• The Korean Journal of Mycology
• /
• v.40 no.4
• /
• pp.224-228
• /
• 2012

In order to screen interactor(s) of the Aspergillus nidulans ${\alpha}$-COP of COPI vesicle, we performed the yeast two hybrid screening by using the gene for A. nidulans ${\alpha}$-COP as a bait and identified ${\varepsilon}$-COP of the COPI vesicle as an interacting protein. The A. nidualns gene for the ${\varepsilon}$-COP was designated $aneA^+$ ($\underline{A.}$ $\underline{n}$idulans $\underline{e}$psi-lone-COP), which encoded 296 amino acid residues with high level of identity with orthologs from other fungi. Domain analyses with yeast two-hybrid system suggested that the interaction between ${\alpha}$-COP and ${\varepsilon}$-COP relied on the C-terminus of both proteins, and that the N-terminal WD domian of ${\alpha}$-COP and the TPR region of ${\varepsilon}$-COP were not essential but required for the enhancement of the interaction. These results indicate that the interaction mode between ${\alpha}$-COP and ${\varepsilon}$-COP of COPI vesicle is evolutionarily well conserved in eukaryotes.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.60 no.4
• /
• pp.436-441
• /
• 2015

'Superhongmi' is a new rice variety with reddish brown color developed from a crossing a high cyanidin 3-glucoside (C3G) content beeeds including Heugjinju and Suwon 425 and a big grain size breed such as Daeribbyeo. This variety heads on Sep. 5 and has 86.7 cm culm length. The fertility ratio of Superhongmi was about 81.7% and weight of 1,000-grain was about 26.8 g. The 1,000 grain weight of Superhongmi was heavier than that of Superjami. The total essential amino acid content of Superhongmi was lower than those of others. However, Superhongmi was high in tyrosine and arginine. Total polyphenol content of Superhongmi was 2 times higher than that of Heugjinju and flavonoid content was 8 times higher than that of Dongjinbyeo. In addition, hydroxy radical scavenging activity of Superhongmi was significantly higher than that of Dongjinbyeo. However, significant differences were not found in the newly breed colored rice. These results suggest that Superhongmi variety has very high value as a source of various functional food as well as stable food.

• Molecules and Cells
• /
• v.38 no.4
• /
• pp.373-379
• /
• 2015

Pyruvate kinase M2 isoform (PKM2), a rate-limiting enzyme in the final step of glycolysis, is known to be associated with the metabolic rewiring of cancer cells, and considered an important cancer therapeutic target. Herein, we report a novel PKM2 activator, PA-12, which was identified via the molecular docking-based virtual screening. We demonstrate that PA-12 stimulates the pyruvate kinase activity of recombinant PKM2 in vitro, with a half-maximal activity concentration of $4.92{\mu}M$, and effectively suppresses both anchorage-dependent and -independent growth of lung cancer cells in non-essential amino acid-depleted medium. In addition, PA-12 blocked the nuclear translocalization of PKM2 in lung cancer cells, resulting in the inhibition of hypoxia response element (HRE)-mediated reporter activity as well as hypoxia-inducible factor 1 (HIF-1) target gene expression, eventually leading to the suppression of cell viability under hypoxia. We also verified that the effects of PA-12 were dependent on PKM2 expression in cancer cells, demonstrating the specificity of PA-12 for PKM2 protein. Taken together, our data suggest that PA-12 is a novel and potent PKM2 activator that has therapeutic implications for lung cancer.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.6
• /
• pp.997-1006
• /
• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.5
• /
• pp.614-628
• /
• 2019

Objective: The inhibitory leukocyte immunoglobulin-like receptors (LILRBs) play an important role in innate immunity. The present study represents the first description of the cloning and structural and functional analysis of LILRB1 and LILRB3 isolated from two genetically disparate chicken lines. Methods: Chicken LILRB1-3 genes were identified by bioinformatics approach. Expression studies were performed by transfection, quantitative polymerase chain reaction. Signal transduction was analyzed by western blots, immunoprecipitation and flow cytometric. Cytokine levels were determined by enzyme-linked immunosorbent assay. Results: Amino acid homology and phylogenetic analyses showed that the homologies of LILRB1 and LILRB3 in the chicken line 6.3 to those proteins in the chicken line 7.2 ranged between 97%-99%, while homologies between chicken and mammal proteins ranged between 13%-19%, and 13%-69%, respectively. Our findings indicate that LILRB1 and LILRB3 subdivided into two groups based on the immunoreceptor tyrosine-based inhibitory motifs (ITIM) present in the transmembrane domain. Chicken line 6.3 has two ITIM motifs of the sequence LxYxxL and SxYxxV while line 7.2 has two ITIM motifs of the sequences LxYxxL and LxYxxV. These motifs bind to SHP-2 (protein tyrosine phosphatase, non-receptor type 11) that plays a regulatory role in immune functions. Moreover, our data indicate that LILRB1 and LILRB3 associated with and activated major histocompatibility complex (MHC) class I and ${\beta}2-microglobulin$ and induced the expression of transporters associated with antigen processing, which are essential for MHC class I antigen presentation. This suggests that LILRB1 and LILRB3 are transcriptional regulators, modulating the expression of components in the MHC class I pathway and thereby regulating immune responses. Furthermore, LILRB1 and LILRB3 activated Janus kinase2/tyrosine kinase 2 (JAK2/TYK2); signal transducer and activator of transcription1/3 (STAT1/3), and suppressor of cytokine signaling 1 genes expressed in Macrophage (HD11) cells, which induced Th1, Th2, and Th17 cytokines. Conclusion: These data indicate that LILRB1 and LILRB3 are innate immune receptors associated with SHP-2, MHC class I, ${\beta}2-microglobulin$, and they activate the Janus kinase/signal transducer and activator of transcription signaling pathway. Thus, our study provides novel insights into the regulation of immunity and immunopathology.

• Nutrition Research and Practice
• /
• v.16 no.6
• /
• pp.729-744
• /
• 2022

BACKGROUND/OBJECTIVES: 4-Hydroxy-2-nonenal (HNE) is a biomarker for oxidative stress to induce inflammation. Methionine is an essential sulfur-containing amino acid with antioxidative activity. On the other hand, the evidence on whether and how methionine can depress HNE-derived inflammation is lacking. In particular, the link between the regulation of the nuclear factor-κB (NF-κB) signaling pathway and methionine intake is unclear. This study examined the link between depression from HNE accumulation and the anti-inflammatory function of ⳑ-methionine in rats. MATERIALS/METHODS: Male Wistar rats (3-week-old, weighing 70-80 g) were administered different levels of ⳑ-methionine orally at 215.0, 268.8, 322.5, and 430.0 mg/kg body weight for two weeks. The control group was fed commercial pellets. The hepatic HNE contents and the protein expression and mRNA levels of the inflammatory mediators were measured. The interleukin-10 (IL-10) and glutathione S-transferase (GST) levels were also estimated. RESULTS: Compared to the control group, hepatic HNE levels were reduced significantly in all groups fed ⳑ-methionine, which were attributed to the stimulation of GST by ⳑ-methionine. With decreasing HNE levels, ⳑ-methionine inhibited the activation of NF-κB by up-regulating inhibitory κBα and depressing phosphoinositide 3 kinase/protein kinase B. The mRNA levels of the inflammatory mediators (cyclooxygenase-2, interleukin-1β, interleukin-6, inducible nitric oxide synthase, tumor necrotic factor alpha) were decreased significantly by ⳑ-methionine. In contrast, the protein expression of these inflammatory mediators was effectively down regulated by ⳑ-methionine. The anti-inflammatory action of ⳑ-methionine was also reflected by the up-regulation of IL-10. CONCLUSIONS: This study revealed a link between the inhibition of HNE accumulation and the depression of inflammation in growing rats, which was attributed to ⳑ-methionine availability. The anti-inflammatory mechanism exerted by ⳑ-methionine was to inhibit NF-κB activation and to up-regulate GST.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.36 no.11
• /
• pp.1472-1481
• /
• 2007

The purposes of the study were to find refined taste of ancestor through historical research about traditional cooking method and ingredient for the purpose of enriching today#s dietary life and to hand down a particular style of regional dish and excellence of nutritional aspect by providing a standard recipe and nutrition analysis data on #Dong-rae Pajeon#. To collect data about traditional ingredients and cooking method, researcher interviewed seven local natives who have kept a traditional food costumes, visited four restaurants, and reviewed ten cookbooks. The interviewees recalled and demonstrated the cooking procedure. The standard recipe of #Dong-rae Pajeon# was created after three experimental cookings, based on the recipes of the natives, restaurants, and cookbooks. According to the natives# statements, #Dong-rae Pajeon# was a special dish that was offered to the king at #Samzi-nal# (March 3rd of the lunar calendar). It was also a seasonal (before cherry blooming time) and memorial service dish of the province#s high society. The main ingredients were small green onion, dropwort, beef, seafood (large clam, mussel, clam meat, oyster, shrimp, fresh water conch), waxy rice powder, non-wax rice powder, and sesame oil which were abundant in Busan and Kijang region. Energy per 100 g of #Dong-rae Pajeon# was 148 kcal. Protein, lipid, fiber, Ca, and Fe contents were 8.8 g, 2.0 g, 8.6 g, 57.7 mg, and 1.8 mg respectively. Contents of cystine, lysine, leucine, valine, isoleucine which are essential amino acids were high in #Dong-rae Pajeon#. Fatty acids contents are oleic acid (20.5%), linoleic acid (20.1%) and linolenic acid (10.4%) while P/M/S ratio was 0.73/0.67/1.

• Journal of Animal Science and Technology
• /
• v.48 no.5
• /
• pp.747-758
• /
• 2006

A total of 30 pigs (average 60kg) were used to investigate the influence of breeds on meat quality. Pigs were alloted into one of three experimental groups [A (Landrace×Yorkshire×Duroc), B (England Berkshire), and C (Kagoshima Berkshire)]. Pigs were slaughtered at approximately 110kg of live weight for measurement of meat qualities. Crude protein was significantly higher in B than those of other pig breed groups. Shear force value was higher and pH was lower in A than those of other pig breeds. In color, C was significantly higher in a* of meat color and lower in b* of fat color and in brittleness, hardness, springiness, chewiness as compared with other pig breeds. Essential fatty acid were significantly higher in A and lower in B than those of other pig breeds. Aspartic, threonine, serine, glutamic and phenylalanine were higher in C, whereas glycine, alanine, cystine, valine, leucine, tyrosine, histidine, lysine and arginine were significantly higher in A. In sensory evaluation of fresh meat, color was significantly higher in C. However, moisture, crude fat, crude ash and drip loss, marbling score, overall acceptability of fresh meat and all items in sensory evaluation of cooked meat were not significant difference among the pig groups. In conclusions, loin from Kagoshima Berkshire had the lowest shear force and the highest a* value. In the current study few effects of pig breeds were observed. Further studies are needed on the relationship between physico-chemical properties and the pig breeds.

• Korean Journal of Microbiology
• /
• v.42 no.2
• /
• pp.111-115
• /
• 2006

A feeding trial was conducted to test the use of marine yeasts isolated from seawaters and sediments as a dietary source in cultivating a Cladocera, Moina macrocopa which is available as an alternative live food for fish larvae. The marine yeast-fed M. macrocopa had similar essential amino acid profiles to the documented values for Rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. $S{\pi}-1$ lacked ${\omega}-3$ high unsaturated fatty acids (HUFAs), $20:5{\omega}-3$ (EPA) and $22:6{\omega}-3$ (DHA), which were also poor but detected in both the marine yeasts. An increase in the $20:5{\omega}-3$ and $22:6{\omega}-3$ levels, compared with the levels in marine yeast strains themselves, was more pronounced in the $22:6{\omega}-3$ level of Moina fed the Candida sp. Y-16, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their ${\delta}^{13}C$ values shifted gradually toward the values of the switched diets. Diet switch from Erythrobacter sp. $S{\pi}-1$to Candide sp. Y.16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the ${\delta}^{13}C$ values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the ${\delta}^{13}C$ values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly Candida sp. Y-16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietaty values than the commercial diet (Erythrobacter sp. $S{\pi}-1$).

• Journal of Animal Science and Technology
• /
• v.47 no.6
• /
• pp.1087-1094
• /
• 2005

The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose transport protein. In the glucose transport protein family, GLUT4 plays a key role in cellular glucose uptake stimulated by insulin in skeletal muscles and adipose tissue in rodents and human. In this studies, we reported the identification, characterization, and expression of Hanwoo GLUT4 gene. The Hanwoo GLUT4 cDNA includes a 1527 bp open reading frame encoding a protein of 509 amino acids. The GLUT4 amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The highest mRNA expression of GLUT4 was detected in heart and lower expression was detected in rib meat, sirloin, and colon. We confirmed the expression of GLUT4 in the subcutaneous and small intestinal adipose tissue using RT-PCR. To investigate the expression of GLUT4 in the bovine intramuscular adipose differentiation, fibroblast-like cells were isolated from the sirloin of Hanwoo bull aged 12 months by collagenase digestion of minced tissue and cultured with activators of PPAR gamma. We identified that GLUT4 mRNA expression decreased during differentiation of preadipocytes into adipocyte in Korean cattle. These results indicated that function of GLUT4 in bovine adipose tissue was different from that of mouse and human.