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Glutamate-rich 4 Binds to Kinesin Superfamily Protein 5A (Glutamate-rich 4와 kinesin superfamily protein 5A와의 결합)

  • Se Young Pyo;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Sang Jin Kim;Mooseong Kim;Jung Goo Lee;Dae-Hyun Seog
    • Journal of Life Science
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    • v.33 no.1
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    • pp.1-7
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    • 2023
  • Intracellular cargo transport is mediated by molecular motor proteins, such as kinesin and cytoplasmic dynein. Kinesins make up a large subfamily of molecular motors. Kinesin-1 is a plus-end-directed molecular motor protein that moves various cargoes, such as organelles, protein complexes, and mRNAs, along a microtubule track. It consists of the kinesin superfamily protein (KIF) 5A, 5B, and 5C (also called kinesin heavy chains) and kinesin light chains (KLCs). Kinesin-1 interacts with many different binding proteins through its carboxyl (C)-terminal region of KIF5s and KLCs, but their binding proteins have not yet been fully identified. In this study, a yeast two-hybrid assay was used to identify the proteins that interact with the KIF5A specific C-terminal region. The assay revealed an interaction between KIF5A and glutamate-rich 4 (ERICH4). ERICH4 bound to the KIF5A specific the C-terminal region but did not interact with the C-terminal region of KIF5B or KIF3A (a motor protein of kinesin-2). In addition, KIF5A did not interact with another isoform, ERICH1. Glutathione S-transferase (GST) pull-downs showed that KIF5A interacts with GST-ERICH4 and GST-ERICH4-amino (N)-terminal but not with GST-ERICH4-C or GST alone. When co-expressed in HEK-293T cells, ERICH4 co-localized with KIF5A and co-immunoprecipitated with KIF5A and KLC but not KIF3B. Together, our findings suggest that ERICH4 is capable of binding to KIF5A and that it may serve as an adaptor protein that links kinesin-1 with cargo.

LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma (황련 추출물의 LC-MS/MS 분석 및 항염증 효과)

  • Min-Jung, Kim;Ye-Jin, Yang;Kwang-Youn, Kim;Hun Hwan, Kim;Jae Dong, Son;Ju-Hye, Yang;Dong bin, Lee;Woo Hyun, Kim;Hu-Jang, Lee;Seon Been, Bak;Kwang-Il, Park
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.1-10
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    • 2023
  • Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

Lysophosphatidic Acid Stimulates SKOV-3 Cell Migration through the Generation of Reactive Oxygen Species via the mTORC2/Akt1/NOX Signaling Axis (리소포스타티드산은 SKOV-3 난소암세포의 mTORC2/Akt1/NOX 신호전달 기전을 통해 활성산소를 형성하고 이를 통해 세포의 이동을 촉진)

  • Eun Kyoung Kim;Seo Yeon Jin;Jung Min Ha;Sun Sik Bae
    • Journal of Life Science
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    • v.33 no.2
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    • pp.129-137
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    • 2023
  • Reactive oxygen species (ROS) play an essential role in a variety of cellular physiological phenomena. The present study assessed the signaling axis that mediates the lysophosphatidic acid (LPA)-induced migration of SKOV-3 cells. Insulin-like growth factor-1 (IGF-1) stimulated SKOV-3 cell migration in a time- and dose-dependent manner. Similarly, LPA stimulated SKOV-3 cell migration and the phosphorylation of Akt in a time- and dose-dependent manner. The pharmacological inhibition of LPA receptors (LPA1/LPA3) significantly suppressed LPA-induced SKOV-3 cell migration. However, IGF-1-induced SKOV-3 cell migration was not affected by the inhibition of LPA1 and LPA3. Pharmacological inhibition of phosphoinositide 3-kinase (PI3K) or Rho-associated kinase (ROCK) significantly suppressed LPA-induced migration, whereas the inhibition of MAPK kinase (MEK) had no effect. Inhibition of PI3K or ROCK completely suppressed LPA-induced ROS generation, and suppression of nicotinamide adenine dinucleotide phosphate oxidase (NOX) or chelation of ROS by N-acetylcysteine (NAC) blocked LPA-induced SKOV-3 cell migration. LPA-induced ROS generation was suppressed by silencing Rictor or Akt1 but not Raptor or Akt2. Silencing Rictor or Akt1 significantly suppressed LPA-induced SKOV-3 cell migration, whereas silencing Raptor or Akt2 had no effect. Finally, the overexpression of the constitutively active form Akt1 (CA-Akt1) significantly enhanced the LPA-induced migration of SKOV-3 cells. Given these results, we suggest that LPA stimulates SKOV-3 cell migration by ROS generation, which is mediated by the mTORC2/Akt1/NOX signaling axis.

Current Status of Sericulture and Insect Industry to Respond to Human Survival Crisis (인류의 생존 위기 대응을 위한 양잠과 곤충 산업의 현황)

  • A-Young, Kim;Kee-Young, Kim;Hee Jung, Choi;Hyun Woo, Park;Young Ho, Koh
    • Korean journal of applied entomology
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    • v.61 no.4
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    • pp.605-614
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    • 2022
  • Two major problems currently threaten human survival on Earth: climate change and the rapid aging of the population in developed countries. Climate change is a result of the increase in greenhouse gas (GHG) concentrations in the atmosphere due to the increase in the use of fossil fuels owing to economic and transportation development. The rapid increase in the age of the population is a result of the rise in life expectancy due to the development of biomedical science and technology and the improvement of personal hygiene in developed countries. To avoid irreversible global climate change, it is necessary to quickly transition from the current fossil fuel-based economy to a zero-carbon renewable energy-based economy that does not emit GHGs. To achieve this goal, the dairy and livestock industry, which generates the most GHGs in the agricultural sector, must transition to using low-carbon emission production methods while simultaneously increasing consumers' preference for low-carbon diets. Although 77% of currently available arable land globally is used to produce livestock feed, only 37% and 18% of the proteins and calories that humans consume come from dairy and livestock farming and industry. Therefore, using edible insects as a protein source represents a good alternative, as it generates less GHG and reduces water consumption and breeding space while ensuring a higher feed conversion rate than that of livestock. Additionally, utilizing the functionality of medicinal insects, such as silkworms, which have been proven to have certain health enhancement effects, it is possible to develop functional foods that can prevent or delay the onset of currently incurable degenerative diseases that occur more frequently in the elderly. Insects are among the first animals to have appeared on Earth, and regardless of whether humans survive, they will continue to adapt, evolve, and thrive. Therefore, the use of various edible and medicinal insects, including silkworms, in industry will provide an important foundation for human survival and prosperity on Earth in the near future by resolving the current two major problems.

Differential Effects of Transforming Growth Factor-β on NKG2D Ligands Expression and NK Cell-mediated Immune Responses in Primary and Metastatic Colon Cancer (원발성 및 전이성 대장암에서 TGF-beta가 NKG2D 리간드 발현과 NK 세포 매개 면역반응에 미치는 영향)

  • Eun-Jung Yun;Yu-Rim Kim;Seong Jun Park;Sang-Yull Lee;Jaeho Bae
    • Journal of Life Science
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    • v.33 no.2
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    • pp.149-157
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    • 2023
  • Transforming growth factor-β (TGF-β) is a multifunctional cytokine that affects not only the survival and growth of cancer cells but also the activity of immune cells. Although it has been generally accepted that cancer cell-derived TGF-β could promote the survival and growth of early cancer cells and have immunosuppressive roles, it has been known that TGF-β has differential effects according to the type or stage of cancer cells. Therefore, it is hard to clearly define its role in cancer progression and immune responses. This study investigated the effects of TGF-β signaling on the expression of five NKG2D ligands and the NK cell-mediated anticancer immune response in the primary colon cancer cell line KM12C and its two metastatic cell lines, KM12SM and KM12L4A. At the surface protein level, exogenous TGF-β decreased the expression of MICA, MICB, ULBP1, and ULBP2, and galunisertib increased the expression of MICA, MIAB, ULBP1, ULBP2, and ULBP3 in KM12C. However, KM12SM and KM12L4A showed no significant changes in the expression of NKG2DLs after treatment with TGF-β or galunisertib. TGF-β signaling inhibition via galunisertib improved the NK cell-mediated anticancer immune response against KM12C but did not show a significant response to KM12SM and KM12L4A. Therefore, the suppression of TGF-β signaling could improve the NK cell-mediated anticancer immune response against KM12C. However, an increase in NKG2DLs expression and an enhanced NK cell-mediated cancer immune response is hard to expect due to the alteration of TGF-β signaling in KM12SM and KM12L4A.

Tyrosinase Inhibition-mediated Anti-melanogenic Effects by Catechin Derivatives Extracted from Ulmus parvifolia (참느릅나무에서 추출된 catechin 유도체 화합물의 멜라닌 생성 억제 효과)

  • Taehyeok Hwang;Hyo Jung Lee;Dong-Min Kang;Kyoung Mi Moon;Jae Cheal Yoo;Mi-Jeong Ahn;Dong Kyu Moon;Dong Kyun Woo
    • Journal of Life Science
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    • v.33 no.2
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    • pp.169-175
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    • 2023
  • As a protective defensive mechanism against ultraviolet (UV) light exposure in skin tissue, melanocytes produce the pigment melanin. Tyrosinase plays a key role in melanin production in melanocytes. However, the overproduction of melanin can lead to lesions, such as freckles and dark spots. Thus, it is clinically important to find a modulating molecule to control melanogenesis by regulating tyrosinase expression and/or activity. It is known that catechin, a plant flavonoid, can reduce melano- genesis through the downregulation of tyrosinase expression. Here, we tested whether catechin derivatives isolated from the stem bark of Ulmus parvifolia have an effect on melanin production by regulating tyrosinase in mouse melanoma cells and in vitro mushroom tyrosinase. The catechin derivatives used in this study included C5A, C7A, C7G, and C7X. Treatments using these catechin derivatives reduced melanin production in mouse melanoma B16F10 cells in which melanogenesis was stimulated by α-MSH. Notably, the anti-melanogenic effects of catechin derivatives were similar to those of kojic acid, a well-known anti-melanogenic molecule. Both C5A and C7A directly inhibited the activity of tyrosinase isolated from mushrooms in vitro. Furthermore, our in silico computational simulation showed that these two compounds were expected to bind to the active site of tyrosinase, which is similar to kojic acid. In addition, all four catechin derivatives reduced tyrosinase protein expression. In summary, our results showed that catechin derivatives can reduce melanogenesis by regulating tyrosinase activity or expression. Thus, this study suggests that catechin derivatives isolated from U. parvifolia can be novel modulators of melanin production.

Morphometric Characterization of Newly Defined Subspecies Apis cerana koreana (Hymenoptera: Apidae) in the Republic of Korea (국내 토종벌(Apis cerana koreana) 아종의 형태적 특성 분석)

  • Olga, Frunze;Jung-Eun, Kim;Dongwon, Kim;Eun-Jin, Kang;Kyungmun, Kim;Bo-Sun, Park;Yong-Soo, Choi
    • Korean journal of applied entomology
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    • v.61 no.3
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    • pp.399-408
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    • 2022
  • There has been much debate on the morphometric divergence between the recently identified Apis cerana koreana and Apis cerana honey bees. The aim of this study was to obtain phenotypic information that can be used to compare A. c. koreana data with other A. cerana subspecies data from open resources and determine breeding results on the basis of morphometric traits. To differentiate A. c. koreana, we investigated 22 classic morphological characteristics; royal jelly secretion; and the weight of workers, queens, and drones of A. c. koreana bred in Korea. To define the selection results, we used the geometric morphometric method. The artificially selected A. c. koreana secreted significantly more royal jelly (1.18 times) than the naturally selected A. c. koreana, which positively influenced the health of the colonies. These honey bees were identified more clearly with the geometric morphometric method than with the classic morphometric method, which is traditionally used to determine the subspecies. Large trends were noted for A. c. koreana on the basis of our results and literature from the 1980s regarding A. cerana sizes in Korea (tarsal index, length of forewing, and cubital index were measured). The cluster analysis revealed the proximity of A. c. koreana, A. cerana in China, and A. c. indica on the basis of eight classic characters, which, perhaps, relay the origin of the honey bees. The results of this study defined the morphometric responses of A. c. koreana honey bees to geographic isolation, climate change, and selection, which are important to identify, protect, and preserve honey bee stock in Korea.

Developmental and Reproductive Characteristics of Mythimna loreyi (Noctuidae) Reared on Artificial Diets (인공사료로 사육한 뒷흰가는줄무늬밤나방(Mythimna loreyi ) (밤나방과)의 발육과 생식 특성)

  • Eun Young, Kim;I Hyeon, Kim;Jin Kyo, Jung
    • Korean journal of applied entomology
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    • v.61 no.3
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    • pp.423-434
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    • 2022
  • The two previously developed artificial diets (N4 and N6) used for rearing Spodoptera frugiperda (Noctuidae) larvae, were selected as highly-fit ones for rearing Mythimna loreyi larvae. Almost all biological characteristics were not significantly different between the colonies reared on the two diets at 25℃ and 15:9 h (light:dark) photoperiod. The developmental periods were 4.9-5.2 days for eggs, and 22.3-23.2 days for larvae. The pupal period and weight were different between the sexes in each diet colony. The pupal periods in females and males showed 12.6-12.8 days and 14.1-14.5 days, respectively. The pupal weights were ca. 345 mg for females and ca. 380 mg for males. The pupation and emergence rates were ca. 91-94%, and ca. 91-95%, respectively, without significant differences between the two colonies. The pre-oviposition and oviposition periods were 3.4 days and 4.7-4.8 days, respectively. The adult longevity was 8.2 days in females and 10.3-12.4 days in males. Total offsprings produced were found to be 724-847 larvae on an average with ca. 1,400 maximum larvae. In the life table analysis, the intrinsic rates of increases (0.1181 for N4 and 0.1253 for N6) were not significantly different between the two colonies. Individual differences in the larval instar number 5 and 6 were found within a diet colony. The ratios of 5-instar larvae were ca. 22% in N4 colony and ca. 7% in N6 colony. The larval period of 6-instar larvae was longer than that of 5-instar larvae. Width of head capsule in larvae varied from ca. 309 ㎛ for 1st instar to ca. 3,065 ㎛ for 6th instar. Body lengths measured from ca. 2.0 mm for 1st instar to ca. 29.1 mm for 6th instar. Larvae of M. loreyi and M. separata were found at the same time in a maize field during June and July, 2020.

Effects of the Aroma Roll-on Inhalation Method on Stress, Pain, Depression and Blood Pressure in the Elderly in a Senior Citizen Clubs (아로마 롤온 흡입법이 경로당 이용 노인의 스트레스, 통증, 우울 및 혈압에 미치는 효과 )

  • Eun-Kyoung Han;Soo-Ah Jo;Ga-Eul Park;Ji-Eun Kim;Soo-Yeon Lee;Eun-Kyung Choi;Yeon-Ju Chae;Eun-Bin Oh;Jo-Hyeon Kim;Ha-Young Lee;Ji-Woo Yang;Yeong-Ju Jeong;Soo-Ah Park;Jin-Ah Kwon;Min-Jung Kim;Eun-Seo Choi;Ju-Eon Yang;Ga-Eun Jo;Ji-Woo Yang
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.1
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    • pp.13-23
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    • 2023
  • This study is an experimental study on the single group pre-post test design for the effects of the aroma inhalation method with roll-on on stress, pain, depression and blood pressure of the elderly. 33 elderly people who met the selection criteria participated among the elderly using the senior citizen clubs located in S city, Gyeonggi-do. The data collection period was from November 14, 2022 to December 19, 2022. A questionnaire consisting of stress, pain, depression, and blood pressure was measured before and after the aroma roll-on inhalation intervention. The components of aroma essential oil included Marjoram, Geranium, Peppermint and Orange sweet that were diluted in Jojoba oil which was the carrier oil. The aroma inhalation method with roll-on was carried out 3 times day for 4 weeks. In the result of this study, the aroma inhalation method with roll-on were statistically significant differences in stress(t=2.63, p=.013), pain(t=2.46, p=.019), and systolic blood pressure(t=3.01, p=.005) of the participants. Therefore, aroma roll-on therapy is expected to be available as a useful nursing intervention method for reducing stress, relieving pain, and managing blood pressure in the elderly using the senior citizen clubs.

Deep Learning Approaches for Accurate Weed Area Assessment in Maize Fields (딥러닝 기반 옥수수 포장의 잡초 면적 평가)

  • Hyeok-jin Bak;Dongwon Kwon;Wan-Gyu Sang;Ho-young Ban;Sungyul Chang;Jae-Kyeong Baek;Yun-Ho Lee;Woo-jin Im;Myung-chul Seo;Jung-Il Cho
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.25 no.1
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    • pp.17-27
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    • 2023
  • Weeds are one of the factors that reduce crop yield through nutrient and photosynthetic competition. Quantification of weed density are an important part of making accurate decisions for precision weeding. In this study, we tried to quantify the density of weeds in images of maize fields taken by unmanned aerial vehicle (UAV). UAV image data collection took place in maize fields from May 17 to June 4, 2021, when maize was in its early growth stage. UAV images were labeled with pixels from maize and those without and the cropped to be used as the input data of the semantic segmentation network for the maize detection model. We trained a model to separate maize from background using the deep learning segmentation networks DeepLabV3+, U-Net, Linknet, and FPN. All four models showed pixel accuracy of 0.97, and the mIOU score was 0.76 and 0.74 in DeepLabV3+ and U-Net, higher than 0.69 for Linknet and FPN. Weed density was calculated as the difference between the green area classified as ExGR (Excess green-Excess red) and the maize area predicted by the model. Each image evaluated for weed density was recombined to quantify and visualize the distribution and density of weeds in a wide range of maize fields. We propose a method to quantify weed density for accurate weeding by effectively separating weeds, maize, and background from UAV images of maize fields.