CatSper is a unique Ca2＋ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we stu...CatSper is a unique Ca2＋ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we studied the mechanism of regulation of CatSper2-dependent Ca〉 influx, extracellular and intracellular Ca2＋on sperm hyperactivated motility. The siRNA duplexes were transfected into the sperm cells by electroporation （EP） to silence the expression of CatSper2. The results for targeted disruption of CatSper2 showed the highest silence efficiency 77.7% （P〈0.05）, the hyperactivated sperm rate calculated by computer-assisted semen analysis （CASA） analysis of interferenced sperm was significantly lower 11.1% than the control 99.2%. Flow cytometry （FCM） detection of the intracellular Ca2＋ concentration of interferenced sperm was 50 nmol L-t higher than the normal. It was remarkably lower than hyperactivated sperm with 200-1 000 nmol L-1 （P〈0.05）. It was not sufficient to evoke hyperactivation. To trigger hyperactivation, the sperm were incubated in 50 pmol L-t thimerosal or 5 mmol L-1 procaine, it sharply increased the intracellular Ca2＋ via two different channels. CASA and FCM detection indicated that intracellular Ca〉 is required for initiating hyperactivation while extracellular Ca2＋ is essential to maintain the process. We concluded that to mediate sperm hyperactivation, it is essential to inhibit Ca〉peak present with targeted disruption of CatSper2 expression. This provides important prospective for further exploration of signal channel of sperm hyperactivated motility, potential factors for male infertility and provide further reference to the exploration of male contraceptive drug targets of male reproduction.展开更多
Introduction: Men without sperm (azoospermic) make up about 15% of all infertile men in reproductive age of fatherhood. Male infertility is a health and social problem in many communities. Subjects and Methods: This r...Introduction: Men without sperm (azoospermic) make up about 15% of all infertile men in reproductive age of fatherhood. Male infertility is a health and social problem in many communities. Subjects and Methods: This retrospective study was carried from 2010 to 2015. Initially, 907 men were included in the study among whom 109 (12.02%), 346 (38.15%) and 452 (49.83%) were azoospermic, oligospermic and normospermic, respectively. This study only investigated the association between age, BMI, semen volume, liquefaction time and pus cells among normospermic and azoospermic men. Standard semen analysis was performed and subjects were categorized by age into Results: The 562 subjects of the study had means (±sd) age of 42.6 (±7.10) years and BMI of 27.0 (4.1) kg/m2 respectively. In all, 109 (19.4%) were azoospermic. Overweight and obese men were, respectively, more than 1? and about 2? times more likely to be azoospermic compared to normal weight men. Azoospermic men were significantly heavier than men with normal sperm count (t = -0.34;P-value = 0.003). Among those with normal weight, liquefaction time was significantly shorter (t = 5.49, P-value = 0.000001) in azoospermic (28.70 min.) than in men with normal sperm count (31.82 min.). Obese azoospermic men were about 4 times as likely to have high pus cells in semen than normal weight azoospermic men (OR = 3.82;95% CI: 0.39, 37.01). Multivariate regression analysis shows a strong but negative coefficient correlation between sperm concentration and BMI (coef. = -0.48, Std Err. = 0.25, P-value = 0.05, 95% CI: -0.96, 0.10). Conclusion: Our findings suggest that high BMI is associated with azoospermia. Azoospermia also did not seem to negatively impact liquefaction time.展开更多
Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by ...Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.展开更多
The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinc...The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages,with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup forrepeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providingpractitioners with a comprehensive analysis of each option’s strengths and limitations. Comparative studies indicate that, althoughfresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthawoutcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparablerates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular spermextraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and proceduralconvenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individualneeds and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes andprovide personalized reproductive care.展开更多
Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities...Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities.PCD is typically resulted from variants in genes encoding assembly or structural proteins that are indispensable for the movement of motile cilia.Here,we identified a novel nonsense mutation,c.466G>T,in cilia-and flagella-associated protein 300(CFAP300)resulting in a stop codon(p.Glu156*)through whole-exome sequencing(WES).The proband had a PCD phenotype with laterality defects and immotile sperm flagella displaying a combined loss of the inner dynein arm(IDA)and outer dynein arm(ODA).Bioinformatic programs predicted that the mutation is deleterious.Successful pregnancy was achieved through intracytoplasmic sperm injection(ICSI).Our results expand the spectrum of CFAP300 variants in PCD and provide reproductive guidance for infertile couples suffering from PCD caused by them.展开更多
Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been sh...Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been shown that patients with sperm parameters abnormalities and abnormal morphology have a high rate of fragmentation and sperm DNA decondensation[2,3],and that sperm DNA fragmentation analysis could be used as a predictor factor of fertility potential[4].展开更多
The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm ...The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm fertilizing capacity.Among these,sperm concentration and motility are the first parameters to be evaluated through an estimation carried out by expert examiners.展开更多
Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in ...Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in antidepressant consumption.Antidepressants are associated with sexual dysfunction in both men and women;however,their role in male fertility has been scarcely studied.Fluoxetine and sertraline,two serotonin reuptake inhibitors(SSRIs),are among the most prescribed antidepressants worldwide.To determine their possible effects,human sperm cells were exposed to either sertraline or fluoxetine at concentrations previously found in blood and seminal fluid of patients undergoing treatment.Spermatozoa were incubated for up to 24 h at 37℃ and 5%CO_(2),and important functional parameters such as sperm motility,viability,mitochondrial membrane potential,cellular reactive oxygen species(ROS)production,chromatin/DNA integrity,acrosome status,and tyrosine phosphorylation were assessed.At low levels,fluoxetine consistently decreased progressive motility throughout time while promoting fluctuations in ROS levels and sperm capacitation.Nevertheless,it did not affect viability,mitochondrial membrane potential,acrosome reaction nor chromatin/DNA integrity.Sertraline,on the other hand,had little to nonsignificant impact at low doses,but affected almost all tested parameters at supratherapeutic concentrations.Altogether,our results suggest that both antidepressants may impair sperm function,possibly through different mechanisms of action,but fluoxetine is the only exhibiting mild negative effects at doses found in vivo.展开更多
The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary p...The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary possibility of azoospermic men fathering their own genetic offspring.展开更多
High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,b...High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,but the 6th edition of the World Health Organization(WHO)manual for semen analysis included the SDF assessment in the chapter on extended semen examinations.展开更多
Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm D...Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm DNA.This oxidative damage is associated with male infertility.On the other hand,when RoS are produced at low levels,they participate in the redox signaling necessary for sperm capacitation.Capacitation-associated RoS are produced by the sperm oxidase,whose identity is still elusive,located in the plasma membrane of the spermatozoon.Ros,such as superoxide anion,hydrogen peroxide,nitric oxide,and peroxynitrite,activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events.Peroxiredoxins(PRDXs),antioxidant enzymes that fight against oxidative stress,regulate redox signaling during capacitation.Among them,PRDX6,which possesses peroxidase and calcium-independent phospholipase A,(iPLA,)activities,is the primary regulator of redox signaling and the antioxidant response in human spermatozoa.The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)pathway,and it is regulated by PRDX6 iPLA2,protein kinase C(PKC),and receptor-type protein tyrosine kinase.The understanding of redox signaling is crucial to pave theway fornovel diagnostic tools and treatments of male infertility.展开更多
Kamil Gill,PhD,Kamil Gill is a biologist who graduated from the University of Szczecin(Poland)with a Master degree.Dr.Gill earned his scientific degree from the Pomeranian Medical University in Szczecin(PUM).His docto...Kamil Gill,PhD,Kamil Gill is a biologist who graduated from the University of Szczecin(Poland)with a Master degree.Dr.Gill earned his scientific degree from the Pomeranian Medical University in Szczecin(PUM).His doctoral thesis was entitled:"Molecular markers of male infertility-evaluation of selected human sperm chromatin parameters and its influence on the zygote and embryo development in assisted reproduction conditions".展开更多
Except in cases of hypogonadotropic hypogonadism,the use of medical therapy before microsurgical testicular sperm extraction(micro-TESE)is controversial.In some studies,hormone therapy has been shown to improve the po...Except in cases of hypogonadotropic hypogonadism,the use of medical therapy before microsurgical testicular sperm extraction(micro-TESE)is controversial.In some studies,hormone therapy has been shown to improve the possibility of sperm retrieval during micro-TESE and even lead to the presence of sperm in the ejaculate in some cases,thereby obviating the need for micro-TESE.However,their routine use before micro-TESE in cases of nonobstructive azoospermia(NOA)being associated with hypergonadotropic hypogonadism and eugonadism(normogonadotropic condition)has not been supported with robust evidence.In this review,we discuss different types of medical therapy used before micro-TESE for NOA,their risks and benefits,and the available evidence surrounding their use in this setting.展开更多
Sperm cryopreservation is an essential technique for male fertility preservation,especially in men who are undergoing medical treatment.Conventional cryopreservation methods face limitations such as oxidative stress,D...Sperm cryopreservation is an essential technique for male fertility preservation,especially in men who are undergoing medical treatment.Conventional cryopreservation methods face limitations such as oxidative stress,DNA fragmentation,and cytotoxicity associated with traditional cryoprotectants like dimethyl sulfoxide(DMSO).Recent breakthroughs have focused on improving post-thaw sperm viability with novel cryoprotectants and innovative freezing strategies.Prospective approaches include the use of amino acid-based cryoprotectants,deep eutectic solvents,and antioxidants that have been described to prevent oxidative damage and maintain DNA integrity.Vitrification,a high-speed freezing technique that prevents ice crystal formation,has demonstrated superior outcomes compared to conventional slow freezing.Moreover,the Direct Dropping Method,a cryoprotectant-free approach,has been introduced as a contamination-minimizing technique that preserves sperm functionality.Multiomics tools are also utilized to determine biomarkers for protocol optimization.Despite these advancements,cryoprotectant toxicity is a central challenge,emphasizing the necessity for safer agents.Future research must focus on long-term sperm functionality and individualized cryopreservation strategies to maximize reproductive outcomes.The current review highlights the challenges associated with sperm cryopreservation,explores innovative strategies and novel cryoprotectants,underscores the significance of maintaining DNA integrity,and proposes future research directions to improve fertility preservation outcomes.展开更多
Medically assisted reproduction(MAR)techniques are highly dependent on the sperm quantity and quality.Low sperm concentrations can be bypassed at least to some point by the usage of more sophisticated MAR techniques l...Medically assisted reproduction(MAR)techniques are highly dependent on the sperm quantity and quality.Low sperm concentrations can be bypassed at least to some point by the usage of more sophisticated MAR techniques like intracytoplasmic sperm injection(ICSI).Compared to this,disruptions in established indicators of sperm quality like motility or morphology pose greater challenges for the therapy of couple infertility.展开更多
Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed a...Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed at the time of sperm retrieval.Onco-micro-TESE addresses the dual objectives of treating both infertility and the testicular tumor simultaneously.The technique is intricate,necessitating a comprehensive understanding of testicular anatomy,physiology,tumor biology,and advanced microsurgical methods.It aims to carefully extract viable spermatozoa while minimizing the risk of tumor dissemination.This review encapsulates the procedural intricacies,evaluates success determinants,including tumor pathology and spermatogenic tissue health,and discusses the implementation of imaging techniques for enhanced surgical precision.Ethical considerations are paramount,as the procedure implicates complex decision-making that weighs the potential oncological risks against the profound desire for fatherhood using the male gametes.The review aims to provide a holistic overview of onco-micro-TESE,detailing methodological advances,clinical outcomes,and the ethical landscape,thus offering an indispensable resource for clinicians navigating this multifaceted clinical scenario.展开更多
Objective:To assess the effects of azoxystrobin on sperm quality,hormone levels and testicular structure in Swiss albino mice.Methods:48 Swiss albino male mice were divided into 7 groups containing 6 mice in each grou...Objective:To assess the effects of azoxystrobin on sperm quality,hormone levels and testicular structure in Swiss albino mice.Methods:48 Swiss albino male mice were divided into 7 groups containing 6 mice in each group except Group桏in which 12 mice were taken.Group栺served as the control group and treated with vehicle(distil water for 30 and 60 days).Azoxystrobin at three different doses(125,250,500 mg/kg body weight)was orally administered to Group栻,栿and桇for 30 days and Group桋,桍and桏for 60 days.After dose completion,sperm parameters,hormonal profile and testis histology were analysed.Half animals of group桏(500 mg/kg body weight)were left untreated for next 30 days to assess the recovery from reproductive toxicity.Results:At azoxystrobin 125 and 250 mg/kg,sperm viability,count,and motility remained largely unaffected,but a significant decline was observed at azoxystrobin 500 mg/kg after 30 days.After 60 days,all dose levels led to a significant decrease in sperm parameters.Morphological abnormalities in sperm,such as globular heads,bent or coiled tails,and headless or tailless sperm,were noted to be increased in a dose dependent manner.Antioxidant parameters,serum level of reproductive hormones and steroidogenic enzymes followed a similar trend with non-significant changes at low and medium doses but a marked decline at the high dose after both 30 and 60 days of exposure.Histopathology of testis also revealed degenerative changes in seminiferous tubules and Leydig cell.Conclusions:Azoxystrobin exposure at high dose(500 mg/kg)affects sperm quality and disrupts testicular function via potentially impairing antioxidant defences,deregulating steroidogenesis in Swiss albino male mice.展开更多
Background:Testicular sperm aspiration(TESA)is a minimally invasive testicular sperm retrieval technique that has been utilized in the treatment of male factor infertility.We sought to evaluate sperm retrieval outcome...Background:Testicular sperm aspiration(TESA)is a minimally invasive testicular sperm retrieval technique that has been utilized in the treatment of male factor infertility.We sought to evaluate sperm retrieval outcomes of primary and redo TESA in men with severe oligoasthenoteratozoospermia(OAT)and obstructive azoospermia(OA).Methods:This is a retrospective analysis of consecutive TESAs(primary and redo)for men with severe OAT and OA performed between January 2011 and August 2022 at a high-volume infertility center.We compared TESA outcomes in men with severe OAT to those with OA and compared outcomes of men who underwent primary and redo TESA on the same testicular unit.Results:439 TESAs(366 primary and 73 redo)in men with severe OAT(n=133)and OA(n=306)were included.Men with OA had significantly higher sperm retrieval rate(SRR)and motile SRR compared to men with severe OAT(99%vs.95%and 98%vs.83%,respectively,p<0.05).The requirement for multiple biopsies and the total number of aspirates were significantly lower in men with OA compared to those with severe OAT(15%vs.32%and 1.2±0.5 vs.1.4±0.7,respectively,p<0.05).In both groups,SRR,motile SRR,the requirement for multiple biopsies,and the total number of aspirates were not significantly different in primary compared to redo cases.Conclusion:Our data demonstrate that TESA retrieval rates are significantly higher in men with OA compared to those with severe OAT.The data also demonstrate that a redo TESA in these men is as effective as a primary TESA,suggesting that areas of active spermatogenesis are preserved 6 months after TESA.展开更多
Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia(OA)and nonobstructive azoospermia(NOA).For men with NOA,testicular sperm extraction(TESE)is the onl...Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia(OA)and nonobstructive azoospermia(NOA).For men with NOA,testicular sperm extraction(TESE)is the only method to obtain sperm for assisted reproductive technology(ART).Given the rarity of these sperm and the unpredictable success of subsequent retrieval attempts,cryopreservation of microdissection-TESE-obtained sperm is essential.Effective cryopreservation prevents the need for repeated surgical procedures and supports future ART attempts.After first delving into the physiological and molecular aspects of sperm cryopreservation,this review aims to examine the current methods and devices for preserving small numbers of sperm.It presents conventional freezing and vitrification techniques,evaluating their respective strengths and limitations in effectively preserving rare sperm,and compares the efficacy of using fresh versus cryopreserved testicular sperm.展开更多
基金supported by the National Key Technology R&D Program of China (2006BAD04A12)
文摘CatSper is a unique Ca2＋ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we studied the mechanism of regulation of CatSper2-dependent Ca〉 influx, extracellular and intracellular Ca2＋on sperm hyperactivated motility. The siRNA duplexes were transfected into the sperm cells by electroporation （EP） to silence the expression of CatSper2. The results for targeted disruption of CatSper2 showed the highest silence efficiency 77.7% （P〈0.05）, the hyperactivated sperm rate calculated by computer-assisted semen analysis （CASA） analysis of interferenced sperm was significantly lower 11.1% than the control 99.2%. Flow cytometry （FCM） detection of the intracellular Ca2＋ concentration of interferenced sperm was 50 nmol L-t higher than the normal. It was remarkably lower than hyperactivated sperm with 200-1 000 nmol L-1 （P〈0.05）. It was not sufficient to evoke hyperactivation. To trigger hyperactivation, the sperm were incubated in 50 pmol L-t thimerosal or 5 mmol L-1 procaine, it sharply increased the intracellular Ca2＋ via two different channels. CASA and FCM detection indicated that intracellular Ca〉 is required for initiating hyperactivation while extracellular Ca2＋ is essential to maintain the process. We concluded that to mediate sperm hyperactivation, it is essential to inhibit Ca〉peak present with targeted disruption of CatSper2 expression. This provides important prospective for further exploration of signal channel of sperm hyperactivated motility, potential factors for male infertility and provide further reference to the exploration of male contraceptive drug targets of male reproduction.
文摘Introduction: Men without sperm (azoospermic) make up about 15% of all infertile men in reproductive age of fatherhood. Male infertility is a health and social problem in many communities. Subjects and Methods: This retrospective study was carried from 2010 to 2015. Initially, 907 men were included in the study among whom 109 (12.02%), 346 (38.15%) and 452 (49.83%) were azoospermic, oligospermic and normospermic, respectively. This study only investigated the association between age, BMI, semen volume, liquefaction time and pus cells among normospermic and azoospermic men. Standard semen analysis was performed and subjects were categorized by age into Results: The 562 subjects of the study had means (±sd) age of 42.6 (±7.10) years and BMI of 27.0 (4.1) kg/m2 respectively. In all, 109 (19.4%) were azoospermic. Overweight and obese men were, respectively, more than 1? and about 2? times more likely to be azoospermic compared to normal weight men. Azoospermic men were significantly heavier than men with normal sperm count (t = -0.34;P-value = 0.003). Among those with normal weight, liquefaction time was significantly shorter (t = 5.49, P-value = 0.000001) in azoospermic (28.70 min.) than in men with normal sperm count (31.82 min.). Obese azoospermic men were about 4 times as likely to have high pus cells in semen than normal weight azoospermic men (OR = 3.82;95% CI: 0.39, 37.01). Multivariate regression analysis shows a strong but negative coefficient correlation between sperm concentration and BMI (coef. = -0.48, Std Err. = 0.25, P-value = 0.05, 95% CI: -0.96, 0.10). Conclusion: Our findings suggest that high BMI is associated with azoospermia. Azoospermia also did not seem to negatively impact liquefaction time.
文摘Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.
文摘The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages,with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup forrepeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providingpractitioners with a comprehensive analysis of each option’s strengths and limitations. Comparative studies indicate that, althoughfresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthawoutcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparablerates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular spermextraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and proceduralconvenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individualneeds and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes andprovide personalized reproductive care.
基金supported by the China Postdoctoral Science Foundation Grant(2023M734294)Jiangsu Provincial Medical Key Discipline Cultivation Unit(JSDW202215)the National Natural Science Foundation of China(No.82001618).
文摘Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities.PCD is typically resulted from variants in genes encoding assembly or structural proteins that are indispensable for the movement of motile cilia.Here,we identified a novel nonsense mutation,c.466G>T,in cilia-and flagella-associated protein 300(CFAP300)resulting in a stop codon(p.Glu156*)through whole-exome sequencing(WES).The proband had a PCD phenotype with laterality defects and immotile sperm flagella displaying a combined loss of the inner dynein arm(IDA)and outer dynein arm(ODA).Bioinformatic programs predicted that the mutation is deleterious.Successful pregnancy was achieved through intracytoplasmic sperm injection(ICSI).Our results expand the spectrum of CFAP300 variants in PCD and provide reproductive guidance for infertile couples suffering from PCD caused by them.
文摘Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been shown that patients with sperm parameters abnormalities and abnormal morphology have a high rate of fragmentation and sperm DNA decondensation[2,3],and that sperm DNA fragmentation analysis could be used as a predictor factor of fertility potential[4].
文摘The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm fertilizing capacity.Among these,sperm concentration and motility are the first parameters to be evaluated through an estimation carried out by expert examiners.
文摘Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in antidepressant consumption.Antidepressants are associated with sexual dysfunction in both men and women;however,their role in male fertility has been scarcely studied.Fluoxetine and sertraline,two serotonin reuptake inhibitors(SSRIs),are among the most prescribed antidepressants worldwide.To determine their possible effects,human sperm cells were exposed to either sertraline or fluoxetine at concentrations previously found in blood and seminal fluid of patients undergoing treatment.Spermatozoa were incubated for up to 24 h at 37℃ and 5%CO_(2),and important functional parameters such as sperm motility,viability,mitochondrial membrane potential,cellular reactive oxygen species(ROS)production,chromatin/DNA integrity,acrosome status,and tyrosine phosphorylation were assessed.At low levels,fluoxetine consistently decreased progressive motility throughout time while promoting fluctuations in ROS levels and sperm capacitation.Nevertheless,it did not affect viability,mitochondrial membrane potential,acrosome reaction nor chromatin/DNA integrity.Sertraline,on the other hand,had little to nonsignificant impact at low doses,but affected almost all tested parameters at supratherapeutic concentrations.Altogether,our results suggest that both antidepressants may impair sperm function,possibly through different mechanisms of action,but fluoxetine is the only exhibiting mild negative effects at doses found in vivo.
文摘The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary possibility of azoospermic men fathering their own genetic offspring.
文摘High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,but the 6th edition of the World Health Organization(WHO)manual for semen analysis included the SDF assessment in the chapter on extended semen examinations.
基金supported by The Canadian Institutes of Health Research(PJT165962).
文摘Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm DNA.This oxidative damage is associated with male infertility.On the other hand,when RoS are produced at low levels,they participate in the redox signaling necessary for sperm capacitation.Capacitation-associated RoS are produced by the sperm oxidase,whose identity is still elusive,located in the plasma membrane of the spermatozoon.Ros,such as superoxide anion,hydrogen peroxide,nitric oxide,and peroxynitrite,activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events.Peroxiredoxins(PRDXs),antioxidant enzymes that fight against oxidative stress,regulate redox signaling during capacitation.Among them,PRDX6,which possesses peroxidase and calcium-independent phospholipase A,(iPLA,)activities,is the primary regulator of redox signaling and the antioxidant response in human spermatozoa.The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)pathway,and it is regulated by PRDX6 iPLA2,protein kinase C(PKC),and receptor-type protein tyrosine kinase.The understanding of redox signaling is crucial to pave theway fornovel diagnostic tools and treatments of male infertility.
文摘Kamil Gill,PhD,Kamil Gill is a biologist who graduated from the University of Szczecin(Poland)with a Master degree.Dr.Gill earned his scientific degree from the Pomeranian Medical University in Szczecin(PUM).His doctoral thesis was entitled:"Molecular markers of male infertility-evaluation of selected human sperm chromatin parameters and its influence on the zygote and embryo development in assisted reproduction conditions".
文摘Except in cases of hypogonadotropic hypogonadism,the use of medical therapy before microsurgical testicular sperm extraction(micro-TESE)is controversial.In some studies,hormone therapy has been shown to improve the possibility of sperm retrieval during micro-TESE and even lead to the presence of sperm in the ejaculate in some cases,thereby obviating the need for micro-TESE.However,their routine use before micro-TESE in cases of nonobstructive azoospermia(NOA)being associated with hypergonadotropic hypogonadism and eugonadism(normogonadotropic condition)has not been supported with robust evidence.In this review,we discuss different types of medical therapy used before micro-TESE for NOA,their risks and benefits,and the available evidence surrounding their use in this setting.
文摘Sperm cryopreservation is an essential technique for male fertility preservation,especially in men who are undergoing medical treatment.Conventional cryopreservation methods face limitations such as oxidative stress,DNA fragmentation,and cytotoxicity associated with traditional cryoprotectants like dimethyl sulfoxide(DMSO).Recent breakthroughs have focused on improving post-thaw sperm viability with novel cryoprotectants and innovative freezing strategies.Prospective approaches include the use of amino acid-based cryoprotectants,deep eutectic solvents,and antioxidants that have been described to prevent oxidative damage and maintain DNA integrity.Vitrification,a high-speed freezing technique that prevents ice crystal formation,has demonstrated superior outcomes compared to conventional slow freezing.Moreover,the Direct Dropping Method,a cryoprotectant-free approach,has been introduced as a contamination-minimizing technique that preserves sperm functionality.Multiomics tools are also utilized to determine biomarkers for protocol optimization.Despite these advancements,cryoprotectant toxicity is a central challenge,emphasizing the necessity for safer agents.Future research must focus on long-term sperm functionality and individualized cryopreservation strategies to maximize reproductive outcomes.The current review highlights the challenges associated with sperm cryopreservation,explores innovative strategies and novel cryoprotectants,underscores the significance of maintaining DNA integrity,and proposes future research directions to improve fertility preservation outcomes.
文摘Medically assisted reproduction(MAR)techniques are highly dependent on the sperm quantity and quality.Low sperm concentrations can be bypassed at least to some point by the usage of more sophisticated MAR techniques like intracytoplasmic sperm injection(ICSI).Compared to this,disruptions in established indicators of sperm quality like motility or morphology pose greater challenges for the therapy of couple infertility.
基金supported by the National Natural Science Foundation of China(No.82371633)Peking University Clinical Scientist Training Program and the Fundamental Research Funds for the Central University(BMU2023PYJ H012).
文摘Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed at the time of sperm retrieval.Onco-micro-TESE addresses the dual objectives of treating both infertility and the testicular tumor simultaneously.The technique is intricate,necessitating a comprehensive understanding of testicular anatomy,physiology,tumor biology,and advanced microsurgical methods.It aims to carefully extract viable spermatozoa while minimizing the risk of tumor dissemination.This review encapsulates the procedural intricacies,evaluates success determinants,including tumor pathology and spermatogenic tissue health,and discusses the implementation of imaging techniques for enhanced surgical precision.Ethical considerations are paramount,as the procedure implicates complex decision-making that weighs the potential oncological risks against the profound desire for fatherhood using the male gametes.The review aims to provide a holistic overview of onco-micro-TESE,detailing methodological advances,clinical outcomes,and the ethical landscape,thus offering an indispensable resource for clinicians navigating this multifaceted clinical scenario.
基金The research was conducted without any financial support from external funding agency.
文摘Objective:To assess the effects of azoxystrobin on sperm quality,hormone levels and testicular structure in Swiss albino mice.Methods:48 Swiss albino male mice were divided into 7 groups containing 6 mice in each group except Group桏in which 12 mice were taken.Group栺served as the control group and treated with vehicle(distil water for 30 and 60 days).Azoxystrobin at three different doses(125,250,500 mg/kg body weight)was orally administered to Group栻,栿and桇for 30 days and Group桋,桍and桏for 60 days.After dose completion,sperm parameters,hormonal profile and testis histology were analysed.Half animals of group桏(500 mg/kg body weight)were left untreated for next 30 days to assess the recovery from reproductive toxicity.Results:At azoxystrobin 125 and 250 mg/kg,sperm viability,count,and motility remained largely unaffected,but a significant decline was observed at azoxystrobin 500 mg/kg after 30 days.After 60 days,all dose levels led to a significant decrease in sperm parameters.Morphological abnormalities in sperm,such as globular heads,bent or coiled tails,and headless or tailless sperm,were noted to be increased in a dose dependent manner.Antioxidant parameters,serum level of reproductive hormones and steroidogenic enzymes followed a similar trend with non-significant changes at low and medium doses but a marked decline at the high dose after both 30 and 60 days of exposure.Histopathology of testis also revealed degenerative changes in seminiferous tubules and Leydig cell.Conclusions:Azoxystrobin exposure at high dose(500 mg/kg)affects sperm quality and disrupts testicular function via potentially impairing antioxidant defences,deregulating steroidogenesis in Swiss albino male mice.
文摘Background:Testicular sperm aspiration(TESA)is a minimally invasive testicular sperm retrieval technique that has been utilized in the treatment of male factor infertility.We sought to evaluate sperm retrieval outcomes of primary and redo TESA in men with severe oligoasthenoteratozoospermia(OAT)and obstructive azoospermia(OA).Methods:This is a retrospective analysis of consecutive TESAs(primary and redo)for men with severe OAT and OA performed between January 2011 and August 2022 at a high-volume infertility center.We compared TESA outcomes in men with severe OAT to those with OA and compared outcomes of men who underwent primary and redo TESA on the same testicular unit.Results:439 TESAs(366 primary and 73 redo)in men with severe OAT(n=133)and OA(n=306)were included.Men with OA had significantly higher sperm retrieval rate(SRR)and motile SRR compared to men with severe OAT(99%vs.95%and 98%vs.83%,respectively,p<0.05).The requirement for multiple biopsies and the total number of aspirates were significantly lower in men with OA compared to those with severe OAT(15%vs.32%and 1.2±0.5 vs.1.4±0.7,respectively,p<0.05).In both groups,SRR,motile SRR,the requirement for multiple biopsies,and the total number of aspirates were not significantly different in primary compared to redo cases.Conclusion:Our data demonstrate that TESA retrieval rates are significantly higher in men with OA compared to those with severe OAT.The data also demonstrate that a redo TESA in these men is as effective as a primary TESA,suggesting that areas of active spermatogenesis are preserved 6 months after TESA.
文摘Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia(OA)and nonobstructive azoospermia(NOA).For men with NOA,testicular sperm extraction(TESE)is the only method to obtain sperm for assisted reproductive technology(ART).Given the rarity of these sperm and the unpredictable success of subsequent retrieval attempts,cryopreservation of microdissection-TESE-obtained sperm is essential.Effective cryopreservation prevents the need for repeated surgical procedures and supports future ART attempts.After first delving into the physiological and molecular aspects of sperm cryopreservation,this review aims to examine the current methods and devices for preserving small numbers of sperm.It presents conventional freezing and vitrification techniques,evaluating their respective strengths and limitations in effectively preserving rare sperm,and compares the efficacy of using fresh versus cryopreserved testicular sperm.
