Exploring novel function of Gpx5 antioxidant activity: Assisting epididymal cells secrete functional extracellular vesicles.

Glutathione peroxisomal-5 (Gpx5) promotes the elimination of H2O2 or organic hydrogen peroxide, and plays an important role in the physiological process of resistance to oxidative stress (OS). To directly and better understand the protection of Gpx5 against OS in epididymal cells and sperm, we studied its mechanism of antioxidant protection from multiple aspects. To more directly investigate the role of Gpx5 in combating oxidative damage, we started with epididymal tissue morphology and Gpx5 expression profiles in combination with the mouse epididymal epithelial cell line PC1 (proximal caput 1) expressing recombinant Gpx5. The Gpx5 is highly expressed in adult male epididymal caput, and its protein signal can be detected in the sperm of the whole epididymis. Gpx5 has been shown to alleviate OS damage induced by 3-Nitropropionic Acid (3-NPA), including enhancing antioxidant activity, reducing mitochondrial damage, and suppressing cell apoptosis. Gpx5 reduces OS damage in PC1 and maintains the well-functioning extracellular vesicles (EVs) secreted by PC1, and the additional epididymal EVs play a role in the response of sperm to OS damage, including reducing plasma membrane oxidation and death, and increasing sperm motility and sperm-egg binding ability. Our study suggests that GPX5 plays an important role as an antioxidant in the antioxidant processes of epididymal cells and sperm, including plasma membrane oxidation, mitochondrial oxidation, apoptosis, sperm motility, and sperm-egg binding ability.

Analysis of morphology, histology characteristics, and circadian clock gene expression of Onychostoma macrolepis at the overwintering period and the breeding period.

Fish typically adapt to their environment through evolutionary traits, and this adaptive strategy plays a critical role in promoting species diversity. Onychostoma macrolepis is a rare and endangered wild species that exhibits a life history of overwintering in caves and breeding in mountain streams. We analyzed the morphological characteristics, histological structure, and expression of circadian clock genes in O. macrolepis to elucidate its adaptive strategies to environmental changes in this study. The results showed that the relative values of O. macrolepis eye diameter, body height, and caudal peduncle height enlarged significantly during the breeding period. The outer layer of the heart was dense; the ventricular myocardial wall was thickened; the fat was accumulated in the liver cells; the red and white pulp structures of the spleen, renal tubules, and glomeruli were increased; and the goblet cells of the intestine were decreased in the breeding period. In addition, the spermatogenic cyst contained mature sperm, and the ovaries were filled with eggs at various stages of development. Throughout the overwintering period, the melano-macrophage center is located between the spleen and kidney, and the melano-macrophage center in the cytoplasm has the ability to synthesize melanin, and is arranged in clusters to form cell clusters or white pulp scattered in it. Circadian clock genes were identified in all organs, exhibiting significant differences between the before/after overwintering period and the breeding period. These findings indicate that the environment plays an important role in shaping the behavior of O. macrolepis, helping the animals to build self-defense mechanisms during cyclical habitat changes. Studying the morphological, histological structure and circadian clock gene expression of O. macrolepis during the overwintering and breeding periods is beneficial for understanding its unique hibernation behavior in caves. Additionally, it provides an excellent biological sample for investigating the environmental adaptability of atypical cavefish species.

Exosomes from porcine serum as endogenous additive maintain function of boar sperm during liquid preservation at 17 °C in vitro.

The supplementation of sperm culture media with serum is quite common, and improves both sperm survival and motility. However, the link between serum and sperm remains poorly understood. The present study is the first investigation of the effects on sperm quality and function of endogenous porcine serum exosomes in medium used for culturing boar sperm. Scanning electron microscopy (SEM) confirmed that serum-derived exosomes from both castrated boars (cbsExos) and sows (ssExos) exhibited typical nanostructural morphology and expressed CD63, CD9, and Alix, as shown by Western blotting. At 17 °C, the progressive motility and membrane integrity of sperm were significantly increased after incubation of fresh boar semen for 7 days with cbsExos-4 (8 × 1010 particles/mL) or ssExos-16 (32 × 1010 particles/mL). Moreover, cbsExos-4 and ssExos-16 were found to be effective sperm additives, improving mitochondrial transmembrane potential (ΔΨm) and adenosine triphosphate (ATP) content, total antioxidant activity (T-AOC), superoxide dismutase (SOD) activity, and glutathione peroxidase (GPx) activity while reducing reactive oxygen species (ROS) levels, and malondialdehyde (MDA) content following preservation at 17 °C after a 5-day incubation. Both fluorescence and SEM showed that the serum exosomes bound directly to the sperm membrane, suggesting an interaction that could influence sperm-zona pellucida binding. Overall, this study provides new insights into the potential benefits of adding cbsExos and ssExos to enhance the quality of boar sperm during ambient temperature preservation, which may lead to advancements in sperm preservation strategies.

Selenium nanoparticles in aquaculture: Unique advantages in the production of Se-enriched grass carp (Ctenopharyngodon idella).

The production of selenium-enriched fish can contribute to alleviating selenium deficiency in human diets. However, it is still unclear which selenium source, as an additive, can efficiently and cost-effectively produce high-quality selenium-enriched fish. This study evaluated the effects of selenium nanoparticles (SeNP), selenite, and selenomethionine (SeMet) on the growth, antioxidant capacity, selenium content, selenium speciation, and meat quality of grass carp. Ten diets were prepared, including a basal diet (BD) and three concentrations (0.1, 0.3, and 0.9 mg/kg) of SeNP, selenite, and SeMet. A total of 600 fish (250.79 ± 1.57 g) were randomly assigned to 30 tanks (3 tanks/group). Fish were fed the experimental diet three times daily for 60 d. In this study, SeNP most significantly promoted the growth and antioxidant capacity of grass carp, with 0.3 mg/kg SeNP identified as the optimal additive concentration. Additionally, SeNP demonstrated equally excellent bioavailability as SeMet and significantly increased the content of SeMet in grass carp (Ctenopharyngodon idella) muscle. Furthermore, compared to SeMet and selenite, dietary SeNP could more significantly enhance the content of selenocysteine (SeCys2) and methylselenocysteine (MeSeCys) in grass carp muscle tissue. In addition, we have demonstrated that SeCys2 and MeSeCys promote apoptosis of cancer cells (HeLa) through the mitochondrial apoptotic pathway (involving Bax and Bcl-2). Furthermore, as an additive, 0.3 mg/kg SeNP significantly improved the flesh quality of grass carp by reducing crude fat and heavy metal content, as well as increasing the levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and the ratio of n-3/n-6 polyunsaturated fatty acid (PUFA). In summary, SeNP is the most suitable additive for producing selenium-enriched fish.

Selenium nanoparticles improve fish sperm quality by enhancing glucose uptake capacity via AMPK activation.

Appropriate additives can provide a suitable physiological environment for storage of fish sperm and facilitate the large-scale breeding of endangered species and commercial fish. Suitable additives for fish sperm storage in vitro are required for artificial insemination. This study evaluate the effects of 0.1, 0.5, 1.5, and 4.5 mg/L selenium nanoparticles (SeNPs) on the quality of Schizothorax prenanti and Onychostoma macrolepis sperm storage in vitro at 4 °C for 72 h. We found that 0.5 mg/L SeNPs was a suitable concentration for maintaining the normal physiological state of O. macrolepis sperm during storage at 4 °C (p < 0.05). Higher adenosine triphosphate (ATP) content of O. macrolepis sperm before and after activation was present at that concentration. To further explore the potential mechanism of action of SeNPs on O. macrolepis sperm, western blotting and glucose uptake analyses were performed. The results implied that after 24 h of in vitro preservation, 0.5 mg/L SeNPs significantly improved p-AMPK levels and glucose uptake capacity of O. macrolepis sperm, while compound C (CC), the inhibitor of activated AMP-activated protein kinase (p-AMPK), significantly restricted the function of SeNPs on stored sperm. Similar effects of 0.5 mg/L SeNPs were found on Schizothorax prenanti sperm. Our study demonstrates that SeNPs maintained ATP content and O. macrolepis and Schizothorax prenanti sperm function during storage in vitro for 72 h, possibly because SeNPs enhanced the glucose uptake capacity of sperm by maintaining the level of p-AMPK.

Metformin improves fish sperm quality by regulating glucose uptake capacity during in vitro storage.

A suitable additive for fish sperm storage in vitro is necessary for artificial reproduction. In this study, we evaluated the effects of different concentrations (100, 200, 400, and 800 µmol/L) of metformin (Met) on Schizothorax prenanti and Onychostoma macrolepis sperm under storage in vitro for 72 h. Compared with the control group, 400 µmol/L Met was more effective at improving the quality and fertilization capacity of S. prenanti sperm by increasing the adenosine triphosphate (ATP) content within the sperm. Further study found that Met stabilized the ATP level by enhancing the glucose uptake in S. prenanti sperm, and this effect might be associated with the activation of AMP-activated protein kinase (AMPK) in sperm. In this study, we also found that glucose could be absorbed by the sperm of S. prenanti, which was mainly accumulated in the midpiece of S. prenanti sperm, where mitochondria were located. In addition, Compound C significantly inhibited the beneficial effects of Met on the quality and glucose uptake capacity of S. prenanti sperm by inhibiting AMPK phosphorylation. These results revealed that AMPK played an important role in vitro sperm storage, and Met maintained ATP content and increased the storage time of S. prenanti sperm in vitro for 72 h, possibly due to Met enhanced glucose uptake capacity of sperm by activating AMPK. Similarly, the beneficial effects of Met on S. prenanti sperm were also found in O. macrolepis sperm, suggesting that Met may hold great promise for the practice of storing fish in vitro.

Generally, what matters most to fish sperm is a good storage condition in vitro that can both ensure its high motility after fresh water activation and avoid the damage of the nonactivated state. An appropriate additive can provide suitable storage conditions for sperm. Therefore, a suitable additive of fish sperm is necessary for fish to promote effective artificial reproduction. In this study, we evaluated the effects of different concentrations of metformin (Met) on fish sperm under storage in vitro. Compared with the control group, Met was more effective at improving the quality and fertilization capacity of fish by increasing the adenosine triphosphate (ATP) content within the sperm. Further study found that Met stabilized the ATP level by enhancing the glucose uptake in fish sperm, and this effect might be associated with the activation of AMP-activated protein kinase (AMPK) in sperm. The beneficial effects of Met on fish sperm suggest that Met may hold great promise for the practice of storing fish in future vitro storage.

Linking self-disgust, negative affect, and PTSD in sexual assault: An ecological momentary assessment approach.

OBJECTIVE: Although self-disgust has been implicated in sexual assault-related posttraumatic stress disorder (PTSD), empirical evidence of this association is limited. METHOD: Participants with sexual assault-related PTSD (n = 19), sexual assault without PTSD (n = 20), and healthy controls (n = 21) completed "trait" measures of disgust proneness, self-disgust, and negative affect (NA; i.e., anxiety and depression). An ecological momentary assessment approach was also employed, where participants reported "state" levels of PTSD symptoms, self-disgust, and NA in the morning, afternoon, and evening for 1 week. RESULTS: Those with PTSD reported more trait disgust proneness, self-disgust, and NA than those who experienced sexual assault without PTSD and controls. However, those experiencing sexual assault without PTSD and controls did not differ from each other. Participants with a history of sexual assault also experienced higher self-disgust and NA during the week than controls. Those with higher PTSD symptoms during the week also experienced more self-disgust and NA. Similarly, changes in PTSD symptoms were associated with changes in self-disgust and NA during the week. Although a temporal association was found where NA predicted subsequent PTSD symptoms (and vice versa) during the week, a temporal link between self-disgust and PTSD symptoms (or vice versa) was not found. CONCLUSIONS: These findings suggest that the link between self-disgust and sexual assault-related PTSD is more correlational than causal. The implications of these findings for understanding how trait and state self-disgust fits within the broader emotional network of sexual assault-related PTSD are discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Inhibition of breast cancer cell survival by Xanthohumol via modulation of the Notch signaling pathway in vivo and in vitro.

Natural compounds derived from plants have been an important source of numerous clinically useful anticancer agents. Nevertheless, limited studies indicate that xanthohumol (XN), a major prenylated flavonoid in hop plants (Humulus lupulus), may possess anticarcinogenic properties. The purpose of the present study was to clarify the antitumorigenic effects and the underlying mechanism of XN on breast cancer in vivo and in vitro. A 4T1 breast tumor mouse model was used in the present study to investigate XN suppression of tumor growth as detected by tumorigenicity assays in vivo. In addition, in vitro studies revealed that XN significantly decreased cell viability, induced G0/G1 cell cycle arrest and apoptosis in MCF-7 and MDA-MB-231 cells, as confirmed by an MTT assay, flow cytometry and western blot analysis, indicating anticarcinogenic activity of XN against breast cancer. Furthermore, immunohistochemistry was performed to confirm the inactivation of the Notch signaling pathway, Notch 1 and Ki-67, in vivo; consistently, XN caused decreased activation of the Notch signaling pathway and apoptotic regulators B-cell lymphoma-2 (Bcl-2), Bcl-extra large and caspase 3, as determined by western blot analysis in vitro. This study suggests that XN may potentially be useful as a chemopreventive agent during breast hyperplasia and carcinogenesis, acting via the regulation of Notch associated apoptotic regulators in vivo and in vitro.

Feasibility of ultrasound-guided capsule-sheath space block combined with anterior cervical cutaneous nerves block for thyroidectomy: an observational pilot study.

BACKGROUND: We evaluated the efficacy of a new anesthetic technique termed ultrasound-guided capsule-sheath space block (CSSB) combined with anterior cervical cutaneous nerve block (CCNB) for thyroidectomy. METHODS: The study included two parts: Part one was an imaging study to determine technique feasibility. The CSSB was performed on five healthy volunteers by introducing the needle 0.5 cm lateral to the probe under in-plane needle ultrasound guidance. After puncture of the false capsule and its subsequent contraction with the true capsule of thyroid, 10 mL of contrast medium was deposited slowly in the capsule-sheath space. The CCNB was performed bilaterally as follows: Under ultrasound guidance, a subcutaneous injection was made along the sternocleidomastoid using 10 mL of contrast medium which was followed by a girdle-shaped picchu raised from the cricoid cartilage to supraclavicular region. The spreading pattern of contrast medium was imaged using computed tomographic scanning. In part two (a clinical case series) the technique efficacy was evaluated. Seventy-eight patients undergoing thyroidectomy had ultrasound-guided CSSB and CCNB with local anesthetics. The sensory onset of CCNB, intraoperative hemodynamic parameters, and analgesic effect were assessed and complications were noted. RESULTS: The distribution of contrast medium was well defined. In part two the onset time of CCNB was 2.2 ± 0.7 min, and the hemodynamic parameters remained stable intraoperatively. The recall of visual analogue scale scores during surgery was 2 [1-4] for median (range). The patients' and surgeons' satisfaction scores were 2 [1-4] and 1 [1-3] for median (range). No serious complications occurred. CONCLUSIONS: Combining ultrasound-guided CSSB and CCNB is a feasible, effective and safe technique for thyroidectomy. TRIAL REGISTRATION: Current Controlled Trials ChiCTR-ONC-12002025. Registered 19 March 2012.