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Optimization of Protoplast Isolation and Ribonucleoprotein/Nanoparticle Complex Formation in Lentinula edodes (표고버섯의 원형질체 분리 최적화와 RNPs/나노파티클 복합체 형성)

  • Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.178-182
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    • 2022
  • Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

Antioxidant efficacy of Jeju crop extracts using Jeju lava seawater as a solvent (제주 용암해수를 용매로 한 제주 작물 추출물의 항산화 효능)

  • Areumi Park;Yeon-Ji Lee;Nalae Kang;Do-Hyung Kang;Soo-Jin Heo
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.347-355
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    • 2022
  • Jeju lava seawater's abundant minerals are known to exert antioxidant effects that remove the free radicals responsible for aging. Therefore, lava seawater reportedly possesses high commercial value as a functional food material. This study compared and analyzed the antioxidant activities of extracts from crops produced in Jeju (carrots, blueberries, and mandarins) using distilled water and lava seawater as solvents. Lava seawater extracts exhibited higher total polyphenol and flavonoid contents of blueberries and mandarins than distilled water extracts. Furthermore, the antioxidant enzymatic and 2, 2-diphenyl-1-picrylhydrazyl radical-scavenging activities of these crops were higher in lava seawater extracts than in distilled water extracts. In particular, using Vero cells, the ROS-scavenging efficacies of blueberries and mandarins were found to be higher in lava seawater extracts. Meanwhile, the antioxidant activities of carrots were higher in lava seawater extracts, despite no difference in total polyphenol and flavonoid contents. These results suggest that lava seawater exhibits favorable potential as a solvent in the functional food industry, and lava seawater-based Jeju crop extracts are potentially useful as functional food ingredients.

The Effect of Blue Light Interception and SPF Boosting of Sunscreen Prepared with Bandgap-controlled TiO2 (밴드갭이 제어된 TiO2 를 이용한 자외선 차단제의 블루라이트 차단 및 SPF 부스팅 효과)

  • Sung Eun Wang;Jung Kyung Yoon;Gui Su Chung;Sung Bong Kye;Ho Sik Rho;Dae Soo Jung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.2
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    • pp.159-167
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    • 2023
  • Titanium dioxide (TiO2) is commonly used in sunscreen formulations to protect the skin surface and prevent the penetration of harmful ultraviolet (UV) rays by the physical scattering action of light. However, a disadvantage of using TiO2 is that it can cause white turbidity when used on skin due to its inactive mineral ingredient. In addition, when TiO2 particles are reduced to nanosize to eliminate opacity, they can increase the transmittance of visible light and reduce whitening, but may lead to serious skin problems, such as allergic inflammation. To overcome these issues, the bandgap of TiO2 was controlled by adjusting the amount of oxygen defect and nitrogen amount, resulting in color TiO2 tailored to the skin. This innovative technology can reduce the whitening phenomenon and effectively block blue light, which is known to cause skin aging by inducing active oxygen. The bandgap controlled TiO2 compounds proposed in this study are hypoallergenic, broad-spectrum, and environmentally friendly. Furthermore, these compounds have been shown to significantly enhance sun protection factor (SPF) of sunscreens, demonstrating their compatibility with blue light blocking products.

Elderly Sarcopenia and Vitamin B Deficiency: A Relationship? (비타민 B 결핍에 의한 노인성 근감소증)

  • Kisang Kwon;Hye-Jeong Jang;Sun-Nyoung Yu;Soon-Cheol Ahn;O-Yu Kwon
    • Journal of Life Science
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    • v.33 no.7
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    • pp.574-585
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    • 2023
  • Sarcopenia is a leading cause of increased medical and nursing care costs among the elderly. In Korea, preventive measures for sarcopenia are mostly targeted toward the general elderly population without specific diseases. However, it is also necessary to implement measures for elderly individuals living in nursing homes and hospitals, where the prevalence of sarcopenia is high. Currently, computed tomography and/or magnetic resonance imaging are considered standard diagnostic tools. However, their complexity and time-consuming nature make them unsuitable for clinical use. The exact pathophysiological mechanisms of sarcopenia are unclear, as they involve various molecular biological pathways, including decreased exercise, protein and nutrient intake, changes in testosterone and growth hormone, and inflammation. Sarcopenia symptoms can lead to several diseases, such as osteoporosis, fractures, dementia, diabetes, and cardiovascular disease. Vitamin B deficiency is a significant factor in sarcopenia induction, with B vitamins being directly involved in energy and protein metabolism and nerve function. Vitamin B deficiency can lead to neuromuscular and neurogenic disorders, which often overlap with sarcopenia. Suboptimal intake of B vitamins, malabsorption, and anorexia are common among the elderly. This study aims to provide information on the role of water-soluble B vitamins in preventing and controlling muscle mass loss and deterioration among the elderly with sarcopenia. In addition, we discuss the potential of myokines from the B vitamin family in modulating sarcopenia.

Strategies for Development of Seafloor Polymetallic Sulphides in Consideration of International Progress (해저열수광상 개발동향과 우리나라의 대응방안)

  • Park, Seong-Wook;Yang, Hee-Cheol;Jeong, Hyeong-Su
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.13 no.3
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    • pp.271-279
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    • 2008
  • Polymetallic sulphides means hydrothermally formed deposits of sulphide minerals which contain concentrations of metals including, inter alia, copper, lead, zinc, gold and silver. Nautilus is the first company to commercially explore the seafloor polymetallic sulphide deposits. The Company holds exploration licences and exploration applications for more than 370,000 $km^2$ in the jurisdictional seas of Papua New Guinea, Fiji, Tonga, the Solomon Islands and New Zealand along the western Pacific Ocean's Rim of Fire. Neptune Minerals is also a leading explorer and developer in this field, with exploration licences awarded totalling more than 270,000 $km^2$ in the territorial seas or EEZ of New Zealand, Papua New Guinea and the Federated States of Micronesia. These two companies now carry out the most active investment activities for seafloor polymetallic sulphide deposits with a goal of commercial production by 2010. China and Japan carry out exploration activities for the seafloor polymetallic sulphide deposits to secure supplies of strategic metals. China carries out national R&D projects relating to deep sea mineral resources in the world ocean through China Ocean Mineral Resources R&D Association(COMRA). And Japan investigates her own EEZ for exploration of the seafloor polymetallic sulphide deposits. In consideration of aforementioned international activities of coastal nations as well as private companies for exploring the sulphide deposits, Korea shall prepare strategic plans : First, consolidation of the authorities concerned and legislative support; second, determination of main entity of the project; third, securing government's decisive investment of sufficient budget; and lastly, establishment of the mid, long-term plan for development of seafloor polymetallic sulphides deposits.

Prodigiosin Production From Serratia sp. PDGS120915 Isolated From Daeyeon Stream Water in Busan (하천에서 분리한 Serratia sp. PDGS120915의 프로디지오신 생산)

  • Keunho Ji;Young Tae Kim
    • Journal of Life Science
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    • v.34 no.6
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    • pp.377-384
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    • 2024
  • Prodigiosin is a red pigment characterized by a common pyrrolylpyrromethane skeleton. It is produced by Serratia marcescens, Vibrio psychroerythrus, Hahella chejuensis, etc. Prodigiosin has been reported to possess anticancer, immunosuppressant, antifungal antimalarial, and algicidal activities. However, despite prodigiosin's diverse range of activities, its production rate is significantly low and biosynthesis conditions are difficult. Consequently, the selling price is high, and its usability is limited. This study aimed to increase the efficiency of prodigiosin production according to the culture conditions of Serratia. In this study, a bacterial strain PDGS120915 producing prodigiosin was isolated from lightly contaminated stream water in Busan and identified as a strain of Serratia sp. based on 16S rDNA gene sequence analysis and physiological characteristics. The reddish pigment from PDGS120915 was directly extracted using acidified ethanol, and a characterization analysis confirmed that it was a prodigiosin compound. The optimal conditions for pigment production were 25℃, pH 7, and 0% NaCl concentration for a duration of 14 hr. Furthermore, by treating carbon and nitrogen sources, such as fructose and beef extract, respectively, prodigiosin production increased approximately six-fold and four-fold. Among the minerals tested, 0.1% KCl was found to be the most effective for prodigiosin production. Moreover, casein was identified as the most suitable source for prodigiosin production.

Antioxidative Activity of Aralia elata Shoot and Leaf Extracts (두릅 순 및 잎 추출물의 항산화 효과)

  • Cha, Jae-Young;Ahn, Hee-Young;Eom, Kyung-Eun;Park, Bo-Kyung;Jun, Bang-Sil;Cho, Young-Su
    • Journal of Life Science
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    • v.19 no.5
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    • pp.652-658
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    • 2009
  • The comparative activities of aqueous, ethanol, and methanol extracts from Aralia elata shoot (AES) and leaf (AEL) were tested by in vitro experimental models of linoleic acid peroxidation by thiocyanate and thiobarbituric acid (TBA) methods and scavenging activities of free radicals by DPPH (${\alpha}$,${\alpha}$'-diphenyl-${\beta}$-picrylhydrazyl). In addition, bio-active materials (phenolic compounds and minerals) were also measured. The extract yield of each solvent extracted from AES and AEL was 3.08% and 3.13% in aqueous, 0.58% and 0.66% in ethanol, and 0.81% and 1.73% in methanol, respectively. The highest extract yield was found in the aqueous extract from AEL. Major mineral contents (mg%) of AES and AEL were 575.7 and 759.3 in Ca, 353.5 and 330.0 in K, and 31.3 and 31.0 in Mg, respectively. The highest free radical scavenging activity was found in the aqueous extract by 28.69% at 0.1% additional level from AES and in the methanol extract by 92.36% at 0.1% additional level from AEL. Free radical scavenging activity was stronger in AEL than in AES. In antioxidative activities determined by thiocyanate and TBA methods against lipid peroxidation using linoleic acid, ethanol extracts from AEL showed the highest antioxidative activity at all treatment concentrations. These results may provide the basic data to understand the biological activities of bio-active materials derived from AES and AEL.

AMP-activated Kinase Regulates Adipocyte Differentiation Process in 3T3-L1 Adipocytes Treated with Selenium (AMP-activated protein kinase가 셀레늄으로 처리된 3T3-L1 지방세포의 분화과정 억제에 관한 연구)

  • Park, Song-Yi;Hwang, Jin-Taek;Lee, Yun-Kyoung;Kim, Young-Min;Park, Ock-Jin
    • Journal of Life Science
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    • v.19 no.4
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    • pp.423-428
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    • 2009
  • Selenium was investigated using human origin preadipocytes to see whether it possesses preventive or therapeutic effects for obesity. Unveiling the potential of selenium in the reduction of adipogenesis can help predict the therapeutic capabilities of selenium in obesity. In the present study, the molecular mechanism of the inhibition of adipogenesis by selenium was explored to unravel the involvement of the AMP-activated protein kinase. There is emerging evidence that AMPK, a sensor of cellular energy status, is a possible molecular target of controlling adipocyte differentiation on the basis of discovery that AMPK is responsible for the major metabolic responses to exercise, and integration of nutritional and hormonal signals to modulate feeding behavior or energy expenditure in the hypothalamus. Treatment of selenium resulted in inhibition of the adipocyte differentiation process and induction of mature apoptosis in 3T3-L1 adipocytes. We hypothesized that selenium may exert anti-adipogenic potential though modulating AMPK. We have found that selenium significantly activated AMPK and phosphorylated its substrate acetyl-CoA carboxylase ($ACC-serine^{79}$) during the inhibitory process of adipocytes. Also, the inhibition process of adipocyte differentiation by selenium was comparable to either reveratrol or a synthetic AMPK activator, AICAR (5-aminoimidazole-4-carboxamide-1-${\beta}$-D-ribofuranoside). To evaluate the involvement of AMPK in anti-lipogensis, we applied AICAR and Compound C, an AMPK inhibitor, to 3T3-L1-adipocytes and found that AMPK is required for the adipocyte differentiation blocking process. These results suggest that selenium has a potential to control adipogenesis and that this effect is mediated by AMPK, an essential kinase for both inhibition of adipocyte differentiation and apoptosis of mature adipocytes.

Effect of 2-D DBP/PLGA Hybrid Films on Attachment and Proliferation of Intervertebral Disc Cells (2차원적 DBP/PLGA 하이브리드 필름이 디스크 세포의 부착과 증식에 미치는 영향)

  • Ko, Youn-Kyung;Jeong, Jae-Soo;Kim, Soon-Hee;Lim, Ji-Ye;Rhee, John-M.;Kim, Moon-Suk;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.2
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    • pp.109-115
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    • 2008
  • Because demineralized bone particle (DBP) contains various bioactive molecules such as cytokines, it is widely used biomaterials in the field of tissue engineering. In this study, we investigated the effect of 2-dimensional DBP/PLGA hybrid films on adhesion, proliferation and phenotype maintenance of intervertebral disc cells. PLGA films incorporated with different amount (0, 10, 20, 40 and 80 wt%) of DBP were prepared by the solvent evaporation method and characterized by scanning election microscopy (SEM). PLGA film has a flat and smooth surface. According to the increase of content of DBP, the surface of DBP/PLGA film exhibited few agglomerates and increased the roughness of the surface. Annulus fibrosus (AF) and nucleus pulposus (NP) cells were cultured on PLGA and DBP/PLGA film surface, and then examined the cell adhesion and proliferation by the cell count and SEM observation. The result of cell count and SEM observation revealed that 10 and 20% DBP in DBP/PLGA films were superior to adhesion and proliferation of both AF and NP cells. We confirmed that specific gene expression of disc cells on DBP/PLGA film based on the cell count result. Disc cells seeded on 20% DBP/PLGA film expressed the gene of type I and II collagen continuously. Therefore, pertinent content of biomaterials could provide more appropriate condition on adhesion and proliferation of cell. And this results may be used as a basic data for the intervertebral disc regeneration using tissue engineering.

Evaluation of Environment-friendly Control Agents for the Management of Powdery Mildew Infection during Seedling Stage of Three Cucurbitaceae Vegetables (친환경 육묘시 세 가지 박과채소의 흰가루병에 대한 친환경 제제의 방제효과)

  • Yeo, Kyung-Hwan;Jang, Yoon Ah;Kim, Su;Um, Young Chul;Lee, Sang Gyu;Rhee, Han Cheol
    • Journal of Bio-Environment Control
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    • v.22 no.4
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    • pp.413-420
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    • 2013
  • The purpose of this study was to evaluate the control efficacy of major environment-friendly control agents against powdery mildew, as affected by the application condition such as disease-developing stage and microclimate, as compared with the efficacy of pesticides in plug seedling of three Cucurbitaceae vegetables, including cucumber, melon, and oriental melon. Single or combined application of major six environment-friendly control agents was used in the experiment: two biofungicide (Ampelomyces quisqualis 94013 and Bacillus subtilis Y1336), two plant extracts (neem oil and extracts of Rheum undulatum), and two mineral materials (wettable sulfur powder and lime bordeaux mixture). These control agents were treated to the plug seedlings for preventing powdery mildew and curative applications for managing powdery mildew. In all treatments, the disease incidence declined as daily average temperatures increased to $30^{\circ}C$ for consecutive 6~8 days with maximum temperature over $40^{\circ}C$. In preventative application, the control efficacy against powdery mildew was the highest in the treatment of wettable sulfur powder, and lowest in the B. subtilis Y1336, with values of 20~40%. In cucumber seedlings, the preventive single application of neem oil or wettable sulfur powder was more effective than curative application of fungicides, while the control efficacy of these agents was similar to those of fungicides in melon seedlings. The single application of R. undulatum extracts was also effective in preventing the disease for both cucumber and melon seedlings, showing a higher control efficacy than those of biofungicides during seedling stage. The treatment of water spray was not effective and showed a higher disease incidence than the untreated control plot in the oriental melon and melon seedlings. The curative application with environment friendly control agents, after powdery mildew was first detected, could not successfully controlled the disease at the middle stage (5~10% of disease incidence) of disease development. The curative combined application of [R. undulatum extracts (1st application) + wettable sulfur powder (2nd) + neem oil (3rd)] showed the highest control efficacy among the other treatments, with control value over 80% at the early stage (less than 1% of disease incidence) of disease development.