Search Results (24)

Osteoporosis is a common skeletal disease characterized by decreased bone density, leading to bone fragility and fractures, especially in menopausal women. The purpose of this study is to confirm the anti-osteoporosis activity of stem cell extracellular vesicles (EVs) as a material of regenerative medicine. Mesenchymal stem cells have a potential to differentiate into osteocytes, so directly reconstruct bone tissue or facilitate bone regeneration via paracrine effects. Paracrine effects are mediated by functional molecules delivered in EVs released from stem cells. EVs containing high concentrations of growth factors (GFs) and neurotrophic factors (NFs) were attained via hypoxia culture of human amniotic membrane stem cells (AMSCs). From the EVs with a mean diameter of 77 nm, 751 proteins and 15 species of lipids were identified. Sprague-Dawley rats were ovariectomized, and eight weeks later, intravenously injected with EVs at doses of 1 × 10⁸, 3 × 10⁸ or 1 × 10⁹ particles/100 μL/body, weekly for eight weeks. One week after the final administration, the serum and bone parameters related to bone density were analyzed. Serum 17β-estradiol, alkaline phosphatase, and calcium levels that decreased in ovariectomized rats were restored by EVs in a dose-dependent manner. Bone parameters such as bone mineral density, bone mineral content, bone volume/tissue volume ratio, trabecular number, trabecular space, and bending strength were also improved by treatment with EVs. Such effects were confirmed by morphological findings of micro-computed tomography. Taken together, it is suggested that AMSC-EVs containing high concentrations of GFs and NFs preserve bone soundness by promoting bone regeneration and inhibiting bone resorption. Full article

Abnormal sexual maturity exhibits significant detrimental effects on adult health outcomes, and previous studies have indicated that targeting histone acetylation might serve as a potential therapeutic approach to regulate sexual maturity. However, the mechanisms that account for it remain to be further elucidated. Using the mouse model, we showed that Trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, downregulated the protein level of Hdac1 in ovaries to promote the apoptosis of granulosa cells (GCs), and thus arrested follicular development and delayed sexual maturity. Using porcine GCs as a cell model, a novel sexual maturity-associated lncRNA, which was named as the stimulatory factor of follicular development (SFFD), transcribed from mitochondrion and mediated by HDAC1, was identified using RNA sequencing. Mechanistically, HDAC1 knockdown significantly reduced the H3K27ac level at the −953/−661 region of SFFD to epigenetically inhibit its transcription. SFFD knockdown released miR-202-3p to reduce the expression of cyclooxygenase 1 (COX1), an essential rate-limited enzyme involved in prostaglandin synthesis. This reduction inhibited the proliferation and secretion of 17β-estradiol (E2) while promoting the apoptosis of GCs. Consequently, follicular development was arrested and sexual maturity was delayed. Taken together, HDAC1 knockdown-mediated SFFD downregulation promoted the apoptosis of GCs through the miR-202-3p-COX1 axis and lead to delayed sexual maturity. Our findings reveal a novel regulatory network modulated by HDAC1, and HDAC1-mediated SFFD may be a promising new therapeutic target to treat delayed sexual maturity. Full article

Women are at a higher risk of cognitive impairments and Alzheimer’s disease (AD), particularly after the menopause, when the estrous cycle becomes irregular and diminishes. Numerous studies have shown that estrogen deficiency, especially estradiol (E2) deficiency, plays a key role in this phenomenon. Recently, a novel polymeric drug, hyaluronic acid–17β-estradiol conjugate (HA-E2), has been introduced for the delivery of E2 to brain tissues. Studies have indicated that HA-E2 crosses the blood–brain barrier (BBB) and facilitates a prolonged E2 release profile while lowering the risk of estrogen-supplement-related side effects. In this study, we used ovariohysterectomy (OHE) rats, a postmenopausal cognitive deficit model, to explore the effect of a 2-week HA-E2 treatment (210 ng/kg body weight, twice a week) on the cholinergic septo-hippocampal innervation system, synaptic transmission in hippocampal pyramidal neurons and cognitive improvements. Our study revealed an 11% rise in choline acetyltransferase (ChAT) expression in both the medial septal nucleus (MS nucleus) and the hippocampus, along with a 14–18% increase in dendritic spine density in hippocampal pyramidal neurons, following HA-E2 treatment in OHE rats. These enhancements prompted the recovery of cognitive functions such as spatial learning and memory. These findings suggest that HA-E2 may prevent and improve estrogen-deficiency-induced cognitive impairment and AD. Full article

Given the dramatic increase in the L. sceleratus population in the southeastern Aegean Sea, there is growing interest in assessing the toxicity of this pufferfish and the factors controlling its tetrodotoxin (TTX) content. In the present study, liver, gonads, muscle and skin of 37 L. sceleratus specimens collected during May and June 2021 from the island of Rhodes, Greece, were subjected to multi-analyte profiling using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in order to quantitate TTX and evaluate whether this biotoxin interrelates with hormones. TTX and its analogues 4-epiTTX, 11-deoxyTTX, 11-norTTX-6-ol, 4,9-anhydroTTX and 5,11/6,11-dideoxyTTX were detected in all tissue types. Liver and gonads were the most toxic tissues, with the highest TTX concentrations being observed in the ovaries of female specimens. Only 22% of the analyzed muscle samples were non-toxic according to the Japanese toxicity threshold (2.2 μg TTX eq g⁻¹), confirming the high poisoning risk from the inadvertent consumption of this species. Four steroid hormones (i.e., cortisol, testosterone, androstenedione and β-estradiol) and the gonadotropin-releasing hormone (GnRH) were detected in the gonads. Androstenedione dominated in female specimens, while GnRH was more abundant in males. A positive correlation of TTX and its analogues with β-estradiol was observed. However, a model incorporating sex rather than β-estradiol as the independent variable proven to be more efficient in predicting TTX concentration, implying that other sex-related characteristics are more important than specific hormone-regulated processes. Full article

The seed production of small yellow croaker (SYC) is constrained by reproductive dysfunction in captive-reared females. Reproductive dysfunction is closely linked to endocrine reproductive mechanisms. To better understand the reproductive dysfunction in captive broodstock, functional characterization of gonadotropins (GtHs: follicle stimulating hormone β subunit, fshβ; luteinizing hormone β subunit, lhβ; and glycoprotein α subunit, gpα) and sex steroids (17β-estradiol, E2; testosterone, T; progesterone; P) was performed using qRT-PCR, ELISA, in vivo, and in-vitro assay. The pituitary GtHs and gonadal steroids levels were significantly higher in ripen fish of both sexes. However, changes in lhβ and E2 levels in females were not significant in the developing and ripen stages. Furthermore, GtHs and steroids levels were lower in females compared to males throughout the reproductive cycle. In vivo administration of gonadotropin releasing hormone analogue (GnRHa) significantly increased the expression of GtHs in both dose- and time-related manners. The lower and higher doses of GnRHa led to successful spawning in male and female SYC, respectively. Sex steroids in vitro significantly inhibited the expression of lhβ in female SYC. Overall, GtHs were shown to play a vital role in final gonadal maturation, while steroids promoted negative feedback in the regulation of pituitary GtHs. Lower levels of GtHs and steroids might be key components in the reproductive dysfunction of captive-reared female SYC. Full article

Sex reversal induced by 17β-estradiol (E₂) has shown the potential possibility for monoculture technology development. The present study aimed to determine whether dietary supplementation with different concentrations of E₂ could induce sex reversal in M. nipponense, and select the sex-related genes by performing the gonadal transcriptome analysis of normal male (M), normal female (FM), sex-reversed male prawns (RM), and unreversed male prawns (NRM). Histology, transcriptome analysis, and qPCR were performed to compare differences in gonad development, key metabolic pathways, and genes. Compared with the control, after 40 days, feeding E₂ with 200 mg/kg at PL25 (PL: post-larvae developmental stage) resulted in the highest sex ratio (female: male) of 2.22:1. Histological observations demonstrated the co-existence of testis and ovaries in the same prawn. Male prawns from the NRM group exhibited slower testis development without mature sperm. RNA sequencing revealed 3702 differentially expressed genes (DEGs) between M vs. FM, 3111 between M vs. RM, and 4978 between FM vs. NRM. Retinol metabolism and nucleotide excision repair pathways were identified as the key pathways for sex reversal and sperm maturation, respectively. Sperm gelatinase (SG) was not screened in M vs. NRM, corroborating the results of the slice D. In M vs. RM, reproduction-related genes such as cathepsin C (CatC), heat shock protein cognate (HSP), double-sex (Dsx), and gonadotropin-releasing hormone receptor (GnRH) were expressed differently from the other two groups, indicating that these are involved in the process of sex reversal. Exogenous E₂ can induce sex reversal, providing valuable evidence for the establishment of monoculture in this species. Full article

Chub mackerel, Scomber japonicus, is heavily farmed and harvested due to its demand as a high-quality protein source rich in fatty acids. However, the effects of environmental cues on sexual maturation of the fish remain understudied. We aim to elucidate the effect of light manipulation on the hormones related to reproduction and on the stress response in the species. Mackerel were exposed to different photoperiods (12 h light:12 h dark or 14 h light:10 h dark) and light wavelengths (provided by white fluorescent bulbs or green LEDs). Total RNA extracted from the brain was assayed with quantitative polymerase chain reaction (a powerful technique for advancing functional genomics) and blood plasma was analyzed via immunoassay using ELISA kits. The mRNA expression of gene-encoding gonadotropin-releasing hormone, gonadotropin hormone, follicle-stimulating hormone, and luteinizing hormone were significantly increased through the use of an extended photoperiod and green wavelength, which also increased testosterone and 17β-estradiol plasma levels. Plasma levels of cortisol and glucose, which are indicators of a stress response, were significantly decreased through green LED exposure. Our results indicate that environmental light conditions affect the production of pituitary and sex hormones, and reduce the stress response in S. japonicus. Full article

The production and use of manufactured chemicals have risen significantly in the last few decades. With interest in preserving and improving the state of the environment, there is also growing interested in new technologies for water purification and wastewater treatment. One frequently discussed technological group is advanced oxidation processes (AOPs). AOPs using sulphur-based radicals appear to reduce the volume of organic contaminants in wastewater significantly. The use of persulfate has excellent potential to successfully eliminate the number of emerging contaminants released into the environment. The main disadvantage of sulphur-based AOPs is the need for activation. We investigated an economically and environmentally friendly solution based on hydrodynamic cavitation, which does not require heating or additional activation of chemical substances. The method was evaluated for emerging contaminant removal research, specifically for the group of steroid estrogens. The mixture of estrone (E1), 17β-estradiol (E2), estriol (E3), and 17α-ethinylestradiol (EE2) was effectively eliminated and completely removed during a treatment that lasted just a few seconds. This novel method can be used in a broad spectrum of water treatment processes or as the intensification of reactions in chemical engineering technologies. Full article

The aim of this study was to apply biochemical parameters to Onychostoma barbatulum to understand whether Wuling Farm activities have caused organic pollution from xenoestrogens to flow into surrounding streams and thereby affected area aquatic ecosystems. Individuals of the Taiwan shovel-jaw carp (O. barbatulum) were collected by fyke netting in the Cijiawan, Kaoshan, and Yusheng Rivers in the protected area of Shei-Pa National Park and in the Ikawan River outside the protected areas from 4–5 June 2015 to 20–21 September 2015. The collected male individuals of O. barbatulum were divided into two groups. In the first group monooxygenase (Mon) activity, glutathione S-transferase (GST) activity, and vitellogenin (VTG) concentration in the liver were measured right after they were captured. Those in the second group were acclimated for 14 days and then exposed to 0 (control) or 10 ng/L 17β-estradiol for 10 days and then measured for changes in Mon, GST, and VTG in their livers. The value of VTG concentrations in O. barbatulum individuals collected from the Ikawan River was 2.12 ± 1.62 μg PO₄²⁺/g protein, which was the highest among all samples, followed by those collected from the Yusheng River (0.78 ± 1.00 μg PO₄²⁺/g protein). Individuals collected from the Yusheng River had the highest liver Mon activity (4.16 ± 1.08^△A650/30 min/g protein) and the highest GST activity (1.58 ± 1.13 ^△A340/20 min/g protein), followed by those from the Ikawan River (Mon: 3.63 ± 1.13 ^△A650/30 min/g protein; GST: 1.24 ± 0.73 ^△A340/20 min/g protein). Comprehensive analyses showed that Mon and GST activities and VTG concentrations in livers measured right after being collected were lower than individuals exposed to 10 ng 17β-estradiol/L in the laboratory. Induced VTG concentrations were 9.87 ± 0.89 μg PO₄²⁺/g protein, Mon activity 8.02 ± 1.74 ^△A650/30 min/g protein, and GST activity 3.24 ± 0.62 ^△A340/20 min/g protein, indicating that farming activities have not significantly affected these aquatic organisms. However, pollution sources are still releasing pollutants containing xenoestrogens into the Yusheng River and Ikawan River. The impact of xenoestrogens is worthy of continuous long-term follow-up monitoring. Full article

Spats1 (spermatogenesis-associated, serinerich 1) has been characterized as a male-biased gene which acts an important role in the germ cell differentiation of mammals. Nevertheless, the function of Spats1 in the Chinese soft-shelled turtle (P. sinensis) has not yet been reported. To initially explore the expression of Spats1 in P. sinensis and its response to sex steroid treatment, we cloned the CDS of Spats1 for the first time and analyzed its expression profile in different tissues, including the testes in different seasons. The Spats1 cDNA fragment is 1201 base pairs (bp) in length and contains an open reading frame (ORF) of 849 bp, which codes for 283 amino acids. Spats1 mRNA was highly expressed in the testes (p < 0.01) and barely detectable in other tissues. In P. sinensis, the relative expression of Spats1 also responsive to seasonal changes in testis development. In summer (July) and autumn (October), Spats1 gene expression was significantly higher in the testes than in other seasons (p < 0.05). Spats1 mRNA was found to be specifically expressed in germ cells by chemical in situ hybridization (CISH), and it was mainly located in primary spermatocytes (Sc1), secondary spermatocytes (Sc2) and spermatozoa (St). Spats1 expression in embryos was not significantly changed after 17α-methyltestosterone (MT)and 17β-estradiol (E2) treatment. In adults, MT significantly induced Spats1 expression in male P. sinensis. However, the expression of Spats1 in testes was not responsive to E2 treatment. In addition, the expression of Spats1 in females was not affected by either MT or E2 treatment. These results imply that Spats1 is a male-specific expressed gene that is mainly regulated by MT and is closely linked to spermatogenesis and release in P. sinensis. Full article

New technologies have been developed around the world to tackle current emergencies such as biowaste recycling, renewable energy production and reduction of environmental pollution. The thermochemical and biological conversions of waste biomass for bioenergy production release solid coproducts and byproducts, namely biochar (BC), hydrochar (HC) and digestate (DG), which can have important environmental and agricultural applications. Due to their physicochemical properties, these carbon-rich materials can behave as biosorbents of contaminants and be used for both wastewater treatment and soil remediation, representing a valid alternative to more expensive products and sophisticated strategies. The alkylphenols bisphenol A, octylphenol and nonylphenol possess estrogenic activity comparable to that of the human steroid hormones estrone, 17β-estradiol (and synthetic analog 17α-ethinyl estradiol) and estriol. Their ubiquitous presence in ecosystems poses a serious threat to wildlife and humans. Conventional wastewater treatment plants often fail to remove environmental estrogens (EEs). This review aims to focus attention on the urgent need to limit the presence of EEs in the environment through a modern and sustainable approach based on the use of recycled biowaste. Materials such as BC, HC and DG, the last being examined here for the first time as a biosorbent, appear appropriate for the removal of EEs both for their negligible cost and continuously improving performance and because their production contributes to solving other emergencies, such as virtuous management of organic waste, carbon sequestration, bioenergy production and implementation of the circular economy. Characterization of biosorbents, qualitative and quantitative aspects of the adsorption/desorption process and data modeling are examined. Full article

Pikeperch Sander lucioperca is a piscivorous species considered a promising candidate for the diversification of intensive aquaculture. This study aimed to determine the effect of a sustained-release delivery system incorporating mammalian gonadotropin-releasing hormone agonist (mGnRHa) into poly(lactic-co-glycolic acid) (PLGA) microparticles on the sex steroid levels and aspects of artificial reproduction of pikeperch. Fish were divided into four groups and injected with 20 µg mGnRHa/kg, 5-day release microparticles encapsulated with 5 µg GnRHa/kg BW (PLGA 5), 20 µg GnRHa/kg (PLGA 20), or 1 mL/kg 0.9% NaCl (control). Cumulative percentage ovulation was 100% in the PLGA 5 group, significantly higher than in other tested groups. No differences among groups were observed in latency or fecundity. The level of 11-ketotestosterone (11-KT) peaked at 40 h post-injection, and was sustained during ovulation, in all treated groups. The 17β-estradiol (E2) concentration increased in the mGnRHa-only group immediately after hormone injection, while both PLGA groups showed a reduction in E2 after injection, continuing to decrease until ovulation. A low dose of mGnRHa in PLGA microparticles significantly improves induction of ovulation and results in acceptable reproductive performance, which may positively affect pikeperch production under controlled conditions. Full article

Estrogens can elicit rapid cellular responses via the G-protein-coupled receptor 30 (GPR30), followed by estrogen receptor α (ERα/ESR1)-mediated genomic effects. Here, we investigated whether rapid estrogen signaling via GRP30 may affect ESR1 expression, and we examined the underlying molecular mechanisms. The exposure of human endometrial cancer cells to 17β-estradiol promoted p62 phosphorylation and increased ESR1 protein expression. However, both a GPR30 antagonist and GPR30 silencing abrogated this phenomenon. GPR30 activation by 17β-estradiol elicited the SRC/EGFR/PI3K/Akt/mTOR signaling pathway. Intriguingly, unphosphorylated p62 and ESR1 were found to form an intracellular complex with the substrate adaptor protein KEAP1. Upon phosphorylation, p62 promoted ESR1 release from the complex, to increase its protein expression. Given the critical role played by p62 in autophagy, we also examined how this process affected ESR1 expression. The activation of autophagy by everolimus decreased ESR1 by promoting p62 degradation, whereas autophagy inhibition with chloroquine increased ESR1 expression. The treatment of female C57BL/6 mice with the autophagy inhibitor hydroxychloroquine—which promotes p62 expression—increased both phosphorylated p62 and ESR1 expression in uterine epithelial cells. Collectively, our results indicate that 17β-estradiol-mediated GPR30 activation elicits the SRC/EGFR/PI3K/Akt/mTOR signaling pathway and promotes p62 phosphorylation. In turn, phosphorylated p62 increased ESR1 expression by inducing its release from complexes that included KEAP1. Our findings may lead to novel pharmacological strategies aimed at decreasing ESR1 expression in estrogen-sensitive cells. Full article

The application of nanoparticles has experienced a vertiginous growth, but their interaction with food and medicinal plants in organisms, especially in the control of reproduction, remains unresolved. We examined the influence of copper nanoparticles supported on titania (CuNPs/TiO₂), plant extracts (buckwheat (Fagopyrum esculentum) and vitex (Vitex agnus-castus)), phytochemicals (rutin and apigenin), and their combination with CuNPs/TiO₂ on ovarian cell functions, using cultured porcine ovarian granulosa cells. Cell viability, proliferation (PCNA accumulation), apoptosis (accumulation of bax), and hormones release (progesterone, testosterone, and 17β-estradiol) were analyzed by the Trypan blue test, quantitative immunocytochemistry, and ELISA, respectively. CuNPs/TiO₂ increased cell viability, proliferation, apoptosis, and testosterone but not progesterone release, and reduced the 17β-estradiol output. Plant extracts and components have similar stimulatory action on ovarian cell functions as CuNPs/TiO₂, but abated the majority of the CuNPs/TiO₂ effects. This study concludes that (1) CuNPs/TiO₂ can directly stimulate ovarian cell functions, promoting ovarian cell proliferation, apoptosis, turnover, viability, and steroid hormones release; (2) the plants buckwheat and vitex, as well as rutin and apigenin, can promote some of these ovarian functions too; and (3) these plant additives mitigate the CuNPs/TiO₂’s activity, something that must be considered when applied together. Full article

A novel bio-hybrid drug delivery system was obtained involving a Mg/Al-NO₃ layered double hydroxide (LDH) intercalated either with ibuprofenate anions (IBU) or a phospholipid bilayer (BL) containing a neutral drug, i.e., 17β-estradiol, and then embedded in chitosan beads. The combination of these components in a hierarchical structure led to synergistic effects investigated through characterization of the intermediates and the final bio-composites by XRD, TG, SEM, and TEM. That allowed determining the presence and yield of IBU and of BL in the interlayer space of LDH, and of the encapsulated LDH in the beads, as well as the morphology of the latter. Peculiar attention has been paid to the intercalation process of the BL for which all available data substantiate the hypothesis of a first interaction at the defect of the LDH, as well as on the interaction mode of these components. ¹H, ³¹P and ²⁷Al MAS-NMR studies allowed establishing that the intercalated BL is not homogeneous and likely formed patches. Release kinetics were performed for sodium ibuprofenate as well as for the association of 17β-estradiol within the negatively charged BL, each encapsulated in the LDH/chitosan hybrid materials. Such new bio-hybrids offer an interesting outlook into the pharmaceutical domain with the ability to be used as sustained release systems for a wide variety of anionic and, importantly, neutral drugs. Full article