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Study on skin anti-inflammatory activity of fig (Ficus carica L.) fruit extract fractions (무화과(Ficus carica L.) 열매 추출 분획의 피부 항염증 활성 연구)

  • Hee Joon Kwon;Geun soo Lee;Jin Hwa Kim;Soon Woo Kwon;Hyung seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.416-423
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    • 2023
  • Figs has known to have antioxidant, whitening, anti-inflammatory, and antibacterial effects in their leaves, roots, stems, latex, and fruits. In order to develop cosmetic materials based on natural products, we have studied on the skin activity of the ficin in latex as well as the whitening function of the fruit extract with 70% ethanol, and used it as a raw material for released cosmetic product. However, there is little research on the demand for the development of new eutectic solvent extraction methods and its ability to control skin inflammation and psoriasis regulation. Thus, in this study, we evaluated the effectiveness of fig fruit extracts and fractions using eutectic solvent extraction for skin inflammation control and psoriasis. First, fig fruits were extracted under optimal eutectic solvent conditions and fractionated with n-hexane, dichloromethane, ethyl acetate, and butanol. First, the antioxidant activity and inhibition of nitric oxide (NO) production were confirmed in mouse macrophage RAW264.7 cells. In addition, as a result of observing the mRNA expression through RT-PCR, pro-inflammatory cytokines such as TNF-α, IL1α, and IL-1β were suppressed significantly in the hexane, dichloromethane, and ethyl acetate fractions. In addition, it was confirmed in TNF-α stimulated HaCaT keratinocyte model. Finally, chemokine CC motif ligand 20 (CCL20), marker gene of human psoriasis skin disease, was significantly suppressed in the hexane, dichloromethane, and ethyl acetate fractions. These results suggested its anti-inflammatory and skin soothing effect and the possibility of development as an excellent skin soothing natural cosmetic material in the future through future clinical trials.

A Study of Microbial Contamination in Fresh-Cut and Ready-to-Eat Foods Purchased from Online Markets (온라인 판매 신선편의식품 및 즉석섭취식품의 미생물 오염도 연구)

  • Hye-Sun Hwang;Jae-Hoon Jeong;Young-Hee Kwon;Ye-Jee Byun;Ji-Young Park;Ho-Cheol Yun
    • Journal of Food Hygiene and Safety
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    • v.39 no.4
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    • pp.335-342
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    • 2024
  • This study aimed to examine the delivery conditions and microbial contamination of fresh-cut and ready-to-eat foods purchased from online markets between February and November 2023. Upon arrival, the average surface temperature of the products was 11.3℃. In the fresh-cut foods, the average number of total aerobic bacteria and coliforms was 4.5 log colony-forming units (CFU)/g and 1.2 log CFU/g, respectively, whereas in the ready-to-eat foods, these values were 10.6 log CFU/g and 1.2 log CFU/g, respectively. Pathogens, such as Staphylococcus aureus, Salmonella spp., Clostridium perfringens, Listeria monocytogenes, and pathogenic Escherichia coli were absent from all samples. Bacillus cereus was found in 2.7% of the fresh-cut foods and 0.9% of the ready-to-eat foods, with contamination levels averaging 0.05 log CFU/g and 0.01 log CFU/g, respectively. In the four samples in which B. cereus was detected, genetic testing of the six toxin genes produced by B. cereus revealed the presence of at least one enterotoxin gene, excluding the emetic toxin. L. monocytogenes was absent from ready-to-eat foods but was detected in 0.9% of fresh-cut foods. Analysis of the isolated L. monocytogenes confirmed the presence of six pathogenicity-related genes, including iap, indicating the potential risk of foodborne diseases.

Morphological Characteristics, and Coefficient of Variation, Heritability and Genetic Advance of Major Cultivars of Spray Chrysanthemum (주요 스프레이 국화 품종의 형태적 특성과 변이계수, 유전율 및 유전자 전이율)

  • Shim, Sung-Im;Lim, Ki-Byung;Kim, Chang-Kil;Chung, Mi-Young;Kim, Kyung-Min;Chung, Jae-Dong
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.269-281
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    • 2016
  • The statistical analyses of coefficient of variation, heritability, and genetic advance were carried out to identify differences in morphological characteristics, such as the stem and inflorescence length, of 10 major commercial cultivars of spray chrysanthemum (Chrysanthemum morifolium). For morphological characteristics, stem lengths ranged from 46.4 cm to 54.9 cm, the maximum diameter of stem was 5.6 to 8.5 mm, the hardness of the stem was 0.17 to $0.70kg{\cdot}m^{-2}$, the fresh weight of stem was 7.5 to 17.5 g, the dry weight of the stem was 1.6 to 3.3 g, the ratio of dry weight/fresh weight of stem was 15.9% to 23.1%. Also, the number of leaves on the stem was 8.4 to 12.2, the stem leaf area was 17.8 to $37.8m^2$, the fresh weight stem leaves was 5.3 to 18.6 g, the dry weight was 0.5 to 1.4 g and the ratio of dry weight /fresh weight of stem leaves was 7.6% to 11.5%. The inflorescence length ranged from 10.1 to 18.6 cm, the fresh weight of inflorescence was 7.3 to 26.7 g, the dry weight of inflorescence was 1.2 to 2.8 g, the ratio of dry weight /fresh weight of inflorescence was 10.4% to 17.1%. For flower, the diameter of the flower center was 8.2 to 13.3 mm, the petal width was 5.7 to 14.0 mm, the petal length was 12.9 to 33.1 mm, and the petal thickness was 157.8 to $354.4{\mu}m$. The mean values of each character in each cultivar were very different, and DMRT and LSD values based on morphological characteristics among 10 cultivars were highly significant. For variability and genetic parameters, the lowest CV (coefficient of variation), PCV (phenotypic coefficient of variation), and GCV (genotypic coefficient of variation) were 4.79% to 5.15% in stem length, and the highest variations were 62.97% to 65.21% in leaf area. ECV (error or environmental coefficient of variation) was the lowest for leaf area (1.71%) and it was the highest for leaf dry weight (19.30%). Heritability also significantly differed among the characteristics, ranging from 68.69% to 99.67%, the lowest value was shown in ratio of dry weight /fresh weight of stem and the highest value was for leaf area of stem. The value for genetic advance was the lowest in hardness of stem at 0.30 and the highest in leaf thickness at 156.65. The lowest genetic advance as percentage of mean of stem hardness was 9.17%, while the highest percentage of stem length was 134.27%. Thus the characters which had the highest values indicated above show the influence of additive gene action and may provide useful resources for selection programs for agronomic improvement.

Status and Prospect of Herbicide Resistant Weeds in Rice Field of Korea (한국 논에서 제초제 저항성잡초 발생 현황과 전망)

  • Park, Tae-Seon;Lee, In-Yong;Seong, Ki-Yeong;Cho, Hyeon-Suk;Park, Hong-Kyu;Ko, Jae-Kwon;Kang, Ui-Gum
    • Korean Journal of Weed Science
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    • v.31 no.2
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    • pp.119-133
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    • 2011
  • Sulfonylurea (SU)-resistant weeds include seven annual weeds such as Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and three perennial weeds of Scirpus planiculmis, Sagittaria pigmaea and Eleocharis acicularis as of 2010 since identification Monochoria korsakowii in the reclaimed rice field in 1998. The Echinochloa oryzoides resistant to acetyl CoA carboxylase (ACCase) and acetolactate synthase (ALS) inhibitors has been confirmed in wet-direct seeding rice field of the southern province, Korea in 2009. In the beginning of occurrence of SU-resistant weeds the M. vaginalis, S. juncoides and C. difformis were rapidly and individually spreaded in different fields, however, theses resistant weeds have been occurring simultaneously in the same filed as time goes by. The resistant biotype by weed species demonstrated about 10- to 1,000-fold resistance, base on $GR_{50}$ (50% growth reduction) values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on $GR_{50}$ values. In history of paddy herbicides in Korea, the introduction of SU herbicides including besulfuron-metyl and pyrazosulfuron-ethyl that control many troublesome weeds at low use rates and provide excellent crop safety gave farmers and many workers for herbicide business refreshing jolt. The products and applied area of SU-included herbicides have been rapidly increased, and have accounted for about 69% and 96%, respectively, in Korea. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in direct-seeded rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which are effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity ($I_{50}$) of the SU-resistant M. korsakowii was 14- to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [$^{14}C$]bensulfuron uptake and translocation. ALS genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006, the early period, in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly because the several resistant weeds have been occurring simultaneously in the same filed. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2 leaf stage.

Strain Improvement of the Genus Pleurotus by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯속(屬)의 품종개발(品種開發))

  • Yoo, Young-Bok;You, Chang-Hyun;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.200-211
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    • 1993
  • Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

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Therapeutic Angiogenesis by Intramyocardial Injection of pCK-VEGF165 in Pigs (돼지에서 pCK-VEGF165의 심근내 주입에 의한 치료적 혈관조성)

  • Choi Jae-Sung;Han Woong;Kim Dong Sik;Park Jin Sik;Lee Jong Jin;Lee Dong Soo;Kim Ki-Bong
    • Journal of Chest Surgery
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    • v.38 no.5 s.250
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    • pp.323-334
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    • 2005
  • Background: Gene therapy is a new and promising option for the treatment of severe myocardial ischemia by therapeutic angiogenesis. The goal of this study was to elucidate the efficacy of therapeutic angiogenesis by using VEGF165 in large animals. Material and Method: Twenty-one pigs that underwent ligation of the distal left anterior descending coronary artery were randomly allocated to one of two treatments: intramyocardial injection of pCK-VEGF (VEGF) or intramyocardial injection of pCK-Null (Control). Injections were administered 30 days after ligation. Seven pigs died during the trial, but eight pigs from VEGF and six from Control survived. Echo-cardiography was performed on day 0 (preoperative) and on days 30 and 60 following coronary ligation. Gated myocardial single photon emission computed tomography imaging (SPECT) with $^{99m}Tc-labeled$ sestamibi was performed on days 30 and 60. Myocardial perfusion was assessed from the uptake of $^{99m}Tc-labeled$ sestamibi at rest. Global and regional myocardial function as well as post-infarction left ventricular remodeling were assessed from segmental wall thickening; left ventricular ejection fraction (EF); end systolic volume (ESV); and end diastolic volume (EDV) using gated SPECT and echocardiography. Myocardium of the ischemic border zone into which pCK plasmid vector had been injected was also sampled to assess micro-capillary density. Result: Micro-capillary density was significantly higher in the VEGF than in Control ($386\pm110/mm^{2}\;vs.\;291\pm127/mm^{2};\;p<0.001$). Segmental perfusion increased significantly from day 30 to day 60 after intramyocardial injection of plasmid vector in VEGF ($48.4\pm15.2\%\;vs.\;53.8\pm19.6\%;\;p<0.001$), while no significant change was observed in the Control ($45.1\pm17.0\%\;vs.\;43.4\pm17.7\%;\;p=0.186$). This resulted in a significant difference in the percentage changes between the two groups ($11.4\pm27.0\%\;increase\;vs.\;2.7\pm19.0\%\;decrease;\;p=0.003$). Segmental wall thickening increased significantly from day 30 to day 60 in both groups; the increments did not differ between groups. ESV measured using echocardiography increased significantly from day 0 to day 30 in VEGF ($22.9\pm9.9\;mL\;vs.\;32.3\pm9.1\;mL;\; p=0.006$) and in Control ($26.3\pm12.0\;mL\;vs.\;36.8\pm9.7\;mL;\;p=0.046$). EF decreased significantly in VEGF ($52.0\pm7.7\%\;vs.\;46.5\pm7.4\%;\;p=0.004$) and in Control ($48.2\pm9.2\%\;vs.\;41.6\pm10.0\%;\;p=0.028$). There was no significant change in EDV. The interval changes (days $30\~60$) of EF, ESV, and EDV did not differ significantly between groups both by gated SPECT and by echocardiography. Conclusion: Intramyocardial injection of pCK-VEGF165 induced therapeutic angiogenesis and improved myocardial perfusion. However, post-infarction remodeling and global myocardial function were not improved.

Usefulness of Pulsatile Flow Aortic Aneurysm Phantoms for Stent-graft Placement (스텐트그라프트 장치술을 위한 대동맥류 혈류 팬텀의 유용성)

  • Kim, Tae-Hyung;Ko, Gi-Young;Song, Ho-Young;Park, In-Kook;Shin, Ji-Hoon;Lim, Jin-Oh;Kim, Jin-Hyoung;Choi, Eu-Gene K.
    • Journal of radiological science and technology
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    • v.30 no.3
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    • pp.205-212
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    • 2007
  • To evaluate the feasibility and efficacy of a pulsatile aortic aneurysm phantoms for in-vitro study. The phantoms consisted of a pulsating motor part(heart part) and an aortic aneurysm part, which mimicked true physiologic conditions. The heart part was created from a high-pressured water pump and a pulsatile flow solenoid valve for the simulation of aortic flow. The aortic aneurysm part was manufactured from paper clay, which was placed inside a acrylic plastic square box, where liquid silicone was poured. After the silicone was formed, the clay was removed, and a silicone tube was used to connect the heart and aneurysm part. We measured the change in pressure as related to the opening time(pulse rate, Kruskal-Wallis method) and pressure before and after the stent-graft implantation(n = 5, Wilcoxon's signed ranks test). The changes in blood pressures according to pulse rate were all statistically significant(p<0.05). The systolic/diastolic pressures at the proximal aorta, the aortic aneurysm, and the distal aorta of the model were $157.80{\pm}1.92/130.20{\pm}1.92$, $159.40{\pm}1.14/134.00{\pm}2.92$, and $147.20{\pm}1.480/129.60{\pm}2.70\;mmHg$, respectively, when the pulse rate was 0.5 beat/second. The pressures changed to $161.40{\pm}1.34/90.20{\pm}1.64$, $175.00{\pm}1.58/93.00{\pm}1.58$, and $176.80{\pm}1.48/90.80{\pm}1.92\;mmHg$, respectively, when the pulse rate was 1.0 beat/second, and $159.40{\pm}1.82/127.20{\pm}1.48$, $166.60{\pm}1.67/138.00{\pm}1.87$, and $161.00{\pm}1.22/135.40{\pm}1.67\;mmHg$, respectively, when it was 1.5 beat/second. When pulse rate was set at 1.0 beat/second, the pressures were $143.60{\pm}1.67/90.20{\pm}1.64$, $147.20{\pm}1.92/84.60{\pm}1.82$, and $137.40{\pm}1.52/88.80{\pm}1.64\;mmHg$ after stent-graft implantation. The changes of pressure before and after stent-graft implantation were statistically significant(p<0.05) except the diastolic pressures at the proximal(p =1.00) and distal aorta(p=0.157). The aortic aneurysm phantoms seems to be useful for the evaluation of the efficacy of stent-graft before animal or clinical studies because of its easy reproducibility and ability to display a wide range of pressures.

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Association between Genetic Polymorphisms of Milk Proteins and Milk Compositions in Holstein Cows (Holstein 종(種) 유우(乳牛)의 유단백질(乳蛋白質)의 유전적다형(遺傳的多型)과 유조성분간(乳組成分間)의 연관성)

  • Sang, Byong Chan;Lee, Jo Yoon;Choi, Jong Woo;Sung, Chang Keun
    • Korean Journal of Agricultural Science
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    • v.20 no.1
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    • pp.56-67
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    • 1993
  • To applying of genetic markers of milk proteins as dairy cow registration and selection aids for genetic improvement, genopypes controlling the 4 milk protein loci, ${\alpha}S1$-casein (${\alpha}S1$-CN), ${\beta}$-casein(${\beta}$-CN), ${\kappa}$-casein(${\kappa}$-CN), and ${\beta}$-lactoglobulin(${\beta}$-LG), from a total of 159 Holstein lactating cows reared at National Animal Breeding Station in 1992 were detected by polyacrylamide gel(PAGE) electrophoresis, and associations between genetic polymorphisms of milk proteins and milk compositions were analyzed. The observed distribution of phenotypes for ${\alpha}S1$-CN, ${\beta}$-CN, ${\kappa}$-CN and ${\beta}$-LG were agreement with those expected under the assumption of genetic equilibrium. The observed genotypic frequencies of the ${\alpha}S1$-CN BB, ${\beta}$-CN AA, ${\kappa}$-CN AA and ${\beta}$-LG AB genotypes were founded to be very high as 79.87%, 84.28%, 71.70% and 49.10%, respectively. Gene frequencies were 0.899 and 0.101 for ${\alpha}S1-CN^B$ and ${\alpha}S1-CN^C$, 0.921 and 0.079 for ${\beta}-CN^A$ and ${\beta}-CN^B$, 0.837 and 0.163 for ${\kappa}-CN^A$ and ${\kappa}-CN^B$, 0.378 and 0.622 for ${\beta}-LG^A$ and ${\beta}-CN^B$. According to the results of analysis of variance, the genotypes of the ${\alpha}S1-CN$, ${\beta}-CN$, ${\kappa}-CN$ and ${\beta}-LG$ were significantly difference for fat, protein and total solid percentage in milk compositions. On milk compositions, the ${\kappa}$-CN BB genotype was very high fat and protein percentage more than ${\kappa}$-CN AA and AB genotypes, and ${\beta}$-LG AA genotype was very high fat percentage more than ${\beta}$-LG AB and BB genotype at 5% level of significant difference, respectively. As a consequence, the fat and protein percentage may be improved to select to ${\kappa}$-CN BB and ${\beta}$-LG AA genotypes.

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A Natural L-Arginine Analog, L-Canavanine-Induced Apoptosis is Suppressed by Protein Tyrosine Kinase p56lck in Human Acute Leukemia Jurkat T Cells (인체 급성백혈병 Jurkat T 세포에 있어서 L-canavanine에 의해 유도되는 세포자살기전에 미치는 단백질 티로신 키나아제 p56lck의 저해 효과)

  • Park, Hae-Sun;Jun, Do-Youn;Woo, Hyun-Ju;Rue, Seok-Woo;Kim, Sang-Kook;Kim, Kyung-Min;Park, Wan;Moon, Byung-Jo;Kim, Young-Ho
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1529-1537
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    • 2009
  • To elucidate further the antitumor effects of a natural L-arginine analogue, L-canavanine, the mechanism underlying apoptogenic activity of L-canavanine and its modulation by protein tyrosine kinase $p56^{lck}$ was investigated in human Jurkat T cells. When the cells were treated with 1.25 to 2.5 mM L-canavanine for 36 h, several apoptotic events including mitochondrial membrane potential (${\Delta\Psi}m$) loss, activation of caspase-9, -3, -8, and -7, poly (ADP-ribose) polymerase (PARP) degradation, and DNA fragmentation were induced without alteration in the levels of Fas or FasL. These apoptotic changes were more significant in $p56^{lck}$-deficient Jurkat clone JCaM1.6 than in $p56^{lck}$-positive Jurkat clone E6.1. The L-canavanine-induced apoptosis observed in $p56^{lck}$-deficient JCaM1.6 cells was significantly reduced by introducing $p56^{lck}$ gene into JCaM1.6 cells by stable transfection. Treatment of JCaM1.6/lck cells with L-canavanine caused a transient 1.6-fold increase in the kinase activity of $p56^{lck}$. Both FADD-positive wild-type Jurkat T cell clone A3 and FADD-deficient Jurkat T cell clone I2.1 exhibited a similar susceptibility to the cytotoxicity of L-canavanine, excluding involvement of Fas/FasL system in triggering L-canavanine-induced apoptosis. The L-canavanine-induced apoptotic sub-$G_1$ peak and activation of caspase-3, -8, and -7 were abrogated by pan-caspase inhibitor (z-VAD-fmk), whereas L-canavanine-induced activation of caspase-9 was not affected. These results demonstrated that L-canavanine caused apoptosis of Jurkat T cells via the loss of ${\Delta\Psi}m$, and the activation of caspase-9, -3, -8, and -7, leading to PARP degradation, and that the $p56^{lck}$ kinase attenuated the ${\Delta\Psi}m$ loss and activation of caspases, and thus contributed as a negative regulator to L-canavanine-induced apoptosis.

The Study of anti-cancer mechanism with Cobrotoxin on Human prostatic cancer cell line(PC-3) (전립선 암세포에 대한 Cobrotoxin의 항암(抗癌) 기전(機轉) 연구(硏究))

  • Chae, Sang-jin;Song, Ho-seup
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.169-183
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    • 2005
  • Objective : The purpose of this study was to investigate the anti-caner effect of cobrotoxin on the prostatic cancer cell line (PC-3).The goal of study is to ascertain whether cobrotoxin inhibits tile cell growth and cell cycle of PC-3, or the expression of relative genes and whether the regression of PC-3 cell growth is due to cell death or the expression of gene related to apoptosis. Methods : After the treatment of Pc-3 cells with cobrotoxin, we performed 형광현미경, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify the cell viability, cell death, apoptosis, the changes of cell cycle and the related protein, Adk, MAP kinase. Results : 1. Compared with normal cell, the inhibition of cell growth reduced in proportion with the dose of cobrotoxin(0-16nM) in PC-3. 2. Cell viabilities of 0.1, 1, 4nM cobrotoxin treatment were decreased and those of 8, 16nM were decreased significantly. 3. S phase of cell cycle was decreased at the group of 1, 2, 4, 8, 16nM cobrotoxin, but M phase was increased at 0.1, 1, 2, 4, 8, 16nM cobrotoxin. 4. Cox-2 expression after cobrotoxin was peaked at 12hours and was decreased significantly after 6, 12, 24 hours. 5. The expression of Cdk4 was decreased dose-dependently at 1, 2, 4, 8nM cobrotoxin and was decreased siginificantly at 4, 8nM Cyclin D1 was decreased at 1, 2, 4, 8nM and Cycline E was not changed. Cycline B was decreased at 1, 2, 4, 8nM dose-dependently and was decreased siginificanlty at 2, 4, 8nM. 6. The expression of Akt was decreased at 1, 2, 4, 8nM dose-dependently and was decreased significantly at 2, 4, 8nM. 7. ERK was increased at 1, 2nM and decreased at 4, 8nM, p-ERK was increased at 1, 2, 4 nM, but decreased at 8nM. JNK and p-JNK were increased at 1, 4, 8 nM. p38 was increased at 2nM p-p38 was increased at lnM but decreased significantly at 2, 4, 8nM. 8. The nucli of normal cells were stained round and homogenous in DAPI staining, but those of PC-3 were stained condense and splitted. Apoptosis was increased dose-dependently at 2, 4, 8, 16nM and increased significantly at 2, 4, 8, 16nM. 9. Bax wasn`t changed at 1, 2, 4, 8nM and Bcl-2 was decreased significantly at 1, 2, 4, 8nM. Caspase 3 and 9 weren`t changed at 1, 2, 4nM but were decreased significantly at 8nM. Conclusions : These results indicate that cobrotoxin inhibits the growth of prostate Cancer cells, has anti-cancer effects by inducing apoptosis.

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