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Prions are infectious protein particles known to cause prion diseases. The biochemical entity of the pathogen is the misfolded prion protein (PrPSc) that forms insoluble amyloids to impair brain function. PrPSc interacts with the non-pathogenic, cellular prion protein (PrPC) and facilitates conversion into a nascent misfolded isoform. Several small molecules have been reported to inhibit the aggregation of PrPSc but no pharmacological intervention was well established thus far. We, here, report that acylthiosemicarbazides inhibit the prion aggregation. Compounds 7x and 7y showed almost perfect inhibition (EC50 = 5 µM) in prion aggregation formation assay. The activity was further confirmed by atomic force microscopy, semi-denaturing detergent agarose gel electrophoresis and real-time quaking induced conversion assay (EC50 = 0.9 and 2.8 µM, respectively). These compounds also disaggregated pre-existing aggregates in vitro and one of them decreased the level of PrPSc in cultured cells with permanent prion infection, suggesting their potential as a treatment platform. In conclusion, hydroxy-2-naphthoylthiosemicarbazides can be an excellent scaffold for the discovery of anti-prion therapeutics.
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Doenças Priônicas , Príons , Humanos , Príons/metabolismo , Proteínas Priônicas/metabolismo , Encéfalo , Doenças Priônicas/tratamento farmacológico , Doenças Priônicas/metabolismo , Doenças Priônicas/patologia , Células CultivadasRESUMO
Sirtuin 1 (SIRT1) is a member of the sirtuin family that functions to deacetylate both histones and non-histone proteins. Previous studies have identified significant SIRT1 upregulation in eutopic endometrium from infertile women with endometriosis. However, SIRT1 function in the uterus has not been directly studied. Using immunochemistry analysis, we found SIRT1 to be most strongly expressed at GD4.5 and GD5.5 in decidualized cells and at GD7.5 in secondary decidual cells in mouse. To assess the role of SIRT1 in uterine function, we generated uterine Sirt1 conditional knockout mice (Pgrcre/+Sirt1f/f; Sirt1d/d). A 6-month fertility trial revealed that Sirt1d/d females were subfertile. Implantation site numbers were significantly decreased in Sirt1d/d mice compared with controls at GD5.5. Sirt1d/d implantation sites at GD4.5 could be divided into two groups, Group #1 with luminal closure and nonspecific COX2 expression compared with controls (14/20) and Group #2 with an open lumen and no COX2 (6/20). In Sirt1d/d Group #1, nuclear FOXO1 expression in luminal epithelial cells was significantly decreased. In Sirt1d/d Group #2, nuclear FOXO1 expression was almost completely absent, and there was strong PGR expression in epithelial cells. At GD5.5, stromal PGR and COX2 were significantly decreased in Sirt1d/d uterine in the areas surrounding the embryo compared with controls, indicating defective decidualization. An artificially induced decidualization test revealed that Sirt1d/d females showed defects in decidualization response. All together, these data suggest that SIRT1 is important for decidualization and contributes to preparing a receptive endometrium for successful implantation.
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Infertilidade Feminina , Sirtuína 1 , Animais , Ciclo-Oxigenase 2/metabolismo , Decídua/metabolismo , Implantação do Embrião/fisiologia , Endométrio/metabolismo , Feminino , Humanos , Infertilidade Feminina/genética , Infertilidade Feminina/metabolismo , Camundongos , Camundongos Knockout , Gravidez , Sirtuína 1/genética , Sirtuína 1/metabolismo , Células Estromais/metabolismo , Útero/metabolismoRESUMO
Uterine endometrial differentiation is essential for developmental continuity and female health. A convenient in vitro model mimicking the physiological status is needed to effectively evaluate implantation and uterine response mechanisms. Thus, we developed a promising in vitro model, the FSS (FSH mimic-stimulated synchronized) model, by using primary mouse uterine stromal cells (mUSCs) obtained from equine chorionic gonadotropin (eCG)-primed mice. These mUSCs could be differentiated into decidualized cells with 17 beta-estradiol (E2) and progesterone (P4). The pregnancy day 4 (PD4) model, in which mUSCs are obtained at day 4 of pregnancy, was used as a control. The cell shape index and polyploidy rates were similar between the two models. The staining intensities of lipids and glycogen were significantly higher in the induced groups in both models but stronger in the FSS model than in the PD4 model. The expression levels of AP-TNAP, cathepsin L, Prl8a2, Gja1, Cebpb, and Igfbp1 were increased at 24 h after decidual induction. PR-alpha and PR-beta levels were also increased at 24 h after decidual induction in both models. These results indicate that the FSS model provides a convenient method for obtaining USCs that are usable for various experimental approaches due to their physiological competence and flexibility for triggering induction. This may serve as a model system for the study of pathogeneses originating from the endometrium or communication with other tissues and lead to a better understanding of embryo implantation mechanisms. Furthermore, the results of this study will be integral for further refinements of 3D uterine culture manipulation techniques.
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Implantação do Embrião , Células Estromais , Gravidez , Feminino , Animais , Cavalos , Camundongos , Células Estromais/metabolismo , Implantação do Embrião/fisiologia , Endométrio , Progesterona/farmacologia , Útero , Gonadotropinas Equinas/farmacologia , Decídua/metabolismoRESUMO
Although a few aquaporins (AQPs) expressed in granulosa cells have been postulated to mediate fluid passage into the antrum, the specific expression of AQPs in different follicle cell types and stages and their roles have not been evaluated extensively. The spatiotemporal expression of aquaporin (Aqp) 7, 8, and 9 and the functional roles of Aqp9 in antral growth and ovulation were examined using a superovulation model and 3-dimensional follicle culture. Aqp9 was expressed at a high level in the rapid growth phase (24-48 h post equine chorionic gonadotropin (eCG) for superovulation induction) compared to Aqp7 (after human chorionic gonadotropin (hCG)) and Aqp8 (8-24 h post eCG and 24 h post hCG). A dramatic increase in the expression and localization of Aqp9 mRNA in theca cells was observed, as evaluated using quantitative reverse transcription-polymerase (RT-PCR) coupled with laser capture microdissection and immunohistochemistry. AQP9 was located primarily on the theca cells of the tertiary and preovulatory follicles but not on the ovulated follicles. In phloretin-treated mice, the diameter of the preovulatory follicles and the number of ovulated oocytes decreased. Consistent with these findings, knocking down Aqp9 expression with an Aqp9 siRNA inhibited follicle growth (0.28:1 = siRNA:control) and decreased the number of ovulated follicles (0.36:1 = siRNA:control) during in vitro growth and ovulation induction. Based on these results, the expression of AQPs is under the control of the physiological status, and AQP9 expression in theca during folliculogenesis is required for antral growth and ovulation in a tissue-specific and stage-dependent manner.
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Aquaporinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Folículo Ovariano/metabolismo , Animais , Aquaporinas/genética , Gonadotropina Coriônica/farmacologia , Feminino , Regulação da Expressão Gênica/fisiologia , Camundongos , Transporte Proteico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Superovulação , Técnicas de Cultura de TecidosRESUMO
PURPOSE: Recent studies have shown that improved clinical outcomes can be achieved by transferring blastocysts rather than cleavage-stage embryos. However, blastocyst transfer is not performed in all patients. The aim of this study was to compare clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles using testicular sperm (TE) with those of ICSI cycles using ejaculated sperm (EJ). METHODS: ICSI was performed using EJ in 141 cycles and TE in 37 cycles. Embryos were cultured for 5 days. The quality of embryos was assessed on days 3 and 5 before embryo transfer. RESULTS: Fertilization rate was 77.3% in the EJ group and 69.6% in the TE group (p < 0.05). The good-quality embryos on day 3 and 5 were not different between the EJ and TE groups. Embryos did not develop to blastocyst stage in 7 cycles of the EJ group (5.0%) and 2 cycles of the TE group (5.4%). There were no significant differences in blastocyst formation and blastocyst quality (46.1% vs. 47.5% and 5.7% vs 5.8%, respectively) on day 5 between both groups. Embryos were transferred in all cycles. Implantation (22.8 vs. 24.7%), clinical pregnancy (44.7 vs. 43.2%), miscarriage (21.7 vs. 33.3%), and delivery (76.5 vs. 66.7%) did not differ between EJ group and TE group. Clinical outcomes of ICSI were not different between the EJ and TE groups. CONCLUSIONS: In conclusion, the potential of testicular sperm supporting embryonic development to blastocysts is comparable to that of ejaculated sperm. Therefore, this study suggests that blastocyst transfer can be a very useful assisted reproductive technique in the ICSI cycles that require the use of testicular sperm, and the clinical outcomes of the cycles are comparable to those of ICSI cycles using ejaculated sperm.
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Transferência Embrionária , Resultado da Gravidez , Injeções de Esperma Intracitoplásmicas , Recuperação Espermática , Espermatozoides/fisiologia , Testículo/fisiologia , Adulto , Ejaculação , Desenvolvimento Embrionário , Feminino , Fertilização in vitro , Humanos , Masculino , Gravidez , Testículo/citologiaRESUMO
Secretory leukocyte peptidase inhibitor (SLPI) plays a role in proliferation and differentiation via the autocrine and paracrine systems. SLPI's expression is well-documented in the reproductive tract, but it remains unclear whether it is active during early embryonic development. In this study, the expression and role of Slpi in the early embryo were evaluated. In vitro embryo cultures in chemically defined simple medium resulted in a reduction in developmental speed from the 8-cell stage, as well as implantation rate compared with in vivo embryos. SLPI protein was localized to the membrane or submembrane cytoplasm in an embryonic stage-dependent manner. In vitro cultured embryos exhibited lower levels of Slpi mRNA expression than in vivo embryos. Slpi knockdown by antisense oligonucleotides attenuated the developmental speed and implantation rate compared with Slpi sense oligonucleotide-transfected embryos and in vitro controls. The critical period for the attenuation of developmental speed occurred after the 8-cell stage. SLPI treatment accelerated development, increased implantation rate, and ameliorated the suppressive effects of Slpi knockdown. Slpi knockdown did not induce changes in the total cell number or inner cell number in blastocysts. Meanwhile, SLPI upregulated the expression of the developmental factors matrix metalloproteinase-14, neutrophil elastase, and tissue inhibitor of metalloproteinase-1. Together, these results suggest that SLPI is an effective regulator of developmental speed and implantation competence in an autocrine and paracrine manner, respectively, and plays a role in controlling the expression of embryonic development factors, such as MMP family members.
Assuntos
Diferenciação Celular , Embrião de Mamíferos/citologia , Inibidor Secretado de Peptidases Leucocitárias/metabolismo , Útero/fisiologia , Animais , Proliferação de Células , Células Cultivadas , Implantação do Embrião , Embrião de Mamíferos/metabolismo , Feminino , Elastase de Leucócito/genética , Elastase de Leucócito/metabolismo , Metaloproteinase 14 da Matriz/genética , Metaloproteinase 14 da Matriz/metabolismo , Camundongos , Gravidez , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inibidor Secretado de Peptidases Leucocitárias/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Regulação para CimaRESUMO
Cellular prion protein (PrPC) encoded at Prnp gene is well-known to form a misfolded isoform, termed scrapie PrP (PrPSC) that cause transmissible degenerative diseases in central nervous system. The physiological role of PrPC has been proposed by many studies, showing that PrPC interacts with various intracellular, membrane, and extracellular molecules including mitochondrial inner membrane as a scaffold. PrPC is expressed in most cell types including reproductive organs. Numerous studies using PrPC knockout rodent models found no obvious phenotypic changes, in particular the clear phenotypes in development and reproduction have not demonstrated in these knockout models. However, various roles of PrPC have been evaluated at the cellular levels. In this review, we summarized the known roles of PrPC in various cell types and tissues with a special emphasis on those involved in reproduction.
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Recent studies suggest that epigenetic modifications, such as DNA methylation and histone modification, can alter the differentiation potential of stem cells or progenitor cells. Specifically, coactivator-associated arginine methyltransferase 1 (CARM1) is known to act as a coactivator for various transcription factors and to regulate gene expression by chromatin remodeling through histone methylation. Here, for the first time, we have used direct protein delivery of CARM1 using cell-penetrating peptide (CPP) to regulate the differentiation potential of human mesenchymal stem cells (hMSCs). Immunofluorescence showed that the CPP-CARM1 protein is successfully delivered into the nuclei of hMSCs. Further experiments using immunofluorescence and Western blotting showed that the delivered CARM1 protein can effectively methylate the arginine 17 residue of histone H3 in both bone marrow (BM)- and adipose-derived (AD)-hMSCs, thus suggesting that the CARM1 protein delivered by the CPP system is biologically active in hMSCs. Chromatin immunoprecipitation (ChIP) assay and genome-wide gene expression profiling supported the result that delivered CARM1 protein can cause chromatin remodeling through histone methylation. Finally, the CPP-CARM1 protein efficiently elevated the differentiation efficiency of BM-hMSCs and AD-hMSCs into adipogenic, osteogenic, and myogenic cell lineages in vitro. Altered expression of critical genes after hMSC differentiation was reconfirmed by real-time reverse transcription polymerase chain reaction (qRT-PCR). Collectively, our results suggest that CPP-CARM1 can elevate the differentiation potential of hMSCs into various cell types, and that this system using CPP is a useful tool for exogenous protein delivery in clinical applications of cell-based therapy.
Assuntos
Peptídeos Penetradores de Células/metabolismo , Células-Tronco Mesenquimais/citologia , Proteína-Arginina N-Metiltransferases/metabolismo , Diferenciação Celular/fisiologia , Peptídeos Penetradores de Células/genética , Expressão Gênica , Humanos , Células-Tronco Mesenquimais/enzimologia , Células-Tronco Mesenquimais/metabolismo , Análise em Microsséries , Regiões Promotoras Genéticas , Proteína-Arginina N-Metiltransferases/genética , Transcrição GênicaRESUMO
Previously, we showed that a chronic-low-dose nonylphenol (NP) exposure resulted in histological changes with sexually dimorphic pattern in rat adrenal glands. We hypothesized that such structural changes are closely related to the hormonal secretory patterns. To test this hypothesis, we developed the short-term adrenal incubation method, and measured the levels of catecholamines and cortical steroids using the high-performance liquid chromatography with electrochemical detection (HPLC-ECD) and specific enzyme-linked immunosorbent assay, respectively. The norepinephrine (NE) levels in media from NP-treated female adrenal, except 100 pM NP, were significantly increased [control (CTL) vs 1 nM NP, p<0.001; vs 10 nM NP, p<0.05; vs 100 nM NP, p<0.001; vs 1 µM NP, p<0.01]. The NE secretion from male adrenal was higher when treated with 100 nM and 1 µM NP (CTL vs 100 nM NP, p<0.05; vs 1 µM NP, p<0.05, respectively). The aldosterone level in the female adrenal media treated with 100 pM NP was significantly decreased, on the other hand, that of media treated with 10 nM NP was significantly increased (CTL vs 100 pM NP, p<0.05; vs 10 nM NP, p<0.01). In male adrenal media, the aldosterone levels of 10 nM, 100 nM and 1 µM NP-treated media were significantly declined (CTL vs 10 nM NP, p<0.001; vs 100 nM NP, p<0.001; vs 1 µM NP, p<0.001). These results showed the NP treatment altered secretory pattern of aldosterone from adrenals of both sexes, showing sexual dimorphism. It may be helpful for understanding possible adrenal pathophysiology, and endocrine disrupting chemicals-related sexually dimorphic phenomena in adrenals.
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Exposure to environmental chemicals, including endocrine-disrupting chemicals, during the gestational period can have profound adverse effects on several organs in offspring. Bisphenol A (BPA) can infiltrate the human body through food and drinks, and its metabolites can cross both the placental and the blood-brain barriers. In this study, we investigate the effect of gestational exposure to BPA on epigenetic, biochemical, and histological modifications in the uterine tissues of F1 adult offspring rats. Pregnant rats were exposed to BPA from gestational day 8-15, and changes in global DNA methylation in uterine tissues obtained from adult offspring born to the exposed mothers were analyzed. Global DNA methylation analysis revealed that gestational exposure to BPA resulted in DNA hypomethylation in the uterus. Progesterone receptor (PR) protein expression in uterine tissues was monitored using western blot analysis, which revealed that the PR protein content was considerably higher in all BPA-exposed groups than in the control. Immunohistochemical examination for the PR revealed that intense PR-positive cells were more frequently observed in the BPA-exposed group than in the control group. To date, the evidence that the upregulation of PRs observed in the present study was caused by the non-methylation of specific PR promoter regions is lacking. Conclusively, these results indicate that exposure to BPA during gestation induces epigenetic alterations in the uteri of adult female offspring. We speculate that the global DNA hypomethylation and upregulation of the PR observed simultaneously in this study might be associated with the uterus.
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Acetaminophen [Paracetamol, N-acetyl-para-aminophenol (APAP)] is a common over-the- counter analgesic agent as nonsteroidal anti-inflammatory drugs (NSAIDs). The high doses or the long-term treatment of acetaminophen via usual gavage feeding resulted in damage of testicles that presented recoverable impairment, as well as liver and kidney. The influence of acetaminophen was examined in male golden hamsters treated with acetaminophen- containing diet feeding. They were divided into 5 groups and subjected to this experiment for 4 weeks: animals housed in long photoperiod (LP) as LP control, animals housed in short photoperiod (SP) for 4 weeks as SP control (SP4), and groups of animals treated with low, middle, and high concentrations of acetaminophen (Low, Middle, High groups). Also animals housed in SP for 8 weeks were included (SP8) to contrast testicular activities, if necessary. As results, spermatozoa filled the seminiferous tubules of the testicles of animals in LP control and SP4 groups. The aspects were seen in the animals taken diets of low and middle doses of acetaminophen. The animals who fed high dose of acetaminophen showed large or small testicles. The large testicles displayed all germ cells at the steps of spermatogenesis. The small testicles presented no sperm as the animals housed in SP for 8 weeks. Thus these results indicate that acetaminophen invokes the antigonadal effects and accelerates the regressing process of the testicles in the animals compared to the animals exposed to SP.
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Zinc oxide nanoparticles (ZnO NPs) are used in various fields, including biological ones. ZnO NPs are eventually disposed of in the environment where they may affect natural systems, and there is no international law to regulate their manufacture, usage, and disposal. Hence, this present study is carried out to synthesise a more non-toxic and bioactive ZnO NPs from the marine algae Sargassum polycystum. The ZnO NPs were biologically produced using the marine algae Sargassum polycystum. The dynamic light scattering result describes that synthesised particles' average size is about 100 nm in diameter. Transmission electron microscopy (TEM) analysis demonstrated the rod-like morphology of ZnO NPs. Fourier tranform-infrared spectroscopy (FT-IR) results revealed the presence of functional groups in ZnO NPs. The selected area electron diffraction (SAED) results strongly suggested the ZnO NPs crystallinity. ZnO NPs surface morphology and compositions were identified by scanning electron microscopy (SEM- EDX) values. To analyse the toxicity of synthesised nanoparticles, zebra fish larvae were used, which involved subjecting embryos to various ZnO NPs concentrations at 1 hpf and analysing the results at 96 hpf. The 60 and 80 ppm sub-lethal doses were chosen for further studies based on the LC50 (82.23 ppm). In the ZnO NPs-treated groups, a significant slowdown in pulse rate and a delay in hatching were seen, both of which impacted the embryonic processes. A teratogenic study revealed a dose-dependent increase in the incidence of developmental deformities in the treated groups. Along with increased oxidants and a corresponding reduction in antioxidant enzymes, Na+ K+-ATPase and AChE activity changes were seen in ZnO NPs-treated zebra fish larvae groups. The apoptosis process was increased in ZnO NPs-treated groups revealed by acridine orange staining. These results indicate that the green synthesis process cannot mitigate the oxidative stress induced by ZnO NPs on oxidative signalling.
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OBJECTIVE: Obesity is strongly associated with metabolic syndrome, but not all obese individuals display a clustering of metabolic risk factors. Recent studies have shown that in vitro subcutaneous (SC)-preadipocyte differentiation is negatively associated with obesity. These results suggest that impaired adipogenesis is an important factor linking obesity to metabolic disorders. We examined whether in vitro preadipocyte differentiation is associated with metabolic syndrome, independent of obesity. DESIGN/PATIENTS/MEASUREMENTS: Paired adipose tissue samples were obtained from the 13 nonobese women and the 65 obese women. The CD34(+)/CD31(-) cells were isolated from the stromal-vascular fraction of both SC and omental (OM) fat depots by immune magnetic separation, and the subset was cultured with a differentiation cocktail. Then, we analysed the relationship between the degree of preadipocyte differentiation and metabolic factors. RESULTS: Obese women without metabolic syndrome (n = 37) had significantly higher SC-preadipocyte differentiation than equally obese women with metabolic syndrome (n = 28); however, OM-preadipocyte differentiation was similar in both groups. SC-preadipocyte differentiation was strongly correlated with triglycerides, HDL cholesterol, homoeostasis model assessment of insulin resistance and OM-adipocyte size. However, OM-preadipocyte differentiation was not correlated with any of these parameters. CONCLUSIONS: This study identified that SC-preadipocyte differentiation is associated with metabolic syndrome independent of obesity, whereas OM-preadipocyte differentiation is not. These findings suggest that, in the setting of obesity, an enhanced adipogenic capacity of SC depots could be protective for metabolic syndrome. Our data underscores an interaction between adipose tissue homoeostasis and metabolic disorder.
Assuntos
Adipócitos/citologia , Adipócitos/fisiologia , Distribuição da Gordura Corporal , Síndrome Metabólica/metabolismo , Obesidade/metabolismo , Adulto , Idoso , Diferenciação Celular , Células Cultivadas , Feminino , Humanos , Gordura Intra-Abdominal/citologia , Gordura Intra-Abdominal/metabolismo , Subpopulações de Linfócitos , Pessoa de Meia-Idade , Gordura Subcutânea/citologia , Gordura Subcutânea/metabolismoRESUMO
Since T cells express diverse sex steroid hormone receptors, they might be a good model to evaluate the effects of sex steroid hormones on immune modulation. Porcine testicular extract contains several sex steroid hormones and may be useful to study the effects of sex steroid hormones during T cell activation. We have examined the effects of the porcine testicular extract on T cell activation: proliferation and secretion of cytokines (IL-2 and IFN-γ) by activated T cells were severely decreased after treatment with porcine testicular extract. The extract produced an immunosuppressive effect and inhibited the proliferation of activated T cells by blocking the cell cycle transition from the G(1) phase to S phase. These effects were mediated by a decrease in the expression of cyclin D1 and cyclin E and constitutive expression of p27(KIP1) after T cell activation.
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Ciclo Celular/efeitos dos fármacos , Extratos Celulares/farmacologia , Proliferação de Células/efeitos dos fármacos , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/farmacologia , Linfócitos T/fisiologia , Testículo/química , Animais , Extratos Celulares/isolamento & purificação , Citocinas/metabolismo , Masculino , Suínos , Linfócitos T/efeitos dos fármacosRESUMO
Kisspeptins, products of KISS1 gene, are ligands of the G-protein coupled receptor (GPR54), and the kisspeptin-GPR54 signaling has an important role as an upstream regulator of gonadotropin releasing hormone (GnRH) neurons. Interestingly, extrahypothalamic expressions of kisspeptin/GPR-54 in gonads have been found in primates and experimental rodents such as rats and mice. Hamsters, another potent experimental rodent, also have a kisspeptin-GPR54 system in their ovaries. The presence of testicular kisspeptin-GPR54 system, however, remains to be solved. The present study was undertaken to determine whether the kisspeptin is expressed in hamster testis. To do this, reverse transcription-polymerase chain reactions (RT-PCRs) and immunohistochemistry (IHC) were employed. After the nest PCR, two cDNA products (320 and 280 bp, respectively) were detected by 3% agarose gel electrophoresis, and sequencing analysis revealed that the 320 bp product was correctly amplified from hamster kisspeptin cDNA. Modest immunoreactive (IR) kisspeptins were detected in Leydig-interstitial cells, and the weak signals were detected in germ cells, mostly in round spermatids and residual bodies of elongated spermatids. In the present study, we found the kisspeptin expression in the testis of Syrian hamster. Further studies on the local role(s) of testicular kisspeptin are expected for a better understanding the physiology of hamster testis, including photoperiodic gonadal regression specifically occurred in hamster gonads.
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The Syrian (golden) hamsters are seasonal breeders whose reproductive functions are active in summer and inactive in winter. In experimental facility mimicking winter climate, short photoperiod (SP) induces gonadal regression. The blood-testis barrier (BTB) of the sexually involuted animals have been reported to be permeable, allowing developing germ cells to be engulfed or sloughed off the epithelium of the seminiferous tubules. The expressions of genes related to the tight junction composing of BTB were investigated in the reproductive active and inactive testes. Claudin-11, occludin, and junctional adhesion molecule (JAM) were definitely expressed in the active testes but not discernably detected in the inactive testes. And spermatozoa (sperm) were observed in the whole lengths of epididymides in the active testes. They were witnessed in only cauda region of the epididymides but not in caput and corpus regions in animals with the inactive testes. The results imply that the disorganization of BTB is associated with the testicular regression. The developing germ cells are swallowed into the Sertoli cells or travel into the lumen, as supported by the presence of the sperm delayed in the last region of the epididymis. These outcomes suggest that both apoptosis and desquamation are the processes that eliminate the germ cells during the regressing stage in the Syrian hamsters.
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Phthalates and their metabolites are well-known endocrine disrupting chemicals. Di-(2-ethylhexyl) phthalate (DEHP) has been widely used in industry and the exposing possibility to adult is high. In this study, DEHP was treated (133 µg/L and 1,330 µg/L in drinking water) according to the OECD test guideline 443 to mature female mice and their adrenal gland were examined for histological characteristics and steroidogenic gene expression. The wet weight of the adrenal gland was increased in all administrated groups compared to control. The diameter of zona fasciculata (ZF) was increased by DEHP in both outer ZF and inner ZF but there was no difference in morphology of the cells and arrangements into zona between groups. In addition, the arrangement of extracellular matrix was not different between control and DEHP groups. CYP11B1 was mainly localized at ZF and the intensity was not different between groups. DAX1 was localized in zona glomerulosa (ZG) and ZF, and its expression levels were decreased by DEHP administration. Its level was lower in DEHP133 group than DEHP1330 group. On the other hand, CYP17A1 was localized in ZG of DEHP1330 group. These results suggest that chronic low-dose DEHP exposing may modify the microstructure and function of the adrenal cortical cortex.
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In the present study, a MoS2@Ti3C2Tx MXene hybrid-based electrochemical aptasensor (MEA) was introduced for sensitive and rapid quantification of Thyroxine (T4). T4 is a crucial hormone and plays a key role in various body functions. Therefore, there is high demand for an accurate, sensitive, and rapid method for the detection of T4. To construct the aptasensor, a nano-hybrid (NH) consisting of Ti3C2Tx MXene and MoS2 nanosheets (NS) was synthesized, and applied to a carbon electrode surface, followed by the electroplating of gold nanostructures (GN). The smart combination of Ti3C2Tx MXene and MoS2NS enhanced the physiochemical properties of the electrode surface, as well as provided a building block to form 3D GN. The 3D architecture of the GN offered a unique substrate to capture numerous T4 aptamer molecules, which consequently amplified the signal by nearly 6-fold. The MEA quantified thyroxine with a limit of detection (LOD) of 0.39 pg/mL over a dynamic range ((7.8 × 10-1) to (7.8 × 106)) pg/mL within 10 min. Moreover, the MEA successfully detected T4 in human serum samples. Lastly, the results obtained from the aptasensor were compared with those from the ELISA standard method. The comparative analysis showed good agreement between the two methods.
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Dissulfetos/química , Técnicas Eletroquímicas/métodos , Molibdênio/química , Tiroxina/sangue , Titânio/química , Técnicas Biossensoriais/métodos , Cromatografia Líquida de Alta Pressão , Galvanoplastia , Ensaio de Imunoadsorção Enzimática , Humanos , Limite de Detecção , Tri-Iodotironina/sangueRESUMO
Ovarian reserve and fertility are reduced by aging and a poor energy balance. To date, the relationships of high energy accumulation and aging with the ovarian reserve have not been elucidated. Here, the effects of obesity on the aging ovarian reserve were evaluated in a leptin-deficient (ob/ob) mouse model. Abnormal estrous cyclicity appeared as early as 6 weeks and worsened with aging. The blood level patterns of 17ß-estradiol (E2), testosterone (T), and progesterone (P4) with aging were similar between lean and ob/ob mice. The blood level of E2 but not P4 or T was similar at 24 weeks. Many more atretic follicles but fewer corpora lutea were observed in ob/ob mice than in lean mice within all age groups. Anti-Müllerian hormone (Amh) mRNA levels were similar between genotypes. Dazl, Stra8, and ZP3 mRNAs were highly expressed in ob/ob mice after 12 weeks. Sohlh1 and Ybx2 mRNAs were highly expressed at 24 weeks in ob/ob compared with lean mice. In addition, SOHLH1-positive primordial follicle counts were significantly increased in ob/ob mice at 24 weeks. The proportions of AMH-positive secondary and small antral follicles were similar between genotypes. Together, these results show that the ovarian reserve lasts longer in ob/ob mice than in lean mice, suggesting that the loss of normal physiological or physical status causes decreased fertility at a young age in ob/ob mice and that an increase in adipocytes without leptin, as in ob/ob mice, can improve the ovarian reserve. Such knowledge can be applied to understanding reproductive dysfunction.
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Phthalates have a long industrial history. It is suspected that phthalates and their metabolites have detrimental effects on reproduction and development. They are well-known for their anti-androgenic effects. Several studies have indicated that phthalates and their metabolites are reprotoxic in males and endocrine disruptors. Reproduction and embryogenesis occur in the uterus of female eutherian mammals. A horizontal analytical method is preferred to elucidate the toxic effects of phthalates on human reproduction. Nevertheless, there are vast numbers of known phthalates and not all of their modes of action have been clarified. Di-(2-ethylhexyl) phthalate (DEHP) is a commonly used plasticizer and has been the subject of numerous toxicological studies. However, few of these have reported on the toxic effects of DEHP, its metabolites, other phthalates, or mixtures on female reproduction. Acute and high doses of DEHP adversely affect uterine histology. Recently, it was disclosed that chronic exposures to low doses of DEHP have endocrine disruption efficacy. DEHP induces various cellular responses including modulation of the expression and regulation of steroid hormone receptors and transcription and paracrine factors. Uteri do not respond uniformly to DEHP exposure. The phenotypic manifestations and effects on fertility in response to DEHP and its metabolites may vary with species, developmental stage, and generation. Hence, DEHP exposure may histological alter the uterus and induce endometriosis, endometriosis, hyperplasia, myoma, and developmental and reproductive toxicity.