A novel method for gas mixing and distribution in multi-chamber embryo incubators.

BACKGROUND: High-quality control of the gas environment in incubators is crucial for in vitro embryo development, which requires high accuracy, fast recovery, and low gas consumption. OBJECTIVE: In this study, we propose a novel gas mixing and distribution system and method as an alternative solution for multi-chamber embryo incubators. METHODS: The system-based embryo incubator enables a controllable gas circulation process and a quantitative supply of CO2 and N2. To determine the optimal parameters for the mixing time and flow rate of the circulated gases, we conducted contrast experiments on the system-based incubator. To evaluate the performance of the gas system in the incubator, we conducted tests under four different initial conditions, simulating various practical application scenarios. Furthermore, we performed a mouse embryo assay to assess the system's effectiveness. RESULTS: The results show that the system achieved a gas concentration accuracy of ± 0.2% (volume fraction) after stabilization, a minimum recovery time of 5 minutes, an average consumption of 8.9 L/d for N2 and 0.83 L/d for CO2 during routine operation, and a blastocyst rate exceeding 90% observed after 96 hours of culture in the incubator. CONCLUSION: The system and method demonstrate a significant advantage in terms of low gas consumption compared to existing incubators, while still maintaining high accuracy and fast recovery.

Resveratrol attenuates MPP+-induced mitochondrial dysfunction and cell apoptosis via AKT/GSK-3ß pathway in SN4741 cells.

Oxidative stress and mitochondrial dysfunction play crucial role in the dopaminergic neurons death, which is the pathological feature of Parkinson's disease (PD). Resveratrol (Res), a polyphenol derived from grapes and blueberries, has been reported to reduce oxidative stress injury and to restore mitochondrial function. In this study, we aimed to explore the underlying molecular mechanism of the beneficial effects of Res against MPP+- induced mitochondrial dysfunction and cell apoptosis in SN4741 cells. The data showed that Res significantly alleviated MPP+- induce cytotoxicity and restored MPP+- induced mitochondrial dysfunction in SN4741 cells. Moreover, Res rescued MPP+- induced a decline on the level of p-AKT, p-GSK-3ßand the ratio of Bcl-2/Bax, and an elevation on the expression of Bax and caspase-3, 9. However, inhibition GSK-3ß activity clearly abolished the protective effects of Res. Taken together, these results suggest that Res attenuates MPP+- induced mitochondrial dysfunction and cell apoptosis, and these protections may be achieved through AKT/GSK-3ß pathway. These also indicate that Res could be a promising therapeutic agent for PD.

The Ras/Raf/Erk Pathway Mediates the Subarachnoid Hemorrhage-Induced Apoptosis of Hippocampal Neurons Through Phosphorylation of p53.

Apoptosis plays a crucial role in the pathogenesis of early brain injury (EBI) following subarachnoid hemorrhage (SAH). However, the exact molecular mechanisms underlying neuronal apoptosis in EBI after SAH have not been fully elucidated. The present study showed that EBI induced significantly neuronal apoptosis activation of Ras/Raf/Erk signals in hippocampus after SAH. Intracisternal administration of PD98059, an inhibitor of Erk1/2, decreased the hippocampal neuronal apoptosis and alleviated the cognitive deficits induced by SAH. Interestingly, an increase in phosphorylation of p53 was paralleled with p-Erk, and PD98059 also blocked the level of p-p53. In primary cultures, oxyhemoglobin (OxyHb) treatment significantly increased p-Erk, p-p53, and apoptosis, which was used to mimic the pathological injury of SAH. Both p53 small interfering RNA (siRNA) and PD98059 reduced the OxyHb-induced apoptosis. Moreover, PD98059 significantly decreased the levels of p-Erk and p-p53; however, p53 siRNA had little effect on the level of p-Erk. Taken together, our study implicates that the Ras/Raf/Erk signals contribute to neuronal death through the phosphorylation of p53 in hippocampus after SAH and also suggests Erk/p53 as a potential target for clinical drug treatment of SAH.

Chaperone-mediated autophagy: roles in neuroprotection.

Chaperone-mediated autophagy (CMA), one of the main pathways of lysosomal proteolysis, is characterized by the selective targeting and direct translocation into the lysosomal lumen of substrate proteins containing a targeting motif biochemically related to the pentapeptide KFERQ. Along with the other two lysosomal pathways, macro- and micro-autophagy, CMA is essential for maintaining cellular homeostasis and survival by selectively degrading misfolded, oxidized, or damaged cytosolic proteins. CMA plays an important role in pathologies such as cancer, kidney disorders, and neurodegenerative diseases. Neurons are post-mitotic and highly susceptible to dysfunction of cellular quality-control systems. Maintaining a balance between protein synthesis and degradation is critical for neuronal functions and homeostasis. Recent studies have revealed several new mechanisms by which CMA protects neurons through regulating factors critical for their viability and homeostasis. In the current review, we summarize recent advances in the understanding of the regulation and physiology of CMA with a specific focus on its possible roles in neuroprotection.

Myricitrin alleviates MPP⁺-induced mitochondrial dysfunction in a DJ-1-dependent manner in SN4741 cells.

Oxidative stress and mitochondrial dysfunction have been linked to Parkinson's disease. DJ-1 is a recessive familial PD gene involved in antioxidative function and mitochondrial maintenance. Myricitrin, a flavanoid isolated from the root bark of Myrica cerifera, has potent antioxidative properties. In the present study, we investigated the protective effects of myricitrin against MPP(+)-induced mitochondrial dysfunction in SN4741 cells and attempted to elucidate the mechanisms underlying this protection. The results showed that incubating SN4741 cells with myricitrin significantly reduced cell death induced by the neurotoxin MPP(+). Furthermore, myricitrin protected cells from MPP(+)-induced effects on mitochondrial morphology and function. However, these protective effects were lost under DJ-1-deficient conditions. Thus, our results suggest that myricitrin alleviates MPP(+)-induced mitochondrial dysfunction and increases cell viability via DJ-1, indicating that myricitrin is a potential beneficial agent for age-related neurodegenerative diseases, particularly Parkinson's disease.

Fabrication of graded TiN coatings on nitinol occluders and effects on in vivo nickel release.

A nano-structured TiN/Ti coating with a total thickness of 0.9 mum was deposited on nitinol cardiac occluders using the filtered multi-arc vacuum ion plating technique at less than 300 degrees C. The coating was composed of laminated TiN/Ti layers with thickness of about 100 nm. The cardiac occluders made of a nitinol mesh with and without a graded nano-structured titanium nitride (TiN) coating were implanted into the hearts of rams. The nickel concentration of the whole blood of the animals were measured one week, one month, three months, and six months after implantation and compared to that before operation. The nickel concentration in the neo-endocardium covered occluders was also measured using graphite furnace atomic absorption spectrophotometry. After one week, the nickel content in the blood increased by a factor of three compared to the level before operation and decreased afterwards returning to the normal level after six months when endothelialization was complete. Statistical analyses showed that the TiN coating could mitigate nickel release into blood (P<0.01). For example, the nickel concentration released from the control increased from about 2.65+/-1.20 microg/kg, the normal concentration, to 7.30+/-1.00 microg/kg but just from 2.56+/-1.16 microg/kg to 4.68+/-1.29 microg/kg from the TiN coated occluder after 7 days. The nickel concentration in the neo-endocardium covered and TiN coated occluders reached 17.0+/-8.05 microg/kg in two months after implantation. In comparison, it was 31.0+/-5.72 microg/kg for the occluder without the TiN coating. While normal concentration of nickel in endocardium is also 2.6+/-1.09 microg/kg. Our results demonstrate that the graded TiN coating can significantly reduce nickel release into the endocardium (P<0.01) under in vivo conditions.