High-grade serous carcinoma of the ovary, fallopian tube and peritoneum (HGSC), the most common type of ovarian cancer, ranks among the deadliest malignancies. Many HGSC patients have excess fluid in the peritoneum called ascites. Ascites is a tumour microenvironment (TME) containing various cells, proteins and extracellular vesicles (EVs). We isolated EVs from patients' ascites by orthogonal methods and analyzed them by mass spectrometry. We identified not only a set of 'core ascitic EV-associated proteins' but also defined their subset unique to HGSC ascites. Using single-cell RNA sequencing data, we mapped the origin of HGSC-specific EVs to different types of cells present in ascites. Surprisingly, EVs did not come predominantly from tumour cells but from non-malignant cell types such as macrophages and fibroblasts. Flow cytometry of ascitic cells in combination with analysis of EV protein composition in matched samples showed that analysis of cell type-specific EV markers in HGSC has more substantial prognostic potential than analysis of ascitic cells. To conclude, we provide evidence that proteomic analysis of EVs can define the cellular composition of HGSC TME. This finding opens numerous avenues both for a better understanding of EV's role in tumour promotion/prevention and for improved HGSC diagnostics.
- MeSH
- ascites metabolismus patologie MeSH
- extracelulární vezikuly * metabolismus MeSH
- lidé MeSH
- nádorové mikroprostředí MeSH
- nádory vaječníků * diagnóza MeSH
- proteomika MeSH
- serózní cystadenokarcinom * diagnóza genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Exposure to bisphenols has been found to have adverse effects on male reproductive function in animals. Human exposure to bisphenols is widespread. Bisphenol A (BPA) and its analogues, including bisphenol S (BPS), bisphenol F (BPF), and bisphenol AF (BPAF) are utilized in various consumer products such as food contact materials and dental resins. The effects of these compounds on male fertility and spermatogenesis are unclear and findings from human studies are inconsistent. In this cross-sectional study, we evaluated the influence of BPA, BPS, BPF, BPAF (BPs) measured in semen on number of spermatozoa, total motility, progressive motility, morphology, and DNA fragmentation. We also examined the association of bisphenols (BPs) exposure with patients' occupation. A total of 358 patients aged 17-62 years with BMI 18-42 were included in the study from 2019 to 2021. BPs were extracted using solvent extraction followed by preconcentration step and determined by high-performance liquid chromatography and tandem mass spectrometry (LC/MSMS). Bisphenols were detected in 343 from 349 analysed samples (98.3% of all the samples). In 6 samples, the concentration of all BPs was under the limit of detection and in 20 samples under the limit of quantification. We did not find a statistically significant relationship between occupation and BPs. However, we observed significant correlations between the concentration of BPA and a lower motility and normal morphology. For BPS, a significant correlation with a lower ejaculate volume and a lower total sperm count was found. BPF and BPAF were detected only in 14.3% and 23.9% of samples, respectively. For BPF and BPAF, no significant correlations with spermiogram parameters were observed. Our results show that BPs are widespread in the male population (more than 90% of analysed samples), independently of an occupation and in case of BPA and BPS having a negative impact on spermiogram parameters.
- MeSH
- benzhydrylové sloučeniny * toxicita analýza MeSH
- fenoly * MeSH
- fluorokarbony * MeSH
- lidé MeSH
- průřezové studie MeSH
- sperma * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
BACKGROUND: Human embryonic stem cells (hESCs) have the unique ability to differentiate into any cell type in the human body and to proliferate indefinitely. Cell therapies involving hESC have shown very promising results for the treatment of certain diseases and confirmed the safety of hESC-derived cells for humans. They are used in cell therapy, mainly in targeted therapy of diseases that are currently incurable. OBJECTIVES: The aim of this study was the derivation of clinical-grade hESCs usable in drug development, non-native medicine and cell therapy. MATERIAL AND METHODS: Embryos were thawed, cultivated to the blastocyst stage if necessary, and assisted hatching was subsequently performed. Embryoblasts were mechanically isolated using narrow needles. Each line was kept as a separate batch. The derived hESCs were cultured under hypoxic culture conditions (5% O2, 5% CO2, 37°C) in a NutriStem® hPSC XF Medium with a daily medium change. RESULTS: From January 2018 to July 2020, 138 selected clients were asked for consent to donate embryos, of whom 52 did not respond, 19 terminated the storage of their embryos and 29 extended the storage. Only 38 clients (27.5%) agreed to donate embryos for the derivation of hESCs. At the same time, personal communication with clients took place and another 17 embryo donors were recruited. A total of 160 embryos from 55 donors aged 26-42 years were collected. The embryos were frozen at the blastocyst (33.1%) or morula (46.3%) stage. After the preparation of 64 embryos, embryoblasts were isolated and cultured. Finally, 7 hESC lines were obtained, 4 research-grade and 3 clinical-grade, the first in the Czech Republic. CONCLUSIONS: We established a current good manufacturing practice (cGMP)-defined xeno-free and feeder-free system for the derivation, culture and banking of clinical-grade hESC lines that are suitable for preclinical and clinical trials. The quality control testing with criteria concerning sterility, safety and characterization according to cGMP ensured the clinical-grade quality of hESC lines.
- MeSH
- buněčné linie MeSH
- embryo savčí MeSH
- embryonální kmenové buňky * MeSH
- kvalita života * MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Tento článek, založený na rešerších literatury publikované do června 2023 v databázích Web of Science (WoS), Scopus, PubMed/Medline, se zaměřením na „breast cancer“ a „fertility preservation“, rozebírá nejdůležitější aspekty a kontroverze péče o reprodukční zdraví žen s karcinomem prsu. Protinádorová léčba účinně potlačuje maligní bujení, nicméně i přes pokrok v medicíně může dojít k poškození gonád. Znalost těchto rizik a funkční spolupráce mezi onkology a lékaři centra ochrany reprodukce jsou pro pacientku zásadní. Moderní postupy ochrany reprodukce se z pohledu reprodukční medicíny jeví jako účinné a bezpečné, i když mezi onkology a gynekology není plný konsenzus.
This article, based on a review of the literature until Jun 2023 in Web of Science (WoS), Scopus and PubMed/Medline databases, focused on “breast cancer” and “fertility preservation”, summarizes aspects and reproductive health care controversies during oncological treatment in female breast cancer patients. Current oncological treatment destroys malignant growth effectively, despite advances in medicine ovarian failure can occur. Knowledges about these risks and functional collaboration between oncologists and gynecologists are essential. Modern fertility preservation procedures are effective and safe from a gynecologist's perspective, although there is not a full consensus among oncologists and gynecologists.
The compounds of seminal plasma have great potential as biomarkers of male fertility and can be used as a diagnostic tool for types of azoospermia. Azoospermia occurs in approximately 1% of the male population, and for an effective therapy of this form of male infertility, it is important to distinguish between obstructive and non-obstructive azoospermia. Proteins in seminal plasma can serve as biomarkers for diagnosing azoospermia. Considering the various types of obstructions, a combination of multiple proteins is advisable for diagnostic purposes. In this context, testicular and epididymal proteins are particularly significant, as they are specific to these tissues and typically absent in ejaculate during most obstructions. A combination of multiple biomarkers is more effective than the analysis of a single protein. This group of markers contains TEX101 and ECM1 proteins, combined detections of these two bring a diagnostic output with a high sensitivity and specificity. Similar results were observed for combined detection of TEX101 and SPAG1. The effective using of specific biomarkers from seminal plasma can significantly improve the existing approaches to diagnosis of the causes of male infertility.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Cíl: Shrnutí nových poznatků v oblasti implantace embrya v závislosti na kvalitě endometria. Metodika: Rešerše literatury publikované do srpna 2022 v databázích WoS, Scopus, PubMed/Medline se zaměřením na „endometrial receptivity“, „polycystic ovary syndrome“, „endometriosis“, „SARS-CoV-2“. Výsledky: Receptivní stav endometria je výsledkem fyziologické remodelace, činnosti imunity modulované mikrobiomem. Tuto rovnováhu narušují myomy, polypy, saktosalpingy, adenomyóza, endometrióza, syndrom polycystických vaječníků, infekce. Diskutuje se dopad infekce SARS-CoV-2. Pro úspěšnou implantaci je klíčové časování embryotransferu. Konvenčně se pro tyto účely využívá ultrazvuk. Ve specifických případech hysteroskopie a biopsie endometria. Vzorek se hodnotí histologicky, imunohistochemicky, vyšetřuje se mikrobiom anebo transkriptom. Jako podpora implantace jsou využívány gestageny, u syndromu polycystických ovarií metformin. U opakovaného selhání implantace intrauterinní infuze mononukleárů, plazmy bohaté na destičky, subkutánní aplikace granulocyty stimulujícího růstového faktoru, intravenózní podání atosibanu, intrauterinní aplikace choriogonadotropinu. Závěr: Nejnovější výzkumy na poli transkriptomiky, proteomiky a reprodukční imunologie hlouběji odkrývají implantaci a otvírají novou etapu asistované reprodukce.
Objective: A summary of new knowledge on embryo implantation in dependence on quality of the endometrium. Methods: Literature review from August 2022 of the relevant publications in Web of Science, Scopus and PubMed/Medline databases, focused on “endometrial receptivity”, “polycystic ovary syndrome”, “endometriosis”, “SARS-CoV-2”. Results: The receptive state of the endometrium is a result of physiological remodeling and immune system activity modulated by the microbiome. This balance can be disturbed by myomas, polyps, sactosalpings, adenomyosis, endometriosis, polycystic ovary syndrome, infections. The effect of SARS-CoV-2 infection is being discussed. For a successful implantation, timing of transfer is crucial. The ultrasound examination is used conventionally. In specific cases, hysteroscopy and endometrium biopsy are recommended. Histological and immunohistochemical evaluation is performed together with examination of microbiome or transcriptome. To support the implantation, gestagenes are used, or metformin in the patients with polycystic ovary syndrome. In cases of a repeated implantation failure, the intrauterine infusion of mononuclear cells or platelet rich plasma is used, subcutaneous application of granulocyte colony stimulating growth factor, intravenous application of atosiban or intrauterine application of human chorionic gonadotropin. Conclusion: Recent research in the field of transcriptomics, proteomics and reproductive immunology uncovers the process of implantation more deeply and opens a new stage of the assisted reproduction.
- MeSH
- choriogonadotropin MeSH
- COVID-19 metabolismus MeSH
- endometrióza MeSH
- implantace embrya * fyziologie MeSH
- lidé MeSH
- syndrom polycystických ovarií MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
Exposure to bisphenols is related to negative effects on male reproduction. The bisphenols exposure is associated with several modes of action including negative impact on the blood-testis barrier (BTB) in testes or direct effect on spermatozoa. Bisphenols have been detected in human seminal plasma, but the possible mechanism of seminal transfer of bisphenols is not clear. Some authors consider the transfer through the blood-testis barrier to be crucial. Therefore, in this work, we compared normozoospermic men and men after vasectomy who have interrupted vas deferens and their ejaculate does not contain testicular products. We measured the concentration of bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) in the urine and seminal plasma of these men using liquid chromatography tandem mass spectrometry (LC/MSMS). We found that the ratio of urinary and seminal plasma content of bisphenols did not differ in normozoospermic men or men after vasectomy. From the obtained data, it can be concluded that the pathways of transport of bisphenols into seminal plasma are not primarily through the testicular tissue, but this pathway is applied similarly to other routes of transmission by a corresponding ejaculate volume ratio. To a much greater extent than through testicular tissue, bisphenols enter the seminal plasma mainly as part of the secretions of the accessory glands.
- Publikační typ
- časopisecké články MeSH
Male fertility has been deteriorating worldwide for considerable time, with the greatest deterioration recorded mainly in the United States, Europe countries, and Australia. That is, especially in countries where an abundance of chemicals called endocrine disruptors has repeatedly been reported, both in the environment and in human matrices. Human exposure to persistent and non-persistent chemicals is ubiquitous and associated with endocrine-disrupting effects. This group of endocrine disrupting chemicals (EDC) can act as agonists or antagonists of hormone receptors and can thus significantly affect a number of physiological processes. It can even negatively affect human reproduction with an impact on the development of gonads and gametogenesis, fertilization, and the subsequent development of embryos. The negative effects of endocrine disruptors on sperm gametogenesis and male fertility in general have been investigated and repeatedly demonstrated in experimental and epidemiological studies. Male reproduction is affected by endocrine disruptors via their effect on testicular development, impact on estrogen and androgen receptors, potential epigenetic effect, production of reactive oxygen species or direct effect on spermatozoa and other cells of testicular tissue. Emerging scientific evidence suggests that the increasing incidence of male infertility is associated with the exposure to persistent and non-persistent endocrine-disrupting chemicals such as bisphenols and perfluoroalkyl chemicals (PFAS). These chemicals may impact men's fertility through various mechanisms. This study provides an overview of the mechanisms of action common to persistent (PFAS) and nonpersistent (bisphenols) EDC on male fertility.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Cíl práce: Kryokonzervace spermatu před gonadotoxickou léčbou je základní a nejúčinnější metodou zachování reprodukce, která může být využita i v období adolescence. Sdělení shrnuje 26 let zkušeností provozu onkologické spermabanky, uvádí analýzy spermiogramů onkologických pacientů, posouzení vztahu patologie spermatu s diagnózou, stanovení počtu zemřelých a využití zamrazeného spermatu. Metodika: Za dobu existence CAR 01 (centrum asistované reprodukce) bylo provedeno více než 50 000 spermiogramů. V rámci spermabanky od ledna 1995 do prosince 2020 bylo vyšetřeno 24 729 mužů, z toho 1 448 (5,9 %) mělo onkologickou diagnózu. Spermiogramy byly vyhodnoceny podle aktuálních manuálů Světové zdravotnické organizace (WHO – World Health Organization). Kryokonzervace spermatu prošla zásadním vývojem. Pravidla pro skladování zamražených buněk jsou od roku 2008 stanoveny zákonem č. 296/2008 Sb. V roce 2019 byla aktualizována metodika "Kryokonzervace reprodukčních buněk a tkání u pacientů před onkologickou léčbou". Ve všech případech byla použita standardní technika rozmrazení. Spermie byly zpracovány metodou swim-up. V rámci léčby metodami asistované reprodukce byly oocyty oplozovány mikromanipulační technikou ICSI (intracytoplazmatická injekce spermie). Výsledky: Z 1 448 vyšetřených spermiogramů u mužů s onkologickou diagnózou byl zhoubný nádor varlete u 43,7 % pacientů, maligní onemocnění mízní a krvetvorné tkáně v 24,1 %, z toho Hodgkinův lymfom 70,1 %, non-Hodgkinův lymfom 29,9 %. Leukemie byla zjištěna u 7,9 %, zhoubné nádory kostí a chrupavek u 6,8 % pacientů. Věk klientů celého souboru byl od 13 do 64 let (27,2 ± 6,8 let). Normozoospermii mělo 38,3 % mužů, 54,2 % spermiogramů vykázalo patologický nález v 1–3 hodnocených parametrech a azoospermii mělo 7,5 % pacientů. Těžká astenozoospermie (pohyblivost ≤ 10 %) byla prokázána u 57,2 % mužů a těžká oligozoospermie (koncentrace ≤ 1 × 106 mm3) u 22,3 % pacientů. Nejnižší hodnoty spermiogramu byly zjištěny u mužů s tumorem varlete, nejlepší hodnoty byly u nádorových onemocnění centrální nervové soustavy (CNS). Kryokonzervace spermatu byla provedena v 1 340 případech (92,5 %). Zamrazené sperma dosud celkem využilo 160 mužů (11,9 %), z toho 83 u nás (6,2 %). V těchto 83 případech byla vždy použita technika ICSI, bylo dosaženo 38 klinických těhotenství (45,8 %) a 32 porodů. Máme evidováno 424 ukončených skladování spermatu (31,6 %), z toho 148 z důvodu úmrtí, tzn. u 11,0 % všech onkologických pacientů, ostatní na vlastní žádost. Specifickou otázkou je využití spermatu zemřelých. Závěr: U onkologických pacientů jsou ve vysokém procentu těžké patologie spermatu. Nejnižší hodnoty spermiogramu byly prokázány u mužů se zhoubným nádorem varlete. Je nutné počítat s dlouhodobým skladováním a oplozením mikromanipulačními metodami. Počet mužů, kteří zemřou, je výrazně vyšší než těch, kteří sperma využijí pro léčbu neplodnosti. Kryokonzervace spermatu by měla být nabídnuta každému pacientovi před terapií vedoucí k destrukci spermatogeneze.
Objective: Sperm cryopreservation before gonadotoxic treatment is the basic and most effective method of preserving reproduction, which can be used during adolescence. The communication summarizes 26 years of experience in the operation of an oncological sperm bank, analyzes spermiograms of oncological patients, assesses the relationship between sperm pathology and diagnosis, and determines the number of deaths and the use of frozen sperm. Methods: During the existence of CAR 01 (assisted reproduction center), more than 50,000 spermiograms were performed. From January 1995 to December 2020, a total of 24,729 men were examined within the sperm bank, of which 1,448 (5.9%) had an oncological diagnosis. The spermiograms were evaluated according to current WHO (World Health Organization) manuals. Cryopreservation of sperm has undergone a major development. The rules for the storage of frozen cells have been laid down by Act No. 296/2008 Coll. since 2008. In 2019, the methodology "Cryopreservation of reproductive cells and tissues in patients before cancer treatment" was updated. In all cases, the standard thawing technique was used. The sperms were processed by the swim-up method. As part of the treatment with assisted reproduction methods, oocytes were fertilized by the ICSI (intracytoplasmatic sperm injection) micromanipulation technique. Results: Out of 1,448 examined spermiograms in men with oncological diagnoses, testicular cancer was present in 43.7% of patients and malignant diseases of lymphatic and hematopoietic tissue were found in 24.1%, of which 70,1% included Hodgkin‘s lymphomas and 29,9% were non-Hodgkin‘s lymphomas. Leukemia was found in 7.9%, bone and cartilage cancers in 6.8%. The age of the clients of the whole group ranged from 13 to 64 years (27.2 ± 6.8 years). A total of 38.3% of men had normozoospermia, 54.2% of spermiograms showed pathological findings in 1 to 3 evaluated parameters and 7.5% of patients had azoospermia. Severe asthenozoospermia (mobility ≤ 10%) was detected in 57.2% of men and severe oligozoospermia (concentration ≤ 1 × 106 mm3) in 22.3% of patients. The lowest values of the spermiogram were found in men with testicular cancer; the best values were seen in CNS (central nervous system) cancers. The cryopreservation of sperm was performed in 1,340 cases (92.5%). So far, a total of 160 men (11.9%) have used frozen sperm, of which 6.2% in our center. In these 83 cases, the ICSI technique was always used, 38 clinical pregnancies (45.8%) and 32 births were achieved. We have registered 424 completed storages of semen (31.6%), of which 148 (11.0% of all oncology patients) were made due to death and the others at patients’ request. Using the sperm of the dead is a specific issue. Conclusion: In cancer patients, sperm pathologies occur in high percentage. The lowest spermiogram values were found in men with testicular cancer. It is necessary to take into account long-term storage and fertilization by micromanipulation methods. The number of men who die is significantly higher than the number of those who use sperm to treat infertility. Cryopreservation of sperm should be offered to each patient prior to the therapy leading to the destruction of spermatogenesis. Publication ethics: The Editorial Board declares that the manuscript met the ICMJE “uniform requirements” for bio medical papers. Conflict of interests: The authors declare they have no potential conflicts of interest concerning the drugs, products or services used in the study. Dedication: The work was supported by the Ministry of Health of the Czech Republic – RVO (FNBr: 65269705), IGA No. NR/ 8469-3, AZV MZ ČR No. NV18-01-00544 and NV-18-08-00291.
