laboratories reported a majority transition from abnormal to normal interpretations, i.e. the tipping point, which should coincide with the lower reference limit. RESULTS: The median and range of bias from the assigned values of each sample were determined for the Makler (-3.3%; -88.6% to +332.8%), haemocytometer (10.6%; -93.3% to +645.5%), Kova (+65.3%; -71.7% to +581.8%) and Vetriplast (+72.4%; -100.0% to +709.1) chambers. Laboratory50 values for the Makler (17.3  ×  106/ml), haemocytometer (13.6  ×  106/ml), Kova (10.0  ×  106/ml) and Vetriplast chambers (8.8  ×  106/ml) reflected the under- and overestimation of the chambers and confirmed a shift in the adjusted lower reference limit then used. The proportion of laboratories reporting erroneous interpretations of the four chambers for oligozoospermic samples were 10.9%, 15.1.%, 40.1% and 44.0%, respectively, and rose as the adjusted lower reference limit decreased. CONCLUSIONS: The between-laboratory and within-sample variation for all the chambers was high and remains a concern. The main impact of an increasing bias of the chambers was a lowering of the laboratory50 tipping point, resulting in an under-reporting of abnormal semen samples.

Retrospective study investigating the performance of the SQA-vision analyser compared with manual semen analysis.

The objective of this study was to compare the results of semen analysis using the manual method and the SQA-Vision sperm analyser after four years of practice and with a large cohort of patients. This was a comparative study of 1130 cases collected for semen analysis between October 2019 and October 2023, which were analysed simultaneously and independently by different operators using the manual microscopic method and an SQA-V automated analyser. For each sample, sperm concentration, progressive motility, motility, normal morphology, and round cells count were performed. There was no significant difference between the SQA-V method and manual assessment for all sperm parameters (Mann-Whitney test p > 0.05). According to the parameter studied, there was a strong correlation (rho = 0.81) and a very high correlation (rho = 0.98) between manual assessment and the SQA-V method. In the analysis of sperm concentration, the sensitivity and specificity were 0.90 and 0.99, respectively. The sensitivity and specificity for the analysis of sperm progressive motility were 0.98 and 0.99, respectively, while the sensitivity and specificity for the analysis of sperm motility were 0.87 and 0.99, respectively. The sensitivity and specificity for the analysis of normal morphology were 0.88 and 0.99, respectively. Regarding the analysis of round cells, the sensitivity and specificity were 0.98 and 0.99, respectively. The results of this retrospective study indicate that the SQA-V system offers satisfactory performance for routine sperm analysis.

Performance evaluation of sperm concentration, motility, and morphological analysis for GSA-810 series of sperm quality analysis system.

BACKGROUND: The performance evaluation of each computer-assisted sperm analysis (CASA) system may provide a basis for the interpretation of clinical results and further improvement of the CASA system. METHODS: The accuracy of the GSA-810 CASA system was evaluated by detecting latex bead quality control products. The precision of sperm concentration, morphology, and percentages of progressively motile sperm (PR) were evaluated by coefficient of variation (CV). Three samples with sperm concentration of about 100 × 106 /mL were diluted to evaluate the linear range. RESULTS: The detection values of latex beads were within the range of target values. The CVs of sperm concentration and PR were significantly and negatively correlated with sperm concentration (r = -0.561, p = 0.001) and PR value (r = -0.621, p < 0.001), respectively. The R2 values of the linear range of sperm concentration were ≥0.99. There was no significant difference in sperm motility and PR within 1-10 min at 36.5°C ± 0.5°C. The coincidence rates of sperm morphology and sperm head morphology for 36 semen samples analyzed by the GSA-810 system and manual method were 99.40% and 99.67%, respectively. The CVs of the percentage of sperm with abnormal morphology and percentage of sperm with abnormal head morphology were less than 5%. CONCLUSION: The GSA-810 system can accurately analyze normal semen samples, but the repeatability of the results is poor for oligozoospermia and asthenozoospermia samples. The future CASA system for analyzing sperm morphology should focus on recognizing the middle and tail segments of a spermatozoon.

Salvia officinalis L. Methanolic Extract Reduces Lead and Nicotine-Induced Sperm Quality Degeneration in Male Rats.

Most heavy metals and industrial chemicals such as nicotine and lead cause harm to the reproduction process through a decrease in sperm motility, fertilization process, and sperm binding to the oocyte. Salvia officinalis L. (sage) has been reported to enhance serum testosterone levels and other certain biochemical enzymes. Thus, the current study is aimed at evaluating the potential health benefits of S. officinalis L. methanol extract on lead and nicotine hydrogen tartrate-induced sperm quality degeneration in male rats and also identifying some of the non-polar volatile bioactive compounds that might be attributed to the bioactivity of S. officinalis extract using gas chromatography-mass spectrometry (GC/MS). In the study, fifty-four mature male albino rats of about 220-250 g [were divided randomly and equally into 9 groups (n=6)]. Sperm quality degeneration was induced through the oral administration of 1.5 g/L of lead acetate in drinking water or peritoneal injection of 0.50 mg/kg (animal weight) nicotine hydrogen tartrate for sixty days. Two doses (200 & 400 mg/kg b.w.) of S. officinalis L. were used. The rats were anesthetized after the experimental period and then sacrificed. Blood samples were collected while the epididymis, testicle, and accessory sex organs (prostates and seminal vesical) were taken for histopathological studies. Twelve major compounds were identified through the GC/MS analysis of S. officinalis L. methanol extract. Lead and nicotine toxicity had a great effect on the rats' sperm quality causing a significant (p<0.05) decrease in the quantity of sperm and sperm motility as well as an upsurge in the abnormalities of the sperm and a reduction in the length & diameter of seminiferous tubules and size & weight of sexual organs (accessory sex glands, epididymis, and testis). The administration of S. officinalis L. methanol extract, however, had a positive impact on the sexual organ weights, semen quality & quantity, and rats' fertility, thus, ameliorating the adversative effects of both lead and nicotine. Further evaluation and isolation of the bioactive components are recommended as potential drug leads.

Intracytoplasmic sperm injection versus conventional in-vitro fertilisation in couples with infertility in whom the male partner has normal total sperm count and motility: an open-label, randomised controlled trial.

BACKGROUND: The use of intracytoplasmic sperm injection has increased substantially worldwide, primarily in couples with non-male factor infertility. However, there is a paucity of evidence from randomised trials supporting this approach compared with conventional in-vitro fertilisation (IVF). We aimed to investigate whether intracytoplasmic sperm injection would result in a higher livebirth rate compared with conventional IVF. METHODS: This open-label, multicentre, randomised trial was done at two IVF centres in Ho Chi Minh City, Vietnam (IVFMD, My Duc Hospital and IVFAS, An Sinh Hospital). Eligible couples were aged at least 18 years and the male partner's sperm count and motility (progressive motility) were normal based on WHO 2010 criteria. Couples had to have undergone two or fewer previous conventional IVF or intracytoplasmic sperm injection attempts, have used an antagonist protocol for ovarian stimulation, and agree to have two or fewer embryos transferred. Couples were randomly assigned (1:1) to undergo either intracytoplasmic sperm injection or conventional IVF, using block randomisation with variable block size of 2, 4, or 8 and a telephone-based central randomisation method. The computer-generated randomisation list was prepared by an independent statistician who had no other involvement in the study. Embryologists and couples were not masked to study groups because of the type of interventions and differences in hospital fees, but clinicians performing embryo transfer were unaware of study group allocation. The primary outcome was livebirth after the first embryo transfer from the initiated cycle. Analyses were done on an intention-to-treat basis. The trial is registered with ClinicalTrials.gov, NCT03428919. FINDINGS: Between March 16, 2018, and Aug 12, 2019, we randomly assigned 1064 couples to intracytoplasmic sperm injection (n=532) or conventional IVF (n=532). Livebirth after the first embryo transfer from the initiated cycle occurred in 184 (35%) of 532 couples randomly assigned to intracytoplasmic sperm injection and in 166 (31%) of 532 couples randomly assigned to conventional IVF (absolute difference 3·4%, 95% CI -2·4 to 9·2; risk ratio [RR] 1·11, 95% CI 0·93 to 1·32; p=0·27). 29 (5%) couples in the intracytoplasmic sperm injection group and 34 (6%) couples in the conventional IVF group had fertilisation failure (absolute difference -0·9%, -4·0 to 2·1, RR 0·85, 95% CI 0·53 to 1·38; p=0·60). INTERPRETATION: In couples with infertility in whom the male partner has a normal total sperm count and motility, intracytoplasmic sperm injection did not improve the livebirth rate compared with conventional IVF. Our results challenge the value of the routine use of intracytoplasmic sperm injection in assisted reproduction techniques for this population. FUNDING: My Duc Hospital and Merck Sharp and Dohme.

The influence of the female reproductive tract and sperm features on the design of microfluidic sperm-sorting devices.

Although medical advancements have successfully helped a lot of couples with their infertility by assisted reproductive technologies (ART), sperm selection, a crucial stage in ART, has remained challenging. Therefore, we aimed to investigate novel sperm separation methods, specifically microfluidic systems, as they do sperm selection based on sperm and/or the female reproductive tract (FRT) features without inflicting any damage to the selected sperm during the process. In this review, after an exhaustive studying of FRT features, which can implement by microfluidics devices, the focus was centered on sperm selection and investigation devices. During this study, we tried not to only point to the deficiencies of these systems, but to put forth suggestions for their improvement as well.

Time intervals between semen production, initiation of analysis, and IUI significantly influence clinical pregnancies and live births.

PURPOSE: Does IDEF mapping help monitor the technical process of IUI and explore the potential improvements which might contribute to increased pregnancy and live birth rates? METHOD: Retrospective analysis of 1729 homologous IUI cycles of couples attending a fertility clinic in a university hospital setting. Standardized conventional semen parameters were analyzed and the semen samples prepared via discontinuous density gradient centrifugation. RESULTS: There was no significant association between sperm concentration, motility and morphology (analysis phase), and pregnancy outcome. Only female and male ages were significantly associated with the pregnancy outcome. There was a significant difference in the odds on clinical pregnancies and live births when analysis was ≤ 21 min initiated, and < 107 min between sample production and IUI, adjusted for male and female age. CONCLUSIONS: Adjusting for the couple's age, we could show that time intervals between semen production and analysis and IUI when kept low significantly influenced clinical pregnancies and live births.

Evaluation of the use of SYBR-14 and propidium iodide stain in a fluorescent computer-assisted spermatozoal quantification method in dogs.

The aim of this study was to evaluate the use of SYBR-14/propidium iodide (PI) stain in a computer-assisted spermatozoal quantification (CASQ) method of determining spermatozoal concentration in canine semen. In Experiment A, the spermatozoal concentration was measured (n = 52) with a haemocytometer and by CASQ under fluorescent illumination using green long-pass (G-LP) and red long-pass filters at measurement concentrations of <25 million/ml. For the red filter, the limits of agreement between the haemocytometer and CASQ were -6.3% to 6.8% and -7.5% to 6.2% between the haemocytometer and CASQ for the G-LP filter. For the red filter, the mean precision CVs were 2.21% ± 4.33% (mean ± 95% CI) for the haemocytometer, 2.19% ± 4.29% for CASQ and using the G-LP filter 2.13% ± 4.18% for the haemocytometer and 2.66% ± 5.21% for CASQ. In Experiment B, spermatozoa were also examined with a green spectrum short-pass (G-SP) filter (n = 50) at measurement concentrations of <12.5 million/ml. The limits of agreement between the haemocytometer and CASQ were -5.4% to 7.8% using the red filter, -15.8% to 14.3% using the G-LP filter and -13.1% to 11.3% using the G-SP filter. The mean precision CVs for the haemocytometer and CASQ, respectively, were 2.68% ± 5.26% (mean ± 95% CI) and 1.93% ± 3.72% using the red filter and 2.01% ± 3.95% and 3.55% ± 6.95% using the G-LP filter, and 3.96% ± 7.76% for CASQ using the G-SP filter. Using the red filter, the agreement between the haemocytometer and CASQ and the precision of both haemocytometer methods and CASQ were better than when using green filters. The CASQ method performed using green filters produced acceptable results; however, CASQ using a red filter with PI staining alone was superior to that using green filters and SYBR-14/PI staining.

At-home sperm testing for epidemiologic studies: Evaluation of the Trak male fertility testing system in an internet-based preconception cohort.

BACKGROUND: Semen quality assessment in population-based epidemiologic studies presents logistical and financial challenges due to reliance on centralised laboratory semen analysis. The Trak Male Fertility Testing System is an FDA-cleared and validated at-home test for sperm concentration and semen volume, with a research use only sperm motility test. Here we evaluate the Trak System's overall utility among men participating in Pregnancy Study Online (PRESTO), a web-based study of North American couples planning pregnancy. METHODS: US male participants aged ≥21 years with ≤6 months of pregnancy attempt time at study enrolment were invited to participate in the semen testing substudy after completing their baseline questionnaire. Consenting participants received a Trak Engine (battery-powered centrifuge) and two test kits. Participants shared their test results via smartphone images uploaded to online questionnaires. Data were then linked with covariate data from the baseline questionnaire. RESULTS: Of the 688 men invited to participate, 373 (54%) provided consent and 271 (73%) completed at least one semen test result. The distributions of semen volume, sperm concentration, motile sperm concentration, total sperm count, and total motile sperm count were similar to 2010 World Health Organization (WHO) semen parameter data of men in the general population. The overall usability score for the Trak System was 1.4 on a 5-point Likert scale (1 = Very Easy, 5 = Difficult), and 92% of participants believed they performed the test correctly and received an accurate result. Lastly, men with higher motile sperm count were more likely to report feeling "at ease" or "excited" following testing, while men with low motile sperm count were more likely to report feeling "concerned" or "frustrated." Overall, 91% of men reported they would like to test again. CONCLUSIONS: The Trak System provides a simple and potentially cost-effective means of measuring important semen parameters and may be useful in population-based epidemiologic fertility studies.

Molecular Alterations in Spermatozoa of a Family Case Living in the Land of Fires. A First Look at Possible Transgenerational Effects of Pollutants.

In our previous work, we reported alterations in protamines/histones ratio, in DNA binding of these proteins and their involvement in DNA oxidative damage in 84% of the young men living in the Land of Fires. In the present work, we extended our findings, evaluating any alterations in spermatozoa of a family case, a father and son, living in this area, to also give a first look at the possibility of transgenerational inherited effects of environmental contaminants on the molecular alterations of sperm nuclear basic proteins (SNBP), DNA and semen parameters. In the father and son, we found a diverse excess of copper and chromium in the semen, different alterations in SNBP content and low DNA binding affinity of these proteins. In addition, DNA damage, in the presence of CuCl2 and H2O2, increased by adding both the father and son SNBP. Interestingly, son SNBP, unlike his father, showed an unstable DNA binding and were able to produce DNA damage even without external addition of CuCl2, in line with a lower seminal antioxidant activity than the father. The peculiarity of some characteristics of son semen could be a basis for possible future studies on transgenerational effects of pollutants on fertility.

An update on the implication of physical activity on semen quality: a systematic review and meta-analysis.

PURPOSE: The aim of this study was to clarify whether physical activity may be associated with semen quality, considering the different types of sports, their intensity, and the semen parameters studied in the literature. METHODS: Eligible studies included those that evaluated the impact of physical activity in semen parameters in human population. Outcomes evaluated included the following seminal quality parameters: volume, concentration, total sperm count, progressive motility, total motility, total motile sperm count, morphology, and motile sperm concentration. RESULTS: We identified 32 manuscripts that analyzed this effect. Among them, 20 articles examined the role of general physical activity and 17 analyzed this relationship among specific sports. Although most results point to a lack of major effects of physical activity on semen quality, recreational physical activity could have a positive effect on semen concentration or progressive motility. On the contrary, elite physical activity could be detrimental for some semen parameters, such as progressive motility. Regarding specific sports, a negative effect of cycling on semen concentration is suggested. CONCLUSIONS: In conclusion, recreational physical activity seems to be of benefit for men with infertility issues. However, elite physical activity could have a detrimental effect on semen quality, which should be taken into consideration.

Development of a fluorescent computer-assisted spermatozoal quantification method and a comparison of results for manual counting with a haemocytometer and computer-assisted semen analysis in dogs.

The aim of this study was to develop and validate a novel, computer-assisted spermatozoal quantification (CASQ) method of determining spermatozoal concentration in canine semen. In Experiment A, the spermatozoal concentration was measured (n = 28) with a haemocytometer using light microscopy, CASQ and computer-assisted semen analysis (CASA; MMC sperm), following three independent dilutions. The limits of agreement between the haemocytometer and CASQ were -13.1% to 13.8% and -27.0% to 28.6% between the haemocytometer and CASA. The precision CVs (limits of agreement) were 5.7% (-7.8% to 8.9%) for the haemocytometer, 6.2% (-8.8% to 12.3%) for CASQ and 10.8% (-16.0% to 19.5%) for CASA. In Experiment B, spermatozoa were manually counted (n = 42) with the haemocytometer under fluorescent illumination using the CASQ sample. The limits of agreement between the CASQ and the haemocytometer were satisfactory (-4.6% to 4.6%) and the precision CVs (limits of agreement) were 6.2% (-9.0% to 11.4%) for the haemocytometer and 4.4% (-5.8% to 8.6%) for CASQ. The CASQ method was then clinically applied to compare the haemocytometer (light and fluorescent methods) with CASQ and CASA. Outlying data were removed. These studies demonstrated that CASQ was reliable and that the MMC sperm CASA was unreliable as methods for determining spermatozoal concentration in canine semen. Computer-assisted spermatozoal quantification was also determined to be more precise than manual counting with the haemocytometer. Using the clinical protocol, the agreement between the haemocytometer and CASQ method was acceptable, but it was worse than in the experiments where duplicate samples and a larger volume of semen were analysed. The CASQ method may be a useful method to measure the membrane status of canine spermatozoa; however, further investigation is required. Counting spermatozoa using fluorescent microscopy and the haemocytometer may improve the efficiency of counting and the accuracy of the method.

Measurement of membrane integrity in canine spermatozoa using a fluorescent computer-assisted spermatozoal quantification method and manual counting after eosin-nigrosin staining compared with manual counting after CFDA/PI staining.

The aim of this study was to compare measurement of spermatozoal membrane status using computer-assisted spermatozoal quantification (CASQ) and eosin-nigrosin (EN) staining with manual counting after CFDA/PI staining. Analysis was performed on both fresh and thawed cryopreserved canine semen. Membrane-disrupted spermatozoa (MDS) were counted using CASQ (n = 311) in an untreated sample and a completely membrane-disrupted sample, and the percentage of membrane-intact spermatozoa (MIS) calculated: (Total count - Untreated sample count) ÷ Total count × 100. Spermatozoa were stained with a one-step EN stain (n = 501), and then, at least 100 spermatozoa were manually examined under ×1,000 magnification and classified as MDS (stained with eosin) or MIS (non-stained). Spermatozoa from the same samples were also stained with CFDA/PI, and then, at least 200 spermatozoa were manually examined under ×1,000 magnification and classified as MIS (completely stained by CFDA) or MDS. The percentage of progressively motile spermatozoa (PMS) was determined by both computer-assisted semen analysis (CASA) and subjective methodologies, and the data were subsequently analysed to measure the agreement between the CASQ and EN methods with the CFDA/PI technique using Bland-Altman methodology. Pearson's correlation was measured between the MIS and PMS percentage samples and correlation coefficients compared. The mean MIS percentage was lower for CASQ and higher for EN than in CFDA/PI for all comparisons. The agreement of MIS percentage between CASQ and CFDA/PI was -20.2% to 32.0%, and between EN and CFDA/PI was -32.9% to 14.9%. In all methods, the MIS and PMS percentages were correlated (p < .001). Measurement of CFDA/PI appeared to be the most reliable and accurate method of determining MIS percentage in dogs. Further investigation is required to determine whether the CASQ technique can be improved. Eosin-nigrosin staining also appeared to be unreliable at MIS <80% and overestimated the MIS percentage.

Prevalence of abnormal semen analysis and levels of adherence with fertility preservation in men undergoing therapy for newly diagnosed cancer: A retrospective study in 2906 patients.

BACKGROUND: Sperm cryopreservation (freezing) should be offered to all men with cancer due to risk of infertility. However, many men with cancer already have impaired spermatogenesis prior to sperm cryopreservation. Furthermore, physical ill-health may hinder attendance of freeze visits. Investigating both the distribution of sperm functions and freeze attendance rates in men with newly diagnosed cancer, may identify patients benefiting from targeted reproductive fertility support. METHODS: We performed a retrospective study of 2906 male patients undergoing sperm cryopreservation prior to cancer therapy at a single UK tertiary centre between 1989 and 2013; all patients were asked to attend three hospital semen collection visits prior to cancer therapy. RESULTS: Fifteen per cent (433/2906) of men with newly diagnosed cancer had severely impaired semen quality (i.e., sperm total motile count, TMC < 1 million) during the first semen collection visit. However, patients with severely impaired semen quality had the poorest attendance of subsequent semen collection visits despite being requested to do so (non-attendance in TMC < 1 million: 43.4%; TMC < 1-30 million: 35.7%, P < 0.05 vs. <1 million; TMC > 30 million: 33.2%, P < 0.01 vs. <1 million). CONCLUSIONS: This study expands understanding of the semen quality of men with newly diagnosed cancer, and their ability to adhere to fertility preservation recommendations. Our data suggest that patients with the poorest semen quality paradoxically suffer the poorest attendance rates of sperm cryopreservation appointments prior to commencing cancer therapy. We suggest that additional support may be of clinical benefit to men with newly diagnosed cancer and TMC < 1 million sperm.

Curcumin nanomicelle improves semen parameters, oxidative stress, inflammatory biomarkers, and reproductive hormones in infertile men: A randomized clinical trial.

At least 50% of infertile couple's problems are related to male factor infertility. This Randomized, Double-Blind, Placebo-Controlled Clinical Trial conducted in Urology unit of Infertility Clinic on 60 infertile men. Patients were randomly assigned to one of intervention and placebo (n = 30) groups. Finally, 28 subjects in each group completed the study. Participants in the intervention group took daily 80 mg curcumin nanomicelle and those in the placebo group took daily placebo for 10 weeks. Semen analysis, anthropometric, physical activity assessments, total antioxidant capacity, malondialdehyde, inflammatory factors, and reproductive hormones were measured at the baseline and at the end of the study. At the end of study, statistically significant differences were seen in the total sperm count, sperm concentration, and motility in the intervention group to the control group. In treatment group, the total sperm count, sperm concentration, and motility levels were also statistically increased at the end of study compared to the baseline values. Curcumin nanomicelle supplementation also resulted in a statistically significant improvement in plasma levels of total antioxidant capacity, malondialdehyde, C-reactive protein, and tumor necrosis factor a in comparison to the placebo. Medical therapy of asthenoteratospermia with curcumin nanomicelle supplement could improve quality of semen parameters. However, further investigation is suggested in this regard.

Changes of Sperm Parameters Along Time Among Groups of Different Qualities.

BACKGROUND: Male infertility is solely responsible for approximately 20% of all infertility in couples. Various factors have been proposed as having a negative effect on sperm quality; however, the reasons for the global decline in sperm parameters during the last few decades are still controversial. OBJECTIVES: To investigate the fluctuations of semen parameters (sperm concentration, motility, and morphology) in three sperm quality groups and to examine the trends of those parameters in the same men over time. RESULTS: Our data showed deterioration in all semen parameters assessed in the group of men originally considered as having normal semen values according to the 2010 criteria of the World Health Organization. In contrast, we found significant improvement over time in all semen parameters in the group of men with severe oligo-terato-asthenozoospermia. CONCLUSIONS: Our results suggest that, although there were changes in sperm quality over time in the groups assessed, the clinical significance is negligible and does not necessarily justify a change in the therapeutic approach to infertility or sperm cryopreservation.

Prohibitin involvement in the generation of mitochondrial superoxide at complex I in human sperm.

Prohibitin (PHB), a major mitochondrial membrane protein, has been shown earlier in our laboratoryto regulate sperm motility via an alteration in mitochondrial membrane potential (MMP) in infertile men with poor sperm quality. To test if PHB expression is associated with sperm mitochondrial superoxide (mROS) levels, here we examined sperm mROS levels, high MMP and lipid peroxidation in infertile men with poor sperm motility (asthenospermia, A) and/or low sperm concentrations (oligoasthenospermia, OA). The diaphorase-type activity of sperm mitochondrial complex I (MCI) and PHB expression were also determined. We demonstrate that mROS and lipid peroxidation levels are significantly higher in sperm from A and OA subjects than in normospermic subjects, whereas high MMP and PHB expression are significantly lower. A positive correlation between mROS and lipid peroxidation and a negative correlation of mROS with PHB expression, high MMP, and sperm motility were found in these subjects. The finding of similar diaphorase-type activity levels of sperm MCI in the three groups studied suggests that the catalytic subunits of MCI in the matrix arm may produce mROS on its own. There may be a dysfunction of electron transport at MCI associated with decreased expression of PHB in sperm with poor quality. We conclude that mROS level is increased and associated with decreased PHB expression, and it may regulate sperm motility via increases in low MMP and lipid peroxidation. This is the first report on the involvement of PHB in human sperm motility loss associated with increased generation of mROS at MCI.

IN0523 (Urs-12-ene-3α,24ß-diol) a plant based derivative of boswellic acid protect Cisplatin induced urogenital toxicity.

The limiting factor for the use of Cisplatin in the treatment of different type of cancers is its toxicity and more specifically urogenital toxicity. Oxidative stress is a well-known phenomenon associated with Cisplatin toxicity. However, in Cisplatin treated group, abnormal animal behavior, decreased body weight, cellular and sub-cellular changes in the kidney and sperm abnormality were observed. Our investigation revealed that Cisplatin when administered in combination with a natural product derivative (Urs-12-ene-3α,24ß-diol, labeled as IN0523) resulted in significant restoration of body weight and protection against the pathological alteration caused by Cisplatin to kidney and testis. Sperm count and motility were significantly restored near to normal. Cisplatin caused depletion of defense system i.e. glutathione peroxidase, catalase and superoxide dismutase, which were restored close to normal by treatment of IN0523. Reduction in excessive lipid peroxidation induced by Cisplatin was also found by treatment with IN0523. The result suggests that IN0523 is a potential candidate for ameliorating Cisplatin induced toxicity in the kidney and testes at a dose of 100mg/kg p.o. via inhibiting the oxidative stress/redox status imbalance and may be improving the efflux mechanism.

Polar Quassinoids in Standardized Eurycoma longifolia Extract Formulated into a Lipid-Based Solid Dispersion to Improve Rat Sperm Count.

Eurycoma longifolia Jack is popularly sought in Southeast Asian countries for traditional remedies to improve sexual performance and fertility. 13α(21)-Epoxyeurycomanone and eurycomanone, two major quassinoids in a root extract (TAF2) were reported to improve rat spermatogenesis and fertility. Unfortunately, these quassinoids possess low bioavailability because of high aqueous solubility and low lipid membrane permeability. Often, other possible barriers may be P-glycoprotein (P-gp) efflux in the gut and presystemic hepatic metabolism. The present study attempted to solve these problems by formulating a lipid-based solid dispersion (TAF2-SD) of optimized mixture of TAF2 and emulsifiers, which was then orally administered to rats prior to sperm count analysis. The TAF2-SD-treated rats showed significantly twofold (p < 0.001) and fourfold (p < 0.001) higher sperm count than did TAF2-treated and vehicle-treated (control) rats, respectively. The study also demonstrated no significant in vitro ileal absorption changes of the quassinoids by P-gp efflux inhibitors and concentration change or secondary metabolite formation upon in vitro incubation with rat liver homogenates, suggesting that P-gp-mediated efflux and presystemic metabolism were not limiting their bioavailability. Further study on orally TAF2-treated rats confirmed that the area under the curve and bioavailability curve of each quassinoid in the absence and presence of ketoconazole were unchanged. Copyright © 2017 John Wiley & Sons, Ltd.

[Transcutaneous electrical acupoint stimulation for asthenozoospermia].

OBJECTIVE: To study the effect of transcutaneous electrical acupoint stimulation (TEAS) in the treatment of asthenozoospermia. METHODS: We randomly divided 72 asthenozoospermia patients into a 2 Hz TEAS (n = 29), a 100 Hz TEAS (n = 20), and a blank control group (n = 23), those in the former two groups treated by 30 minutes of TEAS at 2 Hz and 100 Hz respectively, applied to the acupoints of bilateral Shenshu, left Zusanli, and Guanyuan, once a day for 60 days, while those in the blank control group left untreated. Using computer-assisted sperm analysis (CASA), we examined sperm concentration and motility as well as the percentages of grade a and grade a+b sperm in different groups of the patients. RESULTS: Compared with the baseline, 2 Hz TEAS significantly increased sperm motility (ï¼»12.76 ± 1.39ï¼½ vs ï¼»18.89 ± 2.46ï¼½%, P<0.05) and the percentage of grade a+b sperm ( ï¼»10.68 ± 1.22ï¼½ vs ï¼»16.32 ± 2.10ï¼½%, P<0.05) in the asthenozoospermic patients, while 100 Hz TEAS improved not only sperm motility (ï¼»12.32 ± 2.21ï¼½ vs ï¼»23.81 ± 3.42ï¼½%, P<0.01) and the percentage of grade a+b sperm (ï¼»10.45 ± 1.98ï¼½ vs ï¼»20.25 ± 2.82 ï¼½%, P<0.01), but also the percentage of grade a sperm (ï¼»6.44 ± 1.16ï¼½ vs ï¼»13.31 ± 2.30ï¼½%, P<0.05). Moreover, in comparison with the blank control group, 2 Hz TEAS also remarkably increased sperm motility (ï¼»9.57 ± 1.60ï¼½ vs ï¼»18.89 ± 2.46ï¼½%, P<0.05) and the percentage of grade a+b sperm (ï¼»7.81 ± 1.31ï¼½ vs ï¼»16.32 ± 2.10ï¼½%, P<0.05) in the asthenozoosperma patients, while 100 Hz TEAS improved not only sperm motility (ï¼»9.57 ± 1.60ï¼½ vs ï¼»23.81 ± 3.42ï¼½%, P<0.01) and the percentage of grade a+b sperm (ï¼»7.81 ± 1.31ï¼½ vs ï¼»20.25 ± 2.82ï¼½%, P<0.01) but also the percentage of grade a sperm (ï¼»4.87 ± 1.01ï¼½ vs ï¼»13.31 ± 2.30ï¼½%, P<0.01). Meanwhile, the rate of clinical effectiveness was significantly higher in the 100 Hz TEASthan in the blank control group either in intention-to-treat (ITT) analysis (100% vs 18.18%) orper-protocol (PP) analysis (90% vs 0%), and so was it than in the 2 Hz TEAS group based on the data of ITT (100% vs 33.33%). CONCLUSIONS: Both 2 Hz and 100 Hz TEAS are effective for the treatment of asthenozoospermia by improving sperm motility and vitality.