• Title/Summary/Keyword: sub-G1 DNA

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Effects of Detoxified Sulfur as a Feed Supplement on in Vitro Rumen Fermentation and Methane Mitigation (제독 유황의 반추위 발효성상 및 메탄 저감 효과 연구)

  • Kim, Seon-Ho;Islam, Mahfuzul;Biswas, Ashraf Ali;Cho, Kwang-Keun;Lee, Sang-Suk
    • Journal of Life Science
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    • v.30 no.9
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    • pp.743-748
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    • 2020
  • Sulfate is a reductant that competes for electrons and may lower CH4 production in the rumen. This study was designed to evaluate the beneficial effect of detoxified sulfur powder supplementation on in vitro rumen fermentation and methane mitigation. A ruminally cannulated Holstein Friesian cow was used as a rumen fluid source, and commercial pelleted concentrate was used as a substrate at 1 g dry matter. Treatments included the addition of detoxified sulfur powder at the rate of 0% (Control), 0.2% (T1), 0.4% (T2), 0.6% (T3), 0.8% (T4), and 1.0% (T5) as dry matter (DM) basis. The pH, total gas (TG), methane (CH4) production, DM digestibility, organic matter (OM) digestibility, and volatile fatty acids (VFA) production were analyzed after 12 hr of incubation. The results showed that CH4 production was significantly lowest in T1 (13.78 ml) but highest in the control (20.16 ml). Insignificantly higher total VFA was observed in control and T1 (64.99 and 64.28 mM, respectively) compared to other treatments after 12 hr of incubation. After 12 hr of incubation, the significantly lowest acetate:propionate was observed in T1 (1.90) while the highest was observed in T4 (2.44). However, no significant differences were recorded for pH, TG, DM digestibility, OM digestibility, acetate, propionate, and butyrate between the control and T1. Total number of bacterial DNA copies was significantly lower in the treatment group than the control. Therefore, it can be concluded from this study that detoxified sulfur at 0.2% inclusion level is optimal for production performance and ruminal CH4 mitigation.

Biological Activities of Oenothera laciniata Extracts (Onagraceae, Myrtales) (애기달맞이꽃(Oenothera laciniata Hill) 추출물의 생리활성 탐색)

  • Lee, Jung-A;Kim, Ji-Young;Yoon, Weon-Jong;Oh, Dae-Ju;Jung, Yong-Hwan;Lee, Wook-Jae;Park, Soo-Yeong
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.810-815
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    • 2006
  • The biological activities of Oenothera laciniata extracts were measured, including antioxidant activity and cytotoxic effects. O. laciniata is an endemic species of Jeju Island, Korea with a seaside habitat. The concentration of total polyphenolic compounds from ethanol (EtOH), n-hexane, dichloromethane ($CH_2Cl_2$), ethylacetate (EtOAc), butanol (BuOH), and water fractions of O. laciniata was 63.96, 8.49, 28.11, 172.64, 114.56, and 34.91 mg/g, respectively. The EtOAc fraction contained the highest antioxidative activities ($IC_{50}$), measured as follows: 16.19 ${\mu}g/mL$ in DPPH radical scavenging capacity, 220.37 ${\mu}g/mL$ in xanthine oxidase inhibitory activity, 42.07${\mu}g/mL$ in superoxide radical scavenging capacity, and 421.33 ${\mu}g/mL$ in nitric oxide scavenging capacity. The cytotoxicity of O. laciniata extracts was examined through their effect on the growth of HL-60 cells. Incubation of HL-60 cells with the EtOAc fraction resulted in the greatest inhibition of cell growth; high DNA fragmentation and numerous sub-G1 hypodiploid cells were observed in HL-60 cell cultures treated with the EtOAc fraction. These results suggest that the EtOAc fraction of O. laciniata has potent apoptotic and antioxidative activities in vitro.

Effects of polysaccharides derived from Orostachys japonicus on induction of cell cycle arrest and apoptotic cell death in human colon cancer cells

  • Ryu, Deok-Seon;Baek, Geum-Ok;Kim, Eun-Young;Kim, Ki-Hoon;Lee, Dong-Seok
    • BMB Reports
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    • v.43 no.11
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    • pp.750-755
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    • 2010
  • Crude Orostachys japonicus polysaccharide extract (OJP) was prepared by hot steam extraction. Polysaccharides (OJPI) were separated from OJP by gel filtration chromatography and phenol-sulfuric acid assay. The average molecular weight of the OJPI was 30-50 kDa. The anti-proliferative effect of OJPI on HT-29 human colon cancer cells was investigated via morphology study, cell viability assay, apoptosis assay, cell cycle analysis, and cDNA microarray. OJPI inhibited proliferation and growth of HT29 cells and also stimulated apoptosis in a dose- and time-dependent manner. In cell cycle analysis, treatment with OJPI resulted in a marked increase of cells in the G0 (sub G1) and G2/M phases. To screen for genes involved in the induction of cell cycle arrest and apoptosis, the gene expression profiles of HT-29 cells treated with OJPI were examined by cDNA microarray, revealing that a number of genes were up- or down-regulated by OJPI. Whereas several genes involved in anti-apoptosis, cell proliferation and growth, and cell cycle regulation were down-regulated, expression levels of several genes involved in apoptosis, tumor suppression, and other signal transduction events were up-regulated. These results suggest that OJPI inhibits the growth of HT-29 human colon cancer cells by various apoptosis-aiding activities as well as apoptosis itself. Therefore, OJPI deserve further development as an effective agent exhibiting anticancer activity.

ADFP promoter polymorphism associated with marbling score in Korean cattle

  • Cheong, Hyun-Sub;Yoon, Du-Hak;Bae, Joon-Seol;Kim, Lyoung-Hyo;Kim, Eun-Mi;Kim, Ji-On;Hong, Jin;Kim, Nae-Soo;Shin, Hyoung-Doo
    • BMB Reports
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    • v.42 no.8
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    • pp.529-534
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    • 2009
  • Marbling score (MS) is the major trait that affects carcass quality in beef cattle. In this study, we investigated the association between genetic polymorphisms of the adipose differentiation-related protein gene (ADFP) and carcass traits in Korean cattle (also known as Hanwoo). Using direct DNA sequencing in 24 unrelated Korean cattle, 25 novel polymorphisms were identified within all exons and their flanking regions of ADFP, including the promoter region (1.5 kb). Among them, 21 polymorphic sites were selected for genotyping in the beef cattle (n = 425). Statistical analyses revealed that one promoter polymorphism (c.-56-18A > G) was associated with MS (P = 0.009). The 'A' allele of c.-56-18A > G exerted a lowering effect on MS, e.g., the lowest MS was found in 'A/A' (MS = 2.09 ${\pm}$ 1.23), intermediate in 'A/G' (MS = 2.11 ${\pm}$ 1.31), and the highest in 'G/G' (MS = 2.47 ${\pm}$ 1.47). Our findings suggest that these polymorphisms in ADFP might be important genetic factors involved in carcass quality in beef cattle.

The Effect of Millettia Reticulatas on the Proliferation Inhibition of Human Uterine Leiomyoma Cell and Expression of Apoptosis (계혈등(鷄血藤)이 자궁근종세포(子宮筋腫細胞)의 증식억제(增殖抑制) 및 세포자멸사에 미치는 영향)

  • Lee, Hwa-Kyung;Baek, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.3
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    • pp.135-149
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    • 2006
  • Purpose : This study was aimed to investigate the inhibitory effect of Millettia Reticulatas on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated concentration of Millettia Reticulatas and investigated cell death rate by MTS assay. Furthermore, flow cytometry analyis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the growth of uterine leiomyoma cell treated with Millettia Reticulatas was increased in a concentration proportional. 2) The result of flow cytometry analysis. subG1 phase arrest related3 cell apoptosis was investigated 23.49% in uterine leiomyoma cell treated Millettia Reticulatas and showed the fession of proportional concentration. 3) The gene expression of p27, p53, p21, p16 related cell cycle was increased according to increasing concentration but cyclin E was none exchanged. 4) The character of apoptosis, DNA fragmentation was significantly observed the fession of proportional concentration. 5) The expression of pro-caspase3 and PARP were decreased dependent on treatment concentration. Conclusion : This study showed that Millettia Reticulatas have the inhibitory effect on the proliferation of human uterine leiomyoma cell and the effect was related with apoptosis. The apoptotic mechanism was observed that the gene expression of p27, p53, p21, p16 related cell cycle was increased according to increasing treatment concentration, induced G1 phase arrest and finally cell death was occurred. The decreased expression of pro-caspase 3 and PARP were noted that apoptosis was related with caspase pathway.

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Endogenous Nitric Oxide Strengthens Doxorubicin-induced Apoptosis in Human Colorectal Cell Lines (Doxorubicin에 의한 내인성 산화질소가 인간 대장암 세포주에서의 세포사멸에 미치는 효과)

  • Im, Soon Jae;Kim, Ji Hye;Kim, Min Young
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1137-1143
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    • 2014
  • Doxorubicin is a general chemotherapy drug widely used for a number of cancers. However, the correlation between endogenous nitric oxide ($NO^{\bullet}$) levels and chemoresistance to doxorubicin remains unclear. In this study, we investigated the effect of endogenous $NO^{\bullet}$ on the anticancer activity of doxorubicin in human colon cancer cell lines HCT116 and HT29 with different p53 status. The cells were treated with either doxorubicin alone or in combination with the $NO^{\bullet}$ synthase (NOS) inhibitor $N^G$-monomethyl-L-arginine (NMA). Doxorubicin differentially inhibited the growth of both the HCT116 (p53-WT) and HT29 (p53-MUT) cells, which was mitigated by cotreatment with NMA. Further studies revealed that inhibition of endogenous $NO^{\bullet}$ mitigated doxorubicin-induced apoptosis in the HCT116 and HT29 cells, as evidenced by apoptotic DNA fragmentation and the sub-G1 peak of apoptotic markers. Apoptosis was delayed in the HT29 cells, and its magnitude was greatly reduced, underscoring the importance of the modulation of p53 in the response. RT-PCR analysis revealed that doxorubicin down-regulated levels of inhibitors of the apoptosis family (cellular IAP-1 and-2). Collectively, these data show that induction of apoptosis by doxorubicin in human colon cancer cells is possibly related to modulation of endogenous $NO^{\bullet}$, the expression of the IAP family of genes, and the status of p53. The underlying mechanisms may represent potential targets for adjuvant strategies to improve the efficacy of chemotherapy for colon cancer.

Anti-cancer Effects of Bujeonghangamtang on Human Pancreatic Cancer Cell Line PANC-1 (부정항암탕(扶正抗癌湯)의 사람 췌장암 세포주 PANC-1에 대한 항종양(抗腫瘍) 효과(效果))

  • Kim, Hoon;Won, Jin-Hee;Moon, Goo
    • Herbal Formula Science
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    • v.15 no.1
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    • pp.213-228
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    • 2007
  • Objectives : The purpose of this report was to investigate the chemotherapeutic effect of Bujeonghangamtang against cancer cells. Materials and Methods : Various cancer cell lines including PANC-1, C6 glioma, SH-SY5Y, HepG2, and MCF-7 cells, were used. Apoptosis was determined by DAPI nuclei staining and flow cytometry in PANC-1 cells treated with 1 mg/ml Bujeonghangamtang for 48 hr. Expression of cell cycle arrest mediators including, cdc2p34 and cyclin B1 proteins were measured by Western blot analysis. Mitochondrial membrane potential was measured by fluorescence staining with JC-1, rhodamine 123. Result : Bujeonghangamtang induced the apoptosis of PANC-1, which was characterized as nucleic acid and genomic DNA fragmentation, chromatin condensation, and sub-G0/G1 fraction of cell cycle increase. but not C6 glioma, SH-SY5Y, HepG2, and MCF-7 cells. PANC-1 cells were markedly sensitive to Bujeonghangamtang. Treatment with Bujeonghangamtang resulted in the decreased expression of cdc2p34 and cyclin B1. Treatment with Bujeonghangamtang also increased the ROS production and induced mitochondrial dysfunction. Conclusion : Bujeonghangamtang exerted cytotoxicity against human Pancreatic cancer cells via cell cycle arrest-mediated apoptotic signaling including ROS production and mitochondrial dysfunction. Our data suggest that Bujeonghangamtang may be an important modulator of chemosensitivity of cancer cells against anticancer chemotherapeutic agents.

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Characterization of Acetobacter sp. Strain CV1 Isolated from a Fermented Vinegar (고산도 생성 초산균의 분리 및 발효특성)

  • Baek, Chang-ho;Baek, Seong-yeol;Lee, Se Hee;Kang, Ji-Eun;Choi, Han-Seok;Kim, Jae-Hyun;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.126-133
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    • 2015
  • Ten types of farm-made brewing vinegars were collected and four high acetic acid-producing strains (CV1, CV3, CV5, and CV6) were isolated. Among them strain CV1, exhibiting highly alcohol-resistant and acetic acid-producing properties, was selected and its taxonomic properties were investigated by phenotypic (particularly chemotaxonomic) characterization and phylogenetic inference based on 16S rRNA gene sequence analysis. On SM broth agar, cells of strain CV1 were gram-stainingnegative and formed pale white colonies with smooth to rough surfaces. Strain CV1 produced acetate from ethanol and was resistant to up to 8% (v/v) ethanol in LM broth. Strain CV1 had a G+C content of 61.0 mol%, contained meso-DAP as the cell wall amino acid, and possessed Q-10 as the major ubiquinone. A comparison of 16S rRNA gene sequences showed that strain CV1 was most closely related to Gluconacetobacter saccharivorans (≥99.0% identity). In liquid media, the optimum growth conditions for acetic acid production were 30℃ and pH >3.0 and strain CV1 produced 9.3% and 8.4% acetic acids from 10% and 9% alcohol concentrations, respectively.

G0/G1 Cell Cycle Arrest and Activation of Caspases in Honokiol-mediated Growth Inhibition of Human Gastric Cancer Cells

  • Kang, You-Jin;Chung, Hwa-Jin;Min, Hye-Young;Song, Ja-Young;Park, Hyen-Joo;Youn, Ui-Joung;Bae, Ki-Hwan;Kim, Yeong-Shik;Lee, Sang-Kook
    • Natural Product Sciences
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    • v.18 no.1
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    • pp.16-21
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    • 2012
  • Honokiol, a naturally occurring neolignan mainly found in Magnolia species, has been shown to have the anti-angiogenic, anti-invasive and cancer chemopreventive activities, but the molecular mechanism of actions has not been fully elucidated yet. In the present study, we investigated the effect of honokiol on the growth inhibitory activity in cultured SNU-638 human gastric cancer cells. We found that honokiol exerted potent antiproliferative activity against SNU-638 cells. Honokiol also arrested the cell cycle progression at the G0/G1 phase and induced the apoptotic cell death in a concentration-dependent manner. The cell cycle arrest was well correlated with the downregulation of Rb, cyclin D1, cyclin A, cyclin E, and CDK4 expression, and the induction of cyclin-dependent kinase inhibitor p27. The increase of sub-G1 peak by honokiol was closely related to the induction of apoptosis, which was evidenced by the induction of DNA fragmentation, the cleavage of poly(ADPribose) polymerase, and the sequential activation of caspase cascade. These findings suggest the cell cycle arrest and induction of apoptosis might be one possible mechanism of actions for the anti-proliferative activity of honokiol in human gastric cancer cell.

Synergistic anticancer activity of resveratrol in combination with docetaxel in prostate carcinoma cells

  • Lee, Sang-Han;Lee, Yoon-Jin
    • Nutrition Research and Practice
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    • v.15 no.1
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    • pp.12-25
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    • 2021
  • BACKGROUND/OBJECTIVES: The study was conducted to investigate the efficacy of the combination treatment of phytochemical resveratrol and the anticancer drug docetaxel (DTX) on prostate carcinoma LNCaP cells, including factors related to detailed cell death mechanisms. MATERIALS/METHODS: Using 2-dimensional monolayer and 3-dimensional spheroid culture systems, we examined the effects of resveratrol and DTX on cell viability, reactive oxygen species (ROS) levels, mitochondrial membrane potential, apoptosis, and necroptosis by MTT, flow cytometry, and Western blotting. RESULTS: At concentrations not toxic to normal human prostate epithelial cells, resveratrol effectively decreased the viability of LNCaP cells depending on concentration and time. The combination treatment of resveratrol and DTX exhibited synergistic inhibitory effects on cell growth, demonstrated by an increase in the sub-G0/G1 peak, Annexin V-phycoerythrin positive cell fraction, ROS, mitochondrial dysfunction, and DNA damage response as well as concurrent activation of apoptosis and necroptosis. Apoptosis and necroptosis were rescued by pretreatment with ROS scavenger N-acetylcysteine. CONCLUSIONS: We report resveratrol as an adjuvant drug candidate for improving the outcome of treatment in DTX therapy. Although the underlying mechanisms of necroptosis should be investigated comprehensively, targeting apoptosis and necroptosis simultaneously in the treatment of cancer can be a useful strategy for the development of promising drug candidates.