• Fisheries and Aquatic Sciences
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• v.27 no.5
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• pp.314-328
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• 2024

The mechanism for the elimination of xenobiotics undergoes three different phases of reactions in organisms. Among these, glutathione S-transferases (GSTs) are classified as phase II detoxification enzymes, catalyzing the conjugation of electrophilic substrates to glutathione or reduced hydroperoxides. This study aimed to investigate the molecular characteristics, detoxification functions, and immune responses of GST omega (LhGSTO1) and kappa (LhGSTK1) in redlip mullet. The open reading frames of LhGSTO1 (720 bp) and LhGSTK1 (687 bp) encoded proteins of 239 and 228 amino acids, respectively. Sequence analysis revealed that LhGSTO1 and LhGSTK1 possessed GSH-binding sites in their N-terminal domains. Substrate-binding sites in the C-terminal domain were exclusively identified in LhGSTO1. In the tissue-specific transcription profile analysis, both LhGSTO1 and LhGSTK1 were ubiquitously expressed in all tissues of healthy mullets. Temporal expression analysis of LhGSTO1 and LhGSTK1 in the blood showed that their expression was significantly modulated by polyinosinic:polycytidylic (poly I:C), lipopolysaccharide (LPS), and Lactococcus garvieae. Different chemical and cellular assays were performed to assess the detoxification and cellular protective abilities of the two proteins. A substrate specificity test using the recombinant proteins revealed that both proteins possessed specific activity toward 1-chloro-2,4-dinitrobenzene (CDNB). In the disk diffusion assay, the smallest clearance zones were observed for LhGSTO1 and LGSTK1 against CdCl₂. In the cell protection assay, both LhGSTO1 and LhGSTK1 showed significant Cd detoxification ability compared to the control. Collectively, these results demonstrate that GST omega and kappa are involved in host defense against immune stimulants and xenobiotics in redlip mullet.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.138-144
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• 2024

Background: As the number of households raising companion dogs increases, the pet genetic analysis market also continues to grow. However, most studies have focused on specific purposes or native breeds. This study aimed to collect genomic data through single nucleotide polymorphism (SNP) chip analysis of companion dogs in South Korea and perform genetic diversity analysis and SNP annotation. Methods: We collected samples from 95 dogs belonging to 26 breeds, including mixed breeds, in South Korea. The SNP genotypes were obtained for each sample using an Axiom^TM Canine HD Array. Quality control (QC) was performed to enhance the accuracy of the analysis. A genetic diversity analysis was performed for each SNP. Results: QC initially selected SNPs, and after excluding non-diverse ones, 621,672 SNPs were identified. Genetic diversity analysis revealed minor allele frequencies, polymorphism information content, expected heterozygosity, and observed heterozygosity values of 0.220, 0.244, 0.301, and 0.261, respectively. The SNP annotation indicated that most variations had an uncertain or minimal impact on gene function. However, approximately 16,000 non-synonymous SNPs (nsSNPs) have been found to significantly alter gene function or affect exons by changing translated amino acids. Conclusions: This study obtained data on SNP genetic diversity and functional SNPs in companion dogs raised in South Korea. The results suggest that establishing an SNP set for individual identification could enable a gene-based registration system. Furthermore, identifying and researching nsSNPs related to behavior and diseases could improve dog care and prevent abandonment.

• International Journal of Industrial Entomology and Biomaterials
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• v.48 no.3
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• pp.107-113
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• 2024

The efficiency of protein extraction from Hongjam, a steamed mature silkworm, was quantitatively evaluated using various chemical buffers and physical methods. This study considers the difficulty of protein extraction yield due to the high content of hydrophobic amino acids in Hongjam compared to 5^th instar-3^rd day silkworm larvae. Results indicated that urea buffer enhanced protein yield more effectively than RIPA buffer. Additionally, the application of physical methods such as microwave treatment to samples treated with RIPA buffer increased yields by up to 22%, achieving concentrations as high as 3.9 mg/mL. Circular dichroism (CD) analysis showed that proteins extracted with urea buffer retained their structural integrity, exhibiting deeper and more prominent peaks associated with random coil structure. In addition, physical methods such as vortexing, sonication, microwave and homogenization increased the extraction yield of larger molecules without altering protein structures, suggesting their potential scalability for industrial applications. These results demonstrate the critical role of selecting appropriate extraction methods to optimize the yield and functionality of proteins from Hongjam, with implications for its use in biotechnological applications and nutraceuticals.

• Journal of Microbiology and Biotechnology
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• v.34 no.9
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• pp.1890-1897
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• 2024

Fermentation has been identified as an effective strategy to alter the chemical makeup of tobacco, thereby enhancing its quality. The deliberate introduction of microorganisms can hasten the fermentation process. In this research, microbial consortia harvested from the tobacco surface were utilized to enhance the tobacco quality. This enhancement also elevated several sensory attributes of HnB cigarettes, such as aroma richness, moisture, strength, and reduced irritation, achieving a sensory quality rating of 84.5. This marks a notable improvement compared to the 82 rating of the original, unfermented cigarettes. Untargeted metabolomics analysis revealed a decrease in total polyphenols and unsaturated fatty acids, while the levels of polyacids, alcohols, ketones, furans, and other compounds increased in the fermented tobacco. Additionally, KEGG pathway enrichment analysis indicated that the enhancement in tobacco quality through microbial consortia fermentation is linked to various biological pathways, with pathways related to fatty acid and amino acid degradation playing pivotal roles. The findings of this study will serve as a reference for the commercial production of HnB cigarettes, and the elucidated mechanism offers a theoretical basis for exploring microbial fermentation as a means to improve tobacco quality.

• Molecules and Cells
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• v.27 no.5
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• pp.591-599
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• 2009

Nicotinic acetylcholine receptors (nAChRs) play important roles in nervous system functions and are involved in a variety of diseases. We previously demonstrated that ginsenosides, the active ingredients of Panax ginseng, inhibit subsets of nAChR channel currents, but not ${\alpha}7$, expressed in Xenopus laevis oocytes. Mutation of the highly conserved Leu247 to Thr247 in the transmembrane domain 2 (TM2) channel pore region of ${\alpha}7$ nAChR induces alterations in channel gating properties and converts ${\alpha}7$ nAChR antagonists into agonists. In the present study, we assessed how point mutations in the Leu247 residue leading to various amino acids affect 20(S)-ginsenoside $Rg_3$ ($Rg_3$) activity against the ${\alpha}7$ nAChR. Mutation of L247 to L247A, L247D, L247E, L247I, L247S, and L247T, but not L247K, rendered mutant receptors sensitive to $Rg_3$. We further characterized $Rg_3$ regulation of L247T receptors. We found that $Rg_3$ inhibition of mutant ${\alpha}7$ nAChR channel currents was reversible and concentration-dependent. $Rg_3$ inhibition was strongly voltage-dependent and noncompetitive manner. These results indicate that the interaction between $Rg_3$ and mutant receptors might differ from its interaction with the wild-type receptor. To identify differences in $Rg_3$ interactions between wild-type and L247T receptors, we utilized docked modeling. This modeling revealed that $Rg_3$ forms hydrogen bonds with amino acids, such as Ser240 of subunit I and Thr244 of subunit II and V at the channel pore, whereas $Rg_3$ localizes at the interface of the two wild-type receptor subunits. These results indicate that mutation of Leu247 to Thr247 induces conformational changes in the wild-type receptor and provides a binding pocket for $Rg_3$ at the channel pore.

• Korean Journal of Agricultural Science
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• v.36 no.2
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• pp.185-197
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• 2009

For utilizing Cattail pollen as a raw material for functional foods, the nutrients such as free sugar, free amino acid, fatty acid composition, flavonoid content, and the biological activity within Cattail pollen were measured. The results of proximate analysis within Cattail pollen included the following readings: 12.7-13.2% of moisture, 15.7-17.8% of crude protein, 1.3% of crude fat, 7.5-7.7% of free sugar, 13.7-18.6% of crude fiber, 3.4-4.9% of ash, and 49.7-55.9% of nitrogen free extracts. The composition of free amino acids consisted of 1.923% of T. orientalis, 0.907% of T. angustata, and 0.333% of T. latifolia, which were measurements that varied significantly among different species. However, all species showed considerable portions of GABA alanine, glutamic acid, and proline. Specifically, it was shown that the GABA composition, which is known for increasing immunity while simultaneously lowering blood pressure, exceeded 50%. Therefore, this result implies that Cattail pollens have potential as a powerful utilization for functional foods. The composition of the fatty acids mainly consisted of linoeic, palmitic acid, oleic acid, and linolenic acid, and didn't show many variances across different species. Also, the total contents of unsaturated fatty acid were particularly high with a measured ratio of 67.2-76.0% value. Mineral in Cattail pollen was composed of 0.354-0.492% of K, 0.0516-0.0546% of Mg, 0.045-0.0486% of Ca, and 0.0101-0.0204% of Na. Among the Cattail pollens known as anti-oxidants, flavonoid contains 0.169-0.186% of quercetin, and therefore is the largest constituent followed by rutin making up a measurement of 0.0094-0.0147%. For the purpose of the study, the Cattail pollen and its extracts were fed to SC class rats for a span of 4 weeks. Then, the DPPH radical scavenging activity was measured from the tested rats'serums and the results showed significant variances. Also, the results indicated that the cholesterol and glucose levels in the blood were decreased which in turn led to the conclusion that the cattail pollen can help hyperlipidemia and diabetic treatments.

• Journal of Life Science
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• v.26 no.4
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• pp.419-430
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• 2016

Powdery mildew disease caused by Podosphaera xanthii is a major concern for Cucumis melo production worldwide. Knowledge on genetic behavior of the related genes and their modulating phytohormones often offer the most efficient approach to develop resistance against different diseases. Mildew Resistance Locus O (MLO) genes encode proteins with seven transmembrane domains that have significant function in plant resistance to powdery mildew fungus. We collected 14 MLO genes from ‘Melonomics’ database. Multiple sequence analysis of MLO proteins revealed the existence of both evolutionary conserved cysteine and proline residues. Moreover, natural genetic variation in conserved amino acids and their replacement by other amino acids are also observed. Real-time quantitative PCR expression analysis was conducted for the leaf samples of P. xanthii infected and phyto-hormones (methyl jasmonate and salicylic acid) treated plants in melon ‘SCNU1154’ line. Upon P. xanthii infection using 7 different races, the melon line showed variable disease reactions with respect to spread of infection symptoms and disease severity. Three out of 14 CmMLO genes were up-regulated and 7 were down-regulated in leaf samples in response to all races. The up- or down-regulation of the other 4 CmMLO genes was race-specific. The expression of 14 CmMLO genes under methyl jasmonate and salicylic acid application was also variable. Eleven CmMLO genes were up-regulated under salicylic acid treatment, and 7 were up-regulated under methyl jasmonate treatments in C. melo L. Taken together, these stress-responsive CmMLO genes might be useful resources for the development of powdery mildew disease resistant C. melo L.

• Tuberculosis and Respiratory Diseases
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• v.61 no.3
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• pp.239-247
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• 2006

Background: Culture filtrate proteins secreted by mycobacteria are thought to play an important role in inducing protective immunity and to develop new methods for diagnosing tuberculosis. Methods: A culture filtrate protein of M. avium that was strongly reactive with goat antiserum against M. intracellulare was constructed. Its homologous protein (TB-14) in M. tuberculosis was cloned, expressed and purified. The inductions of IFN-${\gamma}$ stimulated with $10{\mu}g$ of TB-14 recombinant protein and $10{\mu}g$ PPD were estimated by using whole bloods from seven PPD (-) subjects, seven PPD (+) healthy volunteers and nine tuberculosis patients. Results: M. avium culture filtrate protein was confirmed as a hypothetical protein that was termed contig 116. A novel 14-kDa recombinant protein (TB-14) of M. tuberculosis was composed of 148 amino acids, including 30 amino acids of the signal peptide, and it showed 78% homology with M. avium. In the PPD (+) healthy volunteers, recombinant TB-14 protein strongly induced the secretion of IFN-${\gamma}$ in whole blood cultures. Conclusion: These results suggest that TB-14 recombinant protein might play an important role in inducing cell-mediated immunity against tuberculosis. Furthermore, TB-14 protein antigen and its antiserum will be available for the development of new diagnostic tools for tuberculosis.

• Journal of Chest Surgery
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• v.36 no.3
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• pp.131-135
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• 2003

Bovine pericardium fixed in glutaraldehyde solution (GA) has been one of the most popular surgical bioprosthesis, however, late calcific degeneration after implantation remains to be solved. To mitigate calcific degeneration, we posttreated the bovine pericardium with amino acids after GA fixation. Material and Method: 40 small pieces of bovine pericardia were fixed in 0.625% GA solution with 4 g/L $MgCl_26H_2O$as a control group (group 1). 40 pieces fixed in the same GA solution were posttreated with 2% chitosan solution (group 2) and the other 40 pieces posttreated with 8% glutamate (group 3). These were implanted into the belly of forty Fisher 344 rats subdermally and extracted at f month, 2 months, 3 months and 4 months after implantation. Result: With atomic absorption spectrophotometry we measured the deposited calcium amount and the results were as follows; 2.01 $\pm$0.13 mg/g in group 1, 2.34$\pm$0.73 mg/g in group 2, 2.49$\pm$0.15 mg/g in group 3 at 1 month after implantation, and 3.57$\pm$0.15 mg/g in group 1, 3.52$\pm$0.92 mg/g in group 2, 3.46$\pm$0.12 mg/g in group 3 at the second month. But 5.45$\pm$0.42 mg/g in group 1, 3.22 $\pm$1.31 mg/g in group 2 and 4.20$\pm$0.55 mg/g in group 3 at the 3rd month, which have statistical significance in group 2 (p<0.05). Finally at 4th month, 6.01$\pm$1.21 mg/g in group 1, 3.78$\pm$1.82 mg/g in group 2, 3.92$\pm$0.92 mg/g in group 3, which also have statistical significance (p < 0.05). Conclusion: This means posttreatment with 2% chitosan shows meaningful calcium mitigation effects after 3rd month on subcutaneously implanted bovine pericardium in the rat models but 8% glutamate shows mitigation effect after 4months in this experiment.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.8
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• pp.1398-1406
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• 2004

To develop the new type of salt-fermented seafoods, the salt-fermented oysters in olive oil (product SO) were manufactured, and food components and quality characteristics of product SO were examined. The optimum processing condition for product SO is as follows. The raw oyster with no shell was washed off with 3% saline solution. Then dewatered, and dipped in the brine-salting solution made up with saturated saline solution and oyster sauce (2 : 1 v/v) mixture added 1% sodium erythorbic acid and 0.2% polyphosphate. After salt-fermentation it ripened by brine salting at 5$\pm$1$^{\circ}C$ for 15 days. Then dried at 15$^{\circ}C$ for 4 hours with cool-air, and packed in No. 3B hexahedron type can. Finally, poured with olive oil and seamed it by double-seamer. The moisture, crude protein, crude ash and volatile basic nitrogen contents of the product SO were 61.6%, 12.0%, 16.3% and 34.3 mg/100 g, respectively. In taste-active components of the product SO, total amount of free amino acids is 2,335.4 mg/100 g and it has increased by 50% overall during salt-fermentation 15 day. Taurine, glutamic acid, proline, glycine, alanine, $\beta$-alanine and lysine were detected as principal free amino acids. The contents of inorganic ions were rich in Na and K ion, while the amounts of nucleotide and its related compounds and other bases except betaine were small. From the results of this research, the product SO had a superior organoleptic qualities compared with conventional oyster product, and could be reserved in good conditions for storage 90 days at room temperature.