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Mutant p53-Notch1 Signaling Axis Is Involved in Curcumin-Induced Apoptosis of Breast Cancer Cells

  • Bae, Yun-Hee;Ryu, Jong Hyo;Park, Hyun-Joo;Kim, Kwang Rok;Wee, Hee-Jun;Lee, Ok-Hee;Jang, Hye-Ock;Bae, Moon-Kyoung;Kim, Kyu-Won;Bae, Soo-Kyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.4
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    • pp.291-297
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    • 2013
  • Notch1 has been reported to be highly expressed in triple-negative and other subtypes of breast cancer. Mutant p53 (R280K) is overexpressed in MDA-MB-231 triple-negative human breast cancer cells. The present study aimed to determine whether the mutant p53 can be a potent transcriptional activator of the Notch1 in MDA-MB-231 cells, and explore the role of this mutant p53-Notch1 axis in curcumin-induced apoptosis. We found that curcumin treatment resulted in an induction of apoptosis in MDA-MB-231 cells, together with downregulation of Notch1 and its downstream target, Hes1. This reduction in Notch1 expression was determined to be due to the decreased activity of endogenous mutant p53. We confirmed the suppressive effect of curcumin on Notch1 transcription by performing a Notch1 promoter-driven reporter assay and identified a putative p53-binding site in the Notch1 promoter by EMSA and chromatin immunoprecipitation analysis. Overexpression of mutant p53 increased Notch1 promoter activity, whereas knockdown of mutant p53 by small interfering RNA suppressed Notch1 expression, leading to the induction of cellular apoptosis. Moreover, curcumin-induced apoptosis was further enhanced by the knockdown of Notch1 or mutant p53, but it was decreased by the overexpression of active Notch1. Taken together, our results demonstrate, for the first time, that Notch1 is a transcriptional target of mutant p53 in breast cancer cells and suggest that the targeting of mutant p53 and/or Notch1 may be combined with a chemotherapeutic strategy to improve the response of breast cancer cells to curcumin.

Bacterial core community in soybean rhizosphere (콩 근권의 핵심 세균 군집)

  • Lee, Youngmi;Ahn, Jae-Hyung;Choi, Yu-Mi;Weon, Hang-Yeon;Yoon, Jung-Hoon;Song, Jaekyeong
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.347-354
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    • 2015
  • Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.

A Novel Suberoylanilide Hydroxamic Acid Histone Deacetylase Inhibitor Derivative, N25, Exhibiting Improved Antitumor Activity in both Human U251 and H460 Cells

  • Zhang, Song;Huang, Wei-Bin;Wu, Li;Wang, Lai-You;Ye, Lian-Bao;Feng, Bing-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4331-4338
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    • 2014
  • $N^1$-(2, 5-dimethoxyphenyl)-$N^8$-hydroxyoctanediamide (N25) is a novel SAHA cap derivative of HDACi, with a patent (No. CN 103159646). This invention is a hydroxamic acid compound with a structural formula of $RNHCO(CH_2)6CONHOH$ (wherein R=2, 5dimethoxyaniline), a pharmaceutically acceptable salt which is soluble. In the present study, we investigated the effects of N25 with regard to drug distribution and molecular docking, and anti-proliferation, apoptosis, cell cycling, and $LD_{50}$. First, we designed a molecular approach for modeling selected SAHA derivatives based on available structural information regarding human HDAC8 in complex with SAHA (PDB code 1T69). N25 was found to be stabilized by direct interaction with the HDAC8. Anti-proliferative activity was observed in human glioma U251, U87, T98G cells and human lung cancer H460, A549, H1299 cells at moderate concentrations ($0.5-30{\mu}M$). Compared with SAHA, N25 displayed an increased antitumor activity in U251 and H460 cells. We further analyzed cell death mechanisms activated by N25 in U251 and H460 cells. N25 significantly increased acetylation of Histone 3 and inhibited HDAC4. On RT-PCR analysis, N25 increased the mRNA levels of p21, however, decreased the levels of p53. These resulted in promotion of apoptosis, inducing G0/G1 arrest in U251 cells and G2/M arrest in H460 cells in a time-dependent and dose-dependent manner. In addition, N25 was able to distribute to brain tissue through the blood-brain barrier of mice ($LD_{50}$: 240.840mg/kg). In conclusion, our findings demonstrate that N25 will provide an invaluable tool to investigate the molecular mechanism with potential chemotherapeutic value in several malignancies, especially human glioma.

Functional evaluation of sourdough containing lactic acid bacteria isolated from sliced radish kimchi (깍두기로부터 분리된 유산균으로 제조한 사워도우의 기능성 평가)

  • Lim, Eun-Seo;Kim, Young-Mog;Lee, Eun-Woo
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.180-192
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    • 2017
  • The purpose of this study is to investigate the antioxidative and antimicrobial activities of sourdough fermented with the lactic acid bacteria (LAB) isolated from sliced radish kimchi. According to 16S rRNA gene sequence analysis, the isolated lactic acid bacteria were categorized as Leuconostoc dextranicum SRK03, Lactobacillus brevis SRK15, Pediococcus halophilus SRK22, Lactobacillus acidophilus SRK30, Lactobacillus plantarum SRK38, Leuconostoc citreum SRK 42, and Lactobacillus delbrueckii SRK60 with sequence similarity of 99%. After fermentation with L. dextranicum SRK03, L. acidophilus SRK30, L. plantarum SRK38 or L. delbrueckii SRK60 and Saccharomyces cerevisiae KCTC 7246 at $30^{\circ}C$ for 24 h LAB and yeast in sourdough were present at levels of $10^9$ and $10^7CFU/g$, respectively. In particular, the titratable acidity and ethanol and exopolysaccharide contents of sourdough fermented with L. dextranicum SRK03 were also significantly (P < 0.05) higher than those of sourdough fermented with L. acidophilus SRK30, L. plantarum SRK38, or L. delbrueckii SRK60. The sourdough fermented with L. dextranicum SRK03 and L. acidophilus SRK30 showed not only good DPPH radical-scavenging capacity but anti-lipid peroxidation activity. In addition, the viable counts of Bacillus cereus ATCC 11778 and Staphylococcus aureus ATCC 6538 in sourdough during storage for 5 days at $25^{\circ}C$ were significantly (P < 0.05) lower than those of pathogenic bacteria in the control group due to the organic acids and bacteriocin produced by L. acidophilus SRK30 strain.

Antifungal Activity of Bacillus sp. BCNU 2003 against the Human Pathogenic Fungi (인체 병원성 진균에 대한 Bacillus sp. BCNU 2003의 항진균 효과)

  • Choi, Hye-Jung;Yang, Uk-Hee;Kim, Ya-Ell;Choi, Yeon-Hee;Ahn, Cheol-Soo;Jeong, Young-Kee;Kim, Dong-Wan;Joo, Woo-Hong
    • Journal of Life Science
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    • v.20 no.2
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    • pp.269-274
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    • 2010
  • An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

The study on the causal agent of Streptococcicosis (Lactococcus garvieae), isolated from cultured marine fishes (해산 양식어류로부터 분리된 연쇄구균증의 원인균, Lactococcus garvieae에 대한 연구)

  • Lee, Deok-Chan;Lee, Jae-Il;Park, Chan-Il;Park, Soo-Il
    • Journal of fish pathology
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    • v.14 no.2
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    • pp.71-80
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    • 2001
  • The purpose of this study was to investigate the microbiological characteristics and the distributions of the bacteria causing streptococcicosis occurred in marine fish farm, Korea. Many kinds of cultures fishes suffered from the disease accompanied with typical symptoms, including darkening of the skin, exophthalmia, petechiae inside of the opercula and distended abdomen. The isolates from the diseased fishes were compared with Lactococcus garvieae by biochmical, biophysical and serological methods and polymerase chain reaction(PCR) assay. We isolated 35 strains of the geuns Streptococcus from the diseased olive flounder, Paralichthys olivaceus, yellow tail, Seriola quinqueradiata and Korean rockfish, Sebastes schlegeli. 15 strains out of the isolates were identified to L. garvieae and the others were not because of their different biochemical and biophysical charateristics. Seven strains of the isolates were agglutinated by rabbit serum raised against L. garvieae $KG^+$ phenotypic cells(ATCC49156)as a reference strain. Twenty-one strains of the isolates identified to L. garvieae since they were formed the expected band through performing PCR assay using specific primers, pLG-1(5'-CATAACAATGAGATCGC-3') and pLG-2(5'-GCACCCCGCGGTTG-3'). In the present study, it showed that L. garvieae was a dominant strain causing streptococcicosis in the tested area due to occurrence of 21 strains as L. garvieae out of all the isolates, 9 atrains as Streptococcus sp. and 5 strains as Enterococcus sp.

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A Microbiological Analysis and Hazard Factor Evaluation of Food Utensils and Fixtures of Food Service Operations in Schools (학교 집단급식소 내 식기류 및 집기류의 미생물학적 분석 및 위해요인 평가)

  • Park, Sung-Jun;Hong, Sung-Ho;Lee, Anne Ha-Young;Kim, Cheol-Ju;Kim, Su-Jin;Kim, Sung-Kyoon;Ko, Gwang-Pyo
    • Journal of Environmental Health Sciences
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    • v.37 no.5
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    • pp.376-386
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    • 2011
  • Objectives: The aim of this study was to evaluate the microbial hazards posed by food utensils and fixtures in food service operations at selected middle and high schools located in Seoul, Korea. Methods: We collected 200 samples of utensils and fixtures including cups, spoons/chopsticks, food trays and tables from five different schools in Seoul. Target microorganisms of this study were divided into two groups: total bacterial count and total coliform as indicators of microbial contamination and Bacillus cereus and Staphylococcus aureus as pathogens of food poisoning. We used selective media to quantify microbial concentration and 16S rRNA PCR assay for qualitative analysis. In addition, intensive interviews with nutritionists were conducted and observations were made to identify factors that may affect microbial contamination. Logistic regression analysis was employed to examine the relationship between the microbial concentration and operation characteristics of each operation. Results: The level of microbial concentration in school B and C were significantly lower than in school A, D and E (p<0.05). Some samples from school A, D and E showed over 3.4 log CFU/100 $cm^2$ (total bacterial count) and 1.0 log CFU/100 $cm^2$ (total coliform), which requires immediate hygienic action. The number of customers per staff member, periodicity of hygiene education for staff and daily operation time of sterilizers were also found to be important factors related with the microbial contamination of food service operations. Conclusions: These results suggested that not only a HACCP (Hazard Analysis and Critical Control Point) approach, but also efforts to assess internal risk factors within operations be needed to reduce the microbial contamination of food utensils and fixtures. This study is expected to provide preliminary data for assessing microbial hazards in food service operations.

A Study on Bacterial Contamination of Cooking Environments of Food Service Operations at University (대학 구내식당 식품위생환경의 세균오염도 조사연구)

  • Park, SungJun;Yun, Hyun Sun;Lee, Sujin;Yang, Minji;Kwon, Bomi;Lee, Cheonghoon;Ko, GwangPyo
    • Journal of Environmental Health Sciences
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    • v.40 no.2
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    • pp.88-97
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    • 2014
  • Objectives: The aim of this study was to evaluate the occurrence of microbiological contamination of kitchen utensils and environments of food service operations at university located in Seoul, Korea. Methods: We collected swab samples from the surfaces of knives, chopping boards, floors, and drains, as well as drinking water and airborne bacteria samples from 20 food service operations. Three bacterial indicators and five food poisoning bacteria were measured quantitatively and qualitatively, respectively. We used selective culture media and the PCR assay targeting 16S rRNA gene for the microbiological analysis. Results: We detected bacterial indicators on knives or chopping boards in eight different food service operations and, three food service operations (I, M, and O) showed more than 3 log colony forming units $(CFU)/100cm^2$ on their knives, significantly higher than the others. The levels of bacterial indicators on the floors and drains in the cooking areas were much higher than those on the cooking utensils. S. aureus was detected on 10 floors and 8 drains. Culturable bacteria were identified in 5 drinking water samples, and food service operation B ($431.1CFU/m^3$) and C ($551.2CFU/m^3$) showed more than $400CFU/m^3$ of total airborne bacteria. Conclusions: These results suggest that some of food service operations in this study may require additional investigation to secure the microbial safety of cooking environments. In addition, further actions including hygiene education for employees and proper guidelines to maintain clean cooking environments should be prepared.

Probiotic Potential of Plant-Derived Lactic Acid Bacteria with Antihypertensive Activity (항고혈압 활성을 가진 식물유래 젖산균의 생균제 특성)

  • Lee, Ye-Ram;Son, Young-Jun;Park, Soo-Yun;Jang, Eun-Young;Yoo, Ji-Yeon;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.789-798
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    • 2016
  • Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

Effect of magnesium and calcium phosphate coatings on osteoblastic responses to the titanium surface

  • Park, Ki-Deog;Lee, Bo-Ah;Piao, Xing-Hui;Lee, Kyung-Ku;Park, Sang-Won;Oh, Hee-Kyun;Kim, Young-Joon;Park, Hong-Ju
    • The Journal of Advanced Prosthodontics
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    • v.5 no.4
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    • pp.402-408
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    • 2013
  • PURPOSE. The aim of this study was to evaluate the surface properties and in vitro bioactivity to osteoblasts of magnesium and magnesium-hydroxyapatite coated titanium. MATERIALS AND METHODS. Themagnesium (Mg) and magnesium-hydroxyapatite (Mg-HA) coatings on titanium (Ti) substrates were prepared by radio frequency (RF) and direct current (DC) magnetron sputtering.The samples were divided into non-coated smooth Ti (Ti-S group), Mg coatinggroup (Ti-Mg group), and Mg-HA coating group (Ti-MgHA group).The surface properties were evaluated using scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The surface roughness was evaluated by atomic force microscopy (AFM). Cell adhesion, cell proliferation and alkaline phosphatase (ALP) activity were evaluated using MC3T3-E1 cells. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed. RESULTS. Cross-sectional SEM images showed that Mg and Mg-HA depositionson titanium substrates were performed successfully. The surface roughness appeared to be similaramong the three groups. Ti-MgHA and Ti-Mg group had improved cellular responses with regard to the proliferation, alkaline phosphatase (ALP) activity, and bone-associated markers, such as bone sialoprotein (BSP) and osteocalcin (OCN) mRNA compared to those of Ti-S group. However, the differences between Ti-Mg group and Ti-MgHA group were not significant, in spite of the tendency of higher proliferation, ALP activity and BSP expression in Ti-MgHA group. CONCLUSION. Mg and Mg-HAcoatings could stimulate the differentiation into osteoblastic MC3T3-E1 cells, potentially contributing to rapid osseointegration.