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1.
São Paulo; s.n; 2023. 86 p. ilus, tab.
Thesis in Portuguese | LILACS, Inca | ID: biblio-1434705

ABSTRACT

INTRODUÇÃO: A oncofertilidade tem o desafio de buscar estratégias para preservar a função reprodutiva. Este estudo explorou duas possibilidades como implementação para as técnicas de preservação da fertilidade feminina e masculina. OBJETIVO: Analisar a eficiência do cultivo de folículos pré-antrais de camundongos suplementado com lisado de plaquetas humanas e desenvolver um protótipo para criopreservação de sêmen humano. MATERIAIS E MÉTODOS: Os folículos pré-antrais foram isolados mecanicamente de ovários de fêmeas de camundongos e foram cultivados individualmente em sistema entre camadas de óleo mineral. Os folículos foram cultivados divididos em 4 grupos, sendo um controle, sem o uso do lisado de plaquetas e três grupos com diferentes concentrações de lisado de plaquetas humanas (PLTMax®). Foram avaliadas a sobrevivência celular, desenvolvimento folicular e características oocitárias. Para o segundo estudo foi desenvolvido e impresso em 3D com filamentos de acrilonitrilo butadieno-estireno (ABS) um protótipo que suporte 10 palhetas com amostras seminais no vapor de nitrogênio líquido (N2L), etapa essencial para criopreservação de sêmen humano. Para os testes foram utilizadas 40 amostras seminais. A temperatura ambiente e no interior das palhetas de envase das amostras foram medidas e estabelecida a curva de resfriamento. Os parâmetros de motilidade, vitalidade e fragmentação do DNA espermático foram avaliados antes do congelamento e após o descongelamento. Foram realizados dois testes, um de posicionamento das palhetas e outro comparativo entre o protótipo e um dispositivo com suporte em poliestireno expandido (EPS). RESULTADOS: O cultivo de 11 dias induziu um aumento no tamanho folicular em todas as condições, sendo maior no grupo controle, seguido do grupo com 10% de PLTMax®, mas com diferença significativa (p<0,001). O grupo controle apresentou maior número de oócitos intactos (>50%) em relação aos demais (<35%). Todos os 4 grupos apresentaram taxas de vitalidade celular acima de 70%. Quanto aos testes com o protótipo em ABS foi verificado que as curvas de refrigeração foram notavelmente reproduzíveis. O material do protótipo resistiu a inúmeros mergulhos (>300) no N2L, sem demonstrar danos ao material. Diferenças significativas (p<0,001) foram observadas para a taxa de recuperação média da motilidade e vitalidade espermática em relação aos dados da amostra 2 fresca em ambos os testes. A motilidade, a vitalidade e a fragmentação do DNA espermático antes do congelamento e após o descongelamento não mostraram diferenças em relação a posição das palhetas. Também não houve diferença quanto ao índice de fragmentação verificada das amostras criopreservadas com uso do protótipo em ABS e o suporte em EPS, mesmo após o cultivo, após 24 horas de cultivo. Contudo, houve diferença em relação a amostra fresca (p<00,1). Quanto a recuperação das taxas de motilidade e vitalidade não houve diferença entre o ABS e EPS após o descongelamento e 24 horas de cultivo. CONCLUSÃO: O PLTMax®, embora tenha apresentado menor desempenho que o HSA, é um candidato de suplementação para o cultivo de folículos pré-antrais que merece ser mais explorado. O protótipo em ABS demonstrou resistência, praticidade e segurança para criopreservação seminal de forma reprodutível e eficiente.


INTRODUCTION: Oncofertility has the challenge of seeking strategies to preserve reproductive function. This study explored two possibilities as implementations for female and male fertility preservation techniques. PURPOSE: To analyze the efficiency of mouse preantral follicle culture supplemented with human platelet lysate and to develop a prototype for human semen cryopreservation. MATERIAL AND METHODS: Preantral follicles were mechanically isolated from female mouse ovaries and were individually cultured using a mineral oil interlayer system. The follicles were cultured divided into 4 groups, one control, without the use of platelet lysate and three groups with different concentrations of human platelet lysate (PLTMax®). Cell survival, follicular development and oocyte characteristics were evaluated. For the second study, a prototype was developed and printed in 3D with acrylonitrile butadiene styrene (ABS) filaments to support 10 straws with seminal samples in liquid nitrogen (N2L) vapor, an essential step for human semen cryopreservation. For the tests 40 seminal samples were used. Ambient and internal temperatures inside the sample straws were measured and the cooling curve was established. The parameters of motility, vitality and sperm DNA fragmentation were evaluated before freezing and after thawing. Two tests were performed, one for positioning the straws and the other comparing the prototype and a device with expanded polystyrene (EPS) support. RESULTS: The 11-day culture induced an increase in follicular size in all conditions, being higher in the control group followed by the group with 10% PLTMax®, but with significant difference (p<0.001). The control group presented a higher number of intact oocytes (>50%) compared to the others (<35%). All 4 groups presented cell vitality rates above 70%. As for the ABS prototype tests, it was verified that the cooling curves were remarkably reproducible. The prototype withstood numerous dips (>300) in N2L without showing damage to the material. Significant differences (p<0.001) were observed for the mean recovery rate of sperm motility and vitality compared to the fresh sample data in both tests. Motility, vitality and sperm DNA fragmentation before freezing and after thawing showed no differences with respect to the position of the straws. There was also no difference in the fragmentation index verified for samples cryopreserved using the ABS prototype and the EPS support, even after 24 hours of culture. However, there was a difference compared to the fresh 4 sample (p<00.1). As for the recovery of motility and vitality rates there was no difference between ABS and EPS after thawing and 24 hours of culture. CONCLUSION: PLTMax®, although it showed lower performance than HSA, is a supplementation candidate for preantral follicle culture that deserves further exploration. The ABS prototype demonstrated strength, practicality and safety for seminal cryopreservation in a reproducible and efficient manner.


Subject(s)
Humans , Animals , Cryopreservation , Fertility , Semen , Ovarian Follicle , Mice , Neoplasms/complications
2.
Journal of Integrative Medicine ; (12): 130-135, 2023.
Article in English | WPRIM | ID: wpr-971651

ABSTRACT

Male infertility has become a problem worldwide, and recent research has emphasized the development of more effective therapy options. Among natural compounds, rutin has been widely studied for its potential to treat dysfunction related to male infertility, including a reduction in sperm quality, spermatogenesis disruption and structural disruption in the testis. A thorough review of scientific literature published in several databases, including Google Scholar, PubMed/MEDLINE and Scopus, was used to synthesize the present state of research on the role of rutin in male reproductive health. Rutin has been shown to possess antiapoptotic, antioxidant and anti-inflammatory activities, among others, which are crucial in the management of male infertility. Numerous investigations have shown that rutin protects against male infertility and have explored the underlying mechanisms involved. The present review, therefore, assesses the therapeutic mechanisms involved in male infertility treatment using rutin. Rutin was able to mitigate the induced oxidative stress, apoptosis, inflammation, and related physiological processes that can cause testicular dysfunction. Please cite this article as: Rotimi DE, Elebiyo TC, Ojo OA. Therapeutic potential of rutin in male infertility: A mini review. J Integr Med. 2023; 21(2): 130-135.


Subject(s)
Male , Humans , Rutin/analysis , Semen , Testis , Spermatozoa , Oxidative Stress , Infertility, Male/drug therapy
3.
Asian Journal of Andrology ; (6): 240-244, 2023.
Article in English | WPRIM | ID: wpr-971021

ABSTRACT

The effects of the coronavirus disease 2019 (COVID-19) pandemic on male fertility have received considerable attention because human testes contain high levels of angiotensin-converting enzyme-2 receptors, through which severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can enter. Early studies showed decreases in semen quality during and after recovery from COVID-19. However, no semen quality studies have examined the effects of widespread subclinical and mild disease, as well as changes in lifestyle, psychosocial behavior, intake of dietary supplements, and stress. This cross-sectional study compared semen quality parameters in male partners of infertile couples between men who underwent semen analysis before the COVID-19 pandemic (prepandemic group) and men who underwent semen analysis during the pandemic period (pandemic group); the analysis sought to clarify the overall effects of the pandemic. No participants in the pandemic group had experienced clinically overt disease. Among the 239 participants, mean body weight (P = 0.001), mean body mass index (P < 0.001), median sperm concentration (P = 0.014), total sperm count (P = 0.006), and total percentages of motile (P = 0.013) and abnormal cells (P < 0.001) were significantly greater in the pandemic group (n = 137) than those in the prepandemic group (n = 102). Among abnormal cells, the percentages of cells with excess residual cytoplasm (P < 0.001), head defects (P < 0.001), and tail defects (P = 0.015) were significantly greater in the pandemic group than those in the prepandemic group. With the exception of morphology, the overall semenogram results were better in the pandemic group than those in the prepandemic group.


Subject(s)
Humans , Male , Pandemics , Infertility, Male , COVID-19 , Cross-Sectional Studies , Testis , SARS-CoV-2 , Semen , Semen Analysis , Sperm Count
4.
Asian Journal of Andrology ; (6): 119-125, 2023.
Article in English | WPRIM | ID: wpr-971010

ABSTRACT

The hemodynamic characteristics of venous reflux are associated with infertility in patients with varicocele; however, an effective method for quantifying the structural distribution of the reflux is lacking. This study aimed to predict surgical outcomes using a new software for venous reflux quantification. This was a retrospective cohort study of a consecutive series of 105 patients (age range: 22-44 years) between July 2017 and September 2019. Venous reflux of the varicocele was obtained using the Valsalva maneuver during scrotal Doppler ultrasonography before microsurgical varicocelectomy. Using this software, the colored reflux signals were segmented, and the gray scale of the color pixels representing the reflux velocity was comprehensively quantified into the mean reflux velocity of the green layer (MRVG) and the reflux velocity standard deviation of the green layer (RVSDG). Spontaneous pregnancy and changes from baseline in the semen parameters were assessed during a 12-month follow-up period. Data were analyzed using logistic regression analysis. An association of the high MRVG group with impaired progressive motility (odds ratio [OR] = 2.868, 95% confidence interval [CI]: 1.133-7.265) and impaired sperm concentration (OR = 2.943, 95% CI: 1.196-7.239) was found during multivariate analysis. High MRVG (OR = 2.680, 95% CI: 1.086-6.614) and high RVSDG (OR = 2.508, 95% CI: 1.030-6.111) were found to be independent predictors of failure to achieve pregnancy following microsurgical repair. In summary, intense venous reflux is an independent predictor of impaired progressive motility, sperm concentration, and pregnancy outcomes after microsurgical varicocelectomy.


Subject(s)
Pregnancy , Female , Humans , Male , Young Adult , Adult , Varicocele/surgery , Retrospective Studies , Semen , Veins/surgery , Sperm Count , Infertility, Male/surgery , Microsurgery/methods , Sperm Motility
5.
Asian Journal of Andrology ; (6): 252-258, 2023.
Article in English | WPRIM | ID: wpr-971008

ABSTRACT

Wenzhou has improved its environmental quality because of comprehensive environmental remediation; nevertheless, the semen quality of infertile males remains unclear. This study determined whether better environmental quality improved semen quality in this region. We recorded semen quality data from 22 962 infertile males from January 2014 to November 2019 at the Center for Reproductive Health of The First Affiliated Hospital of Wenzhou Medical University (Wenzhou, China). Patients were predominantly 30-35 years old (33.1%) and workers (82.0%), with high school education or lower (77.6%); more than a half of the patients (52.6%) were Wenzhou household registration; and most patients (77.5%) had abnormal semen quality. Patients who were older than 40 years and workers, and those with Wenzhou household registration, had significantly worse semen quality (all P < 0.05). From 2014 to 2019, progressive sperm motility, total sperm motility, and semen volume showed increasing linear trends in all patients (P = 0.021, 0.030, and 0.005, respectively), yet normal sperm morphology showed a linearly decreasing trend (P = 0.046). Sensitivity analyses for subgroups yielded similar results. In conclusion, the improvement of environmental quality and better function of the accessory glands are associated with progressive sperm motility, total sperm motility, and semen volume. Normal sperm morphology is influenced by occupational exposures and personal lifestyle and does not improve with environmental quality.


Subject(s)
Male , Humans , Adult , Semen Analysis , Semen , Sperm Count , Sperm Motility , Infertility, Male , Spermatozoa
6.
Asian Journal of Andrology ; (6): 245-251, 2023.
Article in English | WPRIM | ID: wpr-971006

ABSTRACT

Advanced paternal age has been overlooked, and its effect on fertility remains controversial. Previous studies have focused mainly on intracytoplasmic sperm injection (ICSI) cycles in men with oligozoospermia. However, few studies have reported on men with semen parameters within reference ranges. Therefore, we conducted a retrospective cohort study analyzing the reproductive outcomes of couples with non-male-factor infertility undergoing in vitro fertilization (IVF) cycles. In total, 381 cycles included were subgrouped according to paternal age (<35-year-old, 35-39-year-old, or ≥40-year-old), and maternal age was limited to under 35 years. Data on embryo quality and clinical outcomes were analyzed. The results showed that fertilization and high-quality embryo rates were not significantly different (all P > 0.05). The pregnancy rate was not significantly different in the 35-39-year-old group (42.0%; P > 0.05), but was significantly lower in the ≥40-year-old group (26.1%; P < 0.05) than that in the <35-year-old group (40.3%). Similarly, the implantation rate significantly decreased in the ≥40-year-old group (18.8%) compared with that in the <35-year-old group (31.1%) and 35-39-year-old group (30.0%) (both P < 0.05). The live birth rate (30.6%, 21.7%, and 19.6%) was not significantly different across the paternal age subgroups (<35-year-old, 35-39-year-old, and ≥40-year-old, respectively; all P > 0.05), but showed a declining trend. The miscarriage rate significantly increased in the 35-39-year-old group (44.8%) compared with that in the <35-year-old group (21.0%; P < 0.05). No abnormality in newborn birth weight was found. The results indicated that paternal age over 40 years is a key risk factor that influences the assisted reproductive technology success rate even with good semen parameters, although it has no impact on embryo development.


Subject(s)
Pregnancy , Infant, Newborn , Female , Humans , Male , Adult , Paternal Age , Retrospective Studies , Semen , Fertilization in Vitro , Reproductive Techniques, Assisted , Oligospermia
7.
Asian Journal of Andrology ; (6): 38-42, 2023.
Article in English | WPRIM | ID: wpr-971000

ABSTRACT

The authors performed a comprehensive review of current literature to create a model comparing commonly evaluated variables in male factor infertility, for example, follicle-stimulating hormone (FSH), testicular volume (TV), and testosterone (T), to better predict sperm retrieval rate (SRR). Twenty-nine studies were included, 9 with data on conventional testicular sperm extraction (cTESE) for a total of 1227 patients and 20 studies including data on microdissection testicular sperm extraction (mTESE) for a total of 4760 patients. A weighted-means value of SRR, FSH, T, and TV was created, and a weighted linear regression was then used to describe associations among SRR, type of procedure, FSH, T, and TV. In this study, weighted-means values demonstrated mTESE to be superior to cTESE with an SRR of 51.9% vs 40.1%. Multiple weighted linear regressions were created to describe associations among SRR, procedure type, FSH, T, and TV. The models showed that for every 1.19 mIU ml-1 increase in FSH, there would be a significant decrease in SRR by 1.0%. Seeking to create a more clinically relevant model, FSH values were then divided into normal, moderate elevation, and significant elevation categories (FSH <10 mIU ml-1, 10-19 mIU ml-1, and >20 mIU ml-1, respectively). For an index patient undergoing cTESE, the retrieval rates would be 57.1%, 44.3%, and 31.2% for values normal, moderately elevated, and significantly elevated, respectively. In conclusion, in a large meta-analysis, mTESE was shown to be more successful than cTESE for sperm retrievals. FSH has an inverse relationship to SRR in retrieval techniques and can alone be predictive of cTESE SRR.


Subject(s)
Humans , Male , Follicle Stimulating Hormone , Follicle Stimulating Hormone, Human , Infertility, Male , Linear Models , Semen , Sperm Retrieval , Spermatozoa , Testis/surgery
8.
Asian Journal of Andrology ; (6): 73-77, 2023.
Article in English | WPRIM | ID: wpr-970986

ABSTRACT

Patients with congenital unilateral absence of the vas deferens (CUAVD) manifest diverse symptoms from normospermia to azoospermia. Treatment for CUAVD patients with obstructive azoospermia (OA) is complicated, and there is a lack of relevant reports. In this study, we describe the clinical features and evaluate the treatments and outcomes of CUAVD patients with OA. From December 2015 to December 2020, 33 patients were diagnosed as CUAVD with OA in Shanghai General Hospital (Shanghai, China). Patient information, ultrasound findings, semen analysis, hormone profiles, and treatment information were collected, and the clinical outcomes were evaluated. Of 33 patients, 29 patients were retrospectively analyzed. Vasoepididymostomy (VE) or cross VE was performed in 12 patients, the patency rate was 41.7% (5/12), and natural pregnancy was achieved in one of the patients. The other 17 patients underwent testicular sperm extraction as the distal vas deferens (contralateral side) was obstructed. These findings showed that VE or cross VE remains an alternative treatment for CUAVD patients with OA, even with a relatively low rate of patency and natural pregnancy.


Subject(s)
Pregnancy , Female , Humans , Male , Vas Deferens/abnormalities , Azoospermia/surgery , Epididymis/surgery , Retrospective Studies , Tertiary Care Centers , China , Semen
9.
Asian Journal of Andrology ; (6): 21-28, 2023.
Article in English | WPRIM | ID: wpr-970985

ABSTRACT

In this review, we tried to systematize all the evidence (from PubMed [MEDLINE], Scopus, Cochrane Library, EBSCO, Embase, and Google Scholar) from 1993 to 2021 on the predictors of microsurgical varicocelectomy efficacy in male infertility treatment. Regarding the outcomes of varicocele repair, we considered semen improvement and pregnancy and analyzed them separately. Based on the 2011 Oxford CEBM Levels of Evidence, we assigned a score to each trial that studied the role of the predictor. We systematized the studied predictors based on the total points, which were, in turn, calculated based on the number and quality of studies that confirmed or rejected the studied predictor as significant, into three levels of significance: predictors of high, moderate, and low clinical significance. Preoperative total motile sperm count (TMSC) coupled with sperm concentration can be a significant predictor of semen improvement and pregnancy after varicocelectomy. In addition, for semen improvement alone, scrotal Doppler ultrasound (DUS) parameters, sperm DNA fragmentation index (DFI), and bilateral varicocelectomy are reliable predictors of microsurgical varicocelectomy efficacy.


Subject(s)
Female , Humans , Male , Pregnancy , Infertility, Male/surgery , Microsurgery , Semen , Sperm Count , Sperm Motility , Varicocele/surgery
10.
Asian Journal of Andrology ; (6): 29-37, 2023.
Article in English | WPRIM | ID: wpr-970983

ABSTRACT

Mycoplasma species (spp.) are bacteria that are difficult to detect. Currently, the polymerase chain reaction (PCR) is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinary medicine. There are 13 known species of human Mycoplasma and 15 species of canine Mycoplasma. Owing to the difficulties in identifying the individual species of Mycoplasma, there is a lack of information regarding which species are saprophytic and which are pathogenic. The prevalence of the individual species is also unknown. In addition, in both humans and dogs, the results of some studies on the impact of Mycoplasma are conflicting. The presence of Mycoplasma spp. on the epithelium of reproductive tract is often associated with infertility, although they are also detected in healthy individuals. The occurrence of Mycoplasma spp. is more common in dogs (even 89%) than in humans (1.3%-4%). This is probably because the pH of a dog's genital is more conducive to the growth of Mycoplasma spp. than that of humans. Phylogenetically, human and canine Mycoplasma are related, and majority of them belong to the same taxonomic group. Furthermore, 40% of canine Mycoplasma spp. are placed in common clusters with those of human. This suggests that species from the same cluster can play a similar role in the canine and human reproductive tracts. This review summarizes the current state of knowledge about the impact of Mycoplasma on canine and human male fertility as well as the prospects of further development in this field.


Subject(s)
Humans , Dogs , Male , Animals , Mycoplasma/genetics , Infertility , Semen Analysis , Polymerase Chain Reaction/methods , Prevalence , Semen/chemistry
11.
Chinese Journal of Medical Genetics ; (6): 301-307, 2023.
Article in Chinese | WPRIM | ID: wpr-970923

ABSTRACT

OBJECTIVE@#To explore the genetic basis for 4 patients with globozoospermia.@*METHODS@#Semen and blood samples were collected from the patients for the determination of sperm concentration, viability, survival rate, morphology and acrosome antigen CD46. Meanwhile, DNA was extracted for whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing.@*RESULTS@#All of the four patients were found to harbor variants of the DPY19L2 gene. Patients 1 ~ 3 had homozygous deletions of the DPY19L2 gene. Sanger sequencing confirmed that the DPY19L2 gene in patient 3 was disrupted at a recombination breakpoint area BP2, resulting in nonallelic homologous recombination and complete deletion of the DPY19L2 gene. Patients 2 and 3 respectively harbored novel homozygous deletions of exons 2 ~ 22 and exons 14 ~ 15. Patient 4 harbored heterozygous deletion of the DPY19L2 gene, in addition with a rare homozygous deletion of the 3' UTR region.@*CONCLUSION@#DPY19L2 gene variants probably underlay the globozoospermia in the four patients, which has fit an autosomal recessive pattern of inheritance and the characteristics of genomic diseases.


Subject(s)
Male , Humans , Teratozoospermia/genetics , Homozygote , Semen , Sequence Deletion , 3' Untranslated Regions , Membrane Proteins
12.
Chinese Journal of Medical Genetics ; (6): 148-154, 2023.
Article in Chinese | WPRIM | ID: wpr-970895

ABSTRACT

OBJECTIVE@#To assess the value of single sperm sequencing in preimplantation genetic testing for monogenic disease (PGT-M).@*METHODS@#A Chinese couple with two children whom had died of Spinal muscular atrophy (SMA) and attended the Jiangxi Provincial Maternal and Child Health Care Hospital in June 2020 was selected as the subject. Eleven single sperm samples were isolated by mechanical immobilization and subjected to whole genome amplification. Real-time PCR and Sanger sequencing were used to detect the SMN1 variants in the single sperm samples. Genomic DNA of the wife, her parents and the husband, as well as one single sperm sample harboring the SMN1 variant and two single sperm samples without the variant were used for the linkage analysis. Targeted capture and high-throughput sequencing were carried out to test 100 single nucleotide polymorphisms distributed within 2 Mb up- and downstream the variant site. The haplotypes linked with the SMN1 variants were determined by linkage analysis. Blastocyst embryos were harvested after fertilizing by intracytoplasmic sperm injection. Cells from the trophoblasts of each embryo were biopsied and subjected to whole genome amplification and targeted capture and high-throughput sequencing to determine their carrier status. Chromosomal aneuploidy of wild-type embryos was excluded. An euploid embryo of high quality was transferred. Amniotic fluid sample was taken at 18 weeks of gestation to confirm the status of the fetus.@*RESULTS@#Genetic testing showed that the couple both had deletion of exons 7 ~ 8 of the SMN1 gene. The wife has inherited the deletion from her father, while the husband was de novo. The haplotypes of the husband were successfully constructed by single sperm sequencing. Preimplantation genetic testing has indicated that 5 embryos had harbored the heterozygous variant, 4 embryos were of the wild type, among which 3 were euploid. Prenatal diagnosis during the second trimester of pregnancy has confirmed that the fetus did not carry the deletion.@*CONCLUSION@#By single sperm sequencing and PGT-M, the birth of further affected child has been successfully avoided.


Subject(s)
Humans , Pregnancy , Female , Child , Male , Preimplantation Diagnosis , East Asian People , Semen , Genetic Testing , Muscular Atrophy, Spinal/genetics , Aneuploidy , Blastocyst/pathology , High-Throughput Nucleotide Sequencing , Spermatozoa
13.
Chinese Journal of Medical Genetics ; (6): 26-30, 2023.
Article in Chinese | WPRIM | ID: wpr-970872

ABSTRACT

OBJECTIVE@#To explore the incidence of azoospermia factor c (AZFc) microdeletion among patients with azoospermia or severe oligospermia, its association with sex hormone/chromosomal karyotype, and its effect on the outcome of pregnancy following intracytoplasmic sperm injection (ICSI) treatment.@*METHODS@#A total of 1 364 males with azoospermia or severe oligospermia who presented at the Affiliated Maternity and Child Health Care Hospital of Jiaxing College between 2013 and 2020 were subjected to AZF microdeletion and chromosome karyotyping analysis. The level of reproductive hormones in patients with AZFc deletions was compared with those of control groups A (with normal sperm indices) and B (azoospermia or severe oligospermia without AZFc microdeletion). The outcome of pregnancies for the AZFc-ICSI couples was compared with that of the control groups in regard to fertilization rate, superior embryo rate and clinical pregnancy rate.@*RESULTS@#A total of 51 patients were found to harbor AZFc microdeletion, which yielded a detection rate of 3.74%. Seven patients also had chromosomal aberrations. Compared with control group A, patients with AZFc deletion had higher levels of PRL, FSH and LH (P < 0.05), whilst compared with control group B, only the PRL and FSH were increased (P < 0.05). Twenty two AZFc couples underwent ICSI treatment, and no significant difference was found in the rate of superior embryos and clinical pregnancy between the AZFc-ICSI couples and the control group (P > 0.05).@*CONCLUSION@#The incidence of AZFc microdeletion was 3.74% among patients with azoospermia or severe oligospermia. AZFc microdeletion was associated with chromosomal aberrations and increased levels of PRL, FSH and LH, but did not affect the clinical pregnancy rate after ICSI treatment.


Subject(s)
Child , Humans , Male , Female , Pregnancy , Azoospermia/genetics , Oligospermia/genetics , Incidence , Chromosome Deletion , Chromosomes, Human, Y/genetics , Semen , Infertility, Male/genetics , Chromosome Aberrations , Follicle Stimulating Hormone/genetics
14.
Braz. j. biol ; 83: 1-13, 2023. graf
Article in English | LILACS, VETINDEX | ID: biblio-1468850

ABSTRACT

The cold storage of milt implies potentials alterations in its quality because the storage generates as main process, free radicals that produce spermatozoa membrane lipids damage with the consequent motility and fertilising capacity disruptions. To decrease the damage generated by free radicals the cells have antioxidant defences (proteins, enzymes, and low molecular weight substances). The objective of the present study evaluated the time storage effect and different antioxidants prepared in spermatic diluents on sperm viability of O. mykiss milt stored at 4°C. The two-way ANOVA denoted that the time storage and antioxidant influence have significant effects separated or combined on viability parameters (sperm motility and viability, proteins concentrations and superoxide dismutase enzymatic activity in seminal plasma). In contrast, only the storage time affected the fertilising capacity and catalase enzymatic activity in seminal plasma. The resulting analysis can conclude that the antioxidant presence improves the viability of cold stored milt, especially the transport conditions and the antioxidants allow the fecundity despite motility decrease.


O armazenamento a frio de leite implica potenciais alterações em sua qualidade, pois gera como processo principal radicais livres que provocam danos aos lipídios da membrana dos espermatozoides, com as consequentes alterações na motilidade e na capacidade de fertilização. Para diminuir os danos causados pelos radicais livres, as células têm defesas antioxidantes (proteínas, enzimas e substâncias de baixo peso molecular). O presente estudo avaliou o efeito do tempo de armazenamento e diferentes antioxidantes preparados em diluentes espermáticos no armazenamento de viabilidade de O. mykiss milt a 4°C. A ANOVA de duas vias denotou que o armazenamento no tempo e a influência antioxidante têm efeitos significativos separados ou combinados nos parâmetros de viabilidade (motilidade espermática, viabilidade espermática, concentrações de proteínas e atividade enzimática da superóxido dismutase no plasma seminal), enquanto apenas o tempo de armazenamento afetou a capacidade de fertilização e atividade enzimática da catalase no plasma seminal. A análise resultante pode concluir que a presença de antioxidante melhora a viabilidade do leite frio, especialmente as condições de transporte, e os antioxidantes permitem a fecundidade apesar da diminuição da motilidade.


Subject(s)
Animals , Catalase/analysis , Cryopreservation/methods , Oncorhynchus mykiss , Semen/drug effects , Analysis of Variance
15.
Neotrop. ichthyol ; 21(1): e220071, 2023. ilus, tab, graf
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1418130

ABSTRACT

This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)


Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)


Subject(s)
Animals , Catfishes , Cryopreservation , Dimethyl Sulfoxide/adverse effects , Acetamides/adverse effects , Semen/chemistry
16.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 403-410, 2023.
Article in English | WPRIM | ID: wpr-982711

ABSTRACT

Male infertility is a significant cause of psychosocial and marital distress in approximately 50% of couples who are unable to conceive, with male factors being the underlying cause. Guijiajiao (Colla Carapacis et Plastri, CCP) is a Traditional Chinese Medicine commonly used to treat male infertility. The present study aimed to investigate the potential mechanisms underlying the preventive effects of CCP on male infertility. An infertile male rat model was established using cyclophosphamide (CTX), and CCP was administered for both treatment and prevention. Fecal microbiota transplantation (FMT) was also performed to explore the role of gut microbiota in the CCP-mediated prevention of male infertility in rats. Sperm motility and concentration were determined using a semi-automatic sperm classification analyzer. Subsequently, histopathological analysis using HE staining was performed to examine the changes in the small intestine and testis. Moreover, the serum levels of lipopolysaccharide (LPS) and testosterone were measured by ELISA. In addition, immunohistochemistry was conducted to detect CD3 expression in the small intestine, while RT-qPCR was employed to assess the expressions of interleukin-1 beta (IL-1β), cluster of differentiation 3 (CD3), Monocyte chemoattractant protein-1 (MCP-1), and C-X-C motif chemokine ligand 10 (CXCL-10) in the small intestine and epididymis. Finally, gut microbiota was analyzed by 16S rRNA sequencing. CCP improved sperm motility, number, and concentration in CTX-induced infertile male rats. CCP increased the serum testosterone level, inhibited the immune cell infiltration of the intestinal lamina propria, and promoted the aggregation of CD3+ T cells in CTX-induced male infertility rats. CCP also inhibited the expressions of MCP-1, CXCL-10, and IL-1β in the epididymis of male infertility rats. At the genus level, CTX led to a reduction in the abundance of Lactobacillus, Clostridia_UCG.014, and Romboutsia in the intestinal tract of rats. In contrast, CCP decreased the abundance of Ruminococcus and increased the abundance of Romboutsia in infertile male rats. Additionally, FMT experiments proved that the gut microbiota of CCP-treated rats facilitated testicular tissue recovery and spermatogenesis while also reducing the serum LPS level in infertile male rats. CCP improves the spermatogenic ability of infertile male rats by restoring gut microbiota diversity and inhibiting epididymal inflammation.


Subject(s)
Humans , Rats , Male , Animals , Gastrointestinal Microbiome , Lipopolysaccharides/pharmacology , RNA, Ribosomal, 16S , Semen , Sperm Motility , Infertility, Male/prevention & control , Testosterone
17.
Asian Journal of Andrology ; (6): 426-432, 2023.
Article in English | WPRIM | ID: wpr-981957

ABSTRACT

This study assessed the effects of a simulated high-altitude environment on the reproductive system of prepubertal male rats and the reversibility of these effects upon return to a normal environment. Three-week-old male Wistar rats were randomly allocated to 4 groups that were exposed to different conditions: a normal environment for 6 weeks and 12 weeks, respectively, hypobaric hypoxia for 6 weeks, and hypobaric hypoxia for 6 weeks followed by a normal environment for 6 weeks. Multiple pathophysiological parameters were evaluated at the histological, endocrine, and molecular levels. Hypobaric hypoxia exposure for 6 weeks during the prepubertal phase significantly altered physiological parameters, body functions, blood indices, and reproductive potential. Six weeks after returning to a normal environment, the damaged reproductive functions partially recovered due to compensatory mechanisms. However, several changes were not reversed after returning to a normal environment for 6 weeks, including disorders of body development and metabolism, increased red blood cells, increased fasting blood glucose, abnormal blood lipid metabolism, decreased testicular and epididymis weights, abnormal reproductive hormone levels, excessive apoptosis of reproductive cells, and decreased sperm concentration. In summary, a hypobaric hypoxic environment significantly impaired the reproductive function of prepubertal male rats, and a return to normal conditions during the postpubertal phase did not fully recover these impairments.


Subject(s)
Rats , Male , Animals , Rats, Wistar , Altitude , Semen/metabolism , Hypoxia/pathology , Genitalia, Male
18.
Asian Journal of Andrology ; (6): 350-355, 2023.
Article in English | WPRIM | ID: wpr-981954

ABSTRACT

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenozoospermia categorized by immotile spermatozoa with abnormal flagella in ejaculate. Whole-exome sequencing (WES) is used to detect pathogenic variants in patients with MMAF. In this study, a novel homozygous frameshift variant (c.6158_6159insT) in dynein axonemal heavy chain 8 (DNAH8) from two infertile brothers with MMAF in a consanguineous Pakistani family was identified by WES. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed DNAH8 mRNA decay in these patients with the DNAH8 mutation. Hematoxylin-eosin staining and transmission electron microscopy revealed highly divergent morphology and ultrastructure of sperm flagella in these patients. Furthermore, an immunofluorescence assay showed the absence of DNAH8 and a reduction in its associated protein DNAH17 in the patients' spermatozoa. Collectively, our study expands the phenotypic spectrum of patients with DNAH8-related MMAF worldwide.


Subject(s)
Humans , Male , Consanguinity , Pakistan , Infertility, Male/metabolism , Semen/metabolism , Sperm Tail/metabolism , Spermatozoa/metabolism , Flagella/pathology , Mutation
19.
Asian Journal of Andrology ; (6): 404-409, 2023.
Article in English | WPRIM | ID: wpr-981951

ABSTRACT

Male infertility caused by idiopathic oligoasthenospermia (OAT) is known as idiopathic male infertility. Glutathione S-transferase (GST) and fluoride may play important roles in idiopathic male infertility, but their effects are still unknown. Our study examined the relationship between GST polymorphisms and fluoride-induced toxicity in idiopathic male infertility and determined the underlying mechanism. Sperm, blood, and urine samples were collected from 560 males. Fluoride levels were measured by a highly selective electrode method, and GST genotypes were identified using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). Semen parameters, DNA fragmentation index (DFI), mitochondrial membrane potential (MMP), and oxidative stress (OS) biomarkers were statistically assessed at the P < 0.05 level. Compared with healthy fertile group, semen parameters, fluoride levels, OS biomarkers, sex hormone levels, and MMP and DFI levels were lower in the idiopathic male infertility group. For glutathione S-transferase M1 (GSTM1[-]) and glutathione S-transferase T1 (GSTT1[-]) or glutathione S-transferase P1 (GSTP1) mutant genotypes, levels of semen fluoride, OS, MMP, and DFI were considerably higher, and the mean levels of sperm parameters and testosterone were statistically significant in GSTM1(+), GSTT1(+), and GSTP1 wild-type genotypes. Both semen and blood fluoride levels were associated with oxidative stress in idiopathic male infertility patients. Elevated fluoride in semen with the genotypes listed above was linked to reproductive quality in idiopathic male infertility patients. In conclusion, GST polymorphisms and fluorine may have an indicative relationship between reproductive quality and sex hormone levels, and OS participates in the development of idiopathic male infertility.


Subject(s)
Humans , Male , Fluorides/adverse effects , Semen , Polymorphism, Genetic , Glutathione Transferase/genetics , Glutathione S-Transferase pi/genetics , Infertility, Male/genetics , Genotype , Biomarkers , Genetic Predisposition to Disease , Case-Control Studies
20.
Asian Journal of Andrology ; (6): 339-344, 2023.
Article in English | WPRIM | ID: wpr-981949

ABSTRACT

The process of semen collection plays a key role in the quality of semen specimens. However, the association between semen collection time and semen quality is still unclear. In this study, ejaculates by masturbation from 746 subfertile men or healthy men who underwent semen analysis were examined. The median (interquartile range) semen collection time for all participants was 7.0 (5.0-11.0) min, and the median time taken for semen collection was lower in healthy men than that in subfertile men (6.0 min vs 7.0 min). An increase in the time required to produce semen samples was associated with poorer semen quality. Among those undergoing assisted reproductive technology (ART), the miscarriage rate was positively correlated with the semen collection time. After adjusting for confounders, the highest quartile (Q4) of collection time was negatively associated with semen volume and sperm concentration. A longer time to produce semen samples (Q3 and Q4) was negatively correlated with progressive and total sperm motility. In addition, there was a significant negative linear association between the semen collection time and the sperm morphology. Higher risks of asthenozoospermia (adjusted odds ratio [OR] = 2.06, 95% confidence interval [CI]: 1.31-3.25, P = 0.002) and teratozoospermia (adjusted OR = 1.98, 95% CI: 1.10-3.55, P = 0.02) were observed in Q3 than those in Q1. Our results indicate that a higher risk of abnormal semen parameter values was associated with an increase in time for semen collection, which may be related to male fertility through its association with semen quality.


Subject(s)
Male , Humans , Semen Analysis , Semen , Sperm Motility , Sperm Count , Asthenozoospermia , Spermatozoa
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