Healthy Sperm Research Articles

Sperm-carried IGF2: towards the discovery of a spark contributing to embryo growth and development.

Spermatozoa have been shown to carry key RNAs which, according to animal evidence, seem to play a role in early embryo development. In this context, a potential key growth regulator is insulin-like growth factor 2 (IGF2), a highly conserved paternally expressed imprinted gene involved in cell growth and proliferation which, recent observations indicate, is expressed in human spermatozoa. We herein hypothesized that sperm IGF2 gene expression and transmission at fertilization is required to support early embryo development. To test this hypothesis, we analyzed sperm IGF2 mRNA levels in the same semen aliquot used for homologous assisted reproductive technique (ART) in infertile couples and correlated these levels with embryo morphokinetics. To find a mechanistic explanation for the observed results, the transcriptomes of blastocysts obtained after injection of Igf2 mRNA in mouse parthenotes were analyzed. Sperm IGF2 mRNA negatively correlated with time of 2-cell stage (t2), t3, t4, t5, and time of expanded blastocyst (tEB), independently of maternal age, body mass index, anti-Müllerian hormone levels, and oocyte quality. An IGF2 mRNA index >4.9 predicted the ability of the embryos to reach the blastocyst stage on Day 5, with a sensitivity of 100% and a specificity of 71.6% (AUC 0.845; P < 0.001). In the animal study, transcriptome analysis demonstrated that 65 and 36 genes were, respectively, up- and down-regulated in the experimental group compared to the control group. These genes belong to pathways that regulate early embryo development, thus supporting the findings found in humans. This study has the potential to challenge the longstanding tenet that spermatozoa are simply vehicles carrying paternal DNA. Instead, it suggests that IGF2 mRNA in healthy spermatozoa provides critical support for early embryo development. Pre-ART sperm-carried IGF2 mRNA levels may be used as a marker to predict the chances of obtaining blastocysts to be transferred for infertile couples undergoing ART.

Protective Effects of Chitosan-Loaded Pomegranate Peel Extract Nanoparticles on Infertility in Diabetic Male Rats.

Diabetes Mellitus (DM) is known to have an impact on the health of the male reproductive system. It is linked to low sperm quality, increased oxidative stress, and an increased generation of reactive oxygen species in the seminal fluid. Pomegranate extract has phenolic compounds and significant protective properties against oxidative stress, male sex hormone disruptions, and sperm abnormalities. The current study aimed to evaluate the effectiveness of Pomegranate Peel Extract Nanoparticles (PPENPs) on male fertility in diabetic rats. DM was induced in rats by intraperitoneal injection of streptozotocin (60 mg/kg). Twenty-four rats were divided into four groups, 6 rats in each group: control, DM, DM+empty NPs (60 mg/kg, orally), and DM+PPENPs (60 mg/kg, orally). Administration of PPENPs increased the levels of insulin, FSH, LH, testosterone, catalase, glutathione reduced, and semen fructose. PPENPs also improved sperm quality, as seen by improvements in sperm morphology, motility, count, and the ability of metabolically active spermatozoa to convert blue resazurin dye to pink resorufin. However, PPENPs decreased levels of glucose, malonaldehyde, nitric oxide, and sperm abnormalities. Also, histological investigation of the PPENPs showed improvement in testis tissue architecture and increased the diameter size of seminiferous tubules and germinative layer thickness. Our investigation proved that the treatment of PPENPs has a protective effect on the reproductive system of male diabetic rats, improving fertility parameters, healthy sperm profiles, and the antioxidant system.

A SYSTEMATIC REVIEW OF THE CRUCIAL ROLE OF RASA DHATU IN PRODUCING HEALTHY PROGENY.

Each year in India, there are approximately 28 million pregnancies, in which 26 million live births, a million neonatal deaths &amp; congenital deformities is occurring due to malnutrition of mother and fetus. The classics of Ayurveda detail the crucial role of Rasa dhatu in the production of healthy progeny. During gametogenesis, Rasa dhatu is the foremost requirement for normal, healthy sperm and oocytes (Shukra &amp; Artava). Malformed paternal Rasa dhatu leads to malnourished and malformed gametocytes through mitochondrial dysfunction, epigenetics and defects in meiosis cause Bija dosha, which in turn lead to Garbhshrava (miscarriage), deformed Garbha formation or congenital deformities like Suchi Mukh uterus in female fetus and sterile progeny. After fertilization, the ceased Artava (accumulated nutritional substance in the fluid of decidual cells) acts as a storage nutrition for the developing embryo in the embryonic phase. During organogenesis the Artava which is the Upadhatu of Rasa dhatu forms the maternal part of placenta. After formation of placenta &amp; umbilical cord the Maternal Rasa dhatu provides directly nutrition to growing fetus. The malformed Rasa dhatu or the blocked Rasavaha channels of mother may cause different levels of intra uterine growth retardation such as Nagdhadara, Upavistaka, Lina garbha, Garbhashaya or Mritgarbha (IUD).

An Ayurvedic Management of Bandhyatva Associated with Bilateral Tubal Blockage

In Ayurveda, infertility is addressed under the concept known as "Bandhyatava," which is described in ancient texts such as Charak Samhita, Sushrut Samhita, Ras Ratna Samuchya, and Harit Samhita. Acharya Sushrut described four crucial factors for fertility: Ritu (menstruation), Kshetra (functioning reproductive organs), Ambu (digestive juices) and Beej (healthy sperm and ovum). It occurs due to the vitiation of Vata dosha where Kapha dosha also contributes to the formation of blockage. Beeja Grahana is unable due to Sanga Strotodushti of Artavavaha i.e., tubal blockage which leads to failure of conception. Fallopian tube is an essential part of the Artavavaha Srotas as the Beej Samagam (ovum and sperm) takes place here. In the current case scenario, Bandhyatva is due to Tubal Blockage. A 37-year-old female patient was reported to PTSR OPD with the complaints of irregular menses with decreased duration and wants to conceive irrespective of unprotected sexual intercourse since 3 years. Her HSG scan suggestive of B/L fallopian tube cornual blockage. Aims and Objectives: To maintain the balance between the four factors to conceive through Sanshodhan therapy. Patients has been treated with Sanshodhan Karma followed by 3 consecutive cycles of Sahacharadi Taila Uttarbasti and 3 cycles of Phalghrita Uttarbasti along with Shaman Chikitsa. Results: After completion of treatment Bilateral Tubes were patent observed in Sonohysterosalpingography. Conclusion: This case study found that the treatment administered was highly effective in addressing infertility caused by tubal blockage.

P-058 The role of sperm selection in improving embryo euploidy rate: a comparison between Swim-up and Microfluidic Sperm Selection

Abstract Study question Does Microfluidic Sperm Selection (MSS) improve the blastocysts’ development rate and the euploidy rate compared to Swim-up sperm selection? Summary answer MSS doesn’t improve the rate of blastocyst development and euploidy compared to Swim-up. Furthermore, in women age ≥ 37 years D6 aneuploid blastocyst rate increases What is known already Sperm defects have been shown to have a negative impact on embryo quality and development. MSS was introduced as an alternative to traditional centrifugation-based sperm preparation techniques to efficiently isolate highly motile and healthy spermatozoa. Centrifugation has been suggested to compromise membrane integrity, motility, mitochondrial energy production, and DNA structure. Less DNA fragmentation and ROS formation was observed in spermatozoa isolated with MSS compared to standard techniques. Our study aims to analyze whether MSS also improves biological outcomes. Study design, size, duration In a prospective study of 51 ICSI/PGT-A cycles performed between September 2022 and November 2023, freshly ejaculated semen sample was divided into two aliquots and processed by Swim-up and MSS. Split sperm sample was used to inject siblings’ mature oocytes (249 vs 274 oocytes in Swim-up vs MSS groups). Inclusion criteria: retrieved MII oocytes ≥8, oligoteratozoospermic and normozoospermic semen. Results were analyzed according to women’s age &amp;lt;37 years (21 cycles) and ≥37 years (30 cycles). Participants/materials, setting, methods Spermatozoa were selected using either conventional Swim-up or a microfluidics device (ZyMōtTM). Post treatment sperm parameters (sperm count; total sperm motility; progressive motility and hyperactivated motility) were examined for both. Embryo culture was performed in Irvine Continuous Culture-NX media until day5 or day6 and biopsy was performed at the blastocyst stage by next generation sequencing. Primary endpoint was fertilization rate, blastocyst development and usable blastocysts rate. Chi-square analysis was performed and p &amp;lt; 0.05 was considered significant Main results and the role of chance There is no significant differences overall between Swim-up and MSS in fertilization rate (73% vs 75%), blastocyst development rate (47% vs 54%) and ploidy rate: aneuploid (47% vs 50%), usable blastocyst (i.e. euploid+mosaics) (53% vs 50%). When considering female age, no significant difference is observed in the primary endpoints in the &amp;lt;37 years group. However, in women aged ≥37 years, the rate of blastocyst development is similar in the two sperm preparation groups, but a significant difference is observed in the timing of blastocyst development: The percentage of blastocysts formed on day 6 in the MSS group is significantly higher than in the Swim-up group (43% vs. 19% p &amp;lt; 0.05). In addition, the aneuploidy rate of blastocysts on day 6 is also significantly higher than in the Swim-up group (79% vs. 50% respectively, p &amp;lt; 0.05). The concentration of sperm recovered post-treatment was similar in Swim-up vs MSS (23 x106 vs 19 x106); total motility and progressive motility were similar in Swim-up compared to MSS (92.7% vs. 95.3% and 85% vs. 87% respectively). Only hyperactivated motility assessed with the SCASCOPE CASA System (Microptic) was significantly higher in MSS vs Swim-up 41% vs 4% respectively (p &amp;lt; 0.0005). Limitations, reasons for caution Our study included couples with different infertility diagnosis and semen parameters. Further studies with an increased number of patients may provide more comprehensive data to better evaluate biological and clinical outcomes to eventually suggest the use of MSS in laboratory routines Wider implications of the findings Recently, some studies reported that microfluidic-sorted sperm showed significantly less ROS formation and DNA fragmentation compared to those treated with conventional Swim-up method. Nevertheless, our data does not currently show the expected increase in embryo euploidy rate to justify the routine use of these expensive devices in male infertility treatment Trial registration number non applicabile

P-176 A novel electrophoretic sperm isolation system achieves equivalent ICSI outcomes to the combined density gradient centrifugation and swim-up method in a significantly shorter processing time

Abstract Study question Does an electrophoretically sperm isolation method, the Felix™ system, have clinical or operational advantages over the combined density gradient centrifugation and swim-up (DGCS) method? Summary answer The Felix™ system produces results that are comparable to the DGCS method, however, Felix™ has the ability to isolate spermatozoa more efficiently. What is known already The isolation of healthy spermatozoa to improve ART outcomes has received enormous attention in the literature recently. Although swim-up and density gradient centrifugation are the most common sperm separation methods, the optimal technique is still unclear. Recently, good sperm selection approaches using various non-centrifugal devices have emerged. Previous reports have shown that a quick and simple sperm separation device, the Felix™ system, can isolate high-quality spermatozoa and improve the results of ART. The Felix™ system combines electrophoresis and separation membranes to collect negatively charged mature normal sperm. Study design, size, duration A prospective oocyte sibling study was conducted between March 2023 and December 2023. The study involved infertile couples (n = 45) with a semen volume of 2.3 mL or more, a motile sperm concentration of 2 million/mL or more, maternal age under 43 years, and at least four mature oocytes at the time of oocyte retrieval. Ejaculated semen was prepared separately using DGCS and Felix™ on the day of oocyte retrieval and subsequent ICSI results were analyzed. Participants/materials, setting, methods In the DGCS group, semen was layered on a 45/90% of sperm grad and centrifuged for 15 minutes, followed by a 30 min swim-up. Subsequently, 0.3 mL of supernatant was collected. In the Felix™ group, 0.3 mL of sperm was collected following six minutes of membrane hydration and electrophoresis. Sperm concentration, total motility, progressive motility and DNA fragmentation (SDFR) were measured. Fertilization, blastocyst development, good-quality blastocyst development, and embryo utilisation rates were also assessed. Main results and the role of chance The average female age at the time of oocyte retrieval was 36.5±4.0 years, and the average male age was 38.1±5.9 years. Of 401 oocytes, 200 and 201 oocytes underwent ICSI using sperm prepared by DGCS and Felix™, respectively. The average processing time of Felix™ was significantly shorter than that of DGCS (54.0±4.3 vs. 7.4±0.2 minutes, p &amp;lt; 0.001, respectively). No significant difference was observed between the DGCS and Felix™ groups for sperm concentration (p = 0.49), motility (p = 0.05), progressive motility (p = 0.34), and SDFR (p = 0.09). Furthermore, there were no significant differences between the two groups with regards to fertilization rate (81.0% vs. 80.6%, p = 0.92), blastocyst development rate (52.9% vs. 58.4%, p = 0.60), good-quality blastocyst development rate (26.1% vs. 35.7%, p = 0.06), and embryo utilisation rate (54.3% vs 58.0%, p = 0.50). Limitations, reasons for caution Although the DGCS method was used as a comparison group, it is necessary to compare the Felix™ system with DGC alone, swim-up alone, or other sperm isolation methods. Larger, more powered studies are required to determine the impact on pregnancy and live birth outcomes. Wider implications of the findings The Felix™ system is a viable option for sperm isolation. While DGCS requires complicated procedures and time-consuming process including centrifugation and incubation; Felix™ requires six minutes only. The Felix™ system reduces processing times significantly and therefore can improve laboratory operations and workflow. Trial registration number Not Applicable

Assessing the Impact of the Novel Sperm Selection Technique 'Annexin-V Coated Polystyrene Bead Technique' on Mouse Assisted Reproductive Techniques Outcomes: Preliminary Findings

ICSI is one of the most commonly used techniques to treat infertility. The sperm selection for the procedure is done ‘randomly’ by the embryologist according to the motility and morphology parameters which is known not to reflect the potential of a sperm for fertilization, pregnancy and a healthy childbearing. Since the apoptosis rate is higher in sperm cells of infertile patients, it is more likely to choose an apoptotic sperm by the 'random selection method'. We recently introduced a novel sperm selection technique namely ‘Annexin-V coated polystrene bead technique’(APB-Tech), for the selection of non-apoptotic sperm cells. The principal of the technique is based on the binding affinity of an apoptotic sperm to ‘Annexin-V covered beads’ enabling to distinguish a viable and a healthy sperm by light microscopy. The aim of the present study was to observe the effects of this technique on ICSI outcomes in mice. Sibling-oocyte trial was conducted and the outcome measures were compared with the results of traditional sperm selection method. Embryo and blastocyst qualities and blastocyst development rates were significantly increased in APB-Tech group, while the other parameters were not affected. Promising results obtained from the technique reflect its promising potential as a new and powerful tool for sperm selection and thus infertility techniques.

Sperm YOLOv8E-TrackEVD: A Novel Approach for Sperm Detection and Tracking.

Male infertility is a global health issue, with 40-50% attributed to sperm abnormalities. The subjectivity and irreproducibility of existing detection methods pose challenges to sperm assessment, making the design of automated semen analysis algorithms crucial for enhancing the reliability of sperm evaluations. This paper proposes a comprehensive sperm tracking algorithm (Sperm YOLOv8E-TrackEVD) that combines an enhanced YOLOv8 small object detection algorithm (SpermYOLOv8-E) with an improved DeepOCSORT tracking algorithm (SpermTrack-EVD) to detect human sperm in a microscopic field of view and track healthy sperm in a sample in a short period effectively. Firstly, we trained the improved YOLOv8 model on the VISEM-Tracking dataset for accurate sperm detection. To enhance the detection of small sperm objects, we introduced an attention mechanism, added a small object detection layer, and integrated the SPDConv and Detect_DyHead modules. Furthermore, we used a new distance metric method and chose IoU loss calculation. Ultimately, we achieved a 1.3% increase in precision, a 1.4% increase in recall rate, and a 2.0% improvement in mAP@0.5:0.95. We applied SpermYOLOv8-E combined with SpermTrack-EVD for sperm tracking. On the VISEM-Tracking dataset, we achieved 74.303% HOTA and 71.167% MOTA. These results show the effectiveness of the designed Sperm YOLOv8E-TrackEVD approach in sperm tracking scenarios.

Association between long-term exposure to ambient ozone and sperm quality in Shandong Province

Objective: To evaluate the association between long-term exposure to ambient ozone (O3) and sperm quality. Methods: From January 1, 2014, to December 31, 2019, healthy sperm donors were recruited through the Human Sperm Bank of Shandong University Affiliated Reproductive Hospital. A total of 37 977 sperm donation data from 2 971 healthy volunteers were analyzed. The average annual O3 concentration (0.01°× 0.01°) was matched according to household address. A multivariate mixed-effect model was used to analyze the exposure-response relationship between the average O3 exposure concentration and sperm quality in the previous year, with each donor as a random intercept. All results were presented as % changes with 95% confidence intervals (CIs) for all sperm parameters associated with 10 μg/m3 increases in O3. The effects of individual characteristics on the association between O3 and sperm quality were evaluated by stratified analysis. Results: The average O3 concentration in the year before semen collection was (107.09±7.50) μg/m3. Each 10 μg/m3 increase in O3 was associated with declined sperm concentration (-3.12%, 95%CI:-4.55%, -1.67%), total sperm count (-5.21%, 95%CI:-7.28%, -3.09%), total sperm motility (-1.49%, 95%CI:-2.37%, -0.61%), progressive motility (-2.53%, 95%CI:-3.78%, -1.26%), total motile sperm count (-5.82%, 95%CI:-8.17%, -3.41%), and progressively motile sperm count (-6.22%, 95%CI:-8.73%, -3.64%). Men aged 30 and above, obese, and with lower education levels might be more susceptible to the influence of O3 on sperm quality, but the difference was not statistically significant (P>0.05). Conclusion: Long-term exposure to O3 in Shandong Province is associated with a decrease in sperm quality.

Investigation of the effects of thymoquinone (TQ) and polyvinylpyrrolidone (PVP) on the motility, viability, and chromatin status of sperm in normozoospermic semen samples

Objective: Adverse effects of polyvinylpyrrolidone (PVP) on sperm membrane and chromatin have been proven in many studies. Among the natural products proposed as an alternative for PVP, thymoquinone (TQ)-a major constituent of Nigella sativa plant- has been suggested as a potential natural spermostasis. Therefore, this study aimed to compare the effects of TQ with PVP for sperms motility, survival, DNA denaturation, and DNA fragmentation in normozoospermic men (men with a normal or healthy sperm profile). Methods: An experimental trial was carried out on 30 normozoospermic men of the Andrology Unit of (Shahid Beheshti Hospital, Isfahan, Iran). Each washed semen samples were divided into four fractions and was randomly treated with TQ (50 μg/ml), %5 PVP, and %10 PVP (M/V) which was compared to untreated fraction (control). Results: There was a significant difference between the four groups in terms of motility, viability, DNA denaturation, and fragmentation (P &lt;0.05). TQ caused sperm immobility, while 5% PVP and 10% PVP decreased (98 and 99%, respectively) sperm motility compared to control. TQ did not affect sperm viability compared to the control group, but PVP decreased it. Besides, TQ did not affect DNA denaturation and fragmentation, but PVP increased it. Conclusion: TQ could be used as an alternative natural spermostasis with less adverse effects rather than PVP which causes more efficient immobilization and isolation of individual sperm cells.

Protective Effect of Heteroxenia fuscescens Extract against Sodium Arsenite-induced Infertility in Male Rats

Background: Arsenic is a potent environmental toxin with dangerous effects on human and animal populations. Heteroxenia fuscescens (H. fuscescens) extract exhibits potential health effects. The purpose of this study was to assess the protective effect of H. fuscescensextract against sodium arsenite-induced infertility and toxicity in rats. Methods: Forty male rats were separated into four groups as follows: control group (2% DMSO, orally), sodium arsenite (10 mg/kg, orally), sodium arsenite + H. fuscescens extract (60 mg/kg in 2% DMSO), and H. fuscescens extract (60 mg/kg in 2% DMSO). Results: GC/MS analysis of H. fuscescens indicates the presence of 10 compounds at retention times of 6.00, 12.53, 19.04, 24.60, 28.56, 35.66, 38.99, 45.29, 48.02, and 52.14. fuscescens extract improves levels of sperm count (53.33 ± 1.52), motility (43.00 ± 1.10), FSH ( 2.17 ± 0.04), LH (2.43 ± 0.06), testosterone (1.87 ± 0.02), and testis weight (0.49 ± 0.01). The antioxidant activity of H. fuscescens extract was reported by a significant decrease in MDA (1.02 ± 0.01) and NO (520.39 ± 14.99) levels, while it significantly increased levels of GSH (1.77 ± 0.07), and CAT (71.38 ± 3.45). Histopathological alterations of the testes, liver, and kidney observed with sodium arsenite have been improved in the treatment group. Conclusion: Heteroxenia fuscescens is beneficial in restoring male sex hormone levels, maintaining a healthy sperm profile, and reducing oxidative stress, all of which lead to an improvement in male rat fertility.

Evaluation of telomere length, reactive oxygen species, and apoptosis in spermatozoa of patients with oligospermia.

50% of cases of infertility are caused by male factor, which acquired or congenital problems may bring on. Male infertility can be caused by oligospermia and asthenozoospermia, which are common. Since the same mutations that cause azoospermia in some people also cause oligozoospermia in others, oligozoospermia may be thought of as a less severe form of azoospermia. Studies have demonstrated telomere length, catalase activity, super oxide dismutase (SOD), and DNA fragmentation can be influential factors for male infertility. The amount of apoptosis, oxidative stress factors, telomere length, and DNA fragmentation were some aspects of healthy sperm that we chose to look into in this study and compare to oligospermia individuals. Oligospermia patients (n = 24) and fertile men (n = 27) semen samples were collected, and the apoptosis rate of sperms in both groups was analyzed (Flow cytometry). Also, gene expression of apoptotic and antiapoptotic markers and telomere length were examined (real-time polymerase chain reaction). The sperm DNA fragmentation kit was used to determine DNA fragmentation and to evaluate catalase and SOD activity; the specific kits and methods were utilized. Higher expression levels of caspase3 (p = .0042), caspase8 (p = .0145), caspase9 (p = .0275), and BAX (p = .0202) mRNA were observed in patients who had oligospermia. In contrast, lower mRNA expression of BCL-2 (p = .0009) was detected in this group. In addition, telomere length was decreased in the oligospermia group (p < .0001) compared to the health group. Moreover, the frequency of apoptosis is induced in patients (p = .0026). The catalase activity is low (p = .0008), but the SOD activity is high (p = .0015) in the patient group. As a result of our findings, we may list the sperm cell apoptosis rate, telomere length, the degree of sperm DNA fragmentation, and lastly, the measurement of significant and efficient oxidative stress markers like SOD and catalase in semen plasma among the principal diagnostic characteristics for oligospermia. Future studies will be better able to treat oligospermia by showing whether these indicators are rising or falling.

Using a Decision Tree Algorithm Predictive Model for Sperm Count Assessment and Risk Factors in Health Screening Population.

Approximately 20% of couples face infertility challenges and struggle to conceive naturally. Despite advances in artificial reproduction, its success hinges on sperm quality. Our previous study used five machine learning (ML) algorithms, random forest, stochastic gradient boosting, least absolute shrinkage and selection operator regression, ridge regression, and extreme gradient boosting, to model health data from 1375 Taiwanese males and identified ten risk factors affecting sperm count. We employed the CART algorithm to generate decision trees using identified risk factors to predict healthy sperm counts. Four error metrics, SMAPE, RAE, RRSE, and RMSE, were used to evaluate the decision trees. We identified the top five decision trees based on their low errors and discussed in detail the tree with the least error. The decision tree featuring the least error, comprising BMI, UA, ST, T-Cho/HDL-C ratio, and BUN, corroborated the negative impacts of metabolic syndrome, particularly high BMI, on sperm count, while emphasizing the link between good sleep and male fertility. Our study also sheds light on the potentially significant influence of high BUN on spermatogenesis. Two novel risk factors, T-Cho/HDL-C and UA, warrant further investigation. The ML algorithm established a predictive model for healthcare personnel to assess low sperm counts. Refinement of the model using additional data is crucial for improved precision. The risk factors identified offer avenues for future investigations.

Bull Sperm SWATH-MS-Based Proteomics Reveals Link between High Fertility and Energy Production, Motility Structures, and Sperm-Oocyte Interaction.

The prediction of male or semen fertility potential remains a persistent challenge that has yet to be fully resolved. This work analyzed several in vitro parameters and proteome of spermatozoa in bulls cataloged as high- (HF; n = 5) and low-field (LF; n = 5) fertility after more than a thousand artificial inseminations. Sperm motility was evaluated by computer-assisted sperm analysis. Sperm viability, mitochondrial membrane potential (MMP) and reactive oxygen species (mROS) of spermatozoa were assessed by flow cytometry. Proteome was evaluated by the SWATH-MS procedure. Spermatozoa of HF bulls showed significantly higher total motility than the LF group (41.4% vs 29.7%). Rates of healthy sperm (live, high MMP, and low mROS) for HF and LF bull groups were 49% and 43%, respectively (p > 0.05). Spermatozoa of HF bulls showed a higher presence of differentially abundant proteins (DAPs) related to both energy production (COX7C), mainly the OXPHOS pathway, and the development of structures linked with the motility process (TPPP2, SSMEM1, and SPAG16). Furthermore, we observed that equatorin (EQTN), together with other DAPs related to the interaction with the oocyte, was overrepresented in HF bull spermatozoa. The biological processes related to protein processing, catabolism, and protein folding were found to be overrepresented in LF bull sperm in which the HSP90AA1 chaperone was identified as the most DAP. Data are available via ProteomeXchange with identifier PXD042286.

The Impact of Obesity on Sperm Parameters in Young Adult Males: A Retrospective Study of Sperm Donors

To utilize a large cohort of healthy sperm bank donors to evaluate the association between body mass index and individual sperm parameters. Sperm parameters from donors across the United States were obtained between 2013 and 2022. Donors were healthy men aged 18-46years old. Semen samples were analyzed in a certified lab following guidelines by the World Health Organization. A multivariable interaction model between age, body mass index, and sperm parameters was conducted. There were 117,357 sperm donations included in our study. In our sample, 98,397 (83.84%) men were classified as young donors (ages 18-32years) and 18,960 (16.16%) were classified as old donors (ages 33-46years). We identified 1032 (0.88%) men as underweight, 76,635 (65.30%) as normal weight, 36,686 (31.26%) as overweight, and 3004 (2.56%) as obese. Participants had a median total motile sperm count (TMSC) of 186 (interquartile ranges [IQR]: 128 million), volume of 3.36 (IQR: 1.82 mL), sperm concentration of 56 (IQR: 34 million/mL) and a progressive motility of 59.84% (IQR: 16.95%). Older obese donor had lower TMSC (β = -22.98 ± 4.66, P < .001), semen volumes (β = -0.85 ± 0.06, P < .001), and progressive motility (β = -3.94 ± 0.56, P < .001) compared to younger, healthy weight donors. We observed lower TMSC, semen volumes, and progressive motility in older obese donors. Although these values are within the normal expected ranges for individual sperm parameters, our ability to detect differences within this healthy population highlights the importance of maintaining a healthy diet and exercise regimen for preserving high sperm counts.

A Dual Architecture Fusion and AutoEncoder for Automatic Morphological Classification of Human Sperm.

Infertility has become a common problem in global health, and unsurprisingly, many couples need medical assistance to achieve reproduction. Many human behaviors can lead to infertility, which is none other than unhealthy sperm. The important thing is that assisted reproductive techniques require selecting healthy sperm. Hence, machine learning algorithms are presented as the subject of this research to effectively modernize and make accurate standards and decisions in classifying sperm. In this study, we developed a deep learning fusion architecture called SwinMobile that combines the Shifted Windows Vision Transformer (Swin) and MobileNetV3 into a unified feature space and classifies sperm from impurities in the SVIA Subset-C. Swin Transformer provides long-range feature extraction, while MobileNetV3 is responsible for extracting local features. We also explored incorporating an autoencoder into the architecture for an automatic noise-removing model. Our model was tested on SVIA, HuSHem, and SMIDS. Comparison to the state-of-the-art models was based on F1-score and accuracy. Our deep learning results accurately classified sperm and performed well in direct comparisons with previous approaches despite the datasets' different characteristics. We compared the model from Xception on the SVIA dataset, the MC-HSH model on the HuSHem dataset, and Ilhan et al.'s model on the SMIDS dataset and the astonishing results given by our model. The proposed model, especially SwinMobile-AE, has strong classification capabilities that enable it to function with high classification results on three different datasets. We propose that our deep learning approach to sperm classification is suitable for modernizing the clinical world. Our work leverages the potential of artificial intelligence technologies to rival humans in terms of accuracy, reliability, and speed of analysis. The SwinMobile-AE method we provide can achieve better results than state-of-the-art, even for three different datasets. Our results were benchmarked by comparisons with three datasets, which included SVIA, HuSHem, and SMIDS, respectively (95.4% vs. 94.9%), (97.6% vs. 95.7%), and (91.7% vs. 90.9%). Thus, the proposed model can realize technological advances in classifying sperm morphology based on the evidential results with three different datasets, each having its characteristics related to data size, number of classes, and color space.

Design of a gradient-rheotaxis microfluidic chip for sorting of high-quality Sperm with progressive motility

Assisted reproductive technology (ART) is an important invention for the treatment of human infertility, and the isolation of high-quality sperm with progressive motility is one of the most critical steps that eventually affect the fertilization rate. Conventional sperm separation approaches include the swim-up method and density gradient centrifugation. However, the quality of isolated sperm obtained from both approaches can still be improved by improving sorted sperm motility, minimizing the DNA fragmentation rate, and removing abnormal phenotypes. Here, we report a Progressive Sperm Sorting Chip (PSSC) for high-quality sperm isolation. Based on the rheotaxis behavior of sperm, a gradient flow field is created in the chip for progressive sperm sorting. Clinical experiment results for 10 volunteers showed that greater than 90% of isolated sperm exhibit high motility (> 25μm/s), high linearity (0.8), and a very low DNA fragmentation rate (< 5%). In addition, the whole process is label and chemical free. These features aid in gentle sperm sorting to obtain healthy sperm. This device uniquely enables the selection of high-quality sperm with progressive motility and might be clinically applied for infertility treatment in the near future.

P-029 Centrifugation-free sperm separation device offers an efficient and standardized protocol to select high quality spermatozoa

Abstract Study question Can centrifugation-free sperm separation device be used to simplify and standardize the selection of high-quality spermatozoa? Summary answer Centrifugation-free CA0 sperm separation device minimizes inter- and intra-operator variability and yields spermatozoa with comparable fertilizing properties in a variety of semen conditions. What is known already Centrifugation-based sperm separation methods have been used in assisted reproductive technology (ART) for many decades. However, the conventional methods are criticized for harmful effects due to centrifugation. To overcome the disadvantage, more noninvasive technologies have been developed and attempted to improve the sperm separation process, e.g., the formation of capillary bridge to select motile spermatozoa, migration-sedimentation technique to sort out functional spermatozoa, microfluidic sorting chip to isolate healthy sperm. While these methods provide alternatives for noninvasive sperm separation, the limitations such as inconsistency of semen quality improvement and lack of standardized procedure remain to be resolved. Study design, size, duration A randomized controlled trial of 76 men who sought ART treatment (Lee Women’s Hospital, Taichung, Taiwan) from June to October 2022 was carried out. Seventy-six neat semen specimens were categorized into 27 normozoospermic specimens, and 49 non-normozoospermic samples (semen quality below any of the WHO 5th Edition lower reference values: concentration &amp;lt;15 million/mL, total motility &amp;lt;40%, or normal morphology &amp;lt;4%). Participants/materials, setting, methods Neat semen samples were separated for three replicates (replicate 1-3) using LensHooke® CA0 sperm separation device (Bonraybio, Taichung, Taiwan). Three operators (operators 1–3) performed CA0 procedures on each sample to test inter-technician variability. Interclass-correlation coefficient (ICC) between replicates as well as operators were evaluated. Pre-selection and post-selection semen quality were evaluated. The parameters included total motility, progressive motility, rapid progressive motility, morphology, DNA fragmentation index (DFI), and acrosome reaction rate (AR). Main results and the role of chance CA0 selects self-propelling spermatozoa within a microenvironment created by a microporous filter membrane. The procedure involves three pipetting steps: loading semen sample, adding sperm washing medium, and recovering the processed sample. Following the standard procedure, CA0 resulted in a low intra- and inter-operator variability and ICC values between replicates and different operators were all greater than 0.9, indicating an excellent reproducibility of CA0. In addition, significant higher levels of motility and normal morphology were observed in post-selection specimens either of normozoospermic or non-normozoospermic samples (pre-selection vs. post-selection, all p &amp;lt; 0.0001). In paired analysis of the advanced semen parameter, our study showed noteworthy results that CA0 significantly improved DFI from 18.2% to 2.6% for normozoospermic samples; such reduction was also found in non-normozoospermic sample processing, from 13.4% to 4.2% (both, p &amp;lt; 0.0001). The levels of AR were significantly reduced in normozoospermic samples (from 14.4% to 5.4%) and non-normozoospermic samples as well (from 12.8% to 5.0%) (both groups p &amp;lt; 0.0001). In conclusion, CA0 provides an efficient, noninvasive, standardized, and reproducible sperm separation model that CA0 diminishes the variations and ensures sperm quality. Limitations, reasons for caution The presented study was a pilot trial examining the sperm quality improvement. Follow-up analysis on IUI/IVF outcomes associated with the improvement in semen quality utilizing CA0 will be assessed in future studies. Wider implications of the findings CA0 provides multifaceted benefits covering consistent clinical outcomes, simplified and standardized procedure, user-friendliness, and cost reduction. We believe CA0 not only allows noninvasive sperm separation of clinically usable but also gives the possibility of standardization on sperm separation procedure. Trial registration number CS2-22039

The role of sperm DNA integrity in seminal plasma of infertile men

One-half of the genome is delivered by the sperm, and a healthy sperm is needed for fertilization, fetal development, and child wellbeing. Abnormal sperm DNA can impact any of these processes. The measurement of sperm DNA fragmentation has been added to some medical andrology laboratories as a crucial addition to the standard analysis of the semen. Sperm DNA fragmentation has also been linked to male infertility including idiopathic male infertility. Some degree of controlled DNA nicking is essential for adequate DNA compaction, but excessive sperm DNA fragmentation is usually associated with reduced male fertility potential, reduced fertilization, poor embryo quality, recurrent pregnancy loss and poor assisted reproductive techniques outcomes. Semen samples from 50 infertile males (as 19 asthenozoospermia, 7 oligozoospermia, 13 normospermia and 11 combined defects) were analyzed for sperm DNA fragmentation. This study followed the World Health Organization 2010 guidelines to assess the sperm parameters. Sample were collected by masturbation and put in a 37 °C incubator to be liquefied, then they were placed in a centrifuge (3000 rpm for 10 min) to extract the pellet, which was used for assessment of DFI% by anillin blue staining. Negative significant correlation (p = 0.023) between morphologically normal sperms and DFI % was found in normozoospermia group. The objective of this study is to examine the correlation of DFI among different seminal fluid categories (normozoospermia, oligozoospermia, asthenozoospermia and, combined).

How to select healthy sperm for intracytoplasmic sperm injection in samples with high sperm DNA fragmentation?

The body of evidence supports the negative impact of increased sperm DNA fragmentation (SDF) on natural fertility as well as assisted reproduction conditions. High SDF has been correlated with low pregnancy and delivery rates following intrauterine insemination. Also, high SDF is accused of reducing the rates of fertilization, implantation, pregnancy, and live birth following in-vitro fertilization (IVF). Despite no impact of high SDF on fertilization or pregnancy rates following intracytoplasmic sperm injection (ICSI), it has been correlated with poor embryo quality and a higher risk of miscarriage. Several methods have been introduced to help select sperm with the best DNA quality to be used in assisted reproductive technology procedures. These include magnetic-activated cell sorting, intracytoplasmic morphologically selected sperm injection, physiologic ICSI, and microfluidic sperm sorters, among others. This article aimed to discuss the impact of high SDF in infertile men on the reproductive outcome of couples undergoing IVF/ICSI. Additionally, this review highlights the principles, advantages, and limitations of different techniques that are currently used for the selection of sperm with intact DNA to be utilized for ICSI.