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Int. braz. j. urol ; 48(3): 471-481, May-June 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1385123


ABSTRACT Purpose: Nonobstructive azoospermia (NOA) associated with primary spermatogenic failure is a common cause of male infertility usually considered untreatable; however, some reports have suggested that hormonal stimulation to boost the intra-testicular testosterone level and spermatogenesis might increase the chance of achieving pregnancy using homologous sperm. Materials and Methods: We report a series of eight NOA males who received long-term treatment with recombinant human chorionic gonadotropin twice a week for spermatogenesis stimulation. Six males received additional recombinant follicle-stimulating hormone (FSH) supplementation 150-225 IU twice weekly. Results: After recombinant gonadotropin therapy, viable spermatozoa were retrieved from the ejaculate in two patients and by testicular sperm aspiration (TESA) in another two subjects. Singleton spermatozoon retrieved from testes were frozen by vitrification on Cell-Sleeper devices. Two live births were obtained after intracytoplasmic sperm injection with ejaculated spermatozoa and one live birth and an ongoing pregnancy using thawed spermatozoa from TESA. Conclusion: Our proof-of-concept study indicates that hormonal therapy with recombinant gonadotropins could be considered in infertile men with NOA as an alternative to sperm donation. Large-scale studies are needed to substantiate hormone stimulation therapy with recombinant gonadotropins in routine clinical practice for this severe form of male infertility.

Asian Journal of Andrology ; (6): 231-237, 2022.
Article in English | WPRIM | ID: wpr-928555


Acephalic spermatozoa syndrome (ASS) is one of the most severe spermatogenic failures of all infertility in men. The cognition of ASS has experienced a tortuous process. Over the past years, with the in-depth understanding of spermatogenesis and the emergence of new genetic research technologies, the unraveling of the genetic causes of spermatogenic failure has become highly active. From these advances, we established a genetic background and made significant progress in the discovery of the genetic causes of ASS. It is important to identify pathogenic genes and mutations in ASS to determine the biological reasons for the occurrence of the disease as well as provide genetic diagnosis and treatment strategies for patients with this syndrome. In this review, we enumerate various technological developments, which have made a positive contribution to the discovery of candidate genes for ASS from the past to the present. Simultaneously, we summarize the known genetic etiology of this phenotype and the clinical outcomes of treatments in the present. Furthermore, we propose perspectives for further study and application of genetic diagnosis and assisted reproductive treatment in the future.

Humans , Infertility, Male/pathology , Male , Membrane Proteins/genetics , Mutation , Spermatogenesis/genetics , Spermatozoa/pathology
Asian Journal of Andrology ; (6): 287-293, 2022.
Article in English | WPRIM | ID: wpr-928534


Intrauterine insemination with donor sperm (IUI-D) is an assisted reproductive technology (ART) offered to couples with definitive male infertility or risk of genetic disease transmission. Here, we sought to evaluate our practice in IUI-D and identify factors that influenced the success rate. We performed a retrospective, single-center study of all IUI-D procedures performed at Lille University Medical Center (Lille, France) between January 1, 2007, and December 31, 2017. Single and multivariate analyses with a mixed logistic model were used to identify factors associated with clinical pregnancies and live births. We included 322 couples and 1179 IUI-D procedures. The clinical pregnancy rate was 23.5%, and the live birth rate was 18.9% per IUI-D. In a multivariate analysis, the women's age was negatively associated with the live birth rate. The number of motile spermatozoa inseminated was the only factor associated with both clinical pregnancies and live births, with a chosen threshold of 0.75 million. The clinical pregnancy and live birth rates were, respectively, 17.3% and 13.0% below the number of motile spermatozoa inseminated threshold and 25.9% and 21.0% at or above the threshold (all P = 0.005). The number of motile spermatozoa inseminated was the only factor that significantly influenced both pregnancies and live-birth rates after IUI-D. Indeed, below a threshold of 0.75 million motile spermatozoa inseminated, those rates were significantly lower. Application of this number of motile spermatozoa inseminated threshold may help centers to allocate donations more effectively while maintaining reasonable waiting times for patients.

Birth Rate , Female , Humans , Insemination , Insemination, Artificial , Male , Pregnancy , Pregnancy Rate , Retrospective Studies , Spermatozoa
Asian Journal of Andrology ; (6): 266-272, 2022.
Article in English | WPRIM | ID: wpr-928525


Gene expression analyses suggest that more than 1000-2000 genes are expressed predominantly in mouse and human testes. Although functional analyses of hundreds of these genes have been performed, there are still many testis-enriched genes whose functions remain unexplored. Analyzing gene function using knockout (KO) mice is a powerful tool to discern if the gene of interest is essential for sperm formation, function, and male fertility in vivo. In this study, we generated KO mice for 12 testis-enriched genes, 1700057G04Rik, 4921539E11Rik, 4930558C23Rik, Cby2, Ldhal6b, Rasef, Slc25a2, Slc25a41, Smim8, Smim9, Tmem210, and Tomm20l, using the clustered regularly interspaced short palindromic repeats /CRISPR-associated protein 9 (CRISPR/Cas9) system. We designed two gRNAs for each gene to excise almost all the protein-coding regions to ensure that the deletions in these genes result in a null mutation. Mating tests of KO mice reveal that these 12 genes are not essential for male fertility, at least when individually ablated, and not together with other potentially compensatory paralogous genes. Our results could prevent other laboratories from expending duplicative effort generating KO mice, for which no apparent phenotype exists.

Animals , CRISPR-Cas Systems/genetics , Fertility/genetics , Gene Editing , Humans , Male , Mice , Mice, Knockout , Testis/metabolism
Asian Journal of Andrology ; (6): 67-72, 2022.
Article in English | WPRIM | ID: wpr-928515


Acephalic spermatozoa syndrome is a rare type of teratozoospermia that severely impairs the reproductive ability of male patients, and genetic defects have been recognized as the main cause of acephalic spermatozoa syndrome. Spermatogenesis and centriole-associated 1 like (SPATC1L) is indispensable for maintaining the integrity of sperm head-to-tail connections in mice, but its roles in human sperm and early embryonic development remain largely unknown. Herein, we conducted whole-exome sequencing (WES) of 22 infertile men with acephalic spermatozoa syndrome. An in silico analysis of the candidate variants was conducted, and WES data analysis was performed using another cohort consisting of 34 patients with acephalic spermatozoa syndrome and 25 control subjects with proven fertility. We identified biallelic mutations in SPATC1L (c.910C>T:p.Arg304Cys and c.994G>T:p.Glu332X) from a patient whose sperm displayed complete acephalia. Both SPATC1L variants are rare and deleterious. SPATC1L is mainly expressed at the head-tail junction of elongating spermatids. Plasmids containing pathogenic variants decreased the level of SPATC1L in vitro. Moreover, none of the patient's four attempts at intracytoplasmic sperm injection (ICSI) resulted in a transplantable embryo, which suggests that SPATC1L defects might affect early embryonic development. In conclusion, this study provides the first identification of SPATC1L as a novel gene for human acephalic spermatozoa syndrome. Furthermore, WES might be applied for patients with acephalic spermatozoa syndrome who exhibit reiterative ICSI failures.

Centrioles/genetics , Homozygote , Humans , Infertility, Male/genetics , Male , Mutation , Spermatogenesis/genetics , Spermatozoa
Asian Journal of Andrology ; (6): 40-44, 2022.
Article in English | WPRIM | ID: wpr-928507


Semen analysis has long been used to evaluate male fertility. Recently, several sperm function tests have been developed. Of those, the sperm DNA fragmentation index (DFI), which describes the status of the sperm DNA, is thought to be a suitable parameter for evaluating male fertility. However, there have been no large-scale studies on the sperm DFI of Japanese men. Therefore, we investigated the feasibility of using an in-house flow cytometry-based sperm DFI analysis based on the sperm DNA fragmentation test of sperm chromatin structure assay (SCSA) to assess male fertility in Japan. This study enrolled 743 infertile and 20 fertile Japanese men. To evaluate reproducibility, inter- and intraobserver precision was analyzed. A receiver operating characteristic curve analysis was used to set a cutoff value for the sperm DFI to identify men who could father children by timed intercourse or intrauterine insemination. The variability of the sperm DFI among fertile volunteers was determined. The relationship between semen parameters and the sperm DFI was assessed by Spearman's rho test. A precision analysis revealed good reproducibility of the sperm DFI. The cutoff value of sperm DNA fragmentation in infertile men was 24.0%. Semen volume had no relationship with the sperm DFI. Sperm concentration, sperm motility, total motile sperm count, and percentage of normal-shaped sperm were significantly and negatively correlated with the sperm DFI. The median sperm DFI was smaller in fertile volunteers (7.7%) than that in infertile men (19.4%). Sperm DNA fragmentation analysis can be used to assess sperm functions that cannot be evaluated by ordinary semen analysis.

Child , Chromatin , DNA Fragmentation , Flow Cytometry , Humans , Infertility, Male/genetics , Japan , Male , Reproducibility of Results , Sperm Motility , Spermatozoa
Braz. J. Pharm. Sci. (Online) ; 58: e19264, 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1374563


Abstract This study investigates the toxic effects of ethanol (Eth) on the reproductive system of male rats and the possible protective role of Silybum marianum seeds-infused solution (SMI) over six consecutive weeks of administration. Animals were divided into the following groups: control, SMI positive control (200 mg/kg/day), Eth1 (1 g/kg/day), Eth2 (2 g/kg/day), Eth1+SMI, and Eth2+SMI. Plasma testosterone concentration, epididymal spermatozoa biology, and testicular and epididymal MDA, GSH and GPx levels were evaluated. The results indicated a significant decrease in testis and epididymis weight, testosterone level, sperm concentration, sperm vitality and sperm motility (total motility, progressive motility, curvilinear velocity, straight-line velocity, velocity average path, beat cross frequency, and lateral head displacement) in both Eth1 and Eth2 compared to the control groups and the combined-treatment groups (Eth1+SMI and Eth2+SMI). Furthermore, results showed a significant elevation in MDA concentration with a significant decrease of testicular and epididymal GSH concentration and GPx activity in theEth1 and Eth2 groups compared to the combined-treatment groups. The administration of SMI succeeded in improving the parameters cited above in the combined-treatment groups compared to the Eth1 and Eth2 groups, and bring them to the levels seen in the control groups. To conclude, SMI has clearly protected reproductive indices against ethanol-induced reprotoxicity in male rats

Rev. colomb. biotecnol ; 23(2): 25-35, jul.-dic. 2021. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1360961


RESUMEN Bocachico Prochilodus magdalenae es una especie endémica y la más importante de la pesquería continental colombiana. No obstante, sus capturas han disminuido aproximadamente el 67% en los últimos cuarenta años, por tanto ha sido categorizada como vulnerable a la extinción. La criopreservación de semen, es una herramienta biotecnológica de conservación por tanto el objetivo del presente estudio fue evaluar la criopreservación de semen de bocachico con etilenglicol (EG) y leche en polvo descremada (LP). La solución crioprotectora estuvo compuesta por EG (6, 8 o 10%), LP (3, 5 o 7%) y glucosa 6%. La calidad del semen descongelado se evaluó con un software tipo CASA (computer assisted semen analysis). El porcentaje de inclusión de EG, no afectó significativamente ninguno de los parámetros de calidad seminal evaluados (p>0,05), a excepción de la tasa de eclosión (p<0,05); mientras que, la LP afectó significativamente el porcentaje de espermatozoides estáticos (p<0,05) y las tasas de fertilización y eclosión (p<0,01). La mayor movilidad total se obtuvo cuando EG se incluyó a 10% y la LP a 7% (38,4±18,4%) (p<0,05); pero las mayores tasas de fertilización (54,3-64,2%) y eclosión (47,7-57,5%) se obtuvieron cuando EG se incluyó a 6 u 8% y la LP se incluyó a la menor concentración evaluada (3%), sin observarse diferencia significativa entre estos tratamientos (p>0,05). Los resultados permiten concluir que la combinación EG 6% con LP 3% permiten la criopreservación de semen de Prochilodus magdalenae de buena calidad y capacidad fecundante.

ABSTRACT Bocachico Prochilodus magdalenae is an endemic species and the most important of the Colombian continental fishery. Its catches have decreased by approximately 67% in the last forty years and, it has been categorized as extinction vulnerable. Semen cryopreservation is a biotechnological conservation tool; therefore, the aim of this study was to evaluate bocachico semen's cryopreservation with ethylene glycol (EG) and skimmed milk powder (LP). The cryoprotective solution was composed of EG (6, 8 or 10%), LP (3, 5 or 7%) and glucose at 6%. The quality of the thawed semen was evaluated with CASA software (computer assisted semen analysis). The inclusion percentage of EG did not significantly affect any of the evaluated semen quality parameters (p>0,05), except for the hatching rate (p <0.05). In contrast, LP presented significant effects on the percentage of static sperm (p <0,05) and on fertilization and hatching rates (p<0,01). The highest total motility was achieved with EG included at 10% and the LP 7% (38,4±18,4%) (p<0,05); but the highest fertility rates (54,3-64,2%) and hatching (47,7-57,5%) were registered when EG included at 6 or 8% and LP included at the lowest rate evaluated (3%), no significant difference was observed between these treatments (p>0,05). The results allow us to conclude that the combination EG 6% with LP 3% allows the cryopreservation of Prochilodus magdalenae semen of good quality and fertilizing capacity.

Int. braz. j. urol ; 47(3): 544-548, May-June 2021. tab
Article in English | LILACS | ID: biblio-1154516


ABSTRACT Introduction: When the vasectomy reversal (VR) fails, and the patient desires natural conception with his sperm, vasectomy re-reversal (VRR) is the only alternative. Purpose: To determine the VRR effectiveness and whether specific parameters can be associated with its success. Materials and Methods: We retrospectively evaluated 18 consecutive vasectomized patients, who had failed their VR through bilateral vasovasostomy, and posteriorly were submitted to VRR. The parameters of the study were: age of the patients, elapsed time between vasectomy and VRR (V-VRRt), elapsed time between VR and VRR (VR-VRRt), presence of spermatozoa in the proximal vas deferens fluid (SptzVDF) in the VRR and results of semen analysis after VRR (SA-VRR). Results: The mean of the age of the patients was 44.11±6.55 years (32.0-57.0), the mean of V-VRRt was 11.76±6.46 years (1.5-25.0) and the mean of VR-VRRt was 2.13±2.27 years (0.5-10.0). SptzVDF in the VRR were found bilaterally in 8 patients, unilaterally in 4 and absent in 6. SA-VRR demonstrated normozoospermia in 9 patients, oligozoospermia in 3 and azoospermia in 6, with patency rate of 66.67%. SA-VRR showed statistically significant dependence only with SptzVDF in the VRR (p <0.01). Conclusions: VRR was effective in restoring the obstruction in more than half of the patients. Furthermore, the presence of spermatozoa in the vas deferens fluid was the parameter associated with the VRR success.

Humans , Male , Adult , Vasectomy , Vasovasostomy , Spermatozoa , Vas Deferens/surgery , Retrospective Studies , Middle Aged
Arq. bras. med. vet. zootec. (Online) ; 73(1): 256-260, Jan.-Feb. 2021. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1153048


As vantagens dos animais transgênicos têm sido demonstradas em diferentes aplicações, entretanto muitas metodologias usadas para gerar animais geneticamente modificados (GM) apresentam baixas taxas de eficiência. O objetivo deste estudo foi avaliar a entrega dos vetores lentivirais (VLs) em zigotos durante a fertilização in vitro (FIV), para gerar embriões GM, com o gene da proteína verde fluorescente (GFP) ou do fator IX de coagulação humana (FIX). Vetores lentivirais com os genes GFP (pLGW-GFP-LV) ou FIX (pLWE2-FIX-LV) foram utilizados na FIV ou na cultura de embriões in vitro (CIV). A coincubação de pLWE2-FIX-LV com espermatozoides e complexos oócitos-células do cumulus (COCs) durante a FIV diminuiu (P<0,05) as taxas de clivagem e de blastocistos, enquanto com pLGW-GFP-LV diminuiu (P<0,05) a taxa de blastocisto quando se comparou ao controle sem VL. A coincubação de pLWE2-FIX-LV e pLGW-GFP-LV com presumíveis zigotos durante a CIV não afetou (P>0,05) o desenvolvimento embrionário. A expressão da proteína GFP não foi detectada em embriões após a coincubação de FIV ou CIV, embora as células do cumulus expressassem a proteína até o dia oito de cultivo in vitro. Reações em cadeia da polimerase (PCR) não detectaram os genes GFP ou FIX em embriões, mas ambos foram detectados em células do cumulus. Assim, a coincubação de VL com espermatozoide bovino e COCs não é eficaz para produzir embriões geneticamente modificados por meio de FIV.(AU)

Animals , Cattle , Zygote , Animals, Genetically Modified/genetics , Transgenes , Embryo, Mammalian , Genetic Vectors/analysis , Fertilization in Vitro/veterinary , Gene Transfer Techniques/veterinary
Article in Chinese | WPRIM | ID: wpr-942257


To explore the genetic causes of 3 male infertility patients with acephalospermia and the outcome of assisted reproductive technology. Clinical diagnosis, sperm morphology examination, sperm transmission electron microscopy examination were performed on 3 patients, and the whole exome sequencing technology was used for screening, Sanger sequencing verification, mutation pathogenicity analysis, and protein sequence homology comparison. Assisted reproductive technology was implemented to assist pregnancy treatment. The 3 patients were all sporadic infertile men, aged 25, 42 and 26 years, and there was no obvious abnormality in the general physical examination. Male external genitalia developed normally, bilateral testicles were normal in volume, and bilateral epididymis and spermatic vein were palpated without nodules, cysts, and tenderness. Repeated semen analysis showed that a large number of immature sperm could be seen, and they had the ability to move. The SUN5 gene of the 3 male infertile patients was a case of homozygous missense mutation c.7C>T (p.Arg3Trp), a case of compound heterozygous missense mutation c.1067G>A (p.Arg356His) and nonsense mutation c.216G>A (p.Trp72*) and a case of homozygous missense mutation c.1043A>T (p.Asn348Ile), of which c.7C>T (p.Arg3Trp) and c.1067G>A (p.Arg356His) were new variants that had not been reported. SIFT, Mutation Taster and PolyPhen-2 software function prediction results were all harmful, the nonsense mutation c.216G>A (p.Trp72*) led to the premature termination of peptide chain synthesis which might have a greater impact on protein function. The homology regions in the protein sequence homology alignment were all highly conserved.The 3 male patients and their spouses obtained 4 biological offspring through intracytoplasmic sperm injection, all of which were boys, and one of them was a twin.Three male infertile patients might be caused by SUN5 gene mutations. Such patients could obtain their biological offspring through assisted reproductive technology. It was still necessary to pay attention to the genetic risk of ASS, it was recommended that both men and women conduct genetic counseling and screening at the same time. In clinical diagnosis, whole exome sequencing technology could be used to perform auxiliary examinations to determine the treatment plan and assisted reproductive methods as soon as possible to reduce the burden on the family and society. The newly discovered mutation sites of SUN5 gene provided clues and directions for elucidating the pathogenic mechanism, and at the same time expanded the pathogenic mutation spectrum of ASS.

Female , Humans , Infertility, Male/genetics , Male , Membrane Proteins/genetics , Mutation , Pregnancy , Sperm Injections, Intracytoplasmic , Spermatozoa
Asian Journal of Andrology ; (6): 363-369, 2021.
Article in English | WPRIM | ID: wpr-888437


Many azoospermic men do not possess mature spermatozoa at the time of surgical sperm extraction. This study is a systematic review and meta-analysis evaluating outcomes following round spermatid injection (ROSI), a technique which utilizes immature precursors of spermatozoa for fertilization. An electronic search was performed to identify relevant articles published through October 2018. Human cohort studies in English involving male patients who had round spermatids identified and used for fertilization with human oocytes were included. Fertilization rate, pregnancy rate, and resultant delivery rate were assessed following ROSI. Meta-analysis outcomes were analyzed using a random-effects model. Data were extracted from 22 studies involving 1099 couples and 4218 embryo transfers. The fertilization rate after ROSI was 38.7% (95% confidence interval [CI]: 31.5%-46.3%), while the pregnancy rate was 3.7% (95% CI: 3.2%-4.4%). The resultant delivery rate was low, with 4.3% of embryo transfers resulting in a delivery (95% CI: 2.3%-7.7%). The pregnancy rate per couple was 13.4% (95% CI: 6.8%-19.1%) and the resultant delivery rate per couple was 8.1% (95% CI: 6.1%-14.4%). ROSI has resulted in clinical pregnancies and live births, but success rates are considerably lower than those achieved with mature spermatozoa. While this technique may be a feasible alternative for men with azoospermia who decline other options, couples should be aware that the odds of a successful delivery are greatly diminished and the prognosis is relatively poor.

Asian Journal of Andrology ; (6): 146-149, 2021.
Article in English | WPRIM | ID: wpr-879737


Varicoceles adversely impact semen quality and sperm DNA fragmentation, which typically improve with surgical repair. Some men with varicoceles have ipsilateral testicular atrophy due to damage from the varicocele. This study assessed semen quality and the sperm DNA fragmentation index (DFI) response to varicocele repair in men with ipsilateral testicular atrophy (TA) versus men with no testicular atrophy (NTA). Semen parameter values and DFI in both groups were compared preoperatively and postoperatively. The Mann-Whitney U test and the Wilcoxon signed-rank test were used where appropriate. There were 20 men in the TA group and 121 men in the NTA group with no difference in age, varicocele grade, or preoperative semen parameter values between the two groups. The NTA group had a higher preoperative DFI than the TA group. Both groups showed improvement in semen quality postoperatively, only the TA group showed a significant improvement in DFI, whereas the NTA group showed significant improvements in several parameter values and DFI. The change from preoperative to postoperative parameter values when comparing the two groups revealed a difference in total sperm motile count and DFI, with a larger mean improvement in the NTA group than in the TA group. Both TA and NTA groups showed improved semen quality and DFI after varicocele repair, but the NTA group had more improvement than the TA group. However, only total motile count (TMC) and DFI had a significantly greater mean change in preoperative to postoperative response in the NTA group than in the TA group.

Orinoquia ; 24(2): 51-78, July-Dec. 2020. tab
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1250435


Resumen La crioconservación es una herramienta biotecnológica que en peces está orientada principalmente a la conservación criogénica de semen como estrategia de preservación del recurso genético y a su uso para la producción de alevinos con fines diferentes. Actualmente, los protocolos de crioconservación seminal en peces de agua dulce establecen una amplia variedad de procedimientos cuya efectividad se basa en aspectos ligados a la calidad seminal post-descongelación y la fertilidad, así como su relación con el desarrollo de la progenie. El efecto de la conservación del semen en nitrógeno líquido por periodos amplios de tiempo también toma importancia en ésta biotecnología. Por lo anterior, el objetivo de la presente revisión es describir aspectos biotecnológicos, celulares y bioquímicos asociados al proceso de crioconservación seminal en peces dulceacuícolas, resaltando los avances, las limitaciones y sus perspectivas.

Abstract Cryopreservation is a biotechnological tool that in fish is mainly aimed at cryogenic conservation of semen as a strategy for preserving the genetic resource and its use for the production of fingerlings with different purposes. Currently, seminal cryopreservation protocols in freshwater fish establish a wide variety of procedures whose effectiveness is based on aspects linked to seminal post-thaw quality and fertility, as well as its relationship with the development of the progeny. The effect of preserving semen in liquid nitrogen for extended periods of time also plays an important role in this biotechnology. Therefore, the objective of this review is to describe biotechnological, cellular and biochemical aspects associated with the seminal cryopreservation process in freshwater fish, highlighting the advances, limitations and perspectives.

Resumo A criopreservação é uma ferramenta biotecnológica que em peixes visa principalmente a conservação criogênica do sêmen como estratégia para a preservação do recurso genético e sua utilização para a produção de alevinos para diferentes fins. Atualmente, os protocolos de criopreservação seminal em peixes de água doce estabelecem uma ampla variedade de procedimentos cuja eficácia se baseia em aspectos relacionados à qualidade e fertilidade pós-descongelamento seminal, bem como sua relação com o desenvolvimento da progênie. O efeito da preservação do sêmen no nitrogênio líquido por longos períodos de tempo também desempenha um papel importante nessa biotecnologia. Portanto, o objetivo desta revisão é descrever aspectos biotecnológicos, celulares e bioquímicos associados ao processo de criopreservação seminal em peixes de água doce, destacando os avanços, limitações e perspectivas.

Rev. chil. obstet. ginecol. (En línea) ; 85(4): 312-323, ago. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1138627


OBJETIVO: El objetivo de este trabajo fue establecer el efecto de la borra de café sobre la movilidad y los parámetros funcionales de los espermatozoides humanos in vitro. MATERIALES Y MÉTODOS: La borra de café, un subproducto obtenido en establecimientos especializados en la preparación de café soluble a base de grano, se diluyo en tampón fosfato salino y se mezcló en proporciones iguales con las muestras de semen de 16 voluntarios aparentemente sanos. A cada muestra se le determinó el efecto sobre la movilidad espermática en función del tiempo (30, 60, 90 y 120 minutos, n=16) y sobre los parámetros funcionales (n=6) por medio de citometría de flujo: potencial de membrana mitocondrial, producción de especies reactivas de oxígeno y lipoperoxidación de la membrana espermática. RESULTADOS: La incubación de los espermatozoides con la borra de café evidencio un cambio positivo en la movilidad espermática. Adicionalmente, la incubación con la borra de café incremento significativamente el potencial de membrana mitocondrial en los espermatozoides. CONCLUSIÓN: La borra de café, seguramente debido a los compuestos antioxidantes, afecta positivamente la movilidad espermática aumentando el potencial de membrana mitocondrial. Por lo tanto, esto es un paso inicial en la búsqueda de un suplemento de origen natural que aumente la calidad seminal.

OBJECTIVE: The objective of this work is to establish the effect of spent coffee grounds on the motility and functional parameters of human spermatozoa, in vitro. MATERIALS AND METHODS: Spent coffee grounds, a by-product obtained in specialized establishments in the preparation of soluble coffee based on grain, was diluted in saline phosphate buffer and mixed in equal proportions with semen samples from 16 apparently healthy volunteers. Each sample was determined the effect on sperm motility as a function of time (30, 60, 90 and 120 minutes, n=16) and on functional parameters (n=6) by means of flow cytometry: mitochondrial membrane potential, reactive oxygen species production and membrane lipoperoxidation. RESULTS: The incubation of the spermatozoa with the spent coffee grounds showed a positive change in sperm motility. Additionally, incubation with spent coffee grounds significantly increased the mitochondrial membrane potential in human sperm cells. CONCLUSION: Spent coffee grounds, probably due to antioxidant compounds, positively affects sperm motility by increasing mitochondrial membrane potential. Therefore, this is an initial step in the search for a supplement of natural origin that increases seminal quality.

Humans , Male , Adult , Young Adult , Semen/drug effects , Sperm Motility/drug effects , Spermatozoa/drug effects , Plant Extracts/pharmacology , Coffee/chemistry , Semen/physiology , Sperm Motility/physiology , Spermatozoa/physiology , Time Factors , In Vitro Techniques
Rev. peru. med. exp. salud publica ; 37(2): 292-296, abr.-jun. 2020. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1127144


RESUMEN Con el objetivo de caracterizar la calidad seminal de hombres en un centro de reproducción asistida de la ciudad Guayaquil (Ecuador), se colectaron 204 muestras de semen de pacientes con problemas de fertilidad de entre 20 y 57 años, atendidos entre mayo de 2017 y septiembre de 2018. Se realizó un espermograma básico a cada muestra, siguiendo las recomendaciones del manual para la examinación y procesamiento de semen humano. El 27,4% de las muestras presentó normozoospermia. Dentro de las alteraciones la teratozoospermia fue de 27,9%, oligoteratozoospermia del 8,8%, evidenciándose mayor número en pacientes de 30 a 39 años. Un alto porcentaje de pacientes presentan una calidad del semen y morfología espermática por debajo los limites de referencia establecidos por la Organización Mundial de la Salud.

ABSTRACT In order to characterize the quality of semen from men in an assisted reproduction center in the city of Guayaquil (Ecuador), 204 semen samples were collected from patients with fertility disorders aged 20 to 57 years, who were admitted between May 2017 and September 2018. A basic spermogram was performed on each sample, following the fabricant recommendations for the examination and processing of human semen. It was found that 27.4% of the samples presented normozoospermia. Among the disorders, it was found that 27.9% had teratozoospermia, 8.8% had oligoteratozoospermia and a higher number of patients were found to be between 30 and 39 years old. A high percentage of patients presented sperm morphology and quality values below the reference limits established by the World Health Organization.

Article | IMSEAR | ID: sea-210908


Dead and damaged spermatozoa cells present universally in the ejaculates of all eutherian mammals exert toxic effect on contemporary healthy cells mostly through generation of excessive free radicals. This is much more evident during extended period of processing, resulting in poor ejaculate quality. The solution lies in depletion of dead/damaged spermatozoa from the neat ejaculates itself. Thus the objective of the study was to evaluate the efficiency of the protocols such as discontinuous PercollTM density gradient centrifugation (PDGC) and glass wool filtration (GWF) for depletion of dead/damaged spermatozoa from fresh semen in buffalo. Random ejaculates (n=6) of Murrah buffalo bulls were divided into two aliquots after quality assessment: PDGC and GWF protocols (Group I and II, respectively). At the end of the purification protocol, efficiency of the protocols in depleting dead/damaged spermatozoa as reflected by certain quality parameters were evaluated. The mean efficiency (%) of purification protocols based on recovery of spermatozoa was 44.68 and 40.02% for PDGC and GWF, respectively. Moreover significantly (p<0.05) greater values for quality parameters was observed in the Group II (26.4+6.8 vs 68.8+4.4 for acrosome integrity (%); 12.68+6.6 vs 57.7+7.5 for functional plasma membrane integrity (%); 20.3+5.8 vs 80.75+6.7 for viability (%) in Group I and II, respectively). It was concluded that GWF is a better technique than PGDC to filter out dead/damaged spermatozoa from fresh semen with improvement in semen quality and can be a valuable tool in assisted reproductive technology

Article | IMSEAR | ID: sea-210906


Spermatozoa undergoes array of signaling and intracellular pathways and ultimately become competent enough to accomplish fertilization. Hormones, ion channels and signaling molecules in both male and female reproductive tract show bidirectional cross play. The recent discovery of endocannabinoids and their receptors in male and female reproductive system opened new vistas for their research in regulating sperm function. Interestingly, endocannabinoids regulate sperm motility, capacitation, hyperactivity and eventually acrosome reaction. However, their complex intracellular pathways are still to be understood in regulating spermatozoa function. The present review highlights the major breakthrough research in the area of endocannabinoids in male reproduction and in more specific in sperm cells, and their association with regulation of sperm fertilizing competence

Pesqui. vet. bras ; 40(4): 306-314, Apr. 2020. tab
Article in English | LILACS, VETINDEX | ID: biblio-1135625


The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P<0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.(AU)

O objetivo deste trabalho foi avaliar a qualidade espermática de gatos domésticos obtidos por eletroejaculação e recuperação da cauda do epidídimo após a refrigeração a -1°C e a 4°C por 24 e 48 horas. Vinte e nove gatos adultos (2 a 6kg) foram utilizados. A colheita de espermatozoides foi realizada por eletroejaculação (EEJ) e, após 48 horas, os gatos foram orquiectomizados, e as amostras espermáticas foram obtidas a partir do ducto deferente e da cauda do epidídimo (EPD). As amostras foram diluídas em ACP-117® e as características espermáticas foram avaliadas em três momentos distintos: fresco, 24 e 48 horas após a refrigeração. Para ser possível comparar as duas temperaturas de refrigeração, a primeira etapa foi analisar se havia diferença entre as técnicas de colheita. Após isto, dois experimentos foram conduzidos: no primeiro, espermatozoides de 14 gatos foram utilizados e a refrigeração foi realizada a -1°C; e no segundo, amostras de 15 gatos foram utilizados e os espermatozoides foram refrigerados a 4°C. A cinética espermática foi avaliada por análise computadorizada (CASA), a concentração por câmara de Neubauer, a morfologia espermática foi avaliada pela coloração de Karras modificada, e a integridade da membrana foi avaliada por eosina nigrosina. Os resultados obtidos foram analisados no software R, versão 3.2.5, utilizando o teste de Mann-Whitney para variáveis com distribuições anormais, considerando significância ao nível de 5%. No ejaculado, maiores valores de defeitos morfológicos totais foram observados após 24 e 48 horas de refrigeração a 4°C (P<0,022) em comparação com refrigeração a -1°C, usando o teste de Friedman. Para quantificar a diminuição na qualidade espermática, as reduções dos parâmetros foram calculadas entre os pontos de tempo (F-24h/F-48h/24h-48h). Na EPD, uma maior redução na qualidade espermática foi detectada após 24 horas de refrigeração a 4°C, tanto na motilidade e na cinética espermática quanto nos índices de movimento e velocidade, em comparação com a refrigeração a -1°C. Com base nos resultados, pode concluir-se que a refrigeração dos espermatozoides felino a -1°C, até 48 horas, foi eficaz na manutenção da qualidade espermático colhidos por EEJ e EPD, e pode ser uma alternativa para a criopreservação de espermatozoides em felinos domésticos.(AU)

Animals , Male , Cats , Semen , Semen Preservation/veterinary , Spermatozoa , Cryopreservation , Reproductive Techniques, Assisted , Reproductive Techniques, Assisted/veterinary , Epididymis
Rev. bras. ciênc. vet ; 27(2): 80-87, abr./jun. 2020. il.
Article in English | LILACS, VETINDEX | ID: biblio-1378276


Some amino acids can protect mammalian sperm cells against oxidation during thermal stress caused by freezing/thawing. Thus, the objective was to evaluate the protective action of the association of the amino acids L-proline (Pro) and L-glutamine (Glu) against the cryoinjury caused to sheep sperm after cryopreservation. Eight ejaculates were collected from four sheep (n=32) and diluted in Tris-Egg Yolk-Glycerol until the final concentration of 200 x106 sptz/mL and kept in a water bath at 32 °C. The amino acids were added as follows: control (without adding amino acids), Pro+Glu 1 (100 µM Pro + 500 µM Glu), Pro+Glu 2 (300 µMPro + 1000 µM Glu), Pro+Glu 3 (500 µM Pro + 1500 µM Glu) and Pro+Glu 4 (700 µM Pro + 2000 µM Glu). Afterwards, the semen was cooled to 5 °C for 2 h, after that period, filled in 0.5 mL straws and then placed under liquid nitrogen vapor (N2L), 8 cm from the liquid sheet for 15min, and then immersed on the N2L. The samples were analyzed for sperm motility, plasma membrane and acrosomal membrane integrity, mitochondrial activity and binding test. The variables were subjected to the normality tests (Lilliefors test) and homoscedasticity tests (Cochran and Bartlett test), afterwards the variables of normal distribution were subjected to analysis of variance and the means compared by the Tukey test with a significance level of 5%. The Pro+Glu 3 group exhibited sperm with a greater (P<0.05) motility after thawing. In addition, the highest percentage of plasma and acrosomal membrane integrity were obtained using Pro+Glu 1, Pro+Glu 2 and Pro+Glu 3; and Pro+Glu 2 and Pro+Glu 3, respectively. Amino acids also kept mitochondrial activity high compared to the control, with Pro+Glu 3 resulting in greater activity (P<0.05). Sperm viability was higher (P<0.05) with the use of Pro+Glu 2 and Pro+Glu 3 than in the control. The number of sperm that showed the ability to bind to the egg yolk perivitelline membrane was higher (P<0.05) in semen treated with amino acids. It is concluded that the addition of synthetic amino acids in the semen of sheep before cryopreservation improves sperm quality and fertilization potential and can thus be added in cryopreservation protocols.

Alguns aminoácidos podem proteger as células espermáticas de mamíferos contra a oxidação durante o estresse térmico causado na congelação/descongelação. Dessa forma, objetivou-se avaliar a ação protetora da associação dos aminoácidos L-prolina (Pro) e L-glutamina (Glu) contra as crioinjúrias causadas aos espermatozoides de ovino após a criopreservação. Foram coletados oito ejaculados de quatro carneiros (n=32) e diluídos em Tris-Gema de ovo-Glicerol até a concentração final de 200 x106 sptz/mL e, mantidos em banho maria a 32 °C. Os aminoácidos foram adicionados da seguinte forma: controle (sem adição de aminoácidos), Pro+Glu 1 (100 µM Pro + 500 µM Glu), Pro+Glu 2 (300 µM Pro + 1000 µM Glu), Pro+Glu 3 (500 µM Pro + 1500 µM Glu) e Pro+Glu 4 (700 µM Pro + 2000 µM Glu). Depois, o sêmen foi resfriado a 5 °C por 2 h, após esse período, envasado em palhetas de 0,5 mL e então acondicionado sob vapor de nitrogênio líquido (N2L), a 8 cm da lâmina líquida por 15 min, e depois imersos no N2L. As amostras foram analisadas quanto à motilidade espermática, integridade da membrana plasmática e da membrana acrossomal, atividade mitocondrial e teste de ligação. As variáveis foram submetidas aos testes de normalidade (Teste de Lilliefors) e homocedacidade (Teste de Cochran e Bartlett), posteriormente as variáveis de distribuição normal foram submetidas à análise de variância e as médias comparadas pelo teste de Tukey com nível de significância de 5%. O grupo Pro+Glu 3 exibiu espermatozoides com uma maior (P<0,05) motilidade após o descongelamento. Além disso o maior percentual de integridade da membrana plasmatica e acrossomal foram obtidos utilizando Pro+Glu 1, Pro+Glu 2 e Pro+Glu 3; e Pro+Glu 2 e Pro+Glu 3, respectivamente. Os aminoácidos também mantiveram alta a atividade mitocondrial em comparação com o controle, com Pro+Glu 3 resultando numa maior atividade (P<0,05). A viabilidade dos espermatozoides foi maior (P<0,05) com o uso de Pro+Glu 2 e Pro+Glu 3 do que no controle. O número de espermatozoides que apresentaram à capacidade de ligação a membrana perivitelina da gema de ovo foi maior (P<0,05) no sêmen tratado com aminoácidos. Conclui-se que, a adição dos aminoácidos sintéticos no sêmen de ovinos antes da criopreservação melhora a qualidade espermática e o potencial fecundante, podendo assim serem adicionados em protocolos de criopreservação.

Animals , Spermatozoa/drug effects , Sheep/genetics , Cryopreservation/veterinary , Semen Analysis/veterinary , Fertility/drug effects , Fertility Agents, Male/administration & dosage , Proline/administration & dosage , Glutamine/administration & dosage