Male Attitudes towards Infertility:Results from a Global Questionnaire

PURPOSE: In general, men are less likely to seek health care than women. Infertility is a global disease that afflicts approximately 15% of reproductive age couples and the male contributes to 40% of the diagnosable cause. Remarkably, no large or multi-national population data exist regarding men’s perceptions about their infertility. The purpose of this study was to advance our knowledge about the infertile male’s social experience regarding: (1) how they feel about their infertility, (2) what motivated them to seek health care, (3) how likely are they to talk with others about their infertility, (4) their awareness of male infertility support groups, and (5) what their primary source for information is regarding male infertility? Based on the results from this study, these simple questions now have clearer definition. MATERIALS AND METHODS: An Institutional Review Board-approved, male-directed, anonymous questionnaire translated into 20 languages was made globally available through the Fertility Europe website (https://fertilityeurope.eu). Males (n=1,171) age 20–49 years were invited to complete the online survey after informed consent. RESULTS: Most respondents were European (86%). Of European men, <15.8% were self-motivated to seek medical help. Further, their physician was not the primary source of information regarding their infertility. While most men (59%) viewed their infertility positively, a large majority were not very likely (73%) to talk about it. Most respondents indicated a lack of awareness or absence of male infertility support groups. CONCLUSIONS: These are the first multi-national population data revealing men’s feelings about their infertility, what motivates them to seek help and their awareness of resources for peer support and information. These findings also serve to highlight significant gaps that exist in the provision of male reproductive health care and in supportive resources for men suffering from infertility. We offer recommendations on how to address the problem(s)

Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B

Breast cancer (BC) is the most common female cancer and the leading cause of cancer death in women worldwide. Alterations in epithelial cadherin (E-cadherin) expression and functions are associated to BC, but the underlying molecular mechanisms have not been fully elucidated. We have previously reported a novel human E-cadherin splice variant (E-cadherin variant) mRNA. Stable transfectants in MCF-7 human BC cells (MCF7Ecadvar) depicted fibroblast-like cell morphology, E-cadherin wild-type downregulation, and other molecular changes characteristic of the epithelial-to-mesenchymal transition process, reduced cell-cell adhesion, and increased cell migration and invasion. In this study, a two-dimensional differential gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS) protein identification and bioinformatics analyses were Results: By 2D-DIGE and MS analysis, 50 proteins were found differentially expressed (≥ Δ1.5) in MCF7Ecadvar compared to control cells. Validation of transcript expression was done in the ten most overexpressed and underexpressed proteins. Bioinformatics analyses revealed that 39 of the 50 proteins identified had been previously associated to BC. Moreover, metabolic processes were the most affected, and glycolysis the canonical pathway most altered. The lactate dehydrogenase B (LDHB) was the highest overexpressed protein, and transcript levels were higher in MCF7Ecadvar than in control cells. In agreement with these findings, MCF7Ecadvar conditioned media had lower glucose and higher lactate levels than control cells. MCF7Ecadvar cell treatment with 5 mM of the glycolytic inhibitor 2-deoxy-glucose led to decreased cell viability, and modulation of LDHB expression in MCF7Ecadvar cells with a specific small interfering RNA resulted in decreased cell proliferation. Finally, a positive association between expression levels of the E-cadherin variant and LDHB transcripts was demonstrated in 21 human breast tumor tissues, and breast tumor samples with higher Ki67 expression showed higher LDHB mRNA levels. Conclusions: Results from this investigation contributed to further characterize molecular changes associated to the novel E-cadherin splice variant expression in BC cells. They also revealed an association between expression of the novel variant and changes related to BC progression and aggressiveness, in particular those associated to cell metabolism. Keywords: Breast cancer, Epithelial cadherin, Alternative splicing, Epithelial to mesenchymal transition, Proteomic analysis, 2D-DIGE, Mass spectrometry, Glycolysis, Lactate dehydrogenase

Expresión plasmídica heteróloga en embriones ovinos obtenidos por fertilización in vitro usando el espermatozoide como vector

Los resultados en la obtención de animales de granja genéticamente modificados (AGGM), no están aún a la altura de las expectativas científicas. A nivel mundial, los AGGM patentados y funcionales son muy escasos. Esta lentitud en resultados medido en términos de AGGM nacidos, reproductivamente viables por ensayo, es asignada al bajo rendimiento de las técnicas utilizadas, su costo elevado y el riesgo asociado a la bioseguridad. La hipótesis de trabajo fue que el espermatozoide fértil e intacto, de vertebrados mamíferos superiores, es capaz de portar ADN heterólogo y durante el acto de la fecundación, sea in-vitro o in-vivo, integrarlo funcionalmente al genoma del cigoto. Consecuentemente, será posible su expresión en los blastómeros del embrión y las células diferenciadas del organismo adulto. Asimismo, se postuló que los resultados medidos en términos de tasa de ensayo/éxito de esta técnica superan ampliamente a las otras técnicas utilizadas. Se lograron generar embriones ovinos por fecundación in-vitro (FIV) de acuerdo a los parámetros del tratamiento del espermatozoide (SPZ) previamente establecidos. Para lograr este objetivo se utilizaron 48 complejos cumulus ovocito (CCOs) provenientes de 35 tractos genitales ovinos de matadero, cultivados y madurados in-vitro. Dos muestras, una tratada con semen tratado y la otra (control) con semen no tratado, fueron puestas a fertilizar. Luego de la fertilización y cultivo los embriones fueron retirados y observados bajo microscopia microcaptura láser fluorescente (LCM) y con focal de fluorescencia. Si bien la muestra control debió ser descartada al momento de la observación por excesiva fibrinosis, en la muestra tratada pudieron observarse 11 blastocistos expandidos con intensa señal de fluorescencia en todas las blastómeros, 6 ovocitos no fertilizadosy 4 embriones malformados que no presentaron señal bajo luz fluorescente. Se consideró testigo a los ovocitos no fertilizados que no presentaron fluorescencia alguna; y se los estimó como evidencia válida para demostrar la transfección realizada por parte del SPZ. Solo aquellos que fueron fecundados, y en este caso la totalidad (100%), evidenciaron fluorescencia en todas las blastómeros en las diferentes muestras.Production of genetically modified farm animals (GMFA) is not yet up to scientific expectations.This slowness results measured in terms of GMFA born, reproductively viable (“founders”) by trial, is assigned to the very low yield of the techniques used to incorporate gene constructs, therefore, high cost and bio-security risk of some of them. Our hypothesis, was that properly treated, fertile and intact mammalian sperm is capable of carrying heterologous DNA, and, during the act of fecundation, either in-vitro or in-vivo, functionally integrate it into the resulting zygote genome, allowing subsequent expression in embryo blastomeres and differentiated cells of the adult organism, and, that the results measured as test rate / success of this technique, far outweigh other techniques used with the same objective. As specific goals we proposed and developed sheep embryos from In-Vitro Fertilization (IVF) according to the parameters of sperm treatment (SPZ) previously established. To achieve this goal 48 Cumulus Oocyte Complex (COCs) was isolated from 35 slaughter-house ovine genital tracts, grown and matured in-vitro as described above. Two oocytes samples, one with semen treated and the other (control) with untreated sperm, are submitted to a fecundation. After fertilization and culture, embryos are removed and observed under fluorescent microscopy microcapture laser (LCM) and fluorescence confocal. While the control sample should be discarded at the time of observation by excessive fibrinosis, treated sample revealed 11 expanded blastocysts with intense fluorescence signal in all blastomeres, 6 unfertilized oocytes and 4 malformed embryos showingno signal under fluorescent light.It is considered as control unfertilized oocytes who did not show any fluorescence as valid evidence to prove the transfection carried out by the SPZ, since only those who were fertilized, in this case all (100%) evidenced strong fluorescence signal in all blastomeres in different samples

E-cadherin: A determinant molecule associated with ovarian cancer progression, dissemination and aggressiveness

Ovarian cancer (OC) is the fifth cancer death cause in women worldwide. The malignant nature of this disease stems from its unique dissemination pattern. Epithelial-to-mesenchymal transition (EMT) has been reported in OC and downregulation of Epithelial cadherin (E-cadherin) is a hallmark of this process. However, findings on the relationship between E-cadherin levels and OC progression, dissemination and aggressiveness are controversial. In this study, the evaluation of E-cadherin expression in an OC tissue microarray revealed its prognostic value to discriminate between advanced-and early-stage tumors, as well as serous tumors from other histologies. Moreover, E-cadherin, Neural cadherin (N-cadherin), cytokeratins and vimentin expression was assessed in TOV-112, SKOV-3, OAW-42 and OV-90 OC cell lines grown in monolayers and under anchorage-independent conditions to mimic ovarian tumor cell dissemination, and results were associated with cell aggressiveness. According to these EMT-related markers, cell lines were classified as mesenchymal (M; TOV-112), intermediate mesenchymal (IM; SKOV-3), intermediate epithelial (IE; OAW-42) and epithelial (E; OV-90). M-and IM-cells depicted the highest migration capacity when grown in monolayers, and aggregates derived from M-and IM-cell lines showed lower cell death, higher adhesion to extracellular matrices and higher invasion capacity than E- antigen 125 levels more than 500 U/mL and platinum-free intervals less than 6 months. Altogether, E-cadherin expression levels were found relevant for the assessment of OC progression and aggressiveness.and IE-aggregates. The analysis of E-cadherin, N-cadherin, cytokeratin 19 and vimentin mRNA levels in 20 advanced-stage high-grade serous human OC ascites showed an IM phenotype in all cases, characterized by higher proportions of N-to E-cadherin and vimentin to cytokeratin 19. In particular, higher E-cadherin mRNA levels were associated with cancer antigen 125 levels more than 500 U/mL and platinum-free intervals less than 6 months. Altogether, E-cadherin expression levels were found relevant for the assessment of OC progression and aggressiveness

Current global status of male reproductive health

BACKGROUND: The widespread interest in male reproductive health (MRH), fueled by emerging evidence, such as the global decline in sperm counts, has intensified concerns about the status of MRH. Consequently, there is a pressing requirement for a strategic, systematic approach to identifying critical questions, collecting pertinent information, and utilizing these data to develop evidence-based strategies. The methods for addressing these questions and the pathways toward their answers will inevitably vary based on the variations in cultural, geopolitical, and health-related contexts. To address these issues, a conjoint ESHRE and Male Reproductive Health Initiative (MRHI) Campus workshop was convened.OBJECTIVE AND RATIONALE: The three objectives were: first, to assess the current state of MRH around the world; second, to identify some of the key gaps in knowledge; and, third, to examine how MRH stakeholders can collaboratively generate intelligent and effective paths forward.SEARCH METHODS: Each expert reviewed and summarized the current literature that was subsequently used to provide a comprehensive overview of challenges related to MRH.OUTCOMES: This narrative report is an overview of the data, opinions, and arguments presented during the workshop. A number of outcomes are presented and can be summarized by the following overarching themes: MRH is a serious global issue and there is a plethora of gaps in our understanding; there is a need for widespread international collaborative networks to undertake multidisciplinary research into fundamental issues, such as lifestyle/environmental exposure studies, and high-quality clinical trials; and there is an urgent requirement for effective strategies to educate young people and the general public to safeguard and improve MRH across diverse population demographics and resources.LIMITATIONS REASONS FOR CAUTION: This was a workshop where worldwide leading experts from a wide range of disciplines presented and discussed the evidence regarding challenges related to MRH. While each expert summarized the current literature and placed it in context, the data in a number of areas are limited and/or sparse. Equally, important areas for consideration may have been missed. Moreover, there are clear gaps in our knowledge base, which makes some conclusions necessarily speculative and warranting of further study.WIDER IMPLICATIONS: Poor MRH is a global issue that suffers from low awareness among the public, patients, and heathcare professionals. Addressing this will require a coordinated multidisciplinary approach. Addressing the significant number of knowledge gaps will require policy makers prioritizing MRH and its funding.STUDY FUNDING/COMPETING INTERESTS: The authors would like to extend their gratitude to ESHRE for providing financial support for the Budapest Campus Workshop, as well as to Microptic S.L. (Barcelona) for kindly sponsoring the workshop. P.B. is the Director of the not-for-profit organization Global Action on Men's Health and receives fees and expenses for his work, (which includes the preparation of this manuscript). Conflicts of interest: C.J.D.J., C.L.R.B., R.A.A., P.B., M.P.C., M.L.E., N.G., N.J., C.K., AAP, M.K.O., S.R.-H., M.H.V.-L.: ESHRE Campus Workshop 2022 (Travel support-personal). C.J.D.J.: Cambridge University Press (book royalties-personal). ESHRE Annual Meeting 2022 and Yale University Panel Meeting 2023 (Travel support-personal). C.L.R.B.: Ferring and IBSA (Lecture), RBMO editor (Honorarium to support travel, etc.), ExSeed and ExScentia (University of Dundee), Bill &amp; Melinda Gates Foundation (for research on contraception). M.P.C.: Previously received funding from pharmaceutical companies for health economic research. The funding was not in relation to this work and had no bearing on the contents of this work. No funding from other sources has been provided in relation to this work (funding was provided to his company Global Market Access Solutions). M.L.E.: Advisor to Ro, Doveras, Next, Hannah, Sandstone. C.K.: European Academy of Andrology (Past president UNPAID), S.K.: CEO of His Turn, a male fertility Diagnostic and Therapeutic company (No payments or profits to date). R.I.M.: www.healthymale.org.au (Australian Government funded not for profit in men's health sector (Employed as Medical Director 0.2 FET), Monash IVF Pty Ltd (Equity holder)). N.J.: Merck (consulting fees), Gedeon Richter (honoraria). S.R.-H.: ESHRE (Travel reimbursements). C.N.: LLC (Nursing educator); COMMIT (Core Outcomes Measures for Infertility Trials) Advisor, meeting attendee, and co-author; COMMA (Core Outcomes in Menopause) Meeting attendee, and co-author; International Federation of Gynecology and Obstetrics (FIGO) Delegate Letters and Sciences; ReproNovo, Advisory board; American Board of Urology Examiner; American Urological Association Journal subsection editor, committee member, guidelines co-author Ferring Scientific trial NexHand Chief Technology Officer, stock ownership Posterity Health Board member, stock ownership. A.P.: Economic and Social Research Council (A collaborator on research grant number ES/W001381/1). Member of an advisory committee for Merck Serono (November 2022), Member of an advisory board for Exceed Health, Speaker fees for educational events organized by Mealis Group; Chairman of the Cryos External Scientific Advisory Committee: All fees associated with this are paid to his former employer The University of Sheffield. Trustee of the Progress Educational Trust (Unpaid). M.K.O.: National Health and Medical Research Council and Australian Research Council (Funding for research of the topic of male fertility), Bill and Melinda Gates Foundation (Funding aimed at the development of male gamete-based contraception), Medical Research Future Fund (Funding aimed at defining the long-term consequences of male infertility). M.H.V.-L.: Department of Sexual and Reproductive Health and Research (SRH)/Human Reproduction Programme (HRP) Research Project Panel RP2/WHO Review Member; MRHI (Core Group Member), COMMIT (member), EGOI (Member); Human Reproduction (Associate Editor), Fertility and Sterility (Editor), AndroLATAM (Founder and Coordinator).</p

Spermatozoa capture HIV-1 through heparan sulfate and efficiently transmit the virus to dendritic cells

Semen is the main vector for HIV-1 dissemination worldwide. It contains three major sources of infectious virus: free virions, infected leukocytes, and spermatozoa-associated virions. We focused on the interaction of HIV-1 with human spermatozoa and dendritic cells (DCs). We report that heparan sulfate is expressed in spermatozoa and plays an important role in the capture of HIV-1. Spermatozoa-attached virus is efficiently transmitted to DCs, macrophages, and T cells. Interaction of spermatozoa with DCs not only leads to the transmission of HIV-1 and the internalization of the spermatozoa but also results in the phenotypic maturation of DCs and the production of IL-10 but not IL-12p70. At low values of extracellular pH (∼6.5 pH units), similar to those found in the vaginal mucosa after sexual intercourse, the binding of HIV-1 to the spermatozoa and the consequent transmission of HIV-1 to DCs were strongly enhanced. Our observations support the notion that far from being a passive carrier, spermatozoa acting in concert with DCs might affect the early course of sexual transmission of HIV-1 infection

Anandamide Capacitates Bull Spermatozoa through CB1 and TRPV1 Activation

Anandamide (AEA), a major endocannabinoid, binds to cannabinoid and vanilloid receptors (CB1, CB2 and TRPV1) and affects many reproductive functions. Nanomolar levels of anandamide are found in reproductive fluids including mid-cycle oviductal fluid. Previously, we found that R(+)-methanandamide, an anandamide analogue, induces sperm releasing from bovine oviductal epithelium and the CB1 antagonist, SR141716A, reversed this effect. Since sperm detachment may be due to surface remodeling brought about by capacitation, the aim of this paper was to investigate whether anandamide at physiological concentrations could act as a capacitating agent in bull spermatozoa. We demonstrated that at nanomolar concentrations R(+)-methanandamide or anandamide induced bull sperm capacitation, whereas SR141716A and capsazepine (a TRPV1 antagonist) inhibited this induction. Previous studies indicate that mammalian spermatozoa possess the enzymatic machinery to produce and degrade their own AEA via the actions of the AEA-synthesizing phospholipase D and the fatty acid amide hydrolase (FAAH) respectively. Our results indicated that, URB597, a potent inhibitor of the FAAH, produced effects on bovine sperm capacitation similar to those elicited by exogenous AEA suggesting that this process is normally regulated by an endogenous tone. We also investigated whether anandamide is involved in bovine heparin-capacitated spermatozoa, since heparin is a known capacitating agent of bovine sperm. When the spermatozoa were incubated in the presence of R(+)-methanandamide and heparin, the percentage of capacitated spermatozoa was similar to that in the presence of R(+)-methanandamide alone. The pre-incubation with CB1 or TRPV1 antagonists inhibited heparin-induced sperm capacitation; moreover the activity of FAAH was 30% lower in heparin-capacitated spermatozoa as compared to control conditions. This suggests that heparin may increase endogenous anandamide levels. Our findings indicate that anandamide induces sperm capacitation through the activation of CB1 and TRPV1 receptors and could be involved in the same molecular pathway as heparin in bovines

Epithelial and Neural Cadherin in Mammalian Fertilization: Studies in the Mouse Model

Successful mammalian fertilization requires a well-orchestrated sequence of molecular events leading to gamete fusion. Since this interaction involves Ca2+-dependent adhesion events, the participation of the Ca+2-dependent cell-cell adhesion proteins Epithelial (E-cad) and Neural (N-cad) cadherin is envisaged. We have previously reported the expression of E-cad and N-cad in human gametes and showed evidence of their involvement in sperm-oocyte adhesion events leading to fertilization. To overcome ethical limitations associated with the use of human gametes in fertilization-related studies, the mouse has been selected worldwide as the experimental model for over 4 decades. Herein, we report a detailed study aimed at characterizing the expression of E-cad and N-cad in murine gametes and their involvement in murine fertilization using specific antibodies and blocking peptides towards both adhesion proteins. E-cad and N-cad protein forms, as well as other members of the adhesion complex, specifically &beta;-catenin and actin, were identified in spermatozoa, cumulus cells and oocytes protein extracts by means of Western immunoblotting. In addition, subcellular localization of these proteins was determined in whole cells using optical fluorescent microscopy. Gamete pre-incubation with anti-E-cad (ECCD-1) or N-cad (H-63) antibodies resulted in decreased (p &lt; 0.05) In Vitro Fertilization (IVF) rates, when using both cumulus-oocytes complexes and cumulus-free oocytes. Moreover, IVF assays done with denuded oocytes and either antibodies or blocking peptides against E-cad and N-cad led to lower (p &lt; 0.05) fertilization rates. When assessing each step, penetration of the cumulus mass was lower (p &lt; 0.05) when spermatozoa were pre-incubated with ECCD-1 or blocking peptides towards E-cad or towards both E- and N-cad. Moreover, sperm-oolemma binding was impaired (p &lt; 0.0005) after sperm pre-incubation with E-cad antibody or blocking peptide towards E-cad, N-cad or both proteins. Finally, sperm-oocyte fusion was lower (p &lt; 0.05) after sperm pre-incubation with either antibody or blocking peptide against E-cad or N-cad. Our studies demonstrate the expression of members of the adherent complex in the murine model, and the use of antibodies and specific peptides revealed E-cad and N-cad participation in mammalian fertilization

An Overview of the proacrosin/acrosin system in human spermatozoa

Among all of the sperm acrosomal trypsin-like proteinases studied, acrosin (EC 3.4.21.10) has been identified in all of the species evaluated and has been associated with human sperm fertility potential. In this report, a brief overview of cellular, biochemical, and molecular aspects related to the human proacrosin/acrosin system, has been compiled from studies performed by our research team, as well as contributions by numerous groups from the clinical and basic field of reproductive biology. As a result of these studies, a putative model for the interaction between the proacrosin/acrosin system and the zona pellucida glycoproteins, in association with proenzyme activation and proteinase activity, is presented.Entre totes les proteïnases espermàtiques semblants a la tripsina estudiades, l'acrosina (EC 3.4.21.10) ha estat identificada en totes les espècies estudiades, i ha estat associada amb el potencial fertilitzador de l'esperma. En aquesta breu revisió, volem presentar els principals aspectes cell. ulars, moleculars i bioquímics del sistema proacrosina/acrosina definits pel nostre grup de recerca i per molts altres equips de recerca del camp de la biologia de la reproducció. Com a resultat de tots aquests estudis, presentem el model putatiu de la interacció del sistema proacrosina/ acrosina amb les glicoproteïnes de la zona pell. úcida i la demostració de l'activació del proenzim i la seva activitat com a proteïnasa