Buckwheat hulls, generally discarded as waste, have been known to possess various flavonoids and high antioxidant activities. The objective of this study was to determine effect of extracting solvents [water, ethanol (20%, 50%, 80%, and 100%), methanol, and acetone] on total phenolic content, flavonoid content and composition, and antioxidant activities of common and tartary buckwheat hull extracts. Antioxidative effect of common and tartary buckwheat hull extracts on lipids in mayonnaise was also investigated. Vitexin, isovitexin, isoorientin, orientin, rutin, isoquercetin, and quercetin were identified in the common buckwheat hull extracts, while rutin, quercetin, isoorientin, and isoquercetin were in the tartary buckwheat hull extracts. The methanol and 80% ethanol extracts had more flavonoids than the others, while the aqueous ethanol extracts from both of the hulls had more total phenolics and antioxidant activities. Oxidative stability of lipids in mayonnaises added with common and tartary buckwheat hull extracts (0.02 and 0.08%, w/w) prepared by 50% ethanol were higher than that in the mayonnaise with butylated hydroxytoluene (0.02%) and control. Oxidative stability was not significantly different between the mayonnaises added with the two buckwheat hull extracts.

Chaga mushroom (Inonotus obliquus) has been known to have antioxidant, anti-virus, anti-inflammatory, and anti-tumor effects due to its bioactive compounds such as β-glucans, triterpenoids, phenolic compounds, and melanin complexes. This mushroom has been used as a folk remedy since the 16th century and mainly consumed as extracted. Hot water extraction, which is a traditional method preparing chaga mushroom extract, has limitations requiring a large amount of solvent and long extraction time, resulting in degradation and coagulation of bioactive compounds. The objective of this study was to develop efficient methods extracting bioactive compounds in chaga mushroom, compared with hot water extraction. High II temperature and pressure, enzyme, and ultrasound extractions were used as non-traditional extraction methods. Chaga mushroom extracts were prepared under conditions determined by preliminary experiments. β-Glucans were higher in high temperature and pressure and enzyme extracts than in hot water extract. Total triterpenoids were higher in the enzyme and ultrasound extracts than in the hot water extract. Total phenolic compounds were higher in the high temperature and pressure extract than in the hot water extract. Major phenolic compounds in the chaga mushroom extracts, which were identified by HPLC, LC-MS, and GC-MS, were 3,4-dihydroxybenzaldehyde and coniferyl aldehyde, which were the highest in the high temperature and pressure and enzyme extracts, respectively. Antioxidant activities were the highest in the high temperature and pressure extract which had the highest content of phenolic compounds. In conclusion, high temperature and pressure and enzyme extractions may be efficient methods to produce chaga mushroom extracts with a large amount of bioactive compounds.

Choline is converted to trimethylamine by gut microbiota and further oxidized to trimethylamine-N-oxide (TMAO) by hepatic flavin monooxygenases. Positive correlation between TMAO and chronic diseases has been reported. Polyphenols in black raspberry (BR), especially anthocyanins, possess various biological activities. The objective of this study was to determine the effects of BR extract on the level of choline-derived metabolites, serum lipid profile, and inflammation markers in rats fed high-fat and high-choline diets. Forty female Sprague-Dawley (SD) rats were randomly divided into four groups and fed for 8 weeks as follows: CON (AIN-93G diet), HF (high-fat diet), HFC (HF + 1.5% choline water), and HFCB (HFC + 0.6% BR extract). Serum levels of TMAO, total cholesterol, and low-density lipoprotein (LDL)-cholesterol and cecal trimethylamine (TMA) level were significantly higher in the HFC than in the HFCB. BR extract decreased mRNA expression of pro-inflammatory genes including nuclear factor-κB (NF-κB), interleukin (IL)-1β, IL-6, and cyclooxygenase-2 (COX-2), and protein expression of NF-κB and COX-2 in liver tissue. These results suggest that consistent intake of BR extract might alleviate hypercholesterolemia and hepatic inflammation induced by excessive choline with a high-fat diet via lowering elevated levels of cecal TMA and serum TMAO in rats.

Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison to mammalian cells. Here, we established and characterized a continuous Atlantic mackerel (Scomber scombrus) skeletal muscle cell line (“Mack” cells). The cells were isolated from muscle biopsies of fresh-caught fish, with separate isolations performed from two distinct fish. Mack1 cells (cells from the first isolation) were cultured for over a year and subcultured over 130 times. The cells proliferated at initial doubling times of 63.9 h (± 19.1 SD). After a spontaneous immortalization crisis from passages 37–43, the cells proliferated at doubling times of 24.3 h (± 4.91 SD). A muscle phenotype was confirmed through characterization of muscle stemness and differentiation via paired-box protein 7 and myosin heavy chain immunostaining, respectively. An adipocyte-like phenotype was also demonstrated for the cells through lipid accumulation, confirmed via Oil Red O staining and quantification of neutral lipids. New qPCR primers (HPRT, PAX3B, MYOD1, MYOG, TNNT3A, and PPARG) were tailored to the mackerel genome and used to characterize mackerel cell genotypes. This work provides the first spontaneously immortalized fish muscle cell line for research, ideally serving as a reference for subsequent investigation.

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