Transmission electron microscopy reveals the presence of SARS-CoV-2 in human spermatozoa associated with an ETosis-like response.

BACKGROUND: Severe acute syndrome coronavirus 2 can invade a variety of tissues, including the testis. Even though this virus is scarcely found in human semen polymerase chain reaction tests, autopsy studies confirm the viral presence in all testicular cell types, including spermatozoa and spermatids. OBJECTIVE: To investigate whether the severe acute syndrome coronavirus 2 is present inside the spermatozoa of negative polymerase chain reaction-infected men up to 3 months after hospital discharge. MATERIALS AND METHODS: This cross-sectional study included 13 confirmed moderate-to-severe COVID-19 patients enrolled 30-90 days after the diagnosis. Semen samples were obtained and examined with real-time polymerase chain reaction for RNA detection and by transmission electron microscopy. RESULTS: In moderate-to-severe clinical scenarios, we identified the severe acute syndrome coronavirus 2 inside spermatozoa in nine of 13 patients up to 90 days after discharge from the hospital. Moreover, some DNA-based extracellular traps were reported in all studied specimens. DISCUSSION AND CONCLUSION: Although severe acute syndrome coronavirus 2 was not present in the infected men's semen, it was intracellularly present in the spermatozoa till 3 months after hospital discharge. The Electron microscopy (EM) findings also suggest that spermatozoa produce nuclear DNA-based extracellular traps, probably in a cell-free DNA-dependent manner, similar to those previously described in the systemic inflammatory response to COVID-19. In moderate-to-severe cases, the blood-testes barrier grants little defence against different pathogenic viruses, including the severe acute syndrome coronavirus 2. The virus could also use the epididymis as a post-testicular route to bind and fuse to the mature spermatozoon and possibly accomplish the reverse transcription of the single-stranded viral RNA into proviral DNA. These mechanisms can elicit extracellular cell-free DNA formation. The potential implications of our findings for assisted conception must be addressed, and the evolutionary history of DNA-based extracellular traps as preserved ammunition in animals' innate defence might improve our understanding of the severe acute syndrome coronavirus 2 pathophysiology in the testis and spermatozoa.

Serum lipid profile levels and semen quality: new insights and clinical perspectives for male infertility and men's health.

PURPOSE: Several clinical scenarios regulate the final ejaculated semen, which is pivotal to reproductive success. Sperm motility and plasma membrane fusogenic activity primarily rely on the peculiar sperm lipid composition, influenced by the patient's metabolism, genetics, nutritional, environmental status, and concomitant clinical entities such as varicocele. This study aimed to determine the relationship between serum lipid profile and testicular function (semen quality and testosterone levels). METHODS: This retrospective study uses medical charts of 278 infertile men who attended andrological care between 2000 and 2019. Seminal analysis data, lipid profile, and total serum testosterone were collected. A multiple linear regression analysis was performed to evaluate the influence of the lipid parameters on the seminal variables. Statistical analyses were carried out with p ≤ 0.05 considered statistically significant. RESULTS: Seminal creatine kinase activity (p = 0.024) is negatively related to HDL (p = 0.032) and triglycerides (p = 0.037), while total testosterone (p < 0.0001) and seminal volume (p = 0.046) appeared both to be negatively related to triglycerides (p = 0.030 and p = 0.033, respectively). CONCLUSION: Medical advice commonly advocated to prevent endothelial dysfunction and cardiovascular disease and improve HDL-cholesterol and triglyceride levels in dyslipidemic patients should also be given to infertile men. Physicians should give patients a thorough assessment, including the blood lipid profile, hormonal status, and routine seminal examinations. We propose a more comprehensive men´s health check-up for the infertile male population, not limited to a simple evaluation of basic sperm parameters.

Marijuana Is Associated With a Hormonal Imbalance Among Several Habits Related to Male Infertility: A Retrospective Study.

Marijuana is one of the most consumed drugs worldwide. There is increasing evidence of an association between marijuana and male infertility. This study intends to assess the repercussion of marijuana smoking and other habits (sedentary lifestyle, alcohol, and tobacco use) in the testicular function of infertile men seeking andrological evaluation. A retrospective study was performed using medical records data of men aged 18-59 years from 2009 to 2017. Complete semen analyses, sperm functional tests, SHBG, and hormonal levels, testosterone-to-estradiol ratio (T/E2), and testis volume were evaluated. Exclusion criteria included cryptorchidism, infertility caused by genetic or infectious diseases, and cancer. A multiple linear regression analysis was performed to investigate which habit could predict certain parameters using the software SPSS 23.0 (P < 0.05). In a sample of 153 men, semen parameters, testosterone levels, and testis volume were not significantly influenced. Marijuana use had the broader hormonal changes since it influences estradiol (P = 0.000; B = -11.616), prolactin (P = 0.000; B = 3.211), SHBG levels (P = 0.017; B = 7.489), and T/E2 (P = 0.004; B = 14.030). Sedentary lifestyle (P = 0.028; B = 1.279) and tobacco smoking (P = 0.031; B = -2.401) influenced the prolactin levels. Marijuana is associated with hormonal imbalance in this infertile cohort by lowering estradiol levels and inhibiting aromatase function.

Male sex rather than socioeconomic vulnerability as a determinant for COVID-19 death in Sao Paulo: A population-based study.

Objectives: To determine the role of the male sex as a risk factor for coronavirus disease deaths in Sao Paulo and to what extent socioeconomic vulnerability and individual health issues can interfere in such risk. Methods: The primary cause of death, age, sex, comorbidities, and code of the Human Development Units of the residence of 37,583 individuals in Sao Paulo, Brazil, were obtained from the records on confirmed coronavirus disease resident hospitalizations of the city of Sao Paulo from the National Influenza Surveillance Information System. A social vulnerability index was assigned to each Human Development Unit. Using "death" as the outcome variable and sex, admission to the intensive care unit, obesity, renal and heart diseases, diabetes, and social vulnerability as confounders, the odds of death for males and females were compared via logistic regression. Results: The odds of death for males were 1.242 (confidence interval 95% = 1.237, 1.247) times the corresponding odds for females with the same values for all confounders. We estimated the odds of death for patients living in regions with high social vulnerability as 2.243 (CI 95% = 2.151, 2.339) times the corresponding odds of patients living in regions with very low social vulnerability with the same values of the remaining variables. Conclusion: The male:female death ratio by severe acute respiratory syndrome coronavirus 2 infection in Sao Paulo cannot be attributed only to comorbidities or social vulnerabilities. Our results suggest that the male sex is an independent biological risk factor for coronavirus disease death. Besides sex-specific factors, further research should focus on crucial biological factors in male sex coronavirus disease mortality.

Testicular pathology in fatal COVID-19: A descriptive autopsy study.

BACKGROUND: Multi-organ damage is a common feature of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, going beyond the initially observed severe pneumonia. Evidence that the testis is also compromised is growing. OBJECTIVE: To describe the pathological findings in testes from fatal cases of COVID-19, including the detection of viral particles and antigens, and inflammatory cell subsets. MATERIALS AND METHODS: Postmortem testicular samples were obtained by percutaneous puncture from 11 deceased men and examined by reverse-transcription polymerase chain reaction (RT-PCR) for RNA detection and by light and electron microscopy (EM) for SARS-CoV-2. Immunohistochemistry (IHC) for the SARS-CoV-2 N-protein and lymphocytic and histiocytic markers was also performed. RESULTS: Eight patients had mild interstitial orchitis, composed mainly of CD68+ and TCD8+ cells. Fibrin thrombi were detected in five cases. All cases presented congestion, interstitial edema, thickening of the tubular basal membrane, decreased Leydig and Sertoli cells with reduced spermatogenesis, and strong expression of vascular cell adhesion molecule (VCAM) in vessels. IHC detected SARS-Cov-2 antigen in Leydig cells, Sertoli cells, spermatogonia, and fibroblasts in all cases. EM detected viral particles in the cytoplasm of fibroblasts, endothelium, Sertoli and Leydig cells, spermatids, and epithelial cells of the rete testis in four cases, while RT-PCR detected SARS-CoV-2 RNA in three cases. DISCUSSION AND CONCLUSION: The COVID-19-associated testicular lesion revealed a combination of orchitis, vascular changes, basal membrane thickening, Leydig and Sertoli cell scarcity, and reduced spermatogenesis associated with SARS-CoV-2 local infection that may impair hormonal function and fertility in men.

Radiological patterns of incidental epididymitis in mild-to-moderate COVID-19 patients revealed by colour Doppler ultrasound.

The testis is a potential target organ for SARS-CoV-2 infection. Our study intended to investigate any testicular involvement in mild-to-moderate COVID-19 men. We conduct a cross-sectional study in 18 to 55-year-old men hospitalised for confirmed COVID-19. A senior radiologist executed the ultrasound with multi-frequency linear probe in all participants, regardless of any scrotal complaints. Exclusion criteria involved any situation that could impair testicular function. Statistical analysis compared independent groups, classified by any pathological change. Categorical and numerical outcome hypotheses were tested by Fisher's Exact and Mann-Whitney tests, using the Excel for Mac, version 16.29 (p < .05). The sample size was 26 men (mean 33.7 ± 6.2 years; range: 21-42 years), all without scrotal complaints. No orchitis was seen. Eleven men (32.6 ± 5.8 years) had epididymitis (42.3%), bilateral in 19.2%. More than half of men with epididymitis displayed epididymal head augmentation > 1.2 cm (p = .002). Two distinct epididymitis' patterns were reported: (a) disseminated micro-abscesses (n = 6) and (b) inhomogeneous echogenicity with reactional hydrocele (n = 5). Both patterns revealed increased epididymal head, augmented Doppler flow and scrotal skin thickening. The use of colour Doppler ultrasound in mild-to-moderate COVID-19 men, even in the absence of testicular complaints, might be useful to diagnose epididymitis that could elicit fertility complications.

Viral infections and implications for male reproductive health.

Viral infections have haunted humankind since times immemorial. Overpopulation, globalization, and extensive deforestation have created an ideal environment for a viral spread with unknown and multiple shedding routes. Many viruses can infect the male reproductive tract, with potential adverse consequences to male reproductive health, including infertility and cancer. Moreover, some genital tract viral infections can be sexually transmitted, potentially impacting the resulting offspring's health. We have summarized the evidence concerning the presence and adverse effects of the relevant viruses on the reproductive tract (mumps virus, human immunodeficiency virus, herpes virus, human papillomavirus, hepatitis B and C viruses, Ebola virus, Zika virus, influenza virus, and coronaviruses), their routes of infection, target organs and cells, prevalence and pattern of virus shedding in semen, as well as diagnosis/testing and treatment strategies. The pathophysiological understanding in the male genital tract is essential to assess its clinical impact on male reproductive health and guide future research.

Serum vitamin D content is associated with semen parameters and serum testosterone levels in men.

The present study aimed to evaluate the influence of serum vitamin D levels on semen quality and testosterone levels. This is a cross-sectional study conducted at Androscience, Science and Innovation Center in Andrology and High-Complex Clinical and Andrology Laboratory in Sao Paulo, Brazil, with 508 male patients, aged 18-60 years, from 2007 to 2017. Seminal parameters and serum sexual hormones were correlated with serum vitamin D concentrations in 260 men selected by strict selection criteria. Patients were divided into normozoospermic group (NZG, n = 124) and a group with seminal abnormalities (SAG, n = 136). Evaluation included complete physical examination, past medical history, habits and lifestyle factors, two complete seminal analysis with sperm functional tests, serum levels of 25-hydroxy-vitamin D3(25(OH)VD3), total and free testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), sex hormone-binding globulin (SHBG), total cholesterol, homeostatic model assessment of insulin resistance (HOMA-IR) index, and karyotype. The mean concentration of 25(OH)VD3was significantly lower in the SAG (P < 0.001) and positively correlated with all baseline seminal parameters and total testosterone levels. In addition, serum vitamin D3concentration was found to be positively correlated with sperm concentration (ß= 2.103; P < 0.001), total number of spermatozoa with progressive motility (ß = 2.069; P = 0.003), total number of motile spermatozoa (ß = 2.571; P = 0.015), and strict morphology (ß = 0.056; P = 0.006), regardless of other variables. This is the first comparative study to address the issue of serum vitamin D3content between normozoospermic patients and those with sperm abnormalities. It clearly demonstrates a direct and positive relationship between serum vitamin D level and overall semen quality, male reproductive potential, and testosterone levels.

SARS-CoV-2 and its relationship with the genitourinary tract: Implications for male reproductive health in the context of COVID-19 pandemic.

BACKGROUND: The current outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, named coronavirus disease 19 (COVID-19), is not the first well-known spillover of an animal originated virus to infect humans. However, one of the few to make such a fast jump in a powerful evolutionary shortcut. The incredible pattern of aggressiveness worldwide since the beginning of the outbreak is that up to 20% of those infected need hospitalization and 5% evolve to critical conditions, not limited to respiratory-related issues, but rather to systemic involvement. OBJECTIVE: This study aims to summarize the current knowledge about the effects of SARS-CoV-2 infection on the male genitourinary tract. MATERIALS AND METHODS: A narrative review was carried out to identify articles on the SARS-CoV-2 infection on the male genitourinary system. RESULTS: Considerations were made about the molecular characteristics of SARS-CoV-2 and immune response to coronavirus. We discussed the influence of the virus on the urinary system, potential mechanisms of COVID-19- related acute kidney injury (AKI), and the role of cytokine release syndrome on the renal pathophysiology of the disease. In the male reproductive tract, it was discussed the testis' vulnerability to SARS-CoV-2 invasion and the possible adverse effects on its function and the seminal findings of COVID-19. DISCUSSION AND CONCLUSION: During the COVID-19 pandemic, an international coordinated scientific effort must arise to understand the role of the urogenital system in the SARS-CoV-2 infection in the clinical setting.

Publications generated from abstracts presented at the 28thCongresso Brasileiro de Patologia and the 28th Congreso de la Sociedad Latinoamericana de Patologia, in 2011 / Publicações geradas a partir de resumos apresentados no XXVIII Congresso Brasileiro de Patologia e no XXVIII Congreso de la Sociedad Latinoamericana de Patología, realizados em 2011

ABSTRACTIntroduction:The medical congresses are an efficient way, although restricted, to expose the scientific production of a particular specialty. However, the number of complete articles published resulting from papers presented at congresses and published as abstracts is much lower than the number of papers presented during the meetings.Objective:Scale the number of complete articles published and indexed in online databases, from 2012 to January 2015, originally presented at the 28th Congresso Brasileiro de Patologia (28th Brazilian Congress of Pathology) and the 28th Congreso de la Sociedad Latinoamericana de Patología (28th Latin American Society of Pathology Congress).Material and method:In the meetings mentioned 701 papers were presented and their abstracts published in the congresses annals. An observational retrospective study was conducted consulting the following biomedical databases: Scientific Electronic Library Online (Scielo) and US National Library of Medicine on the National Institutes of Health (PubMed), with the help of the Google search engine, based on the article title and the list of authors.Results:From 701 studies analyzed, 635 (90.6%) were from Brazilian institutions, 60 (8.6%) from foreign institutions, and six (0.8%) did not identified their home institutions. There were found 63 (8.8%) complete articles published and indexed to consulted databases. International journals were the destination of 38 (60.3%) publications.Conclusion:The number of complete articles published in Pathology area arising from the papers presented in the studied meetings in the first three years after the meetings is relatively small. This number is low compared with other international conferences; however, it is higher than some Brazilian medical congresses analyzed in similar studies.

RESUMOIntrodução:Os congressos médicos são um meio eficiente, embora restrito, de expor a produção científica de uma determinada especialidade. Entretanto, o número de artigos completos publicados resultantes dos trabalhos apresentados em congressos e publicados como resumos é muito inferior ao número dos trabalhos apresentados durante os eventos.Objetivo:Dimensionar o número de artigos completos publicados e indexados em bases de dados virtuais, no período de 2012 a janeiro de 2015, apresentados originalmente no XXVIII Congresso Brasileiro de Patologia e no XXVIII Congreso de La Sociedade Latinoamericana de Patología.Material e método:Nos eventos mencionados, foram apresentados 701 trabalhos e seus resumos publicados nos anais do congresso. Efetuou-se um estudo retrospectivo observacional consultando as seguintes bases de dados biomédicas: Scientific Electronic Library Online (Scielo) e US National Library of Medicine do National Institutes of Health (PubMed), com o auxílio do buscador Google, tomando por base o título do trabalho e a lista de autores.Resultados:Dos 701 trabalhos analisados, 635 (90,6%) eram originados de instituições brasileiras, 60 (8,6%), de instituições estrangeiras e em seis (0,8%) não foram identificadas as instituições de origem. Foram encontrados 63 (8,8%) trabalhos completos publicados e indexados às bases de dados consultadas. As revistas internacionais foram o destino de 38 (60,3%) publicações.Conclusão:É relativamente pequeno o número de artigos completos publicados na área de Patologia, resultantes dos trabalhos apresentados nos eventos estudados, nos três primeiros anos após a realização do evento. Esse número é baixo, quando comparado com o de outros congressos internacionais, contudo, é superior ao de alguns congressos médicos brasileiros analisados em estudos semelhantes.