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1.
PLoS One ; 15(12): e0242218, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33370316

RESUMO

Improvements in survival rates with gonad-sparing protocols for childhood and adolescence cancer have increased the optimism of survivors to become parents after treatment. Findings in rodents indicate that chromosomal aberrations can be induced in male germ cells by genotoxic exposures and transmitted to offspring and future generations with effects on development, fertility and health. Thus, there is a need for effective technologies to identify human sperm carrying chromosomal aberrations to assess the germ-line risks, especially for cancer survivors who have received genotoxic therapies. The time-dependent changes in the burden of sperm carrying structural chromosomal aberrations were assessed for the first time in a cancer setting, using the AM8 sperm FISH protocol which simultaneously detects abnormalities in chromosomal structure and number in sperm. Nine Hodgkin lymphoma (HL) patients provided 20 semen samples before, during, and after NOVP therapy (Novantrone, Oncovin, Velban and Prednisone) and radiation therapy that produced scattered gonadal doses from <0.05 to 0.6 Gy. Late meiosis was found to be the most sensitive to NOVP treatment for the production of sperm with chromosomal abnormalities, both in structure and number. Earlier stages of spermatogenesis were less sensitive and there was no evidence that therapy-exposed stem cells resulted in increased frequencies of sperm with abnormalities in chromosomal structure or number. This indicates that NOVP therapy may increase the risks for paternal transmission of chromosomal structural aberrations for sperm produced 32 to 45 days after a treatment with these drugs and implies that there are no excess risks for pregnancies conceived more than 6 months after this therapy. This clinical evaluation of the AM8 sperm FISH protocol indicates that it is a promising tool for assessing an individual's burden of sperm carrying chromosomal structural aberrations as well as aneuploidies after cancer therapy, with broad applications in other clinical and environmental situations that may pose aneugenic or clastogenic risks to human spermatogenesis.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Aberrações Cromossômicas/efeitos dos fármacos , Doença de Hodgkin/terapia , Meiose/efeitos dos fármacos , Análise do Sêmen/métodos , Espermatozoides/efeitos dos fármacos , Adulto , Células-Tronco Germinativas Adultas/efeitos dos fármacos , Células-Tronco Germinativas Adultas/efeitos da radiação , Sobreviventes de Câncer , Quimiorradioterapia/efeitos adversos , Quimiorradioterapia/métodos , Aberrações Cromossômicas/efeitos da radiação , Estudos de Coortes , Preservação da Fertilidade , Humanos , Hibridização in Situ Fluorescente/métodos , Masculino , Meiose/efeitos da radiação , Mitoxantrona/efeitos adversos , Mutagênese/efeitos dos fármacos , Mutagênese/efeitos da radiação , Tratamentos com Preservação do Órgão/efeitos adversos , Tratamentos com Preservação do Órgão/métodos , Órgãos em Risco/efeitos da radiação , Prednisona/efeitos adversos , Dosagem Radioterapêutica , Espermatogênese/efeitos dos fármacos , Espermatogênese/efeitos da radiação , Espermatozoides/fisiologia , Espermatozoides/efeitos da radiação , Testículo/efeitos dos fármacos , Testículo/efeitos da radiação , Fatores de Tempo , Vimblastina/efeitos adversos , Vincristina/efeitos adversos
2.
Zhonghua Nan Ke Xue ; 26(10): 895-899, 2020 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-33382220

RESUMO

Objective: To investigate the application of the self-made semen quality control (QC) product in internal QC of computer-assisted sperm analysis (CASA). METHODS: CASA was calibrated with high- and low-concentration commercially available semen QC product and meanwhile 15 samples of self-made mixed semen QC product were placed in 75 cryotubes containing liquid nitrogen, followed by CASA of the concentration, motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), wobble (WOB) and straightness (STR) of the sperm using standard procedures and 50 days of continuous monitoring. The Makler counting plate was used to measure the concentration and motility of the self-made sperm. RESULTS: The coefficients of variation (CV) of the commercially available semen QC product at high and low concentrations were 6.18% and 7.85%, respectively. CASA showed that the concentration of the self-made QC product was (25.97 ± 1.41) ×106/ml, with a CV of 5.42%, and the sperm motility, VCL, VSL, VAP, LIN, WOB and STR were (22.15 ± 1.75)% (CV = 7.9%), (59.18 ± 2.05) µm/s (CV = 3.46%), (26.79 ± 1.2) µm/s (CV = 4.48%), (34.98 ± 1.4) µm/s (CV = 4.01%), 46.81 ± 1.55 (CV = 3.3%), 60.52 ± 1.3 (CV = 2.15%) and 76.46 ± 1.98 (CV = 2.59%), respectively. The concentration and motility of the self-made sperm detected with the Makler counting plate were (34.39 ± 2.37) ×106/ml (CV = 6.89%) and (38.04 ± 1.69)% (CV = 4.44%), respectively. Levey-Jennings QC charts were plotted for the indicators using the means and standard deviation. CONCLUSIONS: The self-made internal QC product by liquid nitrogen cryopreservation is feasible and effective for monitoring the accuracy and precision of CASA-derived sperm concentration and motion parameters, and it has a smaller CV than the commercially available QC product in measuring sperm concentration.


Assuntos
Controle de Qualidade , Análise do Sêmen/métodos , Análise do Sêmen/normas , Motilidade Espermática , Computadores , Humanos , Masculino , Espermatozoides
5.
Medicine (Baltimore) ; 99(26): e20894, 2020 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-32590798

RESUMO

RATIONALE: Couples with male balanced-translocation carriers may experience recurrent pregnancy loss (RPL). Although the expectant management of RPL has developed over many years, genetic counseling for RPL couples with male balanced-translocation carriers remains challenging. Here, we describe the expectant management of 2 male carriers of balanced translocations. PATIENT CONCERNS: A 32-year-old and a 28-year-old man presented at the clinic with diagnoses of infertility following spontaneous abortions by their wives. DIAGNOSIS: Both patients had normal semen diagnosed by routine semen analysis and underwent cytogenetic diagnoses. INTERVENTIONS: Following genetic counseling and informed consent, both couples voluntarily chose expectant management with natural conception. OUTCOMES: One couple experienced 2 natural pregnancies, the first of which ended in spontaneous abortion and the second produced a phenotypically normal infant. The other couple's first pregnancy resulted in a fetus with a balanced translocation confirmed by amniocentesis and cytogenetic analysis. LESSONS: Expectant management with natural conception may be an alternative to genetic counseling in male balanced-translocation carriers with RPL, especially those who are reluctant to undergo preimplantation diagnosis.


Assuntos
Infertilidade Masculina/genética , Resultado da Gravidez/genética , Translocação Genética/genética , Aborto Espontâneo/epidemiologia , Adulto , China/epidemiologia , Feminino , Humanos , Infertilidade Masculina/complicações , Infertilidade Masculina/epidemiologia , Masculino , Gravidez , Resultado da Gravidez/epidemiologia , Análise do Sêmen/métodos
7.
Fertil Steril ; 113(4): 865-875, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32164925

RESUMO

OBJECTIVE: To assess the association between psychological stress and male factor infertility as well as testicular function (semen quality, serum reproductive hormones) and erectile dysfunction. DESIGN: Cross-sectional study. SETTING: University Hospital-based research center. PATIENTS: Men with impaired semen quality were included from infertile couples, and men with naturally conceived pregnant partners were used as a reference population. INTERVENTIONS: Participants completed a questionnaire on health and lifestyle, including a 14-item questionnaire about self-rated psychological stress symptoms and stressful life event (SLEs), had a physical examination performed, delivered a semen sample and had a blood sample drawn. MAIN OUTCOMES: Differences in stress scores (calculated from self-reported stress symptoms) and SLEs between infertile and fertile men were assessed in crude and fully adjusted linear regression models. Secondary outcomes were semen quality, serum reproductive hormones, and erectile dysfunction. RESULTS: Of 423 men, 176 (41.6%) experienced at least one SLE in the 3 months prior to inclusion (50.4%/36.9%: infertile/fertile men, P = .03); ß-coefficient and 95% confidence interval for the difference between the groups on the transformed scale in fully adjusted linear regression models was 0.18 (0.06, 0.30). However, there were no differences in psychological stress symptoms between the two groups (ß-coefficient and 95% confidence interval) on the transformed scale (0.14; -0.02, 0.30). No association between stress (self-reported stress symptoms and SLEs) and testicular function or with erectile dysfunction was found in any of the men. CONCLUSION: Infertile men reported a higher number of SLEs than fertile men but did not report more psychological stress symptoms. Distress and SLEs were not associated with reduced male reproductive function.


Assuntos
Infertilidade Masculina/diagnóstico , Infertilidade Masculina/psicologia , Análise do Sêmen/métodos , Estresse Psicológico/diagnóstico , Estresse Psicológico/psicologia , Testículo/fisiologia , Adulto , Estudos Transversais , Humanos , Infertilidade Masculina/sangue , Acontecimentos que Mudam a Vida , Masculino , Pessoa de Meia-Idade , Estresse Psicológico/sangue , Inquéritos e Questionários , Adulto Jovem
8.
Biodemography Soc Biol ; 65(1): 73-87, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32065536

RESUMO

Some male infertility biomarkers are etiologically linked to idiopathic infertility in men, the direct cause of which often cannot be determined with conventional sperm count parameters. Open questions remain regarding the universal and generic infertility definitions that cover and combine the clinical, epidemiological, and demographic perspectives. The main effort in the application of these infertility biomarkers are accounted by more or less strict discrimination criteria. For male infertility, beyond classical sperm count assessments, the DNA fragmentation index (DFI) is an adequate biomarker. DFI strongly correlates with pregnancy rates and even strict discrimination criteria for infertility outcomes. Other common biomarkers are reactive oxygen species (ROS) and antisperm antibodies (ASAs), which can explain some biomedical infertility disorders within major constraints. More frequently applied in demographic research, telomere length component analysis is based on identifying the genetic impact of cellular longevity. Sperm telomere length is becoming established as a potential biomarker in infertility research. The aim of this review is to provide an overview of the current status and limitations to the application of novel biomarkers, including TEX101, for infertility research. The review also discusses potential options for the use of biomarkers in population-based studies.Abbreviations: ASAs: antisperm antibodies; DFI: DNA fragmentation index; DNA: deoxyribonucleic acid; ECM1: extracellular matrix protein 1; FSH: follicle stimulating hormone; HS: hypospermatogenesis: IVF: in vitro fertilization; LDHC: L-lactata dehydrogenase C chain; MA: maturation arrest; microTESE: microdissection testicular sperm extraction; NOA: nonobstructive azoospermia; NP: nonprogressive; OA: obstructive azoospermia; pH: potential Hyrogenii (pH-value); PR: progressive; PTGDS: prostaglandin D synthese; ROS: reactive oxygen species; SA: semen analysis; SCO: sertoli cell only; SCSA: sperm chromatin structure assay (SCSA); TL: telomere length; TESE: testicular sperm extraction; TEX101: a glycoprotein that belongs to Ly6/urokinase type plasminogen activator receptor-like protein (uPAR)(LU) superfamily, to be a germ-cell-specific molecular sperm extraction; TUNEL: terminal deoxnucleotidyl dispersion tranferase dUTP nick-end labeling; WHO: World Health Organization.


Assuntos
Biomarcadores/análise , Sêmen/microbiologia , Contagem de Espermatozoides/estatística & dados numéricos , Adulto , Demografia/métodos , Humanos , Infertilidade Masculina , Masculino , Análise do Sêmen/métodos , Contagem de Espermatozoides/métodos
9.
PLoS One ; 15(1): e0227946, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31978160

RESUMO

Sperm cryopreservation by ultra-rapid cooling based on dropping small volumes of sperm suspension directly into liquid nitrogen, has been successful in some wild ruminant species, including the Iberian ibex (Capra pyrenaica). In ultra-rapid cooling, the contents of these droplets are expected to enter a stable, glass-like state, but to the best of our knowledge no information exists regarding the presence or absence of ice formation in the extracellular milieu when using this technique. Different modifications to the extracellular milieu likely inflict different types of damage on the plasmalemma, the acrosome and mitochondrial membranes. The aims of the present work were: 1) to examine the physical state of the extracellular milieu after cryopreservation at slow and ultra-rapid cooling rates-and thus determine whether ultra-rapid cooling vitrifies the extracellular milieu; and 2) to compare, using conventional sperm analysis techniques and scanning and transmission electron microscopy, the damage to sperm caused by these two methods. Sperm samples were obtained by the transrectal ultrasound-guided massage method (TUMASG) from anesthetized Iberian ibexes, and cryopreserved using slow and ultra-rapid cooling techniques. Sperm motility (22.95 ± 3.22% vs 4.42 ± 0.86%), viability (25.64 ± 3.71% vs 12.8 ± 2.50%), acrosome integrity (41.45± 3.73% vs 27.00 ± 1.84%) and mitochondrial membrane integrity (16.52 ± 3.75% vs 4.00 ± 0.65%) were better after slow cooling (P<0.001) than after ultra-rapid technique. Cryo-scanning electron microscopy (Cryo-SEM) suggested that the vitrified state was not achieved by ultra-rapid cooling, and that the ice crystals formed were smaller and had more stretchmarks (P<0.001) than after slow cooling. Scanning electron microscopy revealed no differences in the types of damage caused by the examined techniques, although transmission electron microscopy showed the damage to the plasmalemma and mitochondrial membrane to be worse after ultra-rapid cooling. In conclusion ultra-rapid cooling provoked more membrane damage than slow cooling, perhaps due to the extracellular ice crystals formed.


Assuntos
Cabras/genética , Análise do Sêmen/métodos , Preservação do Sêmen/métodos , Espermatozoides/fisiologia , Acrossomo/efeitos dos fármacos , Acrossomo/ultraestrutura , Animais , Temperatura Baixa , Criopreservação , Crioprotetores/farmacologia , Humanos , Masculino , Motilidade Espermática/efeitos dos fármacos , Vitrificação/efeitos dos fármacos
10.
Cryobiology ; 92: 197-202, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31962103

RESUMO

Semen extender has a vital role in preservation of sperm cells properties in terms of sperm viability, motility, acrosome integrity, and mitochondrial membrane potential. The objective of the present study was to evaluate a new extender, known as Thai native chicken (TNC) extender compared to BHSV-based and modified Sasaki extenders for freezing chicken semen. Semen from Thai native roosters was collected, pooled, and randomly divided into three groups. Semen was frozen with a simple freezing method using nitrogen vapor and dimethylformamide. In the first experiment, post-thaw motion parameters, viability, acrosome integrity, mitochondrial function, and lipid peroxidation levels were analyzed using computer-assisted sperm analysis, propidium iodide, fluorescein isothiocyanate-conjugate peanut agglutinin, JC-1, and the thiobarbituric acid reaction. Results showed that the type of extender had no effect on the percentage of total motile and curvilinear velocity. The percentage of progressive motile, straight-line velocity, and average path velocity of post-thawed semen were significantly lower in TNC compared to the modified Sasaki extender. However, the percentages of post-thawed acrosome integrity and active mitochondria were significantly higher in TNC extender (P < 0.05). For the second experiment, semen was thawed by using each of extenders thereafter, was inseminated to 48-layer breeder hens to determine the fertility rate. Among the three extenders used, the highest fertility rate was found in TNC extender. In conclusion, TNC extender can be recommended as an appropriate and useful cryopreservation media for native chicken semen since it maintains the quality of rooster semen and fertility after freezing and thawing process.


Assuntos
Acrossomo/efeitos dos fármacos , Criopreservação/métodos , Crioprotetores/farmacologia , Preservação do Sêmen/métodos , Animais , Galinhas , Dimetilformamida/farmacologia , Fertilidade , Congelamento , Humanos , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Sêmen/metabolismo , Análise do Sêmen/métodos , Motilidade Espermática/efeitos dos fármacos , Tailândia
11.
Theriogenology ; 144: 112-121, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31927416

RESUMO

Current artificial insemination (AI) laboratory practices assess semen quality of each boar ejaculate to decide which ones to process into AI doses. This decision is aided with two, world-wide used, motility parameters that come available through computer assisted semen analysis (CASA). This decision process, however, still results in AI doses with variable and sometimes suboptimal fertility outcomes (e.g., small litter size). The hypothesis was that the decision which ejaculates to process into AI doses can be improved by adding more data from CASA systems, and data from other sources, in combination with a data-driven model. Available data consisted of ejaculates that passed the initial decision, and thus, were processed into AI doses and used to inseminate sows. Data were divided into a training set (6793 records) and a validation set (1191 records) for model development, and an independent test set (1434 records) for performance assessment. Gradient Boosting Machine (GBM) models were developed to predict four fertility phenotypes of interest (gestation length, total number born, number born alive, and number of stillborn piglets). Each fertility phenotype was considered as a numeric and as a binary outcome parameter, totaling to eight different fertility phenotypes. Data used to further improve the decision process originated from four sources: 1) CASA information, 2) boar ejaculate information, 3) breeding value estimations, and 4) weather information. These data were used to create seven prediction sets, where each new set added parameters to the ones included in the previous set. The GBM models predicted fertility phenotypes with low correlations (for numeric phenotypes) and area under the curve values (for binary phenotypes) on the test data. Hence, results demonstrated that a combination of more data and GBM did not enable further improvement of the AI dose quality checks, resulting in the rejection of our hypothesis. However, our study revealed parameters affecting boar ejaculate fertility which were not used in today's decision process. These parameters (listed in the top 10 in at least four GBM models) included one parameter associated with boar ejaculate information, two with breeding value estimations, five with CASA information, and one with weather information. These parameters, therefore, should be further investigated for their potential value when assessing the quality of boar ejaculates in daily routine AI doses processing.


Assuntos
Inseminação Artificial/veterinária , Análise do Sêmen/veterinária , Preservação do Sêmen/veterinária , Suínos/fisiologia , Animais , Área Sob a Curva , Processamento de Imagem Assistida por Computador , Aprendizado de Máquina , Masculino , Análise do Sêmen/métodos , Preservação do Sêmen/métodos
12.
Reprod Domest Anim ; 55(3): 309-317, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31899824

RESUMO

The present study aimed to evaluate differences in area, volume and testicular weight among groups related to breed and age of boar and establish associations between testicular parameters and seminal characteristics. Seminal quality has been studied using computer-assisted semen analysis (CASA) systems that reduce the subjectivity of kinematic parameters. Based on the data obtained through this system, sperm quality indexes and their derived parameters have been assessed to identify a better estimator for determining movement and velocity parameters. We also established indexes of velocity and sperm movement for boar semen. To this end, we evaluated 191 ejaculate samples from 63 boars. Differences related to the effects of season and age on quality boar semen were found (p ≤ .05) in the animal groups. Indexes were established for velocity and sperm movement based on CASA-Mot, and we proposed a sperm movement index (SMI) and sperm velocity index (SVI) with the CASA-Mot variables. Correlations were found (p < .05) among sperm indexes and CASA-Mot variables. Boar characteristics were found to be associated with some CASA-Mot variables, and the estimated SMI and SVI indexes predicted a greater variation in kinematic characteristics compared to the parameters evaluated separately using the CASA-Mot system.


Assuntos
Análise do Sêmen/veterinária , Motilidade Espermática , Testículo/anatomia & histologia , Fatores Etários , Animais , Processamento de Imagem Assistida por Computador/métodos , Masculino , Tamanho do Órgão , Estações do Ano , Análise do Sêmen/métodos , Espermatozoides , Sus scrofa/anatomia & histologia
13.
Theriogenology ; 142: 67-76, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31581045

RESUMO

The stallion Breeding Soundness Examination (BSE), as proposed by the Society for Theriogenology, recommends that a stallion produce a minimum of one billion progressively motile, morphologically normal sperm (PMMNS) in the second of two ejaculates collected 1 h apart to be classified as a Satisfactory Prospective Breeder. With this in mind, the first objective of this study was to determine if the classification outcome of the traditional BSE differs depending on the methods used to evaluate sperm motility, morphology and concentration. We hypothesized that application of Computer Assisted Sperm Motion Analysis (CASA) and Differential Interference Contrast (DIC) microscopy to stallion semen evaluation would yield a more conservative estimate of the number of PMMNS. If this hypothesis is correct, then the use of CASA and DIC microscopy for semen evaluation would result in significantly fewer stallions meeting the historical standards for classification as a Satisfactory Prospective Breeder. Additionally, we determined whether the use of these modern technologies resulted in more accurate prediction of the actual fertility of a stallion compared to the use of more traditional technologies. Our results support the hypothesis that modern semen analysis techniques (including CASA and DIC microscopy) result in more conservative estimates of the number of PMMNS when compared to standard semen analysis techniques. As a result, the choice of methods used for semen analysis may impact the outcome of the traditional BSE. However, none of the methodologies used in this study reliably predicted different levels of fertility among this group of moderately to highly fertile stallions within the context of the traditional BSE. Additionally, the only individual semen measure that was significantly correlated with fertility was the percentage of morphologically normal sperm as determined using DIC microscopy. These results caution against strict use of the traditional 'cutoff' of 1 billion PMMNS for classification of breeding potential, particularly when attempting to differentiate between moderately and highly fertile stallions and regardless of the laboratory methods employed for semen analysis.


Assuntos
Contagem de Células/veterinária , Análise do Sêmen/veterinária , Motilidade Espermática/fisiologia , Animais , Contagem de Células/instrumentação , Cavalos , Masculino , Sêmen , Análise do Sêmen/métodos , Testículo/anatomia & histologia
15.
Andrologia ; 52(1): e13378, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31691338

RESUMO

The identification of idiopathic infertility cases, actually, is impossible. Among new functional tests, developed to improve the male fertility diagnosis, the evaluation of spermatic myo-inositol (MI) level, known as Andrositol® test (AT), is one of the most interesting, considering its weak economic burden and ease of use. The aim of this study was to evaluate the predictive power of AT and its potential use for a preliminary evaluation of semen samples. To evaluate the predictive power of AT, 87 sperm samples were analysed in comparison with spermiogram and sperm chromatin dispersion (SCD) Test, the gold standard analyses for male fertility evaluation. The application of AT resulted very useful for a preliminary sample evaluation, predicting the absence of DNA fragmentation in case of Low Responder samples precisely, and the presence of DNA fragmentation in case of medium or High Responder samples with abnormal morphology, predicting SCD results with a probability of 80% for Medium Responder sample and of 96.7% for High Responder sample. Considering the predictive power of this method, we could imagine, as preliminary qualitative analysis, its application before SCD test, deepening sperm analysis, improving the daily activities of laboratory operators and maintaining a good reliability of sperm evaluation.


Assuntos
Infertilidade Masculina/diagnóstico , Inositol/análise , Análise do Sêmen/métodos , Espermatozoides/química , Adulto , Cromatina/metabolismo , Fragmentação do DNA , Fertilidade , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Espermatozoides/metabolismo
16.
Andrologia ; 52(1): e13445, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31657071

RESUMO

Teratozoospermia is characterised by the presence of spermatozoa with abnormal morphology. One of the morphological disorders that lead to male infertility is immotile short-tail sperm (ISTS) defect. In this study, we evaluated the levels of chromatin packing and DNA fragmentation in patients with immotile short-tail sperm defect. Semen samples were obtained from 31 infertile men with ISTS as case group and 31 normozoospermic men as a control group. Protamine status was evaluated using chromomycin A3 (CMA3) staining and sperm DNA fragmentation assessed by sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labelling (TUNEL). The percentage of positive CMA3 spermatozoa was significantly higher in patients' samples (22.6 ± 6.9) compared with controls (16.3 ± 4.2) (p < .05) and also mean (±SD) of sperm DNA fragmentation was significantly higher in patients compared with controls, as measured by TUNEL assay (10.45 ± 4.60 vs. 7.03 ± 2.86, p < .05) and SCSA (24.80 ± 13.1 vs. 15.2 ± 7.2, p < .05). According to our study, sperm DNA fragmentation and chromatin packing abnormality are significantly higher in the ISTS samples compared with normal samples. A possible explanation for this relationship is that sperm chromatin condensation and sperm flagellum formation occur during the same phase of spermatogenesis.


Assuntos
Cromatina/metabolismo , Fragmentação do DNA , Cauda do Espermatozoide/patologia , Teratozoospermia/genética , Adulto , Estudos de Casos e Controles , Cromomicina A3/química , Empacotamento do DNA , Corantes Fluorescentes/química , Humanos , Masculino , Pessoa de Meia-Idade , Teste de Papanicolaou , Protaminas/metabolismo , Análise do Sêmen/métodos , Cauda do Espermatozoide/metabolismo , Teratozoospermia/patologia , Adulto Jovem
17.
Andrologia ; 52(1): e13412, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31671225

RESUMO

MicroRNAs modulate male fertility by regulating gene expression. In this study, dynamics of sperm miR-15a, miR-29b and miR-34a from high fertility (HF) and low fertility (LF) bulls using RT-qPCR were evaluated. Bioinformatic tools were employed to ascertain genes of interest of the sperm miRNAs. The expression levels of p53, BCL2, BAX and DNMT1 in bull spermatozoa were determined by immunoblotting. MicroRNA levels of miR-15a and miR-29 were higher in LF sires when compared with those present in HF bulls. Expression levels of miR-34a did not differ between the two groups. We found an inverse correlation between miR-15a and bull fertility. MiR29-b was also negatively associated with fertility scores. BCL2 and DNMT1 were higher in HF bulls while BAX was higher in the LF group. Our data showed a positive correlation between BCL2 and bull fertility. In addition, DNMT1 was positively associated with bull fertility. Furthermore, levels of BAX were negatively linked with bull fertility scores. Identification of miRNAs found in the spermatozoa of sires with different in vivo fertility helps understand the alterations in the fertilising capacity from cattle and other mammals. These potential biomarkers can be used in reproductive biotechnology as fertility markers to assess semen quality and predict male fertility.


Assuntos
Bovinos/fisiologia , Fertilidade/genética , MicroRNAs/metabolismo , Análise do Sêmen/veterinária , Espermatozoides/metabolismo , Animais , Biomarcadores/metabolismo , Cruzamento , Biologia Computacional , DNA (Citosina-5-)-Metiltransferase 1/genética , Regulação da Expressão Gênica/fisiologia , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/genética , Análise do Sêmen/métodos , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
19.
Pituitary ; 23(2): 160-166, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31834539

RESUMO

OBJECTIVE: Growth hormone (GH) activity might be implicated in male reproductive function. One previous study has suggested significantly reduced semen quality in untreated acromegalic patients due to both reduced sperm counts and sperm motility. DESIGN AND METHODS: A retrospective study comprising ten uncontrolled hypogonadal acromegalic patients (median age 29 years) who delivered semen for cryopreservation before initiation of testosterone therapy. Semen variables and hormone concentrations were compared to those of ten non-acromegalic hypogonadal men with pituitary disease (age 31 years) and those of young healthy men. RESULTS: Acromegalic patients vs. non-acromegalic patients had a higher percentage of progressive motile spermatozoa (62 vs. 47%, p = 0.04). Eight of ten acromegalic patients and 82% of controls had total sperm counts above 39 million and progressive motile spermatozoa above 32% (p = 0.55), corresponding to the WHO 2010 reference levels for expected normal fertility for these variables. Non-acromegalic patients vs. healthy controls had reduced percentage of progressive motile spermatozoa (47 vs. 57%, p = 0.02) and only five of ten patients had semen quality above the WHO reference level, which was significantly lower than observed in healthy controls (p = 0.022). Total sperm counts were similar between both patient groups and controls. There were no differences in reproductive hormone levels between acromegalic patients vs. non-acromegalic patients (p-values between 0.10 and 0.61). Compared to healthy controls both patient groups had severely reduced serum testosterone, calculated free testosterone. CONCLUSIONS: Despite severe hypoandrogenism acromegalic patients had semen quality similar to healthy controls based on determination of the number of progressively motile spermatozoa. By contrast non-acromegalic pituitary patients had reduced sperm motility. Our data do not support reduced semen quality in acromegaly.


Assuntos
Acromegalia/fisiopatologia , Análise do Sêmen/métodos , Adulto , Criopreservação , Hormônio do Crescimento/metabolismo , Humanos , Masculino , Reprodução/fisiologia , Estudos Retrospectivos , Espermatozoides/fisiologia
20.
Braz. J. Vet. Res. Anim. Sci. (Online) ; 57(3): e166205, 2020. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1122181

RESUMO

In freshwater fish with external fertilization, sperm sampling can be contaminated with urine, which triggers motility and gives rise to decreased fertilization success. The maintenance of freshwater fish in hyperosmotic conditions may reduce urine production and improve sperm quality. Thus, the aim of this work was to verify if acute exposure to various NaCl concentrations improves sperm quality in the yellowtail tetra Astyanax altiparanae. Spermiation was induced using a single dose of carp pituitary gland (5 mg kg-1) and the males were maintained at various NaCl concentrations: NaCl 0.00% (control), NaCl 0.45% (hypoosmotic), NaCl 0.9% (isosmotic) and NaCl 1.0% (hyperosmotic) for 6 h at 26 °C. Sperm was collected and verified for activation by urine and motility traits. At 0.00%, 0.45%, and 0.90%, the sperm was motile just after sampling, indicating activation by urine. Surprisingly, at hyperosmotic conditions, no activation was observed. Other sperm and motility parameters did not show any statistical differences, including sperm viability (P = 0.7083), concentration (P = 0.9030), total motility (P = 0.6149), VCL (curvilinear velocity; P = 0.1216), VAP (average path velocity; P = 0.1231) and VSL (straight-line velocity; P = 0.1340). Our results indicate that acute maintenance at hyperosmotic conditions eliminates sperm activation by urine and maintains sperm quality. Such a new procedure is interesting for both basic and applied sciences, including reproductive practice in fish.(AU)


Em peixes de água doce com fertilização externa, a amostragem de espermatozoides pode ser contaminada pela urina, o que desencadeia motilidade e gera menor sucesso na fertilização. A manutenção de peixes de água doce em condições hiperosmóticas pode reduzir a produção de urina e melhorar a qualidade do esperma. Assim, o presente trabalho foi delineado para verificar se a exposição aguda a várias concentrações de NaCl melhora a qualidade do esperma no tetra-amarelo Astyanax altiparanae. A espermiação foi induzida usando uma dose única de hipófise da carpa (5 mg kg-1) e os machos foram mantidos em várias concentrações de NaCl: NaCl 0,00% (controle), NaCl 0,45% (hipoosmótico), NaCl 0,9% (isosmótico) e NaCl 1,0% (hiperosmótico) por seis horas a 26 °C. O esperma foi colhido e verificado quanto à ativação por urina e traços de motilidade. Em 0,00%, 0,45%, 0,90% os espermatozóides eram móveis logo após a amostragem, indicando ativação pela urina. Surpreendentemente, em condições hiperosmóticas, nenhuma ativação foi observada. Outros parâmetros espermáticos e de motilidade não mostraram diferenças estatísticas, incluindo viabilidade espermática (P = 0,7083), concentração (P = 0,9030), motilidade total (P = 0,6149), VCL (Velocidade Curvilinear; P = 0,1216), VMD (Velocidade Média de Deslocamento; P = 0,1230) e VLR (Velocidade em linha Reta; P = 0,1340). Nossos resultados indicam que a manutenção aguda em condições hiperosmóticas elimina a ativação do esperma pela urina e mantém a qualidade do esperma. Esse novo procedimento é interessante para as ciências básicas e aplicadas, incluindo a prática reprodutiva em peixes.(AU)


Assuntos
Animais , Osmose , Salinidade , Análise do Sêmen/métodos , Análise do Sêmen/veterinária , Characidae/fisiologia , Motilidade Espermática
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