Optimal conditions for beef tenderization through radiofrequency heating with cold air.

High-temperature (15-37°C) aging can shorten the tenderizing time of beef; however, the use of constant temperature heating can lead to microbial spoilage. This study tested radiofrequency (RF) tenderization (RF-T) to find the appropriate conditions for the aging-like effect of beef without microbial spoilage. After subjecting beef to 22 h RF-T with four different cooling temperatures (15, 5, -10, and -20°C), the proliferated aerobic bacteria on the surface showed a concentration of 6-6.2 log CFU/g at -10 and -20°C, lower than 7-7.5 log CFU/g at 15 and 5°C. When beef was treated with 25 W/kg RF heating power for 48 h RF-T, the estimated reduction rate of the sliced shear force (SSF) and the increase rate of glutamic acid based on the weight before RF-T were 22.6% and 1.51-fold, which were greater than 19.6% and 1.37-fold with 20 W/kg, and 11.0% and 1.11-fold with 15 W/kg. The optimal specific RF heating power was calculated as 30 W/kg from the results' extrapolation. When processed for 48 h under optimal conditions (30 W/kg specific RF heating power, -20°C cooling air), the tenderization rate and the increased rates of free amino acids based on the weight before RF-T of beef reached over 20% and 1.5-fold with 5.22 log CFU/g aerobic bacteria, which was lesser than the Korean regulation value of 6.7 log CFU/g (5 × 106  CFU/g). Therefore, RF-T could be proposed as a promising high-temperature tenderization method with lowered risk of microbial spoilage. PRACTICAL APPLICATION: We showed that lowering the chamber temperature during RF-T was effective in surface drying and inhibiting aerobic bacteria. RF-T for 24-48 h with 30 W/kg specific RF heating power had an aging-like effect given tenderization and increase in FAAs. Moreover, by providing the matching circuit and impedance during RF-T, this method could be industrially reproducible.

A necessary role of DNMT3A in endurance exercise by suppressing ALDH1L1-mediated oxidative stress.

Exercise can alter the skeletal muscle DNA methylome, yet little is known about the role of the DNA methylation machinery in exercise capacity. Here, we show that DNMT3A expression in oxidative red muscle increases greatly following a bout of endurance exercise. Muscle-specific Dnmt3a knockout mice have reduced tolerance to endurance exercise, accompanied by reduction in oxidative capacity and mitochondrial respiration. Moreover, Dnmt3a-deficient muscle overproduces reactive oxygen species (ROS), the major contributors to muscle dysfunction. Mechanistically, we show that DNMT3A suppresses the Aldh1l1 transcription by binding to its promoter region, altering its epigenetic profile. Forced expression of ALDH1L1 elevates NADPH levels, which results in overproduction of ROS by the action of NADPH oxidase complex, ultimately resulting in mitochondrial defects in myotubes. Thus, inhibition of ALDH1L1 pathway can rescue oxidative stress and mitochondrial dysfunction from Dnmt3a deficiency in myotubes. Finally, we show that in vivo knockdown of Aldh1l1 largely rescues exercise intolerance in Dnmt3a-deficient mice. Together, we establish that DNMT3A in skeletal muscle plays a pivotal role in endurance exercise by controlling intracellular oxidative stress.

Supercooling as a potentially improved storage option for commercial kimchi.

The supercooling degree (SD), which refers to the difference between the ice nucleation temperature and freezing point of kimchi, varies depending on the type of kimchi, manufacturer, recipe, and manufacturing season. The aim of this study is to investigate the major influencing factors for the supercooled storage of kimchi and to analyze the possibility of supercooled storage for commercial kimchi. Pearson correlation analysis determined that, in commercial kimchi manufactured between March and July 2018, the SD of kimchi correlated to the number of aerobic bacteria (P < 0.01), however, was not associated with lactic acid bacteria. Moreover, the ice nucleation temperature of saline solution inoculated with aerobic bacteria was reduced from -3.03 ± 0.04 to -6.18 ± 0.11 °C by 10 kGy gamma ray sterilization. Meanwhile, the ice nucleation temperatures of 1.8 kg of commercial red cabbage kimchi and 500 g of white cabbage kimchi manufactured in February 2020 were -3.93 ± 0.06 °C and -3.57 ± 0.06 °C, respectively, and they could be stored at -2.5 °C for 12 weeks without freezing. Additionally, supercooled storage of kimchi at -2.5 °C caused a fermentation delay effect compared to control storage at 1 °C, considering the acidity and amount of lactic acid bacteria. Therefore, if the number of aerobic bacteria is controlled during the manufacturing process of kimchi, supercooled storage at temperatures below -2.5 °C may extend the shelf life of kimchi. PRACTICAL APPLICATION: We have shown that aerobic bacteria are the key influencing factor for ice nucleation of kimchi during supercooled storage. Aside from the initial sterilization process, fermentation of kimchi can also be delayed by lowering the storage temperature below -2.5 °C. Moreover, the method of direct cool refrigeration may have an industrial-level application.

TET1 is a beige adipocyte-selective epigenetic suppressor of thermogenesis.

It has been suggested that beige fat thermogenesis is tightly controlled by epigenetic regulators that sense environmental cues such as temperature. Here, we report that subcutaneous adipose expression of the DNA demethylase TET1 is suppressed by cold and other stimulators of beige adipocyte thermogenesis. TET1 acts as an autonomous repressor of key thermogenic genes, including Ucp1 and Ppargc1a, in beige adipocytes. Adipose-selective Tet1 knockout mice generated by using Fabp4-Cre improves cold tolerance and increases energy expenditure and protects against diet-induced obesity and insulin resistance. Moreover, the suppressive role of TET1 in the thermogenic gene regulation of beige adipocytes is largely DNA demethylase-independent. Rather, TET1 coordinates with HDAC1 to mediate the epigenetic changes to suppress thermogenic gene transcription. Taken together, TET1 is a potent beige-selective epigenetic breaker of the thermogenic gene program. Our findings may lead to a therapeutic strategy to increase energy expenditure in obesity and related metabolic disorders.

Identification and expression pattern of a new digestive invertebrate-type lysozyme from the earthworm.

BACKGROUND: The invertebrate type (i-type) lysozyme not showing a clear homology with the known types of lysozyme was first demonstrated from a marine bivalve, conch and earthworm by N-terminal sequence. An i-type lysozyme isolated from the earthworm found to be up-regulated upon bacterial challenge, suggesting this lysozyme to function as an inducible immune factor. However, information on the i-type lysozyme related with digestive function is very limited in the earthworm. OBJECTIVE: The objective of this study is to investigate the molecular characteristics and function of the new i-type lysozyme from the earthworm. METHODS: To identify a new i-type lysozyme, multiple amino acid sequence alignment and phylogenetic analyses were employed. Its mRNA expression pattern was observed by fluorescent in situ hybridization (FISH). RESULTS: A new i-type lysozyme (Ea-iLys) from an earthworm, Eisenia andrei with the open reading frame of 678 bp (226 amino acid residues) appeared to comprise conserved 14 cysteine residues for disulfide bridges and amino acid residues for the enzyme activities of lysozyme and isopeptidase, of which mRNA expression is mainly localized in the lining of midgut epithelium. No significant expression signal was detected in immune competent sites such as chloragogue tissue, typhlosole region, body coelom and muscle layers. CONCLUSION: Our results suggest that this enzyme primarily acts as a digestive enzyme rather than an innate immune factor.

Differential expression of primary pair-rule genes during bidirectional regeneration in Perionyx excavatus.

The earthworm Perionyx excavatus is a species highly capable of bidirectional regeneration. Pair-rule genes are thought to have an ancestral function in arthropod segmentation. However, orthologs in annelids (i.e. Capitella teleta) do not exhibit segmental expression in the ectoderm or mesoderm. Their role in regeneration is currently unclear. Here, we report the expression profile of primary pair-rule genes (Pex-EvxA, Pex-EvxB, Pex-RuntA, Pex-RuntB, Pex-Hes1A, Pex-Hes1B, Pex-Hes4A, and Pex-Hey) found in P. excavatus using semi-quantitative reverse transcription-polymerase chain reaction. Our results indicated these genes showed variable expression during bidirectional regeneration. Six of these genes might play diverse and potentially critical roles in head and/or tail regeneration.

Expression patterns of duplicated snail genes in the leech Helobdella.

snail gene family members are zinc-finger transcription factors with key roles in morphogenesis. Involvement of snail family genes in mesoderm formation has been observed in insects and mammals. The snail genes are also involved in cell motility, neural differentiation, cell fate, survival decision, and left-right identity. The functions of snail genes have been studied primarily among ecdysozoans and deuterostomes, with relatively little work carried out in lophotrochozoans. In this study, we isolated two snail homologs (Hau-snail1 and Hau-snail2) from the leech Helobdella austinensis. We characterized the temporal and spatial expression patterns of these two genes by semi-quantitative RT-PCR and in situ hybridization. The expression of Hau-snail1 and Hau-snail2 correlates with ventral nerve cord (VNC) development, segmental mesoderm, and with a ring of cells that comes to lie at the base of the leech proboscis, respectively, showing similarity to the divergent expression of duplicated snail genes in polychaetes. Our results do not support the function of lophotrochozoan snail genes in mesoderm specification.

Spatiotemporal expression of a twist homolog in the leech Helobdella austinensis.

Genes of the twist family encode bHLH transcription factors known to be involved in the regulation and differentiation of early mesoderm. Here, we report our characterization of Hau-twist, a twist homolog from the leech Helobdella austinensis, a tractable lophotrochozoan representative. Hau-twist was expressed in segmental founder cells of the mesodermal lineage, in subsets of cells within the mesodermal lineage of the germinal plate, in circumferential muscle fibers of a provisional integument during segmentation and organogenesis stages and on the ventral side of the developing proboscis. Thus, consistent with other systems, our results suggest that twist gene of the leech Helobdella might function in mesoderm differentiation.

A new earthworm cellulase and its possible role in the innate immunity.

A new endogenous cellulase (Ean-EG) from the earthworm, Eisenia andrei and its expression pattern are demonstrated. Based on a deduced amino acid sequence, the open reading frame (ORF) of Ean-EG consisted of 1368 bps corresponding to a polypeptide of 456 amino acid residues in which is contained the conserved region specific to GHF9 that has the essential amino acid residues for enzyme activity. In multiple alignments and phylogenetic analysis, the deduced amino acid sequence of Ean- EG showed the highest sequence similarity (about 79%) to that of an annelid (Pheretima hilgendorfi) and could be clustered together with other GHF9 cellulases, indicating that Ean-EG could be categorized as a member of the GHF9 to which most animal cellulases belong. The histological expression pattern of Ean-EG mRNA using in situ hybridization revealed that the most distinct expression was observed in epithelial cells with positive hybridization signal in epidermis, chloragogen tissue cells, coelomic cell-aggregate, and even blood vessel, which could strongly support the fact that at least in the earthworm, Eisenia andrei, cellulase function must not be limited to digestive process but be possibly extended to the innate immunity.