• Title/Summary/Keyword: sub-G1 DNA

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[ $2{\beta}$ ], $3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic Acid Induces the Apoptosis of Human Hepatoma HepG2 Cells ($2{\beta}$, $3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid처리에 의한 인간 간암세포주 HepG2의 apoptosis 유도)

  • Yoo, Ki-Hyun;Lee, Jong-Min;HwangBo, Jeon;Song, Myoung-Chong;Yang, Hye-Joung;Baek, Nam-In;Kim, Soung-Hoon;Kim, Dae-Keun;Kwon, Byoung-Mok;Park, Mi-Hyun;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.270-275
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    • 2006
  • [ $2{\beta},\;3{\alpha}$ ], 23-trihydroxyrus-12-ene-28-oic acid was isolated from Trapa pseudoincisa S. et Z. It has a common structure of pentacyclic triterpenes and belongs to the amyrin ursolic acid group. The cytotoxic effect of this compound was investigated in human hepatoma cell line HepG2. $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid showed dose-dependent cytotoxicity in HepG2 cells. Confocal microscopy data showed that green fluorescence was increased in $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid treated-HepG2 cells in a time-dependent manner. $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid also increased the sub-G1 cell population of HepG2 cells as well as ladder-like DNA fragmentation. Taken together, our results indicate that $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid induced apoptosis in HepG2 cells.

Combination of Enrichment and PCR in Rapid Semi-Quantification of Bacillus cereus in Fresh-Cut Vegetables

  • Choi, Yukyung;Lee, Sujung;Yoon, Yohan
    • Journal of Food Hygiene and Safety
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    • v.35 no.4
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    • pp.319-325
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    • 2020
  • Widespread consumption of fresh-cut vegetables without cooking results in ingestion of major foodborne pathogens including Bacillus cereus. In this study, we aimed to develop a method to rapidly detect B. cereus in fresh-cut vegetables by combining commercial PCR analysis with enrichment of the pathogenic levels. A mixture of B. cereus strains (KCTC1013, KCTC1014, KCTC1092, KCTC1094, and KCTC3624) was inoculated on the surface of fresh-cut cabbage lettuce (20 g) and baby leafy vegetables (10 g) to concentration 1, 2, 3, 4, and 5 log CFU/g. Eighty milliliters of TSB with 0.15% polymyxin B was used for cabbage lettuce, and 90 mL of medium was used for baby leafy vegetables and incubated at 42℃ for 0, 2, 3, 4, 5, 6, and 7 h. One milliliter of the enriched media was plated on mannitol-egg yolk-polymyxin agar for quantification, and another 1 mL was used for DNA extraction for PCR analysis. Additionally, the minimum number of sub-samples to be tested from a pack of fresh-cut vegetable samples was determined using 5 sub-samples. The results from this study showed that for detecting B. cereus in fresh-cut cabbage lettuce, 3, 4, 5, 6, and 7 h enrichment were required to at least detect 5, 4, 3, 2, and 1 log CFU/g of B. cereus, respectively. B. cereus in fresh-cut baby leafy vegetables could be detected after 2, 3, 4, 5, and 6 h of enrichment at 5, 4, 3, 2, and 1 log CFU/g, respectively, using a combination of enrichment and PCR analysis. To determine if a pack of fresh-cut vegetable is positive, the minimum number of sub-samples should be 3. These results can be used to develop a rapid detection method to semi-quantify B. cereus in fresh-cut vegetable samples combining enrichment and PCR.

Antitumor Activity of Paecilomyces japonica is Mediated by Apoptotic Cell Death

  • Park, Youn-Hee;Moon, Eun-Kyung;Shin, Yong-Kyu;Bae, Myung-Ae;Kim, Jong-Guk;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.16-20
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    • 2000
  • The aqueous extract from the cultural mycelium of Paecilomyces japonica showed cytotoxicity against several tumor cells including Jurkat, U937, HL-60, HepG2, BW5147.G.1.4, and NIH3T3. When the aqueous extract was fractionated by sequential organic solvent extractions using n-hexane and ethyl acetate, the ethyl acetate fraction appeared to contain the most cytotoxic activity, and the $IC_{50}$ values for various tumor cells were in the range from 1.5 to $10.0{\;}\mu\textrm{g}/ml$. To elucidate the cellular mechanism underlying the induced cytotoxicity, the apoptotic DNA fragmentation along with the cell cycle proression was examined in Jurkat T cells following the ethyl acetate fraction treatment. In the presence of $2.5{\;}\mu\textrm{g}/ml$ of the ethyl acetate fraction, apoptotic DNA fragmentation of the cells was detected within 1 h and increased upto 24 h in a time-dependent manner. Under the same conditions, a sub-G1 peak was detectable by flow cytometry. These results indicate that the cytotoxic effect of P. japonica on tumor cells is attributable to the induced apoptosis.

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Dependence of Sub-Cellular Activities of the Blooming and Harmful Dinoflagellate Cochlodinium Polykrikoides on Temperature (수온에 따른 유해성 Cochlodinium polykrikoides 적조생물의 세포생리 변화)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1194-1201
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    • 2008
  • Water temperature-dependent fluctuations of biochemical and molecular activities in the harmful dinoflagellate, Cochlodinium polykrikoides were studied. In terms of genomic DNA concentration, a similar value of 0.6 was observed at $12^{\circ}C$ and $15^{\circ}C$. However, DNA significantly increased beyond $18^{\circ}C$ (p<0.05), to a maximum of 1.8 at $24^{\circ}C$. DNA concentration significantly decreased to 0.6. The concentrations of RNA and total protein were likely at their highest values of 1.7 and 0.07 ${\mu}g$ $ml^{-1}$ at $24^{\circ}C$, respectively. RNA and total protein concentrations began to increase at $15^{\circ}C$. Oxygen availability between lower and higher temperatures was significantly different and increased from $18^{\circ}C$ according to light intensity, regardless of wavelengths (p<0.05). At $24^{\circ}C$, the highest value of the maximum electron transport rate ($ETR_{max}$), ranging from 537.9 (Ch 1) to 602.5 ${\mu}mol$ electrons $g^{-1}$ Chl ${\alpha}s^{-1}$ (Ch 4), was also apparent. Nitrate reductase (NR) and ATPase activities were at their highest values of 0.11 ${\mu}mol$ $NO_{2}^{-}$ ${\mu}g^{-1}$ Chl ${\alpha}h^{-1}$ and 0.78 pmol 100 $mg^{-1}$ at $24^{\circ}C$, respectively. In an analysis of CHN, the concentration of C and N also significantly increased (p<0.05). Most of the measurements for the cellular activities at $27^{\circ}C$, however, were less than at $24^{\circ}C$. These results suggest that the sub-cellular activities of C. polykrikoides are sensitive to changes in water temperature. It may be desirable to estimate at $18^{\circ}C$ the initiation of the massive blooming development of C. polykrikoides. In nature, it will be very difficult to maintain the massive blooms beyond $24^{\circ}C$ because of a possibly significant decrease in molecular activity of C. polykrikoides.

Apoptotic Effect of Vitisin A from Vitis Amurensis against MES-SA Uterine Cancer Cells (왕머루 포도에서 분리한 Vitisin A의 자궁암주에 대한 자멸사 효과)

  • Lim, Jeong-Han;Lee, Hyo-Jeong;Lee, Eun-Ok;Lee, Hyo-Jung;Kwon, Hee-Young;Shim, Bum-Sang;Ahn, Kyoo-Seok;Kim, Sung-Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.290-295
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    • 2008
  • The cytotoxic characteristics of Vitsin A isolated from Vitis amurensis L. were examined in human colorectal, breast, uterine and renal cancer cells. Vitsin A showed good cytotoxicity against various cancer cells with $IC_{50}$ of $1\;{\sim}\;30\;{\mu}M$. Among them, Vitisin A exhibited strongest cytotoxic effect against MES-SA cells with $IC_{50}$ of 1.11 ${\mu}M$ by SRB assay. To verify whether the cytotoxicity of Vitisin A may be associated with apoptosis, TdT-mediated-dUTP Nick-End Labeling (TUNEL) assay and cell cycle analysis were performed in MES-SA cells. Apoptotic bodies were observed in Vitisin A treated MES-SA cells by TUNEL assay. Also, Vitisin A effectively increased the portion of $sub-G_1$ DNA content by flow cytometric analysis. Taken together, these findings suggest that the cytotoxicity of Vitisin A against MES-SA cells is chiefly mediated by apoptosis.

Cisplatin and Extract of Tissue Cultured Mountain Ginseng-Induced Apoptosis in Human Cervical Cancer Cells (인체 자궁암세포에서 cisplatin과 산삼배양근추출물에 의한 apoptosis유도)

  • Lee, Myeong-Seon
    • Applied Microscopy
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    • v.40 no.3
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    • pp.133-138
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    • 2010
  • Mountain ginseng is a perennial crop rarely found in the deep mountains of Korea. The medicinal effect of the mountain ginseng is well known as a panacea in traditional Chinese medicine for a long time. But scientific studies to elucidate the medicinal effect of the mountain ginseng have never been made on account of lack of sample. Recently an improved method of adventitious root culture system through the use of bioreactor has been developed in Panax ginseng that seems to be a reliable way of commercialization of root derived secondary metabolites. This experiment was conducted to evaluated chemotherapeutic effect against human cervical cancer cells by cisplatin (CDDP) and extract of tissue cultured mountain ginseng (ETCMG). CDDP and ETCMG-induced apoptotic cell death in human cervical cancer cell line, HeLa was confirmed by the analysis of cell growth, morphological changes, DNA fragmentation, flow cytometry showed that ETCMG is an inducer of apoptosis and synergizes with CDDP. These results suggest that ETCMG present evidence of anticancer effect and could have a possibly natural therapeutic potential in cervical cancer patients.

Labrenzia callyspongiae sp. nov., Isolated from Marine Sponge Callyspongia elegans in Jeju Island

  • Park, So Hyun;Kim, Ji Young;Heo, Moon Soo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1969-1974
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    • 2019
  • A Gram-staining-negative, aerobic, light brown pigment bacterium, designated strain CE80T was isolated from marine sponge Callyspongia elegans in Jeju Island, Republic of Korea. Strain CE80T grew optimally at 25℃, in the range of pH 5.0-11.0 (optimum 7.0-8.0), and with 1.0-5.0% NaCl (optimum 1-3% (w/v)). Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain CE80T belonged to the genus Labrenzia and was closely related to L. suaedae YC6927T (98.3%), L. alexandrii DFL-11T (96.6%), L. aggregata IAM 12614T (96.6%) L. marina mano18T (96.5%) and L. alba CECT 5094T (96.2%). The major fatty acids of strain CE80T were C18:1 ω7c, and summed feature. The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamin, one unidentified aminolipid, one phospholipid and four unidentified lipids. The DNA G+C content of strain CE80T was 55.9 mol%. The major respiratory quinone was Q-10. DNA-DNA relatedness between strain CE80T and L. suaedae YC6927T was 56.1±2.8%. On the basis of physiological and biochemical characterization and phylogenetic and chemotaxonomic analysis, strain CE80T represents a novel species of the Labrenzia, for which the name Labrenzia callyspongiae sp. nov., is proposed. The type strain is CE80T (=KCTC 42849T =JCM 31309T).

Epigenetic Changes in Asthma: Role of DNA CpG Methylation

  • Bae, Da-Jeong;Jun, Ji Ae;Chang, Hun Soo;Park, Jong Sook;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.83 no.1
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    • pp.1-13
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    • 2020
  • For the past three decades, more than a thousand of genetic studies have been performed to find out the genetic variants responsible for the risk of asthma. Until now, all of the discovered single nucleotide polymorphisms have explained genetic effects less than initially expected. Thus, clarification of environmental factors has been brought up to overcome the 'missing' heritability. The most exciting solution is epigenesis because it intervenes at the junction between the genome and the environment. Epigenesis is an alteration of genetic expression without changes of DNA sequence caused by environmental factors such as nutrients, allergens, cigarette smoke, air pollutants, use of drugs and infectious agents during pre- and post-natal periods and even in adulthood. Three major forms of epigenesis are composed of DNA methylation, histone modifications, and specific microRNA. Recently, several studies have been published on epigenesis in asthma and allergy as a powerful tool for research of genetic heritability in asthma albeit epigenetic changes are at the starting point to obtain the data on specific phenotypes of asthma. In this presentation, we mainly review the potential role of DNA CpG methylation in the risk of asthma and its sub-phenotypes including nonsteroidal anti-inflammatory exacerbated respiratory diseases.

Induction of Apoptosis by Methanol Extract of Endlicheria anomala in Human Lung and Liver Cancer Cells (Endlicheria anomala 메탄올 추출물에 의한 인체 폐암세포주와 간암세포주의 자가사멸 유도)

  • Park, Hyun-jin;Jin, Soojung;Oh, You Na;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.25 no.4
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    • pp.441-449
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    • 2015
  • Endlicheria anomala, a neotropical plant, is found in northern South America and the Amazon region. It is traditionally used to remove poisons and cure gangrene. According to recent data, this plant has diverse biological properties such as anti-oxidative, anti-inflammatory and anti-melanogenic properties. However, the anti-cancer effect of E. anomala and its molecular mechanisms remain unclear. In this study, we examined the anti-cancer effect and the active mechanism of methanol extract of E. anomala (MEEA) in human lung adenocarcinoma cells (A549) and human liver cancer cells (HepG2). Our data revealed that MEEA showed cytotoxic activity in a dose-dependent manner and induced apoptosis both in A549 and HepG2 cells. We verified evidences of apoptosis via formation of chromatin condensation, apoptotic body and accumulation of cells in the subG1 phase. Following observed apoptosis-related phenomena, we found that the induction of apoptosis by MEEA was associated with the increase of tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) expression. Furthermore, MEEA-induced apoptosis was characterized with proteolytic activation of caspase-3, degradation of poly ADP ribose polymerase (PARP), and up-regulation of pro-apoptotic Bax expression. Taken together, these findings indicate that MEEA may have potential cancer therapeutic utility in A549 and HepG2 cells.

A kinetic study of 4-chlorophenol biodegradation by the novel isolated Bacillus subtilis in batch shake flask

  • Sandhibigraha, Sudhansu;Chakraborty, Sagnik;Bandyopadhyay, Tarunkanti;Bhunia, Biswanath
    • Environmental Engineering Research
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    • v.25 no.1
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    • pp.62-70
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    • 2020
  • Here in this work, a 4-chlorophenol (4-CP)-degrading bacterial strain Bacillus subtilis (B. subtilis) MF447840.1 was isolated from the drain outside the Hyundai car service center, Agartala, Tripura, India. 16S rDNA technique used carried out for genomic recognition of the bacterial species. Isolated bacterial strain was phylogenetically related with B. subtilis. This strain was capable of breaking down both phenol and 4-CP at the concentration of 1,000 mg/L. Also, the isolated strain can able to metabolize five diverse aromatic molecules such as 2-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, 4-nitrophenol, and pentachlorophenol for their growth. An extensive investigation was performed to portray the kinetics of cell growth along with 4-CP degradation in the batch study utilizing 4-CP as substrate. Various unstructured models were applied to evaluate the intrinsic kinetic factors. Levenspiel's model demonstrates a comparatively enhanced R2 value (0.997) amongst every analyzed model. The data of specific growth rate (μ), saturation constant (KS), and YX/S were 0.11 h-1, 39.88 mg/L, along with 0.53 g/g, correspondingly. The isolated strain degrades 1,000 mg/L of 4-CP within 40 h. Therefore, B. subtilis MF447840.1 was considered a potential candidate for 4-CP degradation.