• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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• pp.137-144
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• 2021

In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.7
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• pp.834-840
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• 2008

In order to improve the use of pumpkin seed, the present study was performed to isolate compositions of the bitter components which were not seen in pumpkin seed itself but newly biosynthesized during germination of the seed. The compositions isolated were then further purified by TLC and preparative HPLC in which a fraction with Rf 0.73 and RT 10.3 was obtained. Cucurbitacin E with molecular weight of 557 from the fraction was finally identified by subsequent structural analysis of LC-MS/MS. The production of cucurbitacin E peaked with 224.7 mg/kg at 4 days of germination at $20^{\circ}C$ with the water supply at ntervals of 48 hrs in the darkness, while that of cucurbitacin E reached 146.7 mg/kg in the brightness. In vitro-cell based assays demonstrated that the isolated and purified cucurbitacin E inhibited proliferation of A549 lung cancer cells and suppressed expression of the IL-$1{\beta}$- or PMA-induced cyclooxygenase-2, an inflammatory protein in A549 cells, suggesting its anti-proliferative and anti-inflammatory activities.

• Herbal Formula Science
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• v.28 no.3
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• pp.255-269
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• 2020

Objectives : Hemistepta lyrata Bunge (Bunge) is a wild herb that has been used for managing fever and wound in Korean Traditional Medicine. The present study explored the effects of H. lyrata extract on liver X receptor (LXR) α-dependent lipogenic genes in hepatocyte-derived cells. Methods : After HepG2 cells or Huh7 cells were pre-treated with 1-10 ㎍/mL of H. lyrata extract or its fractionated extract for 0.5 h, the cells were subsequently exposed to LXR ligand for 6-24 h. Cell viability, LXR response element (LXRE)-driven luciferase activity, sterol regulatory element binding protein-response element (SREBP-RE)-driven luciferase activity, SREBP-1c expression, and mRNA levels of LXRα and its-dependent target genes were determined. In addition, LC-MS/MS analysis was conducted to explore major compounds in H. lyrata-chloroform fractionated extract #4 (HL-CF4). Results : Of various H. lyrata extracts tested, chloroform extract and its fractionated extract #4, HL-CF4, significantly decreased T0901317-mediated SREBP-1c expression. In addition, HL-CF4 significantly reduced LXRE atransactivation and LXRα mRNA expression without any cytotoxicity. Moreover, HL-CF4 prevented the SREBP-RE-driven luciferase activity and mRNA levels of fatty acid synthase and stearoyl-CoA desaturase-1 induced by T0901317. Results from LC-MS/MS analysis at positive/negative mode indicated that HL-CF4 contained several compounds showing m/z 197.1176 (C₁₁H₁₇O₃), 693.2913/227.1069 (C₃₈H₄₅O₁₂/C₁₅H₁₅O₂), 203.1797 (C₁₅H₂₃), 181.1225 (C₁₁H₁₇O₂), 591.2957 (C₃₅H₄₃O₈), 379.1040 (C₁₈H₁₉O₉), 409.1509 (C₂₀H₂₅O₉), 309.1348 (C₁₆H₂₁O₆), 391.1404 (C₂₀H₂₃O₈), and 669.2924/389.1248 (C₃₆H₄₅O₁₂/C₂₀H₂₁O₈). Conclusion : Based on its inhibition of the LXRα-dependent signaling pathway, H. lyrata chloroform extract and HL-CF4 have prophylactic potentials for managing non-alcoholic fatty liver.

• Journal of Food Hygiene and Safety
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• v.22 no.1
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• pp.23-28
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• 2007

The residue depletion of streptomycin was investigated in the olive flounder (Paralichthys olivaceus), rockfish (Sebastes schlegeli), and red sea bream (Pagrus major) after consecutive three days treatment with dipping water at a dose of 20 g/ton water. Fishes were sampled for muscle on 1st, 2nd, 3rd, 4th, and 5th day after treatment. Streptomycin concentrations were determined by high performance liquid chromatography with tandem mass spectrometry. The recovery rates of streptomycin in muscle samples ranged from 87.2 to 102.3% and from 80.4 to 94.1% for the concentration of 0.05 mg/kg and 0.1 mg/kg, respectively. Streptomycin concentrations detected on the 1st day after treatment were 0.066, 0.058, and 0.073 mg/kg in muscles of olive flounder, rockfish, and red sea bream, respectively. At day 2, residue concentrations of all samples were believed to decrease to lower than 0.05 mg/kg, the detection limit. From results of the present study, a withdrawal period of streptomycin is proposed on 3 days after consecutive three days treatment with dipping administration at a dose of 20 g/ton water to avoid the presence of excessive residues of the edible muscles of olive flounder, rockfish, and red sea bream. The present study showed that residue concentrations of streptomycin decreased to below 0.05 mg/kg after treatment 2nd day.

• Journal of Ginseng Research
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• v.41 no.1
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• pp.103-112
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• 2017

Background: 20(S)-Protopanaxadiol 20-O-D-glucopyranoside, also called compound K (CK), exerts antidiabetic effects that are mediated by insulin secretion through adenosine triphosphate (ATP)-sensitive potassium ($K_{ATP}$) channels in pancreatic ${\beta}$-cells. However, the antidiabetic effects of CK may be limited because of its low bioavailability. Methods: In this study, we aimed to enhance the antidiabetic activity and lower the toxicity of CK by including it with ${\beta}$-cyclodextrin (CD) (CD-CK), and to determine whether the CD-CK compound enhanced pancreatic islet recovery, compared to CK alone, in an alloxan-induced diabetic zebrafish model. Furthermore, we confirmed the toxicity of CD-CK relative to CK alone by morphological changes, mitochondrial damage, and TdT-UTP nick end labeling (TUNEL) assays, and determined the ratio between the toxic and therapeutic dose for both compounds to verify the relative safety of CK and CD-CK. Results: The CD-CK conjugate ($EC_{50}=2.158{\mu}M$) enhanced the recovery of pancreatic islets, compared to CK alone ($EC_{50}=7.221{\mu}M$), as assessed in alloxan-induced diabetic zebrafish larvae. In addition, CD-CK ($LC_{50} =20.68{\mu}M$) was less toxic than CK alone ($LC_{50}=14.24{\mu}M$). The therapeutic index of CK and CD-CK was 1.98 and 9.58, respectively. Conclusion: The CD-CK inclusion complex enhanced the recovery of damaged pancreatic islets in diabetic zebrafish. The CD-CK inclusion complex has potential as an effective antidiabetic efficacy with lower toxicity.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.810-817
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• 2009

The effect of cryoprotectants (maltodextrin+glycerol) and cryoprotectants+antioxidant [ascorbic acid and/or butylated hydroxytoluene (BHT)] mixtures on the survival, electrolyte leakage, and lipid degradation of freeze-dried Weissella paramesenteroides LC11 during storage was investigated and compared with that of the control (cells without additives) over a 90-day storage period at 4 or $20^{\circ}C$ in glass tubes with water activity ($a_w$) of 0.23. The survival, electrolyte leakage, and lipid degradation were evaluated through colony counts, electrical conductivity, and thiobarbituric acid reactive substances (TBARS) content, respectively. The fatty acids composition was determined by gas chromatography, in both the total lipid extract and the polar lipid fraction, and compared with that of the control after the 90-day storage period. As the storage proceeded, increases in leakage value and TBARS content, as well as a decrease in viability, were observed. After 90 days of storage, the major fatty acids found in both the total lipid extract and the polar lipid fraction were palmitic (16:0), palmitoleic (16:1), stearic (18:0), oleic (18:1), linoleic (18:2), and linolenic (18:3) acids. The survival, leakage value, TBARS content and 18:2/16:0 or 18:3/16:0 ratio were the greatest for the protected strain held at $4^{\circ}C$. Cells with the cryoprotectants+BHT mixture showed the highest percentage of survival and 18:2/16:0 or 18:3/16:0 ratio in both lipid extracts, as well as the lowest leakage value and TBARS content after the 90-day storage period. Drying cells with the cryoprotectants+BHT mixture considerably slowed down polar lipid degradation and loss of membrane integrity, resulting in improved viability during storage.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.638-644
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• 2007

Differences of meat qualities between Japanese Black and Holstein have been known in Japan, however, the causative proteins and/or the genetic background have been unclear. The aim of this study was to identify candidate proteins causing differences of the meat qualities between the two breeds. Using technique of two-dimensional gel electrophoresis, protein profiling was compared from samples of the longissimus dorsi muscle and subcutaneous adipose tissue. Five protein spots were observed with different expression levels between breeds. By using LC-MS/MS analysis and Mascot program, three of them were identified as ankyrin repeat protein 2, phosphoylated myosin light chain 2 and mimecan protein. Subsequently, we compared the DNA coding sequences of three proteins between breeds to find any nucleotide substitution. However, there was no notable mutation which could affect pI or molecular mass of the proteins. The identified proteins may be responsible for different characteristics of the meat qualities between Japanese Black and Holstein cattle.

• Environmental Analysis Health and Toxicology
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• v.25 no.2
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• pp.131-143
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• 2010

We performed the tests of acute and subchronic inhalation toxicity of methyl formate, which has limited toxicological data in spite of its widespread use and enhanced hazard consequent on its high volatility. The median lethal concentration ($LC_{50}$) was evaluated to be above 5,000ppm(12.27 mg/L). In the test with subchronic inhalation, there are no deaths, but with reduction of body weight, food intake, organ weight by exposure to 400 (0.98 mg/L) and 1,600 (3.92 mg/L) ppm, dose-dependently. There were statistical differences in some hematological and blood biochemical parameters as compared to control (e.g. neutrophile and lymphocyte in the 1,600 ppm group, calcium and A/G in 1,600 ppm group). Methyl formate under the exposure of 1,600 ppm showed the respiratory findings with nasal, it was confirmed that the chemical has respiratory hazard with 1,600 ppm inhalation exposure, induces nasal epithelial atrophy, olfactory cell degeneration/regeneration and the contraction of olfactory cells, etc. According to the notification with Ministry of Labor (No. 2009-68) for classification, labeling and MSDS of chemicals, it is suggested for methyl formate to be classified as category 4 in acute (10.0$4\leq20.0$ mg/L), category 2 (0.2$\leq$1.0 mg/L/6h, 90 days) in specific target organ-repeated exposure.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.565-573
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• 1992

We isolated Actinomycetes strain No. 1882-5, which produces the inhibitor on the bleb formation of K562 cell induced by phorbol ester, from soil sample. Through solvent extraction, Amberlite XAD-4, silica and Lobar low pressure LC, antifungal antibiotic MT 1882-1 and bleb forming inhibitor MT 1882-II were purified from strain No. 1882-5. MT 1882-1 was identified as piericidin $A_{1}$($C_{25}H_{37}0_4N$, M.W. 415) and MT 1882-11 as glucopiericidin A($C_{31}H_{47}0_9N$, M.W. 577) from the analysis of physico-chemical properties and UV, $^1H-NMR$, $^13C-NMR$, and mass spectra of these compounds.

• Environmental Engineering Research
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• v.20 no.2
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• pp.127-132
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• 2015

Perfluorooctane sulfonate (PFOS) and Perfluorooctanoic acid (PFOA) are persistent environmental pollutants, extremely stable, and possibly adversely affect human health. They are widely used in many industries and consumer goods, including sunscreen products. These substances are stable chemicals made of long carbon chains, having both lipid- and water-repellent qualities. The research objectives are (1) to find the most effective method for the preparation of semi-liquid samples by comparing solid phase extraction (SPE) and centrifugation after Pressurized liquid extraction (PLE), and (2) to determine the contamination levels of PFOS and PFOA in waterproof sunscreen samples. All sunscreen samples were analyzed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Sunscreen samples were purchased from domestic and international brands sold in Thailand. Special chemical properties were considered for the selection of samples, e.g., those found in waterproof, sweat resistant, water resistant, and non-stick products. Considering the factors of physical properties, e.g., operation time, chemical consumption, and recovery percentage for selecting methods to develop, the centrifugation method using 2 mL of extracted sample with the conditions of 12,000 rpm and $5^{\circ}C$ for 1 hour after PLE was chosen. The highest concentrations of PFOS and PFOA were detected at 0.0671 ng/g and 21.0644 ng/g, respectively. Even though present concentrations are found at ng/g levels, the daily use of sunscreen products is normally several grams. Therefore, a risk assessment of PFOS and PFOA contamination in sunscreen products is an important concern, and more attention needs to be paid to the long-term effects on human health.