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2.
Prz Menopauzalny ; 21(3): 197-199, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36254131

RESUMO

Two important studies evaluating the safety profile of oral estrogen-progestogen hormonal therapies conducted in standard clinical practice with respect to the venous system were recently published. A large prospective controlled cohort study (PRO-E2) based on the non-inferiority design has shown that the relative risk of developing venous thrombosis (VTE) in women using combined oral hormonal contraceptives (COHC) containing 17ß-estradiol (1.5 mg) and nomegestrol acetate (2.5 mg) (E2/NOMAC) was not statistically different from that in users of COHC containing ethinylestradiol and levonorgestrel (EE/LNG). The aim of the recently presented study was to compare the risk of VTE in patients treated with a product for oral continuous combined menopausal hormone therapy containing 1 mg of 17ß-estradiol and 100 mg of micronized progesterone (1 mgE2/100 mgP4) with patients taking conjugated equine estrogens and medroxyprogesterone acetate (CEE/MPA). The study was based on an analysis of records retrieved from a US health insurance database, and was therefore concerned the real-life clinical practice. The hazard ratio of VTE when comparing 1 mgE2/100 mgP4 with CEE/MPA was 0.70 (95% CI: 0.53-0.92). The difference was found to be statistically significant (p < 0.05). The rewieved studies provide further evidence that the use of hormones bioidentical with endogenous steroids in oral contraception and menopausal hormone therapy creates an opportunity to combine high efficacy with a favorable safety profile.

3.
J Endocr Soc ; 6(10): bvac128, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-36168587

RESUMO

Context: Hyperandrogenism is a central feature of polycystic ovary syndrome (PCOS). In vitro studies have demonstrated that inflammatory stimuli promote whereas ibuprofen inhibits androgen production by ovarian theca-interstitial cells. Objective: This work aimed to determine the effects of nonselective inhibitor of cyclooxygenases COX-1 and COX-2 on testosterone levels. Methods: A prospective pilot study took place in an academic hospital of women with PCOS defined according to Rotterdam criteria (N = 20). Evaluations were taken at baseline and after 3 weeks of ibuprofen administration (400 mg twice a day or 400 mg 3 times a day, respectively, in women with weight < and ≥ 70 kg). The main outcome measure was total serum testosterone. Results: Ibuprofen administration was associated with a decline of total testosterone from 0.75 ±â€…0.06 ng/mL to 0.59 ±â€…0.05 ng/mL (P = .008). There was no statistically significant change in the levels of other relevant hormones including dehydroepiandrosterone sulfate, gonadotropins, and insulin. Multiple regression analysis identified the greatest decline of testosterone was independently predicted by baseline testosterone level (P = .004) and by baseline insulin sensitivity index (P = .03). Conclusion: Nonselective inhibition of COX-1 and COX-2 leads to selective reduction of testosterone consistent with direct inhibitory effect on ovarian steroidogenesis.

4.
Sex Med ; 10(2): 100474, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35038623

RESUMO

INTRODUCTION: Postorgasmic illness syndrome (POIS) is an extremely rare urogenital disease which significantly and negatively impacts the functioning and sexual activity. AIMS: A 34-year-old male presented with POIS symptoms and confirmed the allergic component of the POIS. Intensified immunotherapy with autologous semen was recommended. METHODS: The treatment lasted 14 months and included 20 visits. Modified and intensified subcutaneous immunotherapy in both forearms significantly shortened the therapy and improved the outcome, with high-tolerance and no adverse effects or hyperactive responses. MAIN OUTCOME MEASURE: Improvement in POIS symptoms through the use of intensified immunotherapy with autologous semen. RESULTS: The improvement was significant enough to allow for higher sexual activity, and gradual resumption of private and professional activity. CONCLUSION: Intensified immunotherapy with autologous semen seems an effective and safe option for treating patients with suspected immune-allergenic POIS. To the best of our knowledge, this has been the first such intensive and effective allergen-specific immunotherapy of POIS. Wrotynska-Barczynska J, Swat. Swat E, Berger A, et al. Intensified Hyposensitization Is an Effective Treatment of Postorgasmic Illness Syndrome (POIS). Sex Med 2022;10:100474.

5.
J Pers Med ; 11(9)2021 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-34575611

RESUMO

(1) Background: there is a steady increase in the number of procedures performed via minimally invasive surgery, which have many benefits, but post-operative nausea and vomiting (PONV) and significant pain are still a common problem (2) Methods: 300 infertile women (18-40 years old) undergoing minimal invasive surgery. Interventions: laparoscopy and hysteroscopy performing, evaluation of postoperative symptoms, serotonin concentrations assessment, identify genetic polymorphisms. (3) Results: serotonin concentrations were significantly lower among women who required opioids (p = 0.006). The presence of the GG genotype in the rs6318 polymorphism of the 5HTR2C gene had a protective effect on PONV (OR = 0.503; C.I. = [0.300-0.841]; p = 0.008), when the GG variant of the rs11214763 polymorphism of the 5HTR3B gene, when the risk of PONV was 1.65-fold higher (OR = 1.652; C.I. = [1.003-2.723]; p = 0.048). Pain intensity was significantly higher among women with GG genotype of the rs6296 polymorphism of the 5HTR1B gene (OR = 1.660; C.I. = [1.052-2.622]; p = 0.029).; (4) Conclusions: the evaluation of serotonin concentration predicts requirement for opioid pain relief medication. The polymorphisms of the serotonin receptors affect the intensity of postoperative complaints.

6.
Genes (Basel) ; 12(9)2021 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-34573352

RESUMO

The function of the immune system extends from defense against external pathogens to the recognition and elimination of mutated or dying cells, aiding elimination of malignant potential and/or maintaining homeostasis. The many cell types of the immune system secrete a broad range of factors to enable cellular signaling that is vital to physiological processes. Additionally, in the ovary, follicular selection and maturation, as well as ovulation, are directly regulated by the nearby immune cells. Additionally, ovulation and rupture of the follicle have been observed to resemble a local inflammatory response. Cells of the cumulus-oocyte complex (COC) show evolving gene expression profiles throughout the oocytes' lifespan, including genes associated with immunological processes. Analysis of these genes allows the identification of useful molecular markers, as well as highlighting gene functions and interactions in these cells. Cumulus cells were obtained from hormonally stimulated patients undergoing an in vitro fertilization procedure and studied under long-term culture conditions. The microarray technique made it possible to compare the level of CCs' gene expression on the 1st, 7th, 15th and 30th day of cultivation. Additionally, RNA microarray analysis was performed to map gene expression in these cells, associated with immunological processes and associated cytokine signaling. Subsequently, the use of DAVID software allowed us to identify the "defense response to other organism", "defense response", "defense response to virus", "cytokine secretion", "cytokine production" and "cytokine-mediated signaling pathway" GO BP terms, as well as allowing further analysis of the most differentially expressed genes associated with these processes. Of the 122 genes involved, 121 were upregulated and only one was downregulated. The seven most upregulated genes related to the abovementioned terms were ANXA3, IFIT1, HLA-DPA1, MX1, KRT8, HLA-DRA and KRT18. Therefore, genes involved in immunological defense processes are upregulated in CC cultures and could serve as useful molecular markers of growth and development in the COC, as well as the proliferation of granulosa and cumulus cells.


Assuntos
Células do Cúmulo/imunologia , Citocinas/metabolismo , Imunidade/genética , Oócitos/imunologia , Ovulação/imunologia , Adulto , Proliferação de Células/genética , Células Cultivadas , Células do Cúmulo/metabolismo , Feminino , Fertilização in vitro , Perfilação da Expressão Gênica , Humanos , Oócitos/metabolismo , Ovulação/genética , Indução da Ovulação , Cultura Primária de Células , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Regulação para Cima/imunologia
7.
J Clin Med ; 10(13)2021 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-34202365

RESUMO

Polycystic ovary syndrome (PCOS) is the most prevalent endocrine and metabolic disorder, affecting 5-10% of women of reproductive age. It results from complex environmental factors, genetic predisposition, hyperinsulinemia, hormonal imbalance, neuroendocrine abnormalities, chronic inflammation, and autoimmune disorders. PCOS impacts menstrual regularities, fertility, and dermatological complications, and may induce metabolic disturbances, diabetes, and coronary heart disease. Comprehensive metabolic profiling of patients with PCOS may be a big step in understanding and treating the disease. The study aimed to search for potential differences in metabolites concentrations among women with PCOS according to different body mass index (BMI) in comparison to healthy controls. We used broad-spectrum targeted metabolomics to evaluate metabolites' serum concentrations in PCOS patients and compared them with healthy controls. The measurements were performed using high-performance liquid chromatography coupled with the triple quadrupole tandem mass spectrometry technique, which has highly selective multiple reaction monitoring modes. The main differences were found in glycerophospholipid concentrations, with no specific tendency to up-or down-regulation. Insulin resistance and elevated body weight influence acylcarnitine C2 levels more than PCOS itself. Sphingomyelin (SM) C18:1 should be more intensively observed and examined in future studies and maybe serve as one of the PCOS biomarkers. No significant correlations were observed between anthropometric and hormonal parameters and metabolome results.

8.
Cells ; 10(6)2021 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-34198768

RESUMO

The ovarian follicle is the basic functional unit of the ovary, comprising theca cells and granulosa cells (GCs). Two different types of GCs, mural GCs and cumulus cells (CCs), serve different functions during folliculogenesis. Mural GCs produce oestrogen during the follicular phase and progesterone after ovulation, while CCs surround the oocyte tightly and form the cumulus oophurus and corona radiata inner cell layer. CCs are also engaged in bi-directional metabolite exchange with the oocyte, as they form gap-junctions, which are crucial for both the oocyte's proper maturation and GC proliferation. However, the function of both GCs and CCs is dependent on proper follicular angiogenesis. Aside from participating in complex molecular interplay with the oocyte, the ovarian follicular cells exhibit stem-like properties, characteristic of mesenchymal stem cells (MSCs). Both GCs and CCs remain under the influence of various miRNAs, and some of them may contribute to polycystic ovary syndrome (PCOS) or premature ovarian insufficiency (POI) occurrence. Considering increasing female fertility problems worldwide, it is of interest to develop new strategies enhancing assisted reproductive techniques. Therefore, it is important to carefully consider GCs as ovarian stem cells in terms of the cellular features and molecular pathways involved in their development and interactions as well as outline their possible application in translational medicine.


Assuntos
Células do Cúmulo/metabolismo , Neovascularização Fisiológica , Células-Tronco/metabolismo , Feminino , Humanos , Síndrome do Ovário Policístico/metabolismo , Insuficiência Ovariana Primária/metabolismo
9.
Mol Med Rep ; 24(3)2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34296308

RESUMO

Following the publication of this paper, the authors have requested that, on p. 4412 of the above article in the Funding section of the Declarations, the acknowledgement to one of the funding sources should be removed from the paper; essentially, the reference to grant no. 2018/31/B/NZ5/02475, formulated by the Polish National Science Centre (grant providing institution), should be removed from the paper. Therefore, the revised version of the Funding section paragraph should read as follows: Funding: The present study was supported by a grant from Poznan University of Medical Sciences (grant no. 502­14­02227367­10694). The authors confirm that there are no further errors in the study, and all the authors agree to this correction. The authors are grateful to the Editor of Molecular Medicine Reports for granting them this opportunity to publish a Corrigendum, and apologize for any inconvenience caused. [the original article was published in Molecular Medicine Reports 20: 4403-4414, 2019, DOI: 10.3892/mmr.2019.10709].

10.
J Clin Endocrinol Metab ; 106(9): 2566-2573, 2021 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-34050763

RESUMO

CONTEXT: Polycystic ovary syndrome (PCOS) is the most common endocrinopathy affecting women of reproductive age. OBJECTIVE: This study was designed to evaluate effects of lifestyle modifications and synbiotic supplementation on PCOS. DESIGN: A randomized (1:1) double-blind, placebo-controlled trial. SETTING: Academic hospital. PATIENTS OR OTHER PARTICIPANTS: Overweight and obese women with PCOS were identified according to the Rotterdam criteria. Evaluations were performed at baseline and repeated after 3 months of treatment. INTERVENTION: Lifestyle modifications in combination with synbiotic supplementation or placebo. MAIN OUTCOME MEASURES: Change in body mass index (BMI) and testosterone level. RESULTS: In the placebo group, a 5% decrease in BMI was accompanied by significant decreases of the waist, hip, and thigh circumferences. The synbiotic group experienced an 8% decrease in BMI, which was significantly greater than that in the control group (P = 0.03) and was accompanied by decreases in the waist, hip, and thigh circumferences. Testosterone did not decrease significantly in the placebo group (decrease of 6%), whereas in the synbiotic group it decreased by 32% (P < 0.0001). The decrease of testosterone was significantly greater in the synbiotic group than in the placebo group (P = 0.016). CONCLUSIONS: Synbiotic supplementation potentiated effects of lifestyle modifications on weight loss and led to significant reduction of serum testosterone.


Assuntos
Estilo de Vida , Síndrome do Ovário Policístico/terapia , Simbióticos/administração & dosagem , Adulto , Índice de Massa Corporal , Suplementos Nutricionais , Método Duplo-Cego , Feminino , Humanos , Síndrome do Ovário Policístico/sangue , Testosterona/sangue
11.
J Clin Med ; 11(1)2021 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-35011815

RESUMO

In the growing ovarian follicle, the maturing oocyte is accompanied by cumulus (CCs) and granulosa (GCs) cells. Currently, there remain many unanswered questions about the epithelial origin of these cells. Global and targeted gene transcript levels were assessed on 1, 7, 15, 30 days of culture for CCs and GCs. Detailed analysis of the genes belonging to epithelial cell-associated ontological groups allowed us to assess a total of 168 genes expressed in CCs (97 genes) and GCs (71 genes) during long-term in vitro culture. Expression changes of the analyzed genes allowed the identification of the group of genes: TGFBR3, PTGS2, PRKX, AHI1, and IL11, whose expression decreased the most and the group of ANXA3, DKK1, CCND1, STC1, CAV1, and SFRP4 genes, whose expression significantly increased. These genes' expression indicates CCs and GCs epithelialization processes and their epithelial origin. Expression change analysis of genes involved in epithelization processes in GCs and CCs during their in vitro culture made it possible to describe the most significantly altered of the 11 genes. Detailed analysis of gene expression in these two cell populations at different time intervals confirms their ovarian surface epithelial origin. Furthermore, some gene expression profiles appear to have tumorigenic properties, suggesting that granulosa cells may play a role in cancerogenesis.

12.
Artigo em Inglês | MEDLINE | ID: mdl-33322590

RESUMO

BACKGROUND: Polycystic ovary syndrome (PCOS) is characterized by hyperandrogenism, anovulation, infertility, obesity, and insulin resistance, which results in increased concentrations of testosterone (T), which disturbs follicular growth and ovulation. This study aimed to assess PCOS women's clinical, endocrinological, and metabolic parameters concerning hyperandrogenism severity. RESULTS: 314 women (mean age 27.3 ± 4.6; mean body mass index (BMI) 25.7 ± 5.6) with PCOS, were divided into terciles according to T concentrations: <0.64 ng/mL (group 1), 0.64 to 0.84 ng/mL (Group 2) and >0.84 ng/mL (group 3). The mean concentration of T in all women was 0.59 ng/mL and correlated negatively with the number of menstrual cycles per year (MPY) (r = -0.36; p < 0.0001) and positively with Ferriman-Gallway score (FG) (r = 0.33; p < 0.0001), luteinizing hormone (LH) (r = 0.19; p < 0.0001) and dehydroepiandrosterone sulfate (DHEAS) (r = 0.52; p < 0.0001). Positive correlation between BMI and hirsutism (r = 0.16; p < 0.0001), total cholesterol (TC) (r = 0.18; p < 0.0001), low-density lipoprotein (LDL) (r = 0.29; p < 0.0001), and triglycerides (TG) (r = 0.40; p < 0.0001) was demonstrated. The division into subgroups confirmed the lowest MPY, highest LH, and hirsutism in group 3. BMI, insulin sensitivity indices, and lipid profile parameters were not different between the three T subgroups. CONCLUSIONS: We found no correlation between testosterone levels and insulin sensitivity or dyslipidemia in women with PCOS. Metabolic abnormalities may contribute more significantly than hyperandrogenemia to PCOS development.


Assuntos
Hormônio Luteinizante/sangue , Síndrome do Ovário Policístico/metabolismo , Adulto , Índice de Massa Corporal , Sulfato de Desidroepiandrosterona/sangue , Dislipidemias , Feminino , Hirsutismo , Humanos , Resistência à Insulina , Testosterona/sangue , Adulto Jovem
14.
Hum Reprod ; 35(10): 2303-2311, 2020 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-32869098

RESUMO

STUDY QUESTION: Is polycystic ovary syndrome (PCOS) associated with an elevation of markers of endotoxemia? SUMMARY ANSWER: In women with PCOS serum levels of lipopolysaccharides (LPS), the LPS to high-density lipoprotein (HDL) ratio and LPS-binding protein (LBP) are significantly greater than those of normal control subjects. WHAT IS KNOWN ALREADY: Mononuclear cells from women with PCOS respond excessively to LPS by releasing pro-inflammatory cytokines. In rat ovarian theca-interstitial cell cultures LPS stimulates androgen production. STUDY DESIGN, SIZE, DURATION: Cross-sectional study comparing markers of endotoxemia in women with PCOS (n = 62), healthy ovulatory women with polycystic ovary morphology (PCOM, n = 39) and a control group of healthy ovulatory women without PCOM [normal (NL), n = 43]. PARTICIPANTS/MATERIALS, SETTING, METHODS: LPS was measured using a chromogenic assay. LBP was measured by ELISA. Total cholesterol and lipids were measured using a homogeneous enzyme colorimetric method. Androgens, gonadotrophins, prolactin, insulin, high-sensitivity C-reactive protein (hs-CRP) and sex hormone-binding globulin were determined by electrochemiluminescence assays. Glucose was measured using an enzymatic reference method with hexokinase. MAIN RESULTS AND THE ROLE OF CHANCE: Women with PCOS, when compared with NL subjects, had a significantly higher mean LPS (P = 0.045), LPS/HDL ratio (P = 0.007) and LBP (P = 0.01). Women with PCOM had intermediate levels of markers of endotoxemia. Comparison among all groups revealed that markers of endotoxemia correlated positively with testosterone level, ovarian volume, number of antral follicles and hirsutism score, but negatively with the number of spontaneous menses per year. In multiple regression analysis, all measures of endotoxemia correlated independently and positively with hs-CRP and with ovarian volume. LIMITATIONS, REASONS FOR CAUTION: This cross-sectional study reveals that markers of endotoxemia are associated with several clinical features observed in women with PCOS. However, responsible mechanisms and causation remain unknown. Steroid quantification was carried out by electrochemiluminescence assays and not by the current gold standard: liquid chromatography-mass spectrometry. Hence, the relationship of endotoxemia with features of PCOS and the extent to which endotoxemia contributes to reproductive and metabolic dysfunction warrants further investigation. WIDER IMPLICATIONS OF THE FINDINGS: This study reveals the novel observation that markers of endotoxemia are elevated in young and otherwise healthy women with PCOS without significant metabolic dysfunction. Moreover, the association of clinical and endocrine markers of PCOS with those of endotoxemia may represent a pathophysiologic link to reproductive dysfunction as well as metabolic and long-term cardiovascular risks associated with this disorder. STUDY FUNDING/COMPETING INTEREST(S): Intramural funding from Poznan University of Medical Sciences. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.


Assuntos
Endotoxemia , Síndrome do Ovário Policístico , Androgênios , Estudos Transversais , Feminino , Humanos , Síndrome do Ovário Policístico/complicações
15.
J Clin Med ; 9(6)2020 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-32604796

RESUMO

Granulosa cells (GCs) have many functions and are fundamental for both folliculogenesis and oogenesis, releasing hormones and communicating directly with the oocyte. Long-term in vitro cultures of GCs show significant stem-like characteristics. In the current study, RNA of human ovarian granulosa cells was collected at 1, 7, 15 and 30 days of long-term in vitro culture. Understanding the process of differentiation of GCs towards different cell lineages, as well as the molecular pathways underlying these mechanisms, is fundamental to revealing other possible stemness markers of this type of cell. Identifying new markers of GC plasticity may help to understand the aetiology and recurrence of a wide variety of diseases and health conditions and reveal possible clinical applications of the ovarian tissue cells, affecting not only the reproductive ability but also sex hormone production. Granulosa cells were the subject of this study, as they are readily available as remnant material leftover after in vitro fertilisation procedures and exhibit significant stem-like characteristics in culture. The change in gene expression was investigated through a range of molecular and bioinformatic analyses. Expression microarrays were used, allowing the identification of groups of genes typical of specific cellular pathways. This candidate gene study focused on ontological groups associated with muscle cell morphogenesis, structure, development and differentiation, namely, "muscle cell development", "muscle cell differentiation", "muscle contraction", "muscle organ development", "muscle organ morphogenesis", "muscle structure development", "muscle system process" and "muscle tissue development". The results showed that the 10 most upregulated genes were keratin 19, oxytocin receptor, connective tissue growth factor, nexilin, myosin light chain kinase, cysteine and glycine-rich protein 3, caveolin 1, actin, activating transcription factor 3 and tropomyosin, while the 10 most downregulated consisted of epiregulin, prostaglandin-endoperoxide synthase 2, transforming growth factor, interleukin, collagen, 5-hydroxytryptmine, interleukin 4, phosphodiesterase, wingless-type MMTV integration site family and SRY-box 9. Moreover, ultrastructural observations showing heterogeneity of granulosa cell population are presented in the study. At least two morphologically different subpopulations were identified: large, light coloured and small, darker cells. The expression of genes belonging to the mentioned ontological groups suggest the potential ability of GCs to differentiate and proliferate toward muscle lineage, showing possible application in muscle regeneration and the treatment of different diseases.

16.
Cells ; 9(5)2020 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-32455542

RESUMO

In the ovarian follicle, maturation of the oocyte increases in the presence of somatic cells called cumulus cells (CCs). These cells form a direct barrier between the oocyte and external environment. Thanks to bidirectional communication, they have a direct impact on the oocyte, its quality and development potential. Understanding the genetic profile of CCs appears to be important in elucidating the physiology of oocytes. Long-term in vitro culture of CCs collected from patients undergoing controlled ovarian stimulation during in vitro fertilization procedure was conducted. Using microarray expression analysis, transcript levels were assessed on day 1, 7, 15, and 30 of culture. Apoptosis and aging of CCs strictly influence oocyte quality and subsequently the outcome of assisted reproductive technologies (ART). Thus, particular attention was paid to the analysis of genes involved in programmed cell death, aging, and apoptosis. Due to the detailed level of expression analysis of each of the 133 analyzed genes, three groups were selected: first with significantly decreased expression during the culture; second with the statistically lowest increase in expression; and third with the highest significant increase in expression. COL3A1, SFRP4, CTGF, HTR2B, VCAM1, TNFRSF11B genes, belonging to the third group, were identified as potential carriers of information on oocyte quality.


Assuntos
Técnicas de Cultura de Células/métodos , Senescência Celular/genética , Células do Cúmulo/citologia , Células do Cúmulo/metabolismo , Perfilação da Expressão Gênica , Adulto , Biomarcadores/metabolismo , Morte Celular/genética , Forma Celular/genética , Regulação da Expressão Gênica , Ontologia Genética , Redes Reguladoras de Genes , Humanos , Análise de Componente Principal , Reprodutibilidade dos Testes , Fatores de Tempo
17.
Mol Med Rep ; 21(4): 1749-1760, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32319615

RESUMO

The process of neural tissue formation is associated primarily with the course of neurogenesis during embryonic life. The source of neural­like cells is stem cells, which, under the influence of appropriate differentiating factors, may differentiate/transdifferentiate towards a neural­like lineage. The present study suggested that, under long­term in vitro culture conditions, human ovarian granulosa cells (GCs), obtained from granulosa­rich follicular fluid, acquired new properties and expressed genes characteristic of the ontological groups 'neurogenesis' (GO:0022008), 'neuronal precursor cell proliferation' (GO:0061351) and 'nervous system development' (GO:0007399), which are closely related to the formation of neurons. The present study collected GCs from 20 women referred for the procedure of in vitro fertilization. Cells were maintained in long­term in vitro culture for 30 days, and RNA was isolated after 1, 7, 15 and 30 days of culture. The expression profile of individual genes was determined using the Affymetrix microarray method. The 131 genes with the highest expression change in relation to day 1 of culture were then selected; the 10 most affected genes found to be primarily involved in nerve cell formation processes were chosen for consideration in this study: CLDN11, OXTR, DFNA5, ATP8B1, ITGA3, CD9, FRY, NANOS1, CRIM1 and NTN4. The results of the present study revealed that these genes may be considered potential markers of the uninduced differentiation potential of GCs. In addition, it was suggested that GCs may be used to develop a cell line showing neuronal characteristics after 30 days of cultivation. In addition, due to their potential, these cells could possibly be used in the treatment of neurodegenerative diseases, not only in the form of 'cultured neurons' but also as producers of factors involved in the regeneration of the nervous system.


Assuntos
Transdiferenciação Celular/genética , Regulação da Expressão Gênica no Desenvolvimento , Células da Granulosa/citologia , Células da Granulosa/metabolismo , Células-Tronco Neurais/citologia , Neurogênese/genética , Neurônios/citologia , Adolescente , Adulto , Proliferação de Células/genética , Forma Celular/genética , Feminino , Perfilação da Expressão Gênica , Ontologia Genética , Humanos , Células-Tronco Neurais/metabolismo , Neurônios/metabolismo , Adulto Jovem
18.
Histochem Cell Biol ; 154(1): 77-95, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32189110

RESUMO

Genes influencing oocyte maturation may be valuable for predicting their developmental potential, as well as discerning the mechanistic pathways regulating oocyte development. In the presented research microarray gene expression analysis of immature and in vitro matured porcine oocytes was performed. Two groups of oocytes were compared in the study: before (3 × n = 50) and after in vitro maturation (3 × n = 50). The selection of viable oocytes was performed using the brilliant cresyl blue (BCB) test. Furthermore, microarrays and RT-qPCR was used to analyze the transcriptome of the oocytes before and after IVM. The study focused on the genes undergoing differential expression in two gene-ontology groups: "Cellular response to hormone stimulus" and "Cellular response to unfolded protein", which contain genes that may directly or indirectly be involved in signal transduction during oocyte maturation. Examination of all the genes of interest showed a lower level of their expression after IVM. From the total number of genes in these gene ontologies ten of the highest change in expression were identified: FOS, ID2, BTG2, CYR61, ESR1, AR, TACR3, CCND2, EGR2 and TGFBR3. The successful maturation of the oocytes was additionally confirmed with the use of lipid droplet assay. The genes were briefly described and related to the literature sources, to investigate their potential roles in the process of oocyte maturation. The results of the study may serve as a basic molecular reference for further research aimed at improving the methods of oocyte in vitro maturation, which plays an important role in the procedures of assisted reproduction.


Assuntos
Hormônios/metabolismo , Técnicas de Maturação in Vitro de Oócitos , Lipídeos/análise , Oócitos/metabolismo , Animais , Células Cultivadas , Amarelo de Eosina-(YS)/química , Feminino , Hematoxilina/química , Hormônios/genética , Oócitos/crescimento & desenvolvimento , Oxazinas/química , Transdução de Sinais , Suínos
19.
Mol Med Rep ; 21(3): 1537-1551, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32016446

RESUMO

Oocyte maturation is essential for proper fertilization, embryo implantation and early development. While the physiological conditions of these processes are relatively well­known, its exact molecular mechanisms remain widely undiscovered. Oocyte growth, differentiation and maturation are therefore the subject of scientific debate. Precious literature has indicated that the oocyte itself serves a regulatory role in the mechanisms underlying these processes. Hence, the present study performed expression microarrays to analyze the complete transcriptome of porcine oocytes during their in vitro maturation (IVM). Pig material was used for experimentation, as it possesses similarities to the reproductive processes and general genetic proximities of Sus scrofa to human. Oocytes, isolated from the ovaries of slaughtered animals were assessed via the Brilliant Cresyl Blue test and directed to IVM. A number of oocytes were left to be analyzed as the 'before IVM' group. Oocyte mRNA was isolated and used for microarray analysis, which was subsequently validated via RT­qPCR. The current study particularly focused on genes belonging to 'positive regulation of transcription, DNA­dependent', 'positive regulation of gene expression', 'positive regulation of macromolecule metabolic process' and 'positive regulation of transcription from RNA polymerase II promoter' ontologies. FOS, VEGFA, ESR1, AR, CCND2, EGR2, ENDRA, GJA1, INHBA, IHH, INSR, APP, WWTR1, SMARCA1, NFAT5, SMAD4, MAP3K1, EGR1, RORA, ECE1, NR5A1, KIT, IKZF2, MEF2C, SH3D19, MITF and PSMB4 were all determined to be significantly altered (fold change, >|2|; P<0.05) among these groups, with their downregulation being observed after IVM. Genes with the most altered expressions were analyzed and considered to be potential markers of maturation associated with transcription regulation and macromolecule metabolism process.


Assuntos
Diferenciação Celular/genética , Metabolismo Energético , Regulação da Expressão Gênica no Desenvolvimento , Oócitos/citologia , Oócitos/metabolismo , Oogênese/genética , Animais , Biomarcadores , Células Cultivadas , Biologia Computacional/métodos , Feminino , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Imuno-Histoquímica , Metabolômica , Ovário/metabolismo , Suínos , Transcrição Gênica , Transcriptoma
20.
Reprod Sci ; 27(1): 432-442, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-32046411

RESUMO

Women with polycystic ovary syndrome (PCOS) are at increased risk of psychological distress including anxiety and depressive symptoms. However, less is known about sexual satisfaction and self-esteem as well as the relationship of these aspects of psychological function with clinical and hormonal profiles associated with PCOS. This cross-sectional study compared women with PCOS (N = 96) and healthy controls (N = 47). This study assessed sexual function (primary outcome), self-esteem, anxiety, and depression as well as evaluation of clinical, endocrine, and metabolic parameters. Overall, sexual satisfaction scores were comparable among women with and without PCOS. However, psychosexual function of women with PCOS exhibited distinguishing characteristics. The unconscious aspect of sexuality: frequency of erotic dreams, significantly correlated with free testosterone (ρ = 0.24, P = 0.03) and DHEAS (ρ = 0.31, P = 0.004) only in the PCOS group. In contrast, in women with PCOS, the frequency of masturbation did not correlate with endocrine profiles, but correlated with trait anxiety (ρ = 0.21, P = 0.049) and depression (ρ = 0.21, P = 0.05). Only one aspect of self-esteem (body appearance) was reduced in the PCOS group (P = 0.02) and was related to BMI and androgen. Women with PCOS had greater state anxiety (P = 0.02) and depression (P < 0.001); these scores correlated with BMI. However, anxiety and depression correlated with testosterone only in women without PCOS. The above findings indicate that PCOS is associated with a broad range of alterations of psychological function including psychosexual aspects; these alterations are in complex relationship with BMI and androgen levels.


Assuntos
Ansiedade/psicologia , Depressão/psicologia , Satisfação Pessoal , Síndrome do Ovário Policístico/psicologia , Autoimagem , Comportamento Sexual/psicologia , Adulto , Estudos Transversais , Sulfato de Desidroepiandrosterona/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Humanos , Lipídeos/sangue , Hormônio Luteinizante/sangue , Síndrome do Ovário Policístico/sangue , Testosterona/sangue
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