PINK1/Parkin pathway-mediated mitophagy by AS-IV to explore the molecular mechanism of muscle cell damage.

BACKGROUND: Functional disorders of mitochondria are closely related to muscle diseases. Many studies have also shown that oxidative stress can stimulate the production of a large number of reactive oxygen species (ROS), which have various adverse effects on mitochondria and can damage muscle cells. PURPOSE: In this study, based on our previous research, we focused on the PINK1/Parkin pathway to explore the mechanism by which AS-IV alleviates muscle injury by inhibiting excessive mitophagy. METHODS: L6 myoblasts were treated with AS-IV after stimulation with hydrogen peroxide (H2O2) and carbonyl cyanide m-chlorophenylhydrazone (CCCP). Then, we detected the related indices of oxidative stress and mitophagy by different methods. A PINK1 knockdown cell line was established by lentiviral infection to obtain further evidence that AS-IV reduces mitochondrial damage through PINK1/Parkin. RESULTS: After mitochondrial damage, the expression of malondialdehyde (MDA) and intracellular ROS in L6 myoblasts significantly increased, while the expression of superoxide dismutase (SOD) and ATP decreased. The mRNA and protein expression levels of Tom20 and Tim23 were decreased, while those of VDAC1 were increased. PINK1, Parkin, and LC3 II mRNA and protein expression increased, and P62 mRNA and protein expression decreased·H2O2 combined with CCCP strongly activated the mitophagy pathway and impaired mitochondrial function. However, abnormal expression of these factors could be reversed after treatment with AS-IV, and excessive mitochondrial autophagy could also be reversed, thus restoring the regulatory function of mitochondria. However, AS-IV-adjusted function was resisted after PINK1 knockdown. CONCLUSION: AS-IV is a potential drug for myasthenia gravis (MG), and its treatment mechanism is related to mediating mitophagy and restoring mitochondrial function through the PINK1/Parkin pathway.

Study on microbial community of "green-covering" Tuqu and the effect of fortified autochthonous Monascus purpureus on the flavor components of light-aroma-type Baijiu.

"Green-covering" Tuqu (TQ), as one of Xiaoqu, is a special fermentative starter (also known as Jiuqu in Chinese) that originated in southern China and is characterized by a layer of green mold covering (Aspergillus clavatus) the surface and (sometimes) with a red heart. It plays a vital role in producing light-aroma-type Baijiu (LATB). However, to date, the microbiota that causes red heart of TQ remain largely unexplored, and it is still unclear how these microbiota influence on the quality of LATB. In this study, two types of TQ, one with a red heart (RH) and another with a non-red heart (NRH), were investigated by high throughput sequencing (HTS) and directional screening of culture-dependent methods. The obtained results revealed the differences in the microbial communities of different TQ and led to the isolation of two species of Monascus. Interestingly, the results of high performance liquid chromatography (HPLC) detection showed that citrinin was not detected, indicating that Monascus isolated from TQ was no safety risk, and the contents of gamma-aminobutyric acid in the fermented grains of RH were higher than that of NRH during the fermentation. Selecting the superior autochthonous Monascus (M1) isolated from the TQ to reinoculate into the TQ-making process, established a stable method for producing the experimental "red heart" Tuqu (ERH), which confirmed that the cause of "red heart" was the growth of Monascus strains. After the lab-scale production test, ERH increased ethyl ester production and reduced higher alcohols production. In addition, Monascus had an inhibitory effect on the growth of Saccharomyces and Aspergillus. This study provides the safe, health-beneficial, and superior fermentation strains and strategies for improving the quality of TQ and LATB.

Mitochondrial dynamics and biogenesis indicators may serve as potential biomarkers for diagnosis of myasthenia gravis.

Due to challenges in diagnosing myasthenia gravis (MG), identifying novel diagnostic biomarkers for this disease is essential. Mitochondria are key organelles that regulate multiple physiological functions, such as energy production, cell proliferation and cell death. In the present study, Mfn1/2, Opa1, Drp1, Fis1, AMPK, PGC-1α, NRF-1 and TFAM were compared between patients with MG and healthy subjects to identify potential diagnostic biomarkers for MG. Blood samples were collected from 50 patients with MG and 50 healthy subjects. The participants' demographic information and routine blood test results were recorded. Mitochondrial dynamics were evaluated and levels of Mfn1/2, Opa1, Drp1, Fis1, AMPK, PGC-1α, NRF-1 and TFAM were determined in peripheral blood mononuclear cells using western blotting and reverse transcription-quantitative PCR, respectively. Receiver operating characteristic curve analysis was used to evaluate the diagnostic accuracy of these indicators. The areas under the curve values of Mfn1/2, Opa1, Drp1, Fis1,AMPK, PGC-1α, NRF-1 and TFAM were 0.5408-0.8696. Compared with control subjects, mRNA expression levels of Mfn1/2, Opa1, AMPK, PGC-1α, NRF-1 and TFAM were lower, while those of Drp1 and Fis1 were higher in patients with MG. The protein expression levels of all these molecules were lower in patients with MG than in control subjects. These results suggested that mitochondrial dynamics and biogenesis indicators may be diagnostic biomarkers for MG.

Bioaccessibility and Bioavailability of (-)-Epigallocatechin Gallate in the Bread Matrix with Glycemic Reduction.

Bread has a high glycemic index (GI) and rich contents of quickly digestible carbohydrates, which is associated with insulin resistance and the risk of chronic diseases. (-)-Epigallocatechin Gallate (EGCG) is the primary catechin component that inhibits starch hydrolases, while the low release and absorption rates limit its utilization. In this study, EGCG was added to the bread matrix for fortification to reduce its glycemic index compared to white bread. EGCG fortification at 4% decreased the starch digestion rate of baked bread by 24.43% compared to unfortified bread and by 14.31% compared to white bread, with an identical amount of EGCG outside the matrix. Moreover, the predicted GI (pGI) was reduced by 13.17% compared to white bread. Further, 4% EGCG-matched bread enhanced the bioaccessibility and bioavailability of EGCG by 40.38% and 47.11%, respectively, compared to the control. The results of molecular docking demonstrated that EGCG had a higher binding affinity with α-amylase than with α-glucosidase, indicating that EGCG may effectively inhibit the accumulation of carbs during starch digestion. Thus, EGCG can be used as a functional ingredient in bread to reduce its glycemic potential, and the bread matrix can be used as a carrier for EGCG delivery to enhance its bioaccessibility and bioavailability.

Qiangji Jianli Decoction Alleviates Hydrogen Peroxide-Induced Mitochondrial Dysfunction via Regulating Mitochondrial Dynamics and Biogenesis in L6 Myoblasts.

Oxidative stress can cause the excessive generation of reactive oxygen species (ROS) and has various adverse effects on muscular mitochondria. Qiangji Jianli decoction (QJJLD) is an effective traditional Chinese medicine (TCM) that is widely applied to improve muscle weakness, and it has active constituents that prevent mitochondrial dysfunction. To investigate the protective mechanism of QJJLD against hydrogen peroxide- (H2O2-) mediated mitochondrial dysfunction in L6 myoblasts. Cell viability was determined with MTT assay. Mitochondrial ultrastructure was detected by transmission electron microscope (TEM). ROS and mitochondrial membrane potential (MMP) were analyzed by fluorescence microscope and flow cytometry. The superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity, and malondialdehyde (MDA) level were determined by WST-1, TBA, and DTNB methods, respectively. The mRNA and protein levels were measured by quantitative real-time PCR (qRT-PCR) and Western blot. The cell viability was decreased, and the cellular ROS level was increased when L6 myoblasts were exposed to H2O2. After treatment with QJJLD-containing serum, the SOD and GSH-Px activities were increased. MDA level was decreased concurrently. ROS level was decreased while respiratory chain complex activity and ATP content were increased in L6 myoblasts. MMP loss was attenuated. Mitochondrial ultrastructure was also improved. Simultaneously, the protein expressions of p-AMPK, PGC-1α, NRF1, and TFAM were upregulated. The mRNA and protein expressions of Mfn1/2 and Opa1 were also upregulated while Drp1 and Fis1 were downregulated. These results suggest that QJJLD may alleviate mitochondrial dysfunction through the regulation of mitochondrial dynamics and biogenesis, the inhibition of ROS generation, and the promotion of mitochondrial energy metabolism.

Qiangji Jianli Decoction promotes mitochondrial biogenesis in skeletal muscle of myasthenia gravis rats via AMPK/PGC-1α signaling pathway.

The Qiangji Jianli Decoction (QJJLD) is an effective Chinese medicine formula for treating Myasthenia gravis (MG) in the clinic. QJJLD has been proven to regulate mitochondrial fusion and fission of skeletal muscle in myasthenia gravis. In this study, we investigated whether QJJLD plays a therapeutic role in regulating mitochondrial biogenesis in MG and explored the underlying mechanism. Rats were experimentally induced to establish autoimmune myasthenia gravis (EAMG) by subcutaneous immunization with R97-116 peptides. The treatment groups were administered three different dosages of QJJLD respectively. After the intervention of QJJLD, the pathological changes of gastrocnemius muscle in MG rats were significantly improved; SOD, GSH-Px, Na+-K+ ATPase and Ca2+-Mg2+ ATPase activities were increased; and MDA content was decreased in the gastrocnemius muscle. Moreover, AMPK, p38MAPK, PGC-1α, NRF-1, Tfam and COX IV mRNA and protein expression levels were also reversed by QJJLD. These results implied that QJJLD may provide a potential therapeutic strategy through promoting mitochondrial biogenesis to alleviate MG via activating the AMPK/PGC-1α signaling pathway.

Alterations in vaginal temperature during the estrous cycle in dairy cows detected by a new intravaginal device-a pilot study.

Estrus identification is important in dairy cow production. At present, estrus identification is automated with a pedometer or accelerometer and the results remain unsatisfactory. It was previously reported that body temperature changes during estrus. In the present study, dairy cow vaginal temperature (VT) was monitored during various seasons, and an increase in VT of 0.3 °C was suggested for the onset of estrus, using an automated VT monitoring system developed in-house. Natural and synchronized estrus were measured simultaneously. The VT was determined to be in circadian rhythm and significantly higher in summer than in either autumn or winter (P < 0.05). VT difference (between estrus VT and average VT 7 days earlier) gradually increased, reached a peak of 0.56 °C ± 0.17 at 4 h before the end of estrus, and then decreased to the normal. The VT of cows in estrus and the duration of their estrus were significantly affected by seasons and estrus types (P < 0.05). VT gradually decreased in response to prostaglandin (PG) injection and was significantly lower (0.15-0.35 °C) from 9 to 33 h after the drug administration than the average VT at the same time 7 days earlier (P < 0.05). Changes in circadian and seasonal VT and in the estrous cycle can be monitored to assess the physiological status of cows and will help in developing an effective automated estrus identification technique. Results of this pilot study should be validated in further studies.

Early pregnancy diagnoses based on physiological indexes of dairy cattle: a review.

With the development of standardization and scaling in the dairy farming industry, timely and accurate pregnancy diagnosis is required to improve the benefits of breeding by shortening the calving interval. However, the current pregnancy diagnostic methods cannot meet the requirements of the industry. Here, we review changes in the physiological indexes and in the function and morphological status of the reproductive organs of dairy cows at the early stages of pregnancy. In addition, the corresponding pregnancy diagnostic methods based on certain indexes are well development, and the pregnancy diagnostic approaches based on remote sensing and automation technology have become inevitable trends in the industry. These applications will reveal physiological regularities in pregnancy and benefit the detailed management of dairy cows during early pregnancy.

Ultrasound Treatment on Stability of Total and Individual Anthocyanin Extraction from Blueberry Pomace: Optimization and Comparison.

Blueberry pomace is abundant in anthocyanins. This work characterized the anthocyanins in blueberry pomace, discussed the stability of anthocyanins under ultrasound treatment, and compared the extraction conditions for different anthocyanin compositions. Thirteen anthocyanins were identified, and malvidin-3-galactoside (18.56%), which represented the most abundant anthocyanin, was selected as the individual analyte. The general linear model univariate analysis revealed that ultrasound-assisted extraction (UAE) resulted in higher recoveries of both total anthocyanins (TA) and individual anthocyanins (IA) when compared with conventional solvent extraction. The optimized extraction conditions for TA and IA were UAE in pure methanol (12.49 mg/g dry weight) at 25 °C for 30 min and UAE in 70% ethanol (3.57 mg/g dry weight) at 40 °C for 40 min, respectively. Moreover, IA was more vulnerable to degradation compared with TA. Therefore, a specific extraction process of IA is significant for monomer preparation, and harsh conditions should be avoided in UAE.

Effects of heat stress on body temperature, milk production, and reproduction in dairy cows: a novel idea for monitoring and evaluation of heat stress ­ A review

Heat stress exerts a substantial effect on dairy production. The temperature and humidity index (THI) is widely used to assess heat stress in dairy operations. Herein, we review the effects of high temperature and humidity on body temperature, feed intake, milk production, follicle development, estrous behavior, and pregnancy in dairy cows. Analyses of the effects of THI on dairy production have shown that body temperature is an important physiological parameter in the evaluation of the health state of dairy cows. Although THI is an important environmental index and can help to infer the degree of heat stress, it does not reflect the physiological changes experienced by dairy cows undergoing heat stress. However, the simultaneous measurement of THI and physiological indexes (e.g., body temperature) would be very useful for improving dairy production. The successful development of automatic detection techniques makes it possible to combine THI with other physiological indexes (i.e., body temperature and activity), which could help us to comprehensively evaluate heat stress in dairy cows and provide important technical support to effectively prevent heat stress.

Chemical and Cellular Assays Combined with In Vitro Digestion to Determine the Antioxidant Activity of Flavonoids from Chinese Bayberry (Myrica rubra Sieb. et Zucc.) Leaves.

Oxidative stress is highly associated with the development of cardiovascular diseases and cancer and has drawn great attention. Natural products suggest a potential role in prevention of these disorders. The aim of this study was to investigate the antioxidant and anti-cancer properties of Chinese bayberry leaves (Myrica rubra Sieb. et Zucc.) flavonoids (BLF) comprehensively through the combination of in vitro digestion, chemical and cellular antioxidant assays. Based on the LC/MS data, the major flavonoids of BLF were myricitrin and quercetin 3-rhamnoside. BLF owned strong chemical and cellular antioxidant activity (CAA) with its CAA value at 4253.884 ± 435.366 µmol of QE/100 g DW. After the in vitro digestion, the total flavonoids content, myricitrin and quercetin 3-rhamnoside decreased significantly (P < 0.05). Lower levels of the total flavonoid content and cellular uptake of myricitrin and quercetin 3-rhamnoside might contribute to the lower CAA value of digested BLF (DBLF). However, DBLF still owns considerable chemical antioxidant activities and CAA compared with many plants. Furthermore, both BLF and DBLF exhibited dose-dependent relationship against HepG2 proliferation. Taken together, BLF has a great potential to be developed as a natural antioxidant for promoting public health.