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Effect of Pre-harvest Irradiation of UV-A and UV-B LED in Ginsenosides Content of Ginseng Sprouts (새싹 인삼의 수확 전 UV-A 및 -B LED의 조사에 의한 진세노사이드의 영향)

  • Jang, Seong-Nam;Lee, Ga-Oun;Sim, Han-Sol;Bae, Jin-Su;Lee, Ae-Ryeon;Cho, Du-Yong;Cho, Kye-Man;Son, Ki-Ho
    • Journal of Bio-Environment Control
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    • v.31 no.1
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    • pp.28-34
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    • 2022
  • This study was conducted to determine the changes in ginsenosides content according to additional UV-A, and UV-B LED irradiation before harvesting the ginseng sprouts. One-year-old ginseng seedlings (n=100) were transplanted in a tray containing a ginseng medium. The ginseng sprouts were grown for 37 days at a temperature of 20℃ (24h), a humidity of 70%, and an average light intensity of 80 µmol·m-2·s-1 (photoperiod; 24h) in a container-type plant factory. Ginseng sprouts were then transferred to a custom chamber equipped with UV-A (370 nm; 12.90 W·m-2) and UV-B (300 nm; 0.31 W·m-2) LEDs and treated for 3 days. Growth parameters and ginsenoside contents in shoot and root were conducted by harvesting on days 0 (control), 1, 2, and 3 of UV treatments, respectively. The growth parameters showed non-significant differences between the control and the UV treatments (wavelengths or the number of days). Ginsenoside contents of the shoot was highly improved by 186% in UV-A treatment compared to the control in 3 days of the treatment time. The ginsenoside contents of the roots was more improved in UV-A 1-day treatment and UV-B 3-day treatment, compared to the control by 171% and 160%, respectively. As a result of this experiment, it is thought that UV LED irradiation before harvesting can produce sprout ginseng with high ginsenoside contents in a plant factory.

Study on the Safety of Firefly Luciferase in Human as a Transient Reporter Gene of Oncolytic Virotherapy (항암 바이러스 치료제의 보고유전자로써 반딧불이 루시퍼레이즈의 인체 내 안전성에 대한 연구)

  • Hong, Young Mi;Yoon, Woong Hee;Lee, You Ra;Kim, Soo Ji;Ngabire, Daniel;Narayanasamy, Badrinath;Ornella, Mefotse Saha Cyrelle;Kim, Myunghee;Cho, Euna;Lee, Bora;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.31 no.11
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    • pp.1028-1036
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    • 2021
  • Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

Inhibitory Effect of Protaetiamycine 9 Derived from Protaetia brevitarsis seulensis Larvae on LPS-mediated Inflammation in RAW264.7 Cells (LPS로 자극한 RAW264.7 대식세포에서 흰점박이꽃무지 유충 유래 Protaetiamycine 9의 항염증 효과)

  • Choi, Ra-Yeong;Seo, Minchul;Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Hwang, Jae-Sam
    • Journal of Life Science
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    • v.31 no.11
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    • pp.987-994
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    • 2021
  • Our previous studies have reported that antimicrobial peptides (AMPs) derived from the larvae of white-spotted flower chafer (Protaetia brevitarsis seulensis) exert anti-inflammatory and neuroprotective activities. This study explored the anti-inflammatory effects of protaetiamycine 9 (CVLKKAYFLTNLKLRG-NH2), a novel AMP, derived from P. b. seulensis against lipopolysaccharide (LPS)-mediated inflammatory response in RAW264.7 macrophage cells. Protaetiamycine 9 (25, 50, 75, and 100 ㎍/ml) did not cause cytotoxic effects against RAW264.7 cells. The RAW264.7 cells were pre-treated with various concentrations of protaetiamycine 9 (25-100 ㎍/ml) for 1 hr and then exposed to LPS (100 ng/ml) for 24 hr. Protaetiamycine 9 treatments decreased the LPS-induced secretion of inflammatory mediators, such as nitric oxide (NO), in a dose-dependent manner. Protaetiamycine 9 (25-100 ㎍/ml) effectively downregulated the LPS-induced increase in mRNA and the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), which are involved in the production of inflammatory mediators. Protaetiamycine 9 also suppressed the production and gene expression of pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, compared to the presence of LPS alone. Furthermore, protaetiamycine 9 inhibited the degradation of inhibitory kappa B alpha (IκB-α) and the phosphorylation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. In conclusion, these results suggest that protaetiamycine 9 exhibits LPS-mediated inflammatory responses by blocking IκB-α degradation and MAPK phosphorylation.

Antioxidative Effects of Tenebrio molitor Larvae Extract Against Oxidative Stress in ARPE-19 Cells (ARPE-19 세포에서 산화적 스트레스에 대한 갈색거저리 추출물의 항산화 효과)

  • Bong Sun, Kim;Ra-Yeong, Choi;Eu-Jin, Ban;Joon Ha, Lee;In-Woo, Kim;Minchul, Seo
    • Journal of Life Science
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    • v.32 no.11
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    • pp.865-871
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    • 2022
  • Tenebrio molitor larvae is well known as edible insect. Then, although it has been widely studied that Tenebrio molitor larvae has various bioactive functions such as antioxidant, anti-wrinkle, and anticancer. Nevertheless, antioxidant effects of Tenebrio molitor larvae water extract (TMH) has not been well described in Adult Retina Pigment Epithelial cell line (ARPE-19). In this study, we demonstrated that antioxidant effects of TMH against H2O2-induced oxidative stress in ARPE-19. Thus, we selected for our studies and performed a series of dose-response assay to determine the working concentration that lead to a consistent and high degree of cytotoxicity, which we defined as the level of H2O2 that killed 40% of the ARPE-19 cells. ARPE-19 cells were pre-treated with various concentrations of TMH (0.1 up to 2 mg/ml) before exposure to 300 µM H2O2. As we expected, TMH effectively prevented ARPE-19 cells from 300 µM H2O2-induced cell death in a dose-dependent manner. Furthermore, TMH inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs) such as extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. Overall, the inhibitory effects of TMH on H2O2-induced apoptosis and oxidative stress were associated with the protection cleaved caspase-3, Bax, Bcl-2, and HO-1. The TMH suppressed H2O2-induced cell membrane leakage and oxidative stress in ARPE-19 cells. Thus, these results suggest that the TMH plays an important role in antioxidant effect in ARPE-19.

Clinical Outcomes of Ultrasound-Guided Transmeniscal Injection in Medial Compartment Knee Osteoarthritis (무릎 내측 구획 골관절염에서 초음파 유도하 경 반월상 연골 주사의 임상적 효과)

  • Jung, Eui Yub;Wang, Joon Ho;Lee, Eui-Sub;Lee, Sung-Sahn;So, Sang-Yeon
    • Journal of the Korean Orthopaedic Association
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    • v.55 no.5
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    • pp.418-425
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    • 2020
  • Purpose: The purpose of this study was to introduce the ultrasound-guided transmeniscal injection in medial compartment knee osteoarthritis and analyze the clinical outcomes. Materials and Methods: The electronic medical records of 36 patients with medial compartment knee osteoarthritis who were treated with an ultrasound-guided transmeniscal injection from March 2019 to July 2019 were accessed for this retrospective review. Using an ultrasound guided spinal needle, the patients received an intra-articular steroid injection at the medial compartment of the knee. A physical examination was conducted at the initial visit (pre-injection), and at one week, four weeks, and eight weeks after the injection. The numeric pain rating scale (NRS), Lequesne index, and Western Ontario and McMaster Universities Osteoarthritis (WOMAC) score were measured at each visit and analyzed over time. The percentage change of the patients who revealed substantial improvement was analyzed. The NRS, Lequesne index, and percentage of patients, who revealed substantial improvement over time classified by osteoarthritis grade, were analyzed. Results: The NRS and Lequesne index decreased at one week, four weeks, and eight weeks after the injection compared to the initial baseline, and the pain-relief effect continued without change until eight weeks. The percentage of patients who showed substantial improvement at one, four, and eight weeks was 50.0%, 47.2%, and 52.8%, respectively. The WOMAC scores decreased at one, four, and eight weeks compared to the initial baseline, and the decrease was continued without any difference until eight weeks. The percentage of patients with osteoarthritis stage 1 or 2 who revealed more than substantial improvement was significantly higher at one, four, and eight weeks than those with osteoarthritis stages 3 or 4 (p<0.05). Conclusion: In patients with medial compartment knee osteoarthritis, the pain reduction and functional improvement persisted for at least eight weeks after the ultrasound-guided transmeniscal injection at the medial compartment. In particular, patients with medial compartment osteoarthritis stage 1 or 2 showed more effective pain reduction.

Inhibitory Effects of Rosa rugosa Crude Extract and Solvent Fractions on Adipogenic Differentiation of 3T3-L1 Preadipocytes (해당화 추출물이 3T3-L1 지방세포 분화에 미치는 영향)

  • Lee, Hyunjung;Yang, Jiho;Choi, Mi-na;Jeon, Seongeun;Zhou, Xianrong;Oh, Jung Hwan;Karadeniz, Fatih;Seo, Youngwan;Kong, Chang-Suk
    • Journal of Life Science
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    • v.32 no.12
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    • pp.979-988
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    • 2022
  • Halophytes have been reported to possess a variety of physiological activities, such as anti-cancer, anti-oxidant, anti-diabetes, anti-inflammatory, and anti-obesity. Studies on the roots of the halophyte Rosa rugosa, in particular, have shown a variety of physiological activities and are known to be effective for nursing diabetic complications in traditional Korean medicine. In this study, the effect of R. rugosa on adipogenesis was investigated in 3T3-L1 pre-adipocytes treated with crude extract and solvent fractions (H2O, n-BuOH, 85% aq. MeOH, and n-Hex) obtained from R. rugosa roots. Treatment with extract and the solvent fractions inhibited the formation of intracellular lipid droplets in differentiated 3T3-L1 adipocytes compared to the untreated group. In particular, n-BuOH and 85% aq. MeOH fractions effectively decreased the expression of adipogenic transcription factors: peroxisome proliferator activated receptor-γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα), and sterol regulatory element-binding protein 1c (SREBP1c) in both mRNA and protein levels. In conclusion, these results suggest that R. rugosa contains anti-adipogenic molecules that can be utilized as a nutraceutical against obesity. Further refining of n-BuOH and 85% aq. MeOH fractions and analysis of their action mechanisms could yield potential therapeutic agents with anti-adipogenic effects.

A Protocol of Ludox Treatment for Physiological and Molecular Biological Research of Freshwater Cyanobacteria (퇴적층 남조류 휴면세포의 생리적-분자생물학적 연구를 위한 Ludox 처리법)

  • Keonhee Kim;Kyeong-eun Yoo;Hye-in Ho;Chaehong Park;Hyunjin Kim;Soon-Jin Hwang
    • Korean Journal of Ecology and Environment
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    • v.56 no.1
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    • pp.94-103
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    • 2023
  • Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria's early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.

Evaluation of HbA1c Levels Via the Latex Immunoturbidimetric Method by Using Chemistry Autoanalyzer (자동화학분석기에서의 라텍스 면역비탁법의 Autolab HbA1c 평가)

  • Jo, Yongjun;Lee, So-young;Park, Hae-il;Kim, YeongSic;Lee, Jehoon;Kim, Yonggoo;Han, Kyungja
    • Laboratory Medicine Online
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    • v.2 no.1
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    • pp.10-14
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    • 2012
  • Background: Measurement of HbA1c levels is widely used to diagnose diabetes mellitus and to evaluate and monitor plasma-glucose concentrations over 6-8 weeks. In this study, we evaluated the diagnostic performance of the newly developed latex immunoturbidimetric method by using Autolab HbA1c. Methods: We analyzed and compared the diagnostic performance of Autolab HbA1c with that of Toshiba 200FR between April 2009 and July 2009. According to guidelines (EP5-A2, EP6-P, EP9-A2) of the clinical and laboratory standards institute (CLSI), we compared linearity, precision and correlation of Autolab HbA1c with those of G7 (Tosoh Corp., Kyoto, Japan) by using high-performance liquid chromatography (HPLC) method. Results: Data obtained using Autolab HbA1c showed good linearity in mixtures of samples with low (3.1%) and high (15.1%) levels of HbA1c (r2=0.9997). In the analysis of within-run precision of the samples with HbA1c levels of 5.1% and 12.1%, the SDs were 0.04 and 0.06 and covariances of these samples were 0.8% and 0.5%, respectively. In the Deming regression model, the regression equation was as follows: Autolab HbA1c=1.0859×Tosoh HPLC-0.6957. Conclusions: In this study, Autolab HbA1c method showed better performance characteristics than Tosoh G7 did. In reference review, there was no interference of variant hemoglobin. The data acquisition time of Autolab HbA1c was lower than that of Tosoh G7. The advantages of Autolab HbA1c are that it can be used as an autoanlyzer in routine chemical analysis, it does not require pre-analytical treatment, and the samples are automatically treated with distilled water for hemolysis.

Inhibitory Effect on Kaempferia Parviflora Ethanol Extract of IL-1β Induced Inflammation and MMP Expression in CHON-001 Cells (흑생강 추출물의 CHON-001 세포에서의 IL-1β로 유도된 염증과 MMPs 발현)

  • Jeong Ah Lee;Hye Min Seol;Seong Un Jeong;Jae Hyeon Yoon;Jeong Soo Bae;Tae Hee Kim;Hyeong Soo Kim
    • Journal of Life Science
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    • v.34 no.8
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    • pp.558-566
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    • 2024
  • The potential therapeutic effects of Kaempferia Parviflora ethanol extract (KPE) on osteoarthritis were investigated using the human chondrocyte cell line (CHON-001) to explore its application in functional foods. The CHON-001 cells were pre-treated with either 1 ㎍/ml or 5 ㎍/ml of KPE before exposure to 10 ng/ml of IL-1β to induce osteoarthritis. Results showed that KPE treatment significantly suppressed IL-1β-induced TNF-α production by 66% and 50% at concentrations of 1 ㎍/ml and 5 ㎍/ml KPE, respectively. In addition, COX-2 protein expression was reduced by 26% and 34% compared to control levels. The preservation of chondrocyte-specific matrix proteins, aggrecan, and collagen type II, was notable, with aggrecan and mRNA levels maintained by 5% and 8%, and collagen II levels preserved by 62% and 47% at the same KPE concentrations. This preservation is likely due to the reduced expression of MMP1 and MMP13, enzymes responsible for matrix protein degradation. Overall, the current results suggest that KPE may protect chondrocytes from IL-1β-induced osteoarthritis by suppressing TNF-α production and COX-2 expression while preserving critical matrix proteins like aggrecan and collagen II by suppressing the expressions of their degrading enzymes (MMP-1 and MMP-13). Therefore, KPE holds promise as a candidate for developing functional foods aimed at reducing osteoarthritis.

Long Term Follow Up of Interferon-alpha Treatment in Children with Chronic Hepatitis B (만성 B형간염 환아에 대한 Interferon-alpha 치료결과의 장기 추적관찰)

  • Baek, Seoung-Yon;Eom, Ji-Hyun;Chung, Ki-Sup
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.6 no.2
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    • pp.140-151
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    • 2003
  • Purpose: We tried to evaluate the long term efficacy and positive predictive factors of interferon-alpha treatment in children with chronic hepatitis B. Methods: The study population included 113 children who received interferon therapy between May 1982 and July 2002 (20 years) for chronic hepatitis B in Department of Pediatrics, Yonsei University College of Medicine. Male to female ratio was 2.3 : 1 and the mean age at diagnosis was $11.1{\pm}4.1$ years old. Response to treatment was defined as normalization of alanine aminotransferase (ALT), disappearance of HBeAg and HBV-DNA Eighty two children responded while 32 did not. Interferon-alpha was given intramuscularly for 6 months at a dosage of $3{\times}10^6$ unit, 3 times weekly. In relapsed cases, lamivudine or interferon retreatment was done. Results: Seroconversion rate was 77.0% in terms of HBeAg, 74.3% in terms of HBV-DNA, and 80.5% in terms of ALT normalization after treatment. Seroconversion rate of both HBeAg and HBV-DNA was 72.6%. Analyzed by life table method, the effect of the treatment had been maintained over 10 years after cessation of therapy. Pre-treatment ALT level was the only significant positive predictive factor of response. Eleven cases (13.4%) relapsed, and 2 out of 3 showed response when treated with lamivudine and 1 out of 3 with interferon retreatment. Conclusion: Interferon-alpha showed significant efficacy in the treatment of chronic hepatitis B in our study. Further studies about the effect of interferon therapy on complications of hepatitis such as hepatocarcinoma, cirrhosis are warranted.

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