• Journal of fish pathology
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• v.21 no.1
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• pp.1-11
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• 2008

We report the existence of new type of phosphatidylcholine-hydrolyzing phospholipase D (PLD), which has been characterized and partially purified in the scuticociliate, Uronema marinum. The enzyme from partial purification showed that it was existed in membrane fraction and was a neutral PLD, which catalyzed both transphosphatidylation and hydrolysis reaction. The activity of partially purified membrane-bound PLD was also found to be optimal at pH 7.0-7.5 for 2 hours at 37℃ and depended strictly on the presence of Ca2+ (2.5 mM) and Mg2+ (1.6 mM). Immunoblot analysis indicated that the enzyme was distinct from hPLD1 (human PLD1) and hPLD2 (human PLD2) because it was not recognized by a polyclonal antibody raised to the 12 terminal amino acid of these enzymes. We also found that the membrane-bound PLD is a PIP2-dependent PLD and that GTP-binding proteins are not implicated in the regulation of this enzyme: This enzyme activity is markedly stimulated by phosphatidylinositol 4,5-bisphosphate (PIP2) but not by the small G-protein Arf and GTPrS. In addition, this enzyme was capable of hydrolyzing phosphatidylcholine (PC) but not phosphatidylethanolamine (PE), implying that PC was a preferred substrate.

• Biomolecules & Therapeutics
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• v.28 no.5
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• pp.482-489
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• 2020

G protein-coupled receptor kinase 5 (GRK5) has been considered as a potential target for the treatment of heart failure as it has been reported to be an important regulator of pathological cardiac hypertrophy. To discover novel scaffolds that selectively inhibit GRK5, we have identified a novel small molecule inhibitor of GRK5, KR-39038 [7-((3-((4-((3-aminopropyl)amino)butyl)amino)propyl)amino)-2-(2-chlorophenyl)-6-fluoroquinazolin-4(3H)-one]. KR-39038 exhibited potent inhibitory activity (IC₅₀ value=0.02 µM) against GRK5 and significantly inhibited angiotensin II-induced cellular hypertrophy and HDAC5 phosphorylation in neonatal cardiomyocytes. In the pressure overload-induced cardiac hypertrophy mouse model, the daily oral administration of KR-39038 (30 mg/kg) for 14 days showed a 43% reduction in the left ventricular weight. Besides, KR-39038 treatment (10 and 30 mg/kg/day, p.o.) showed significant preservation of cardiac function and attenuation of myocardial remodeling in a rat model of chronic heart failure following coronary artery ligation. These results suggest that potent GRK5 inhibitor could effectively attenuate both cardiac hypertrophy and dysfunction in experimental heart failure, and KR-39038 may be useful as an effective GRK5 inhibitor for pharmaceutical applications.

• Development and Reproduction
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• v.27 no.4
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• pp.205-211
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• 2023

INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.710-724
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• 2024

Flavobacterium can synthesize xanthophyll, particularly the pigment zeaxanthin, which has significant economic value in nutrition and pharmaceuticals. Recently, the use of carotenoid biosynthesis by bacteria and yeast fermentation technology has shown to be very efficient and offers significant advantages in large-scale production, cost-effectiveness, and safety. In the present study, JSWR-1 strain capable of producing xanthophyll pigment was isolated from a freshwater reservoir in Wanju-gun, Republic of Korea. Based on the morphological, physiological, and molecular characteristics, JSWR-1 classified as belonging to the Flavobacterium species. The bacterium is strictly aerobic, Gram-negative, rod-shaped, and psychrophilic. The completed genome sequence of the strain Flavobacterium sp. JSWR-1 is predicted to be a single circular 3,425,829-bp chromosome with a G+C content of 35.2% and 2,941 protein-coding genes. The optimization of carotenoid production was achieved by small-scale cultivation, resulting in zeaxanthin being identified as the predominant carotenoid pigment. The enhancement of zeaxanthin biosynthesis by applying different light-irradiation, variations in pH and temperature, and adding carbon and nitrogen supplies to the growth medium. A significant increase in intracellular zeaxanthin concentrations was also recorded during fed-batch fermentation achieving a maximum of 16.69 ± 0.71 mg/l, corresponding to a product yield of 4.05 ± 0.15 mg zeaxanthin per gram cell dry weight. Batch and fed-batch culture extracts exhibit significant antioxidant activity. The results demonstrated that the JSWR-1 strain can potentially serve as a source for zeaxanthin biosynthesis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.4
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• pp.359-366
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• 2016

Exosome, a small vesicle secreted from cells, has diverse functions depending on cell origins and tissue types and plays a important role in cell viability and intercellular communication. Recently, many researchers have demonstrated the use of exosomes for the treatment of cancers and immune diseases, and the development of diagnostic biomarker. However, the secretion mechanism of exosome from skin cell and its physiological functions in skin remain unclear. Thus, this study aimed to explore whether keratinocyte-derived exosome affects proliferation and migration in HaCaTs. Exosomes were isolated from HaCaTs by ExoQuick-TC and then boiled or unbolied. Boiled and unboiled exosome induced proliferation in HaCaTs in a dose-dependant manner ($0.1{\sim}20{\mu}g/mL$), respectively. Boiled and unboiled exosome at concentration of $20{\mu}g/mL$ increased proliferation level in HaCaTs by $186.96{\pm}3.87%$ and $193.48{\pm}10.48%$ compared with control group. Unboiled exosome stimulated migration in HaCaTs in a dose-dependent manner ($0.1{\sim}20{\mu}g/mL$), which reached a maxium at concentration of $20{\mu}g/mL$ ($179.39{\pm}4.89%$ of control), but boiled exosome did not affect HaCaT migration. In addition, unboiled exosome ($0.1{\sim}20{\mu}g/mL$) dose-dependently stimulated sprout outgrowth in HaCats. These results demonstrate that in exosome from HaCaTs, heat-stable components such as lipid may induce HaCaT proliferation and heat-unstable components such as protein may stimulate migration and sprout outgrowth in HaCaTs, thereby leading to reepithelialization and skin-wound healing activities. It is concluded that exosomes from HaCaTs may be used as cosmetic materials.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.4
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• pp.355-362
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• 1989

For the effective utilization of mackerel as a food sauce, the processing conditions of the frozen seasoned mackerel meat and the changes in taste compounds during its frozen storage were investigated. To prepare the frozen seasoned mackerel meat, the mackerel was headed, gutted manually, washed with tap water and deboned with the meat seperator. Then it was mixed with additives such as emulsion curd(32.1%, w/w), table salt(0.5%, w/w), sugar(2.0%, w/w), sodium bicarbonate(0.4%, w/w), polyphosphate(0.2%, w/w), monosodium glutamate(0.2%, w/w), onion powder(0.3%, w/w), garlic powder(0.1%, w/w), ginger powder(0.1%, w/w), soybean protein(3.0%, w/w) and sodium erythorbate(0.1%, w/w). This seasoned fish meat was frozen with contact freezer, packed In a carton box, and then stored at $-25^{\circ}C$. The moisture and lipid contents in the products were 70.8-71.7% and 10.9-11.3%, respectively. The taste compounds of the frozen seasoned mackerel meat were free amino acids(1625.0-1692.0mg/100g), nucleotides and their related compounds(316.6-366.8 mg/100g) as well as total creatinine(270.2-311.8 mg/100g), and small amount of betaine and TMAO. In free amino acids, the predominant ones were histidine, lysine, glutamic acid and arginine. It was supposed from the results that principal taste compounds of frozen seasoned mackerel meat were free amino acids, and that total creatinine, TMAO, TMA and betaine as well as nucleotides and their related compounds also played an assistant role.

• Korean Chemical Engineering Research
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• v.62 no.1
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• pp.87-98
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• 2024

This study was conducted to investigate the efficiency of the filtering sets composed of fiberboards, which were fabricated with lignocellulosic fiber and cork oak bark-based activated carbon (COA), as well as traditional Korean paper handmade from mulberry trees (KP) for the filtration of PM, TVOC and HCHO. Three-layers fiberboards (WRF) were fabricated with wood fiber in its surface layers and recycled fiber/COA in its core layer using a protein-based adhesive with the resin content of 8%. Filtering sets were composed of three WRF and one sheet of KP. Concentrations of PM, TVOC and HCHO generated with the combustion of a incense in a sealed laboratory hood were reduced efficiently with the operation of air-purifier installed the filtering sets. Except for the WRF fabricated with 4%/4% resin contents, other WRF were prepared with 5%/3% and 6%/2% resin contents in surface/core layers, and then the WRF were used with KP for the fabrication of filtering sets. Filtration efficiency of the filtering sets was improved as the core-layer resin content applied in the fabrication of WRF decreased. In addition, filtration efficiency of the WRF-based filtering set fabricated with KP of 25 g/m² basis weight was higher than that with KP of 45 g/m² basis weight. Filtering sets composed of three-layers fiberboards (RWF) that recycled fiber and wood fiber/COA were used in its surface and core layers, respectively, and KP-25g showed higher filtration efficiency than those of WRF-based filtering sets. Air-inhalation equipment installed the RWF-based, WRF-based filtering sets and without filtering set were operated in small indoor and large outdoor spaces. Efficiency for filtering PM and TVOC of the RWF-based filtering sets was higher than that of other filtering sets. It is concluded that fiberboard-based filtering sets composed of RWF and KP-25g can be used as a filter for reducing the concentrations of PM and TVOC existed in indoor and outdoor spaces.

• Journal of Life Science
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• v.26 no.12
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• pp.1383-1391
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• 2016

Anti-estrogen drugs such as tamoxifen have been used for treating patients with ER-positive, early breast cancer. However, resistance to anti-estrogen treatment is inevitable in most patients. Breast cancer anti-estrogen resistance-3 (BCAR3) has been identified as the protein responsible for the induction of tamoxifen resistance in estrogen-dependent human breast cancer. We have previously reported that BCAR3 regulates the cell cycle progression and the signaling pathway of EGF and insulin leading to DNA synthesis. In this study, we investigated the functional role of BCAR3 in regulating c-Jun transcription in non-tumorigenic human breast epithelial MCF-12A cells. A transient transfection of BCAR3 increased both the mRNA and protein of c-Jun expression, and stable expression of BCAR3 increased c-Jun protein expression. The overexpression of BCAR3 directly activated the promoter of c-jun, AP-1, and SRE but not that of $NF-{\kappa}B$. Furthermore, single-cell microinjection of BCAR3 expression plasmid in the cell cycle-arrested MCF-12A cells induced c-Jun protein expression, and co-injection of dominant negative mutants of Ras, Rac, and Rho suppressed the transcriptional activity of c-Jun in the presence of BCAR3. Furthermore, stable expression of BCAR3 increased the proliferation of MCF-12A cells. The microinjection of inhibitory materials such as anti-BCAR3 antibody and siRNA BCAR3 inhibited EGF-induced c-Jun expression but did not affect IGF-1 induced upregulation of c-Jun. Taken together, we propose that BCAR3 plays a crucial role in c-Jun protein expression and cell proliferation and that small GTPases (e.g., Ras, Rac, and Rho) are required for the BCAR3-mediated activation of c-Jun expression.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.62 no.2
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• pp.143-148
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• 2017

Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrates, isoflavones, and other nutrients for humans and animals. Raffinose and stachyose are the main antinutritional factors in soybean seed. They are carbohydrates belonging to the raffinose family of oligosaccharides, which are not readily digested in humans and cause flatulence or diarrhea. The genetic reduction of the raffinose and stachyose contents in mature soybean seeds will improve the nutritional value of soybean. The objective of this research was to evaluate agronomic traits with 10 $F_6$ strains selected from breeding populations derived from a cross among seven parents. The contents of raffinose and stachyose in mature seeds were detected by high-performance liquid chromatography. Agronomic traits such as flower color, flowering date, harvesting date, lodging, plant height, seed coat color, hilum color, 100 seed weight, and yield were evaluated. Ten intermediate parents showed low raffinose and stachyose contents. The intermediate parent 883-1 had a small seed size, six intermediate parents (15A1, 15D1, RS-5, RS-33, RS-64, and RS-70) had a medium seed size, and two intermediate parents (14G20 and RS-21) had a large seed size. The intermediate parent RS-21 had a black seed coat and a green cotyledon. Four intermediate parents (883-1, 14G20, RS-5, and RS-21) had elite agronomic traits. The new intermediate parents developed through this study will be used to develop improved soybean cultivars with low contents of raffinose and stachyose.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.4
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• pp.353-366
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• 1992

This study was devised to purify the compound from tuna that have cytotoxic activities against various cancer cell lines and to observe its immunopotentiating activities. The cytotoxic compound was partially purified 277 fold, from petroleum ehter extract (crude extract) of tuna by silicic acid column chromatography (fraction D) and thin layer chromatography (Spot I). Cytotoxic activity was monitored using human colon cancer cell, HCT-48. The active compound (Spot I) was composed of seven materials which are fatty acids of four kinds ($C_{14:0},\;C_{16:0},\;C_{17:1},\;and\;C_{18:0}$) and unknown three fat materials. The active compound has cytotoxic activities against various cancer cell lines, that is, murine leukemic lymphocytes (L1210, P388) and human rectal (HRT-18) and colon cancer cells (HCT-48, HT-29). The patterns of size distribution of HCT-48 cells in the medium containing tuna extract were shifted to direction of the small size region. Also, the microscopic shape of HCT-48 cells were shrinked and distracted. The number of plaque forming cell and immunoglobin fraction of serum protein obtained from tuna-treated mice were increased, but natural killer cell activity was not affected.