• Korean Journal of Microbiology
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• v.54 no.4
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• pp.330-342
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• 2018

The Streptavidin and Biotin system has been studied most extensively as the high affinity non-covalent binding of Biotin to STR ($K_D=10^{-14}M$) and four Biotin binding sites in tetrameric Streptavidin makes this system useful for the production of multivalent antibody. For the application of this system, we cloned Streptavidin amplified from Streptomyces avidinii chromosome by PCR and fused to gene of hAY4 single-chain Fv antibody specific to death receptor 4 (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand. The hAY4 single-chain Fv antibody fused to Streptavidin expressed in Escherichia coli showed 43 kDa monomer in heated SDS-PAGE. However, this fusion protein shown in both non-heated SDS-PAGE and Size-exclusion chromatography exhibited 172 kDa as a tetramer suggesting that natural tetramerization of Streptavidin by non-covalent association induced hAY4 single-chain Fv tetramerization. This fusion protein retained a Biotin binding activity similar to natural Streptavidin as shown in Ouchterlony assay and ELISA. Death receptor 4 antigen binding activity of purified hAY4 single-chain Fv fused to Streptavidin was also confirmed by ELISA and Westernblot. In addition, surface plasmon resonance analysis showed 60-fold higher antigen binding affinity of the hAY4-STR than monomeric hAY4 ScFv due to tetramerization. In summary, hAY4 single-chain Fv fused to Streptavidin fusion protein was successfully expressed and purified as a soluble tetramer in E. coli and showed both Biotin and DR4 antigen binding activity suggesting possible production of bifunctional and tetrameric ScFv antibody.

• Journal of Korean Society of Forest Science
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• v.110 no.2
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• pp.254-265
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• 2021

As a result of measuring the polyphenol content of Humulus japonicus (HJ) extract according to the collection time and extraction solvent, bothhot water extract and 70% ethanol extract were collected and extracted in June, and the polyphenol content was high. When the harvesting time was the same, the polyphenol content of the ethanol extract was higher than that of the hot water extract. As a result of measuring the antioxidant activity of HJ extract by measuring electron-donating ability, SOD-like activity, and ABTS radical scavenging ability, HJ6E, which has the highest polyphenol content, showed the highest activity. In addition, in the case of the extract collected in August, the polyphenol content was similar. However, the antioxidant activity of the ethanol extract was high, sothe antioxidant activity remained high when extracted with 70% ethanol. As a result of measuring tyrosinase inhibitory activity for evaluating skin whitening activity, HJ6W did not show any activity. The activity at the highest concentration was 16.18% for HJ8W, 8.07% for HJ6E, and 14.7% for HJ8E. Therefore, the content of ingredients showing skin whitening activity was higher in August than in June. In the elastase inhibitory activity for evaluating the anti-wrinkle activityof the skin, the ethanol extract showed very little activity, and the hot water extract did not. In addition, since all extracts do not show astringent activity, it is judged that it is not appropriate to use HJ as a functional ingredient for preventing wrinkle formation. As a result of measuring the cell viability of HJ6E, which showed the highest polyphenol content and antioxidant activity, it showed a cell proliferation effect at low concentrationsbut strong cytotoxicity at concentrations above 50 ㎍/mL. In the case of the NO production inhibitory ability, as the concentration increased, the NO production of Raw 264.7 was suppressed. Theamount of NO production at 1,000 ㎍/mL decreased to 40.7%. However, whether these results are due to cytotoxicity or the extract's efficacy is a part that requires further research.

• Journal of Life Science
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• v.29 no.11
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• pp.1273-1280
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• 2019

Age-related macular degeneration (AMD) is one of the leading causes of blindness in the elderly population, and damage to retinal pigment epithelial (RPE) cells due to oxidative stress contributes to the development of AMD. Glycyrrhiza uralensis Fischer is one of the most widely used herbal medicines for the treatment of various diseases in Asian countries. Although recent studies indicated that treatment with G. uralensis can protect cells from oxidative stress, its mechanisms in RPE cells remain unknown. We evaluated the effect of a G. uralensis ethanol extract (GU) on $H_2O_2$-induced oxidative injury in ARPE-19 RPE cells. The GU pretreatment attenuated reactive oxygen species (ROS) generation induced by $H_2O_2$, which was associated with induced expression of nuclear factor erythroid-derived-2-like 2 (Nrf2) and heme oxygenase-1 (HO-1). GU also suppressed $H_2O_2$-induced DNA damage and mitochondrial dysfunction. The inhibitory effect of GU on $H_2O_2$-induced apoptosis was associated with the protection of caspase-3 activation. Overall, GU appeared to protect RPE cells from oxidative injury by inhibiting DNA damage and reducing apoptosis. Further studies are needed to determine the regulation of Nrf2-mediated HO-1 expression, but our results suggest the possibility of using GU to reduce the risk of AMD.

• Journal of Life Science
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• v.31 no.8
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• pp.719-728
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• 2021

Melanin pigments are abundantly distributed in mammalian skin, hair, eyes, and nervous system. Under normal physiological conditions, melanin protects the skin against various environmental stresses and acts as a physiological redox buffer to maintain homeostasis. However, abnormal melanin accumulation results in various hyperpigmentation conditions, such as chloasma, freckles, senile lentigo, and inflammatory pigmentation. Tyrosinase, a copper-containing enzyme, plays an important role in the regulation of the melanin pigment biosynthetic pathway. Although several whitening agents based on tyrosinase inhibition have been developed, their side effects, such as allergies, DNA damage, mutagenesis, and cytotoxicity of melanocytes, limit their applications. In this study, we synthesized 4-chromanone derivatives (MHY compounds) and investigated their ability to inhibit tyrosinase activity. Of these compounds, (E)-3-(4-hydroxybenzylidene)chroman-4-one (MHY1294) more potently inhibited the enzymatic activity of tyrosinase (IC₅₀ = 5.1±0.86 μM) than kojic acid (14.3±1.43 μM), a representative tyrosinase inhibitor. In addition, MHY1294 showed competitive inhibitory action at the catalytic site of tyrosinase and had greater binding affinity at this site than kojic acid. Furthermore, MHY1294 effectively inhibited α-melanocyte stimulating hormone (α-MSH)-induced melanin synthesis and intracellular tyrosinase activity in B16F10 melanoma cells. The results of the present study indicate that MHY1294 may be considered as a candidate pharmacological agent and cosmetic whitening ingredient.

• Journal of Life Science
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• v.31 no.6
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• pp.603-608
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• 2021

The purpose of this perspective research is to discuss the potential role of exercise-interventions in COVID-19, terms of prevention and prognosis in the periods of the COVID-19 vaccine. SARCO-CoV-2. COVID-19 was detected as a new virus causing severe cardiovascular and respiratory complications. It emerged as a global public health emergency and national pandemic. It caused more than 1 million deaths in the first 6 months of the pandemic and resulted in huge social and economic fluctuations internationally. Unprecedented stressful situations, such as COVID-19 blue and COVID-19 red impact on many health problems. In healthy individuals, COVID-19 infection may induced no symptoms (i.e., asymptomatic), whereas others may experience flu-like symptoms, such as ARDS, pneumonia, and death. Poor health status, such as obesity and cardiovascular and respiratory complications, are high risk factors for COVID-19 prevention, occurrence, and prognosis. Several COVID-19 vaccines are currently in human trials. However, the efficacy and safety of COVID-19 vaccines, including potential side effects, such as anaphylaxis (a life-threatening allergic reaction) and rare blood clots, still need to be investigated. On the basis of direct and indirect evidence, it seems that regular and moderate physical exercise can be recommended as a nonpharmacological, efficient, and safe way to cope with COVID-19. Physical inactivity and metabolic abnormalities are directly associated with reduced immune responses, including reduced innate, CMI, and AMI responses. Due to prolonged viral shedding, quarantine in inactive, obese and disease people should likely be longer than physical active people. Multicomponent and systemic exercise should be considered for the obese, disease, and elderly people. More mechanism research is needed in this area.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.66 no.4
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• pp.307-317
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• 2021

In recent years, global warming has led to frequent climate change-related problems, and elevated temperatures, among adverse climatic factors, represent a critical problem negatively affecting crop growth and yield. In this context, the present study examined the physiological traits of wheat plants grown under high temperatures. Specifically, the effects of elevated temperatures on seed development after heading were evaluated, and the vegetation indices of different organs were assessed using hyperspectral analysis. Among physiological traits, leaf greenness and OJIP parameters were higher in the high-temperature treatment than in the control treatment. Similarly, the leaf photosynthetic rate during seed development was higher in the high-temperature treatment than in the control treatment. Moreover, temperature by organ was higher in the high-temperature treatment than in the control treatment; consequently, the leaf transpiration rate and stomatal conductance were higher in the control treatment than in the high-temperature treatment. On all measuring dates, the weight of spikes and seeds corresponding to the sink organs was greater in the high-temperature treatment than in the control treatment. Additionally, the seed growth rate was higher in the high-temperature treatment than in the control treatment 14 days after heading, which may be attributed to the higher redistribution of photosynthates at the early stage of seed development in the former. In hyperspectral analysis, the vegetation indices related to leaf chlorophyll content and nitrogen state were higher in the high-temperature treatment than in the control treatment after heading. Our results suggest that elevated temperatures after the booting stage positively affect wheat growth and yield.

• Journal of Life Science
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• v.32 no.1
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• pp.29-35
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• 2022

This study was conducted to analyze the genetic polymorphisms of dopamine receptor D4 (DRD4) in horse breeds and its association with substrate characteristics in Jeju crossbreds (Jeju Horse × Thoroughbred). Polymorphisms in DRD4 are candidate genes associated with temperament in various mammals, including humans. Single nucleotide polymorphism (SNP) G292A in the exon 3 region of the horse DRD4 has a reported association with curiosity and vigilance in thoroughbreds. Sanger sequencing was used to identify polymorphisms of the mutations in DRD4 in three horse breeds. The SNP frequency in Jeju horses was significantly different from the frequency in other breeds. Character evaluation, conducted in the Jeju crossbreds and scored using a temperament test and contact test, revealed a high correlation between each test. Comparison of the polymorphism in the DRD4 of horses and the results of the character evaluation revealed lower scores for all temperaments in horses carrying allele A. Comparison of the SNP of G292A and blood dopamine levels in Jeju crossbreds showed 2.87 times higher levels for the GA type than for the GG type. This study identified an association between DRD4 polymorphism and various test methods for evaluating horse temperament and levels of neurotransmitters. Further research could validate the use of this gene as a genetic marker for character evaluation.

• Journal of Practical Agriculture & Fisheries Research
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• v.11 no.1
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• pp.113-123
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• 2009

This study was carried out to determine effects of long-term supply of Cordyceps militaris mycelia on cordycepin content in milk in dairy cows. Ten Holstein cows in the early stages of lactation were divided into two groups. Control group received no supplement whereas treatment group received 6% of C. militaris mycelia of their dry matter intake for 6 months. Feed intake, milk urea nitrogen and somatic cell counts were not affected by long-term supply of C. militaris mycelia for the whole period. In addition, milk yield and milk composition were not affected by long-term supply of C. militaris mycelia at any time of the periods with the exception of milk protein content and yield. The average of milk protein content and yield from the whole period was higher for C. militaris mycelia supplement group than for the control group. As expected, cordycepin in whole blood and milk was not detected in the control group. The range of cordycepin content in the treatment was 0.31~0.38µ/ml and 0.18~0.26(µ/ml for whole blood and milk, respectively. Individual variation was found to be very high and, furthermore cordycepin was undetected in some milk samples. Thus, no clear pattern could be seen in cordycepin content in milk throughout the whole period. Overall, the results of the present study suggest that the transfer efficiency of cordycepin to milk by supplementing C. militaris mycelia in dairy cows was unpredictable and low.

• Journal of Life Science
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• v.33 no.4
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• pp.315-324
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• 2023

The underlying action of policosanol in lowering cholesterol level has not yet been clearly elucidated. Several recent studies have suggested that sterol regulatory element-binding proteins (SREBP)-1c play a role in the regulation of cholesterol synthesis via the fatty acid synthesis pathway. To date, no study has evaluated the effects of policosanol on SREBP-1c-mediated fatty acid synthesis. Therefore, this study aimed to investigate whether the SREBP-1c-mediated fatty acid biosynthetic pathway is associated with the cholesterol-lowering effects of policosanol. Seven-week-old C57BL/6 male mice were randomly divided into 7 groups (n=7 per group) and treated for 8 weeks as follows: 1) normal diet (normal control), 2) high-fat diet (HFD), 3) HFD+ethanol (Pol-0), 4) HFD+policosanol 1 mg/kg (Pol-1), 5) HFD+policosanol 2 mg/kg (Pol-2), 6) HFD+policosanol 4 mg/kg (Pol-4), and 7) HFD+simvastatin 50 ㎍/kg (positive control). Policosanol and simvastatin were administered at the same time every day while maintaining the HFD. Body weight and food intake were measured weekly for 8 weeks. After 8 weeks, serum cholesterol levels were measured, histological analysis was carried out, and the expressions of SREBP-1c and fatty acid synthase (FAS) in the liver tissues were examined. Policosanol reduced body weight and the amount of food intake in a dose-dependent manner. Serum cholesterol levels were significantly lowered in the Pol-1 and Pol-4 groups. The expression of SREBP-1c and FAS was also significantly decreased in the Pol-4 group. These results suggest that the cholesterol-lowering effects of policosanol can occur due to the inhibition of the expression of SREBP-1c and FAS.

• Journal of the Korean Society of International Agriculture
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• v.31 no.1
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• pp.8-16
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• 2019

Alternatives for increasing the competitiveness of locally manufactured agricultural machinery in domestic and foreign markets has been proposed. This was done by analyzing the major agricultural machinery's price and market share as well as their performance and quality. In the Korean domestic market, the market share of Japanese agricultural machinery has been identified to be 14.5% for tractors, 31.1% for combine harvesters, and 35.8% for rice transplanters, and on track for further increase. Japanese manufacturers' domestic patent shares are 58.5% for tractors, 79.9% for combine harvesters, and 69.8% for rice transplanters, showing the dire need for Korean domestic firms to expand their technological rights. To strengthen the industrial competitiveness of agricultural machinery, therefore, researches that develop the fundamental and elemental technology to reduce the frequency of breakdown should be needed in the short term. To achieve this, it is imperative to establish technology roadmap, promote greater cooperation between academia and industry, and systematically increase research funding. In addition, as a long-term solution for enhancing the competitiveness, an establishment of Agricultural Equipment Technology Institute is strongly recommended to systematically support R&D for developing core technologies, particularly high-quality components that guarantee durability and quality.