P–026 Seminal plasma exosomes: a promising source of biomarkers for fertility evaluation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
P Piomboni ◽  
A Luddi ◽  
C Landi ◽  
A Haxhiu ◽  
F L Presti ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Male Fertility ◽  
Seminal Fluid ◽  
Interaction Analysis ◽  
Secretory Protein ◽  
Sperm Maturation ◽  
University Hospital ◽  
2D Electrophoresis ◽  
Seminal Parameters ◽  
Promising Source

Abstract Study question Do exosomes from seminal plasma have a role in male fertility? Summary answer Exosomes isolated from seminal plasma have a pivotal role during spermatogenesis and sperm maturation and may represent eligible biomarkers for male fertility/infertility. What is known already During their journey along the male reproductive tract, exosomes contained in seminal fluid are involved in the transfer of several molecules to the maturing sperm. Exosomes are extracellular vesicles (EVs) released by all the cells; they carry a cargo of nucleic acids, proteins and lipids. In the male genital tract, they are released at various levels and their composition differs between men of proven fertility and infertile male patients. Recent studies reported the proteomic profile of exosomes, revealing the presence of several proteins with a well know role in sperm maturation and fertilizing ability acquiring. Study design, size, duration This prospective study consisted of 36 Caucasian men; according to seminal parameters (WHO 2010) they were divided in normozoospermic (N; n = 12), oligoasthenoteratozoospermic (OAT: n = 12) and azoospermic (A; n = 12). Semen samples were collected between October 2020 and January 2021 at the Assisted Reproductive Unit, Siena University Hospital (Italy) after institutional ethical approval and signed written consent from all the participants. Participants/materials, setting, methods Ejaculated sperm were analyzed according to WHO–2010 criteria and divided into the three groups: N, OAT and A. Exosomes were isolated by an in-house modified ExoGAG®-polymer precipitation-based protocol and characterized for size and ultrastructure by Nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). The exosomal proteins were extracted and analyzed by 2D-electrophoresis and the identified profiles were examined by applying bioinformatic tools. The expression of selected genes was evaluated by digital droplets PCR (ddPCR). Main results and the role of chance The present work is readily providing an improvement of the standard ExoGAG® protocol and underlines its advantages over more conventional EVs isolation protocols used to date for recovery from seminal fluid: the number of recovered EVs and their size were finely included in the range of exosomes. This isolation protocol provides samples suitable for proteomic analyses, representing the first 2D-electrophoresis reference map of exosome-pay loaded proteins in N respect to OAT/A groups and providing an innovative and comprehensive functional overview of its proteins. Moreover, the STRING protein-protein interaction analysis revealed the deregulation of specific pathways (e.g. signaling proteins, chromatin packaging and/or remodeling, protein folding and apoptosis) in A and OAT in comparison with N group. Gene expression by ddPCR analysis highlighted that most of the analyzed genes are modulated in according to seminal parameters, in particular: GAPDHS (Glyceraldehyde–3-Phosphate Dehydrogenase, Spermatogenic); SPAM1 (Sperm Adhesion Molecule–1) encoding a members of hyaluronidase family; ADAM2 (ADAM Metallopeptidase Domain–2) that plays an important role in sperm-egg interactions; CRISP1,2,3 (Cysteine Rich Secretory Protein 1,2,3) expressed in the epididymis and secreted into the epididymal lumen; CLGN (Calmegin) encoding a testis-specific chaperone protein and PGK2 (Phosphoglycerate Kinase–2) expressed in the later stages of spermatogenesis. Limitations, reasons for caution This study represents a preliminary experiment. We suggest further comparative studies in larger study cohorts. Wider implications of the findings: This pilot study, demonstrating the unique proteomic and transcriptomic pattern of exosomes in N/OAT/A groups, supports the importance of exosomes in sperm production and maturation. This methodological set-up is expected to open new ways for advancement in the use of exosomes as fertility biomarkers, making possible personalized approaches in ART. Trial registration number Not applicable

Detection of prostatic tissue and seminal plasma peptides reacting with human seminal fluid acid phosphatase antibodies

1986 ◽  
Vol 18 (11) ◽  
pp. 1005-1013 ◽  
Author(s):  
Radosława Kuciel ◽  
Izydor Apostoł ◽  
Ewa Wasylewska ◽  
Włodzimierz S. Ostrowski ◽  
Iga Steuden ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Seminal Plasma ◽  
Seminal Fluid ◽  
Prostatic Tissue

Role of Ca2+ in the IVM of spermatozoa from the sterlet Acipenser ruthenus

2017 ◽  
Vol 29 (7) ◽  
pp. 1319 ◽  
Author(s):  
Olga Bondarenko ◽  
Borys Dzyuba ◽  
Marek Rodina ◽  
Jacky Cosson
Keyword(s):  
Sperm Motility ◽  
Seminal Fluid ◽  
Mature Male ◽  
Sperm Maturation ◽  
Wolffian Duct ◽  
Testicular Spermatozoa ◽  
Acipenser Ruthenus ◽  
Sterlet Acipenser Ruthenus ◽  
Motility Parameters

The role of Ca2+ in sturgeon sperm maturation and motility was investigated. Sperm from mature male sterlets (Acipenser ruthenus) were collected from the Wolffian duct and testis 24 h after hormone induction. Testicular spermatozoa (TS) were incubated in Wolffian duct seminal fluid (WDSF) for 5 min at 20°C and were designated ‘TS after IVM’ (TSM). Sperm motility was activated in media with different ion compositions, with motility parameters analysed from standard video microscopy records. To investigate the role of calcium transport in the IVM process, IVM was performed (5 min at 20°C) in the presence of 2 mM EGTA, 100 µM Verapamil or 100 µM Tetracaine. No motility was observed in the case of TS (10 mM Tris, 25 mM NaCl, 50 mM Sucr with or without the addition of 2 mM EGTA). Both incubation of TS in WDSF and supplementation of the activation medium with Ca2+ led to sperm motility. The minimal Ca2+ concentration required for motility activation of Wolffian duct spermatozoa, TS and TSM was determined (1–2 nM for Wolffian duct spermatozoa and TSM; approximately 0.6 mM for TS). Motility was obtained after the addition of verapamil to the incubation medium during IVM, whereas the addition of EGTA completely suppressed motility, implying Ca2+ involvement in sturgeon sperm maturation. Further studies into the roles of Ca2+ transport in sturgeon sperm maturation and motility are required.

scholarly journals Zinc: A Necessary Ion for Mammalian Sperm Fertilization Competency

2018 ◽  
Vol 19 (12) ◽  
pp. 4097 ◽  
Author(s):  
Karl Kerns ◽  
Michal Zigo ◽  
Peter Sutovsky
Keyword(s):  
Male Fertility ◽  
Reproductive Tract ◽  
Zinc Supplementation ◽  
Female Reproductive Tract ◽  
Sperm Maturation ◽  
Cellular Mechanisms ◽  
Human Reproductive ◽  
Semen Processing ◽  
New Research ◽  
And Function

The importance of zinc for male fertility only emerged recently, being propelled in part by consumer interest in nutritional supplements containing ionic trace minerals. Here, we review the properties, biological roles and cellular mechanisms that are relevant to zinc function in the male reproductive system, survey available peer-reviewed data on nutritional zinc supplementation for fertility improvement in livestock animals and infertility therapy in men, and discuss the recently discovered signaling pathways involving zinc in sperm maturation and fertilization. Emphasis is on the zinc-interacting sperm proteome and its involvement in the regulation of sperm structure and function, from spermatogenesis and epididymal sperm maturation to sperm interactions with the female reproductive tract, capacitation, fertilization, and embryo development. Merits of dietary zinc supplementation and zinc inclusion into semen processing media are considered with livestock artificial insemination (AI) and human assisted reproductive therapy (ART) in mind. Collectively, the currently available data underline the importance of zinc ions for male fertility, which could be harnessed to improve human reproductive health and reproductive efficiency in agriculturally important livestock species. Further research will advance the field of sperm and fertilization biology, provide new research tools, and ultimately optimize semen processing procedures for human infertility therapy and livestock AI.

scholarly journals Alteration of transporter activities in the epididymides of infertile initial segment-specific Pten knockout mice†

2018 ◽  
Vol 99 (3) ◽  
pp. 536-545 ◽  
Author(s):  
Bingfang Xu ◽  
Stephen D Turner ◽  
Barry T Hinton
Keyword(s):  
Male Fertility ◽  
Initial Segment ◽  
Molecular Mechanisms ◽  
Fluorescein Isothiocyanate ◽  
Proximal Region ◽  
Sperm Maturation ◽  
Cellular Functions ◽  
Permeability Characteristics ◽  
Protein Levels ◽  
Tensin Homolog

Abstract A fully functional initial segment, the most proximal region of the epididymis, is important for male fertility. Our previous study generated a mouse model to investigate the importance of initial segment function in male fertility. In that model, phosphatase and tensin homolog (Pten) was conditionally removed from the initial segment epithelium, which resulted in epithelial de-differentiation. When spermatozoa progressed through the de-differentiated epithelial duct, they developed angled flagella, suggesting compromised sperm maturation, which eventually resulted in male infertility. To understand the molecular mechanisms, by which PTEN regulates epididymal sperm maturation, we compared the transcriptome profile of the initial segment between controls and initial segment-specific Pten knockouts and revealed that water, ion, and organic solute transporter activities were one of the top molecular and cellular functions altered following loss of Pten. Alteration in protein levels and localization of several transporters following loss of Pten were also observed by immunofluorescence analysis. Epithelial cells of the initial segment from knockouts were more permeable to fluorescein isothiocyanate–dextran (4000 Da) compared to controls. Interestingly, conditional deletion of Pten from other organs also resulted in changes in transporter activity, suggesting a common role of PTEN in regulation of transporter activity. Taken together, our data support the hypothesis that loss of Pten from the initial segment epithelium results in changes in the transporting and permeability characteristics of the epithelium, which in turn altered the luminal fluid microenvironment that is so important for sperm maturation and male fertility.

Seminal Plasma Transferrin Concentration: Relationship with Seminal Parameters and Plasma Hormone Levels

1992 ◽  
Vol 49 (3) ◽  
pp. 158-162 ◽  
Author(s):  
H. Fuse ◽  
S. Satomi ◽  
M. Okumura ◽  
T. Katayama
Keyword(s):  
Seminal Plasma ◽  
Hormone Levels ◽  
Seminal Parameters ◽  
Plasma Hormone

scholarly journals What impact does postgraduate clinical training have on empathy among Japanese trainee dentists?

2020 ◽  
Author(s):  
Toshiko Yoshida ◽  
Sho Watanabe ◽  
Takayuki Kono ◽  
Hiroaki Taketa ◽  
Noriko Shiotsu ◽  
...  
Keyword(s):  
Clinical Training ◽  
Interaction Analysis ◽  
Healthcare Providers ◽  
Japanese Version ◽  
University Hospital ◽  
Simulated Patients ◽  
Patient Anxiety ◽  
Significant Difference ◽  
Education Support ◽  
The Impact

Abstract Background: Enhancing empathy in healthcare education is a critical component in the development of a relationship between healthcare providers and patients that would ensure better patient care; improved patient satisfaction, adherence to treatment, patients’ medication self-efficacy, improved treatment outcomes, and reduced patient anxiety. Unfortunately, however, the decline of empathy among students has been frequently reported. It is especially common when the curriculum transitions to a clinical setting. However, some studies have questioned the significance and frequency of this decline. Thus, the purpose of this study was to determine the impact of postgraduate clinical training on dental trainees’ empathy from cognitive, behavioral, and patients’ perspective.Methods: This study included 64 trainee dentists at Okayama University Hospital and 13 simulated patients (SPs). The trainee dentists carried out initial medical interviews with SPs twice, at the beginning and the end of their clinical training. The trainees completed the Japanese version of the Jefferson Scale of Empathy for health professionals just before each interview. The SPs evaluated the trainees’ communication using an assessment questionnaire immediately after the interviews. All interviews were videotaped and analyzed using the Roter Interaction Analysis System. The hypothesis was that empathy in the trainees would have reduced at the end of their clinical training. The results were compared between the beginning and the end. The data were analyzed utilizing paired t-test and the Wilcoxon signed-ranks test.Results: No significant difference was found in the self-reported empathy of trainees at the beginning and the end of the clinical training (107.73 vs. 108.34, p=0.643). Similarly, there was no difference in the SPs’ evaluation of trainees’ communication (10.73 vs. 10.38, p=0.434). Communication behavior in the emotional responsiveness category for trainees in the beginning was significantly higher than that at the end (2.47 vs. 1.14, p=0.000).Conclusions: Overall, a one-year postgraduate dental training program neither reduced nor increased trainee dentists’ empathy levels. Providing regular education support in this area may help trainees foster their empathy.

scholarly journals Erythropoietin level in normal and abnormal human seminal fluid

2021 ◽  
Vol 21 (4) ◽  
pp. 54-59
Author(s):  
D. Yu. Sosnin ◽  
K. R. Galkovich ◽  
A. V. Krivtsov1
Keyword(s):  
Seminal Plasma ◽  
Seminal Fluid ◽  
Solid Phase ◽  
Human Sperm ◽  
Test System ◽  
Main Group ◽  
Reproductive Age ◽  
Control Group ◽  
Study Objective ◽  
Synthesis Of Hormones

Background. There are not enough publications devoted to the study of erythropoietin in human sperm. According to the results of these studies, the erythropoietin takes part in the regulation of spermatogenesis, affecting the synthesis of hormones, in particular steroid ones. Currently, the physiological and pathogenetic effects of erythropoietin on human ejaculate have not been thoroughly studied. In this regard, the study of this protein in the ejaculate in patients with diseases of the male reproductive system, as well as in their absence, is relevant.The study objective is to determine the concentration of erythropoietin in ejaculate samples of healthy and men with oligoastenozoospermia.Materials and methods. Samples of ejaculate of 52 men of reproductive age were examined. The ejaculate was examined using the SQA-V sperm analyzer (MES, Israel). According to the results of the study, two groups were identified: the main group (n = 18) with reduced fertility and the control group (n = 34) with normal spermogram indicators. In seminal plasma samples, the concentration of erythropoietin was determined by solid-phase enzyme immunoassay using the test system “Erythropoietin-IFA-BEST” (A-8776) (Vector-best LLC, Russia).Results. Erythropoietin was detected in all ejaculate samples, the results ranged from 9.37 to 193.95 mME / ml and varied 20.7 times (p = 0.3). The median concentration in the main group was 64.49 mME / ml (41.96; 118.16 mME / ml) and 1.36 times higher than the results of the comparison group, which were 47.16 mME / ml (18.15; 90.94 mME / ml). No statistically significant regularities were found between the concentration of erythropoietin and the indicators of ejaculate fertility (r <|0,3|).Conclusion. In oligoastenozoospermia, there is a tendency to increase the content of erythropoietin in the seminal plasma, which requires further research, taking into account a more detailed stratification of the groups examined for reasons that caused a decrease in the number of spermatozoa.

scholarly journals Anovel Immunological Technique for Identification of Human Seminal Fluid

2010 ◽  
Vol 7 (2) ◽  
pp. 1023-1027
Author(s):  
Baghdad Science Journal
Keyword(s):  
Seminal Plasma ◽  
Human Body ◽  
Seminal Fluid ◽  
Cross Reactivity ◽  
Body Fluids ◽  
Vaginal Fluid ◽  
Human Semen ◽  
Human Seminal Plasma ◽  
Ear Wax ◽  
Immunological Technique

An immunological technique was investigated for the detection of human semen in forensic analysis.This technique included a preparation of anti-human seminal plasma antibodies, by immunizing rabbits with treated human semen. The human semen was treated with an acid to prevent cross reactivity with other human body fluids. The antibody produced was tested against different animal,s seminal fluid samples (dog, goat ,sheep, cow) and human body fluids( saliva, blood , vaginal fluid, ear wax and human semen). It was found that using this developed technique was only selectively responsed with human semen . The prepered kit was evaluated and tested in Forensic laboratory- Ministry of Health. Finally, results were obtained in a comparison with the recommended techniques.

scholarly journals Male Fertility and Reduction in Semen Parameters: A Single Tertiary-Care Center Experience

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
D. Milardi ◽  
G. Grande ◽  
D. Sacchini ◽  
A. L. Astorri ◽  
G. Pompa ◽  
...  
Keyword(s):  
Reference Values ◽  
Male Fertility ◽  
Ethical Issues ◽  
Semen Analysis ◽  
Care Center ◽  
Tertiary Care ◽  
Healthcare Services ◽  
University Hospital ◽  
Semen Parameters ◽  
Spontaneous Conception

Background. Infertility is both a clinical and a public problem, affecting the life of the couple, the healthcare services, and social environment. Standard semen analysis is the surrogate measure of male fertility in clinical practice.Objective. To provide information about the relationship between semen parameters and spontaneous conception.Methods. We evaluated retrospectively 453 pregnancies that occurred among 2935 infertile couples evaluated at an infertility clinic of a tertiary-care university hospital, between 2004 and 2009.Results. Normal semen analysis was present only in 158 patients; 295 subfertile patients showed alterations in at least one seminal parameter. A reduction in all seminal parameters was observed in 41 patients. Etiological causes of male infertility were identified in 314 patients.Conclusion. Our data highlights the possibility of a spontaneous conception with semen parameters below WHO reference values. Therefore, we support the importance of defining reference values on a population of fertile men. Finally, we analyzed the related ethical issues.

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