• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1499-1506
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• 2016

Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

• Herbal Formula Science
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• v.27 no.4
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• pp.269-284
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• 2019

This study was to investigate whether cultivated ginseng (CG), cultivated wild simulated ginseng (CWG) and cultivated red ginseng (CRG) extracts influences on the obesity. The saponin contents of 3 kinds of ginsengs were analysed by HPLC-ESI-TOF-MS. Total saponin contents were determined in CG on the most contents and since red ginseng has the highest PD (protopanaxadiol type) / PT (protopanaxatriol type) ratio, there may be differences between ginseng, wild ginseng, and red ginseng with respect to their pharmacological effects. Male C57BL/6J mice were fed a normal diet(N), HFD (60% Kcal fat, C), HFD with CG, CWG and CRG extracts (800 mg/kg) for 5 weeks. We observed change of total body weight, degree of hepatic lipid accumulation and immunohistochemical change of GLP-1 and insulin-secreting cells. Also this study attempts to use the physiological analysis method to analyze the changes of blood lipids, insulin and leptin concentration. The change of body weight and size of accumulated lipid droplets in liver lobules decreased in all of the experimental groups than the control(C) group. In the pancreas, the immunohistochemical density of insulin-secreting cells were significantly stronger in the CWG and CRG than C group. The levels of serum insulin and leptin significantly decreased 55.6%, 54.3% respectively in CWG and CRG. The changes of triglyceride, total cholesterol in serum decreased in CRG than the C group. Obesity related CG, CWG and CRG extracts might have contribute to improvement of obesity by regulating the levels of blood lipids and biochemical indicator of fat accumulation.

• Molecules and Cells
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• v.38 no.4
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• pp.318-326
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• 2015

We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.32-39
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• 2016

Coastal plant species Tetragonia tetragonoides (Pall.) Kuntze native to the Dokdo islands was sampled and then 17 endophytic fungi were purely isolated based on morphological differences. The fungal isolates were characterized by their growth properties under NaCl concentration or pH gradient. Culture filtrates of the 17 fungal isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity. As the results, YH103 strain showed the highest plant growth-promoting activity among them. Phylogenetic analysis of the isolates was done by the maximum likelihood method based on partial internal transcribed spacer region (ITS region: contaning ITS1, 5.8S, and ITS2), beta-tubulin (BenA), and calmodulin (CaM) gene sequences. Chromatographic analysis of the strain YH103 culture filtrate showed the existence of gibberellins ($GA_4$, $GA_7$, $GA_8$, and $GA_{19}$). Finally, the strain YH103 was identified as Aspergillus tubingensis by microscopic observation and molecular analysis and, to our knowledge, this is the first report of GAs producing A. tubingensis.

• Journal of Ginseng Research
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• v.42 no.4
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• pp.562-570
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• 2018

Background: Lung cancer is the leading cause of cancer-related death worldwide. In this study, we used a bioactivity-guided isolation technique to identify constituents of Korean Red Ginseng (KRG) with antiproliferative activity against human lung adenocarcinoma cells. Methods: Bioactivity-guided fractionation and preparative/semipreparative HPLC purification were used with LC/MS analysis to separate the bioactive constituents. Cell viability and apoptosis in human lung cancer cell lines (A549, H1264, H1299, and Calu-6) after treatment with KRG extract fractions and constituents thereof were assessed using the water-soluble tetrazolium salt (WST-1) assay and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, respectively. Caspase activation was assessed by detecting its surrogate marker, cleaved poly adenosine diphosphate (ADP-ribose) polymerase, using an immunoblot assay. The expression and subcellular localization of apoptosis-inducing factor were assessed using immunoblotting and immunofluorescence, respectively. Results and conclusion: Bioactivity-guided fractionation of the KRG extract revealed that its ethyl acetate-soluble fraction exerts significant cytotoxic activity against all human lung cancer cell lines tested by inducing apoptosis. Chemical investigation of the ethyl acetatesoluble fraction led to the isolation of six ginsenosides, including ginsenoside Rb1 (1), ginsenoside Rb2 (2), ginsenoside Rc (3), ginsenoside Rd (4), ginsenoside Rg1 (5), and ginsenoside Rg3 (6). Among the isolated ginsenosides, ginsenoside Rg3 exhibited the most cytotoxic activity against all human lung cancer cell lines examined, with $IC_{50}$ values ranging from $161.1{\mu}M$ to $264.6{\mu}M$. The cytotoxicity of ginsenoside Rg3 was found to be mediated by induction of apoptosis in a caspase-independent manner. These findings provide experimental evidence for a novel biological activity of ginsenoside Rg3 against human lung cancer cells.

• Journal of Environmental Health Sciences
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• v.45 no.6
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• pp.675-687
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• 2019

Objective: Methyl bromide (MB) fumigant has been shown to be fatal to human when was exposed. However, it were still used the significant amount in quarantine disinfection sites. The purposes of this study were to assess the MB exposure status and characteristics by fumigation-related workers and to provide supporting data for management plan. Methods: For this study, the three groups related to fumigation work were composed. A total of 107 workers were directly exposed, 20 field inspectors were indirect, and 20 general quarantines were not exposed. The urinary bromide ion concentrations in each group were analyzed by using HPLC/ICP-MS, and the working characteristics were identified using the structured questionnaire. Results: The urinary bromide ion concentration in the exposed group of fumigation workers was higher than the indirect and non-exposed groups. In the work characteristics of workers, there was a significant tendency to increase urinary bromide ion concentrations with higher fumigation work years (≤4 years: 2.84 (1.13-7.11) mg/g cr, >4-15 years: 5.36 (4.37-6.57) mg/g cr, >15-37 years: 6.69 (5.27-8.49) mg/g cr, p=0.034). In the comparison of the average number of working days per month, the more working days, the higher the urinary bromide ion concentration was statistically significant (≤12.5 days: 2.59 (1.19-5.65) mg/g cr, >12.5-19.25 days: 5.46 (4.62-6.44) mg/g cr, >19.25-27.25 days: 7.93 (5.93-10.59) mg/g cr, p=0.002). Conclusion: This study was the first nationwide survey including biological monitoring in workers exposed to methyl bromide. The results of this study were expected to be used as a reference for workers' health rights in relation to fumigation, prevention of addiction accidents, and safe management plan.

• Korean Journal of Medicinal Crop Science
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• v.24 no.2
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• pp.121-128
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• 2016

Background: Turmeric (Curcuma longa L.) that is used as a food material has antioxidant, anticancer and anti-inflammatory properties. Recently the demand for functional foods and drugs has increased. The present study was carried out to determined of contents of residual sulfur dioxide, heavy metals, ash, acid insoluble ash and curcuminoids in turmeric from the Seoul Yak-ryeong market. Methods and Results: A total of 31 samples were obtained. Residual sulfur dioxide was not detected in any samples. Heavy metals (arsenic, cadmium, lead and mercury) were analyzed by ICP-MS (Inductively Coupled Plasma Mass Spectrometer) and a mercury analyzer and were detected in the ranges of 0.00 - 0.28, 0.00 - 0.07, 0.00 - 0.29 and 0.002 - 0.027mg/g respectively. No significant difference were observed between the average heavy metal contents of domestic and imported tumeric. However, average content of ash in domestic samples (7.8%) were significantly higher than that in imported samples (6.1%), whereas that of curcuminoids was significantly higher in imported samples (47.6mg/g) than in domestic samples (11.2mg/g). The average content of acid insoluble ash was not significantly different between two sample types (0.9% in each). Conclusions: There are no specific standards for turmeric used as food materials. Therefore, this study can be provided as basic data for the establishment of quality standards for turmeric.

• Analytical Science and Technology
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• v.18 no.6
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• pp.542-551
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• 2005

This study is to investigate the characteristics of emission concentration according to wallpaper sort and emission time using small chamber method. The target compounds included 45 VOCs and formaldehyde, which were respectively determined by adsorption sampling and thermal desorption coupled with GC/MS method, and by sampling in DNPH cartridge and HPLC method. The emission factor of TVOC and HCHO was detected to $1.1mg/m^2{\cdot}h$ and $0.01mg/m^2{\cdot}h$ respectively, and the wallpapers of 25 satisfied emission standard. TVOC emission factor appeared in order of the concentration of PVC, natural, and Non-PVC wallpaper, while HCHO was detected very low concentration without relation to wallpaper sort. The paraffin hydrocarbons appeared to be the most contributable class of hydrocarbons in terms of their concentrations, followed by aromatics, and olefins, halogenated hydrocarbons was not detected. PVC wallpapers plentifully emitted TVOC above other wallpapers, and toluene was showed higher concentration of 10 times than natural wallpaper. In addition to, emission factor according to elapse was gradually decreased.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.78-89
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• 2020

Tomato fruit quality is determined by the contents of various functional metabolites in addition to fruit appearance. To develop tomato cultivars with higher amounts of functional compounds, an efficient quantification method is required to identify the natural variations in the compounds in the tomato germplasm. In this study, we investigated tomato varieties, which included 23 inbred lines and 12 commercial F₁ cultivars, for their contents of seven watersoluble vitamins (vitamin C, vitamins B1, B2, B3, B5, B6, and B9) and five polyphenolic compounds (quercetin, rutin, kaempferol, myricetin, and naringenin chalcone). The results of high performance liquid chromatography and liquid chromatography-mass spectrometry showed that vitamin C and naringenin chalcone were the major water-soluble vitamins and polyphenolic compounds, respectively, and their abundance was highly variable depending on the cultivar. By contrast, the contents of vitamin B1, quercetin, and kaempferol were lowest among the cultivars. With regard to the relationship between metabolic compounds and fruit characteristics, a significant association was found in fruit size, indicating that cherry tomato varieties contain higher amounts of the compounds compared to large fresh-type varieties. However, no direct association was detected in fruit color, except for naringenin chalcone. The results of this study provide new insights on the quantification of metabolic compounds and the selection of breeding materials, which are prerequisites for the development of functional tomato varieties.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.719-725
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• 2009

Heterocyclic aromatic amines (HCAs) are potent mutagens and possible human carcinogens that are formed during the heating of protein-rich foods. The effects of preheating treatment of beef patties using a microwave prior to frying at $220^{\circ}C$ for 10 min on each side on the reduction of HCAs (amino-carbolines and amino-imidazo-azaarenes) were evaluated. The amount of HCAs was then evaluated by solid-phase extraction and high pressure liquid chromatography (HPLC) analysis. The beef patties were treated by microwaving for various times (0, 1, 1.5, 2, or 3 min) before pan-frying. The results revealed the presence of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b] indole (Trp-P-1), 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1), 2-aminodipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2), 9H-pyrido[3,4-b]indole (Norharman), 1-methyl-9H-pyrido[3,4-b]indole (Harman), 2-amino-9H-pyrido[2,3-b] indole ($A{\alpha}C$), 2-amino-3,8 dimethylimidazo[4,5-f]-quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) in all samples. However, microwave pretreatment for 1 min inhibited the formation of these HCAs by up to 90% when compared to the control.