• Title/Summary/Keyword: Cell viability test

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Physiological Activities of Fermented Gastrodia elata Blume Extracts (발효 천마 추출물의 생리 활성)

  • Park, An Na;Ku, Tae Kyu;Kim, Kyung Sun;Lee, Dong Won;Kim, Sang Jin
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.4
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    • pp.702-711
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    • 2015
  • This study was conducted to determine the feasibility of using Gastrodia elata Blume as a cosmetic raw material by investigating the physiological activities of its extracts, varying the concentration, solvent, and fermentation method (non-fermentation and fermentation using lactic acid bacteria and effective microorganisms). Of the extracts in three different solvents-water, EtOH, and 70% EtOH-at four different concentrations (0.725, 1.25, 2.5, and 5 mg/mL), the EtOH extracts demonstrated the highest contents of antioxidants (flavonoids, polyphenols, and DPPH free radical scavengers). The DPPH free radical scavenging activity in the EtOH extracts of EM-fermented Gastrodia elata Blume increased from $27.08{\pm}0.5%$ at 1.25 mg/mL to $35.89{\pm}0.8%$ at 2.5 mg/mL. The tyrosinase inhibitory activity test was performed to measure skin-whitening capacity and revealed the LB-fermented EtOH extracts to be the most efficacious ($39.1{\pm}0.4%$ at 0.725 mg/mL, $62.8{\pm}1.5%$ at 2.5 mg/mL). Viability was found to exceed 85% in RAW 264.7 cells treated with all extracts (water, EtOH, 70% EtOH at 10, 25, $50{\mu}L$, fermented and non-fermented), thus proving that Gastrodia elata Blume extracts do not cause inflammation. When RAW 264.7 cells were stimulated with lipopolysaccharide as positive controls under the same conditions to determine the antioxidant activity in the presence of reactive oxygen species (ROS), EM-fermentation was found to impart excellent antioxidant capacity. This study verified the physiological activities of fermented Gastrodia elata Blume extracts that are best suited for cosmetic ingredients, such as antioxidants, tyrosinase inhibitors and anti-inflammatory agents.

Antimicrobial, Antioxidant and Cellular Protective Effects against Oxidative Stress of Anemarrhena asphodeloides Bunge Extract and Fraction (지모 뿌리 추출물과 분획물의 항균활성과 항산화 활성 및 세포보호 연구)

  • Lee, Yun Ju;Song, Ba Reum;Lee, Sang Lae;Shin, Hyuk Soo;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.360-371
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    • 2018
  • Extracts and fractions of Anemarrhena asphodeloides Bunge were prepared and their physiological activities and components were analyzed. Antimicrobial activities of the ethyl acetate and aglycone fractions were $78{\mu}g/ml$ and $31{\mu}g/ml$, respectively, for Staphylococcus aureus and $156{\mu}g/ml$ and $125{\mu}g/ml$, respectively, for Pseudomonas aeruginosa. 1,1-Diphenyl-2-picrylhydrazyl free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction, and aglycone fraction of A. asphodeloides extracts were $146.2{\mu}g/ml$, $23.19{\mu}g/ml$, and $71.06{\mu}g/ml$, respectively. The total antioxidant capacity ($OSC_{50}$) in an $Fe^{3+}$-EDTA/hydrogen peroxide ($H_2O_2$) system were $17.5{\mu}g/ml$, $1.5{\mu}g/ml$, and $1.4{\mu}g/ml$, respectively. The cytoprotective effect (${\tau}_{50}$) in $^1O_2$-induced erythrocyte hemolysis was 181 min with $4{\mu}g/ml$ of the aglycone fraction. The ${\tau}_{50}$ of the aglycone fraction was approximately 4-times higher than that of (+)-${\alpha}$-tocopherol (${\tau}_{50}$, 41 min). Analysis of $H_2O_2$-induced damage of HaCaT cells revealed that the maximum cell viabilities for the 50% ethanol extract, ethyl acetate fraction, and aglycone fraction were 86.23%, 86.59%, and 89.70%, respectively. The aglycone fraction increased cell viability up to 11.53% at $1{\mu}g/ml$ compared to the positive control treated with $H_2O_2$. Analysis of ultraviolet B radiation-induced HaCaT cell damage revealed up to 41.77% decreased intracellular reactive oxygen species in the $2{\mu}g/ml$ aglycone fraction compared with the positive control treated with ultraviolet B radiation. The findings suggest that the extracts and fractions of A. asphodeloides Bunge have potential applications in the field of cosmetics as natural preservatives and antioxidants.

Adhesion Characteristics and the High Pressure Resistance of Biofilm Bacteria in Seawater Reverse Osmosis Desalination Process (역삼투 해수담수화 공정 내 바이오필름 형성 미생물의 부착 및 고압내성 특성)

  • Jung, Ji-Yeon;Lee, Jin-Wook;Kim, Sung-Youn;Kim, In-S.
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.51-57
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    • 2009
  • Biofouling in seawater reverse osmosis (SWRO) desalination process causes many problems such as flux decline, biodegradation of membrane, increased cleaning time, and increased energy consumption and operational cost. Therefore biofouling is considered as the most critical problem in system operation. To control biofouling in early stage, detection of the most problematic bacteria causing biofouling is required. In this study, six model bacteria were chosen; Bacillus sp., Flavobacterium sp., Mycobacterium sp., Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhodobacter sp. based on report in the literature and phylogenetic analysis of seawater intake and fouled RO membrane. The adhesion to RO membrane, the high pressure resistance, and the hydrophobicity of the six model bacteria were examined to find out their fouling potential. Rhodobacter sp. and Mycobacterium sp. were found to attach very well to RO membrane surface compared to others used in this study. The test of hydrophobicity revealed that the bacteria which have high hydrophobicity or similar contact angle with RO membrane ($63^{\circ}$ of contact angle) easily attached to RO membrane surface. P. aeruginosa which is highly hydrophilic ($23.07^{\circ}$ of contact angle) showed the least adhesion characteristic among six model bacteria. After applying a pressure of 800 psi to the sample, Rhodobacter sp. was found to show the highest reduction rate; with 59-73% of the cells removed from the membrane under pressure. P. fluorescens on the other hand analyzed as the most pressure resistant bacteria among six model bacteria. The difference between reduction rates using direct counting and plate counting indicates that the viability of each model bacteria was affected significantly from the high pressure. Most cells subjected to high pressure were unable to form colonies even thought they maintained their structural integrity.

Protective effect of Gabjubaekmok (Diospyros kaki) extract against amyloid beta (Aβ)-induced cognitive impairment in a mouse model (아밀로이드 베타(amyloid beta)로 유도된 인지장애 마우스 모델에서 갑주백목(Diospyros kaki) 추출물의 인지기능 및 뇌 신경세포 보호 효과)

  • Yoo, Seul Ki;Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Han, Hye Ju;Park, Hyo Won;Kim, Chul-Woo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.379-392
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    • 2019
  • The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

Newly Synthesized Phosphodiesterase 4 (PDE4) Inhibitor, DWP205505, Inhibits TNF-$\alpha$ Secretion and mRNA Expression

  • Lee, Suk-Kyeong;Lee, Sun-A;Byun, Hye-Sin;Cho, Mi-La;Kim, Wan-Uk;Park, Sung-Hwan;Cho, Chul-Soo;Joo, Young-Shil;Lee, Shin-Seok;Yoo, Eun-Sook;Son, Ho-Jung;Kim, Ho-Youn
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.106-112
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    • 1999
  • The therapeutic potential of phosphodiesterase 4(PDE4) inhibitors in inflammatory diseases including some autoimmune diseases has been explored recently with some hopeful results. These PDE4 inhibitors are thought to show their anti-inflammatory effect by down-regulating tumor necrosis factor-a (TNF-$\alpha$) production in lymphocytes and macrophages. A high concentration of TNF-$\alpha$has been found in rheumatoid arthritis (RA) synovium and reducing TNF-$\alpha$using biological agents was proven to be an effective RA treatment. To test the possibility of using PDE4 inhibitors for RA treatment, the effects of a newly synthesized PDE4 inhibitor, DWP205505, on TNF-$\alpha$ and IL-10 production was tested in cells isolated from normal peripheral blood and rheumatoid arthritis synovial fluid. Cytokine production was assayed at the protein level by sandwich enzyme-linked immunosorbent assay (ELISA) and at the mRNA expression level by semi-quantitative RT-PCR. Another PDE4 inhibitor, RP73401, was used for comparison. DWP205505 and RP73401 had no harmful effect on cell viability up to 10 $\mu$M concentration during the 24 h culture period. DWP205505 as well as RP73401 significantly reduced TNF-$\alpha$ secretion from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (pBMC) and synovial fluid mononuclear cells (SFMC). The effect of DWP205505 or RP73401 treatment on the mRNA expression of TNF-$\alpha$ was also studied in LPS-stimulated PBMC and SFMC. TNF-$\alpha$ mRNA expression was increased by LPS stimulation and both of the PDE4 inhibitors suppressed TNF-$\alpha$ mRNA expression. For interleukin-l0 (IL-l0), a little different results were obtained from PBMC and SFMC; IL-l0 secretion was unaffected by LPS stimulation and only minimally affected by both of the PDE4 inhibitors in PBMC. In unstimulated SFMC, DWP205505 and RP73401 slightly enhanced IL-10 secretion, while they reduced IL-l0 secretion from LPS-stimulated SFMC where IL-l0 secretion was a lot higher than unstimulated SFMC. These results suggest that the newly synthesized PDE4 inhibitor DWP205505 may have anti-rheumatoid arthritis activity.

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Cryopreservation of Bovine IVM/IVF/IVC Hatched Blastocysts (체외생산된 소 완전탈출 배반포기배의 동결보존)

  • Lee, K.S.;Kim, E.Y.;Yi, B.K.;Nam, H.K.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.141-148
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    • 1999
  • This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø>300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p<0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p<0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p<0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.

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