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Apoptotic Cell Death by Pectenotoxin-2 in p53-Deficient Human Hepatocellular Carcinoma Cells (종양억제유전자 p53 결손 인체간암세포에서 Pectenotoxin-2에 의한 Apoptosis 유도)

  • Shin, Dong-Yeok;Kim, Gi-Young;Choi, Byung-Tae;Kang, Ho-Sung;Jung, Jee-H.;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1447-1451
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    • 2007
  • Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.

The effectiveness of sealing technique on in-office bleaching (전문가 미백시 sealing technique의 효율성에 관한 연구)

  • Lee, Yoon;Kwon, So-Ran;Park, Jeong-Won
    • Restorative Dentistry and Endodontics
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    • v.33 no.5
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    • pp.463-471
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    • 2008
  • This study investigated the clinical effectiveness and safety of sealed bleaching compared to conventional in-office bleaching using a randomized clinical trial of split arch design. Ten participants received a chairside bleaching treatment on the upper anterior teeth, and each side was randomly designated as sealed or control side. A mixture of Brite powder (PacDent, Walnut, USA), 3% hydrogen peroxide and carbamide peroxide (KoolWhite, PacDent, Walnut, USA) were used as bleaching agent. The control side was unwrapped and the experimental side was covered with a linear low density polyethylene (LLDPE) wrap for sealed bleaching. The bleaching gel was light activated for 1 hour. The tooth shades were evaluated before treatment, after treatment, and at one week check up by means of a visual shade (VS) assessment using a value oriented shade guide and a computer assisted shade assessment using a spectrophotometer (SP). The data were analyzed by paired t-test. In the control and sealed groups, the visual shade scores after bleaching treatment and at check up showed statistically significant difference from the preoperative shade scores (p<.05). The shade scores of the sealed group were significantly lighter than the control immediately after bleaching and at the check-up appointment (p<0.05). Compared to prebleaching status, the ${\Delta}E$ values at post bleaching condition were $4.35{\pm}1.38\;and\;5.08{\pm}1.34$ for the control and sealed groups, respectively. The ${\Delta}E$ values at check up were $3.73{\pm}1.95\;and\;4.38{\pm}2.08$ for the control and sealed groups. ${\Delta}E$ values were greater for the sealed group both after bleaching (p<.05) and at check up (p<.05). In conclusion, both ${\Delta}E$ and shade score changes were greater for the sealed bleaching group than the conventional bleaching group, effectively demonstrating the improvement of effectiveness through sealing.

Biocontrol of Damping-Off(Rhizoctonia solani) in Cucumber by Trichoderma asperellum T-5 (Trichoderma asperellum T-5를 이용한 오이 모잘록병(Rhizoctonia solani)의 생물학적 제어)

  • Ryu, Ji-Yeon;Jin, Rong-De;Kim, Yong-Woong;Lee, Hyang-Burm;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.4
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    • pp.185-194
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    • 2006
  • A fungal strain of Trichoderma having strong chitinolytic activity was isolated from field soil enriched with crabshell for several years. Based on 5.8S rRNA, partial 18S, 28S rRNA genes, ITS1, ITS2 sequence analysis and morphological characteristics, the fungus was identified as Trichoderma asperellum and named as Trichoderma asperellum T-5 (TaT-5). The fungus released lytic enzymes such as chitinase and ${\beta}$-1, 3-glucanse, and produced six antifungal substances in chitin broth medium. To demonstrate the protective effect of TaT-5 against damping-off in cucumber plant caused by Rhizoctonia solani, TaT-5 culture broth (TA), chitin medium (CM) and distilled water (DW) were applied to each pot at 10 days after sowing, respectively. Then, the homogenized hyphae of R. solani were infected to each pot at 1 week after TaT-5 inoculation. During experimental period, fresh weight of shoot and root in cucumber plant more increased at TA treatment compared to other treatments. PR-proteins (${\beta}$-1, 3-glucanase and chitinase) activities in cucumber leaves markedly increased at CM and DW treatments, but the activity slightly increased and then decreased at TA treatment at 3 days after infection of R. solani. The activity of PR-proteins activities in cucumber roots at all treatments decreased with time where the degree of decrement was more alleviated at TA treatment than CM and DW. These results suggest that the lytic enzymes (chitinase and ${\beta}$-1, 3-glucanse) and antifungal substances produced by TaT-5 can reduce the pathogenic attack by R. solani in cucumber plants.

Characteristics of Flowering and Leaf Emergence in Lycoris Species (상사화류의 개화와 출엽 특성)

  • Park, Yun-Jum;Kim, Hyun-Ju;Seo, Young-Nam;Chon, Sang-Uk;Lee, Beom-Seon;Heo, Buk-Gu
    • Korean Journal of Plant Resources
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    • v.20 no.1
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    • pp.43-49
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    • 2007
  • This study was conducted to examine the pattern and characteristics of flowering and leaf emergence for twenty two kinds of the genus of Lycoris sp. One species of Lycoris was flowered on July to August, twelve species on August, and nine species on September. Shape of flowers were as follows : Nine species belong to L. radiara-shaped flower, seven species L. squamigera-shaped flower, and seven species the intermediate type. Flower color of three kind were an order of descent red and pink. seven kind white, four kind yellow, three kind orange, and two kind purple colors. Ten Lycoris species were good fertile, and two kinds were poor fertile. The average length of pollen grain was ranged from $48.53\;to\;88.62{\mu}m$, and the width from $22.87\;to\;33.67{\mu}m$. Eleven kinds were spring-leaf emergence type, and ten kind autumn-leaf emergence types. Hunter values in leaves were as follows : $L^*$ value was ranged from 31.6 to 45.2; $a^*$ values from -6.74 to -17.46; $b^*$ values from 6.19 to 21.89 Leaf width was ranged from 0.90 to 2.35cm, and its shape was oblong. The length of epidermal cells was 0.33 to 0.75mm, and the width was $38.53\;to\;90.00{\mu}m$. Most of stomata were distributed in the back side of leaves. The length of stomata was ranged from $67.12\;to\;104.89{\mu}m$, and its width was from $14.90\;to\;71.52{\mu}m$.

Preservation of Strawberries and Cucumbers Packaged by Low density polyethylene film impregnated with antimicmbial agent, Scutellariae baicalensis extract (황금추출물을 함유한 항균성 포장필름을 이용한 딸기와 오이의 저장효과)

  • 정순경;조성환
    • Food Science and Preservation
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    • v.9 no.3
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    • pp.271-276
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    • 2002
  • To develop a wrapping film, which suppresses the microbial decay through the storage and prolongs the selflife of fruits and vegetables, the antimicrobial packaging films were prepared and applied to the preservation of strtwberries and cucumbers. Low density polyethylene(LDPE) film of 50㎛ thickness was faricated with 1% of Scutellariae baicalensis extract. The LDPE film impregnated with Scutellariae baicalensis extract showed antimicrobial activity on the disk test against Bacillus cereus, Escherchia coli and Fusarium sp.. The antimicrobial film changed the color and light transmittance, but did not affect heat shrinkage, mechanical tensile strength and wattability. Strawberries and cucumbers were separately wrapped with packaging films in the state of closely-adhered packaging as well as modified atmosphere packaging(MAP). The wrapped strawberries and cucumbers were stored for 21 days at 5$\^{C}$ and for 40 days at l0$\^{C}$, respectively. For the packaged strawberries and cucumbers at 5$\^{C}$ and 10$\^{C}$, the LDPE film impregnated with Scutellariae baicalensis extract showed the reduced growth of total aerobic bacteria, molds and yeasts and did not give any negative effect on other quality attributes during storage in comparison with conttrol film without any additive.

Local Shape Analysis of the Hippocampus using Hierarchical Level-of-Detail Representations (계층적 Level-of-Detail 표현을 이용한 해마의 국부적인 형상 분석)

  • Kim Jeong-Sik;Choi Soo-Mi;Choi Yoo-Ju;Kim Myoung-Hee
    • The KIPS Transactions:PartA
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    • v.11A no.7 s.91
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    • pp.555-562
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    • 2004
  • Both global volume reduction and local shape changes of hippocampus within the brain indicate their abnormal neurological states. Hippocampal shape analysis consists of two main steps. First, construct a hippocampal shape representation model ; second, compute a shape similarity from this representation. This paper proposes a novel method for the analysis of hippocampal shape using integrated Octree-based representation, containing meshes, voxels, and skeletons. First of all, we create multi-level meshes by applying the Marching Cube algorithm to the hippocampal region segmented from MR images. This model is converted to intermediate binary voxel representation. And we extract the 3D skeleton from these voxels using the slice-based skeletonization method. Then, in order to acquire multiresolutional shape representation, we store hierarchically the meshes, voxels, skeletons comprised in nodes of the Octree, and we extract the sample meshes using the ray-tracing based mesh sampling technique. Finally, as a similarity measure between the shapes, we compute $L_2$ Norm and Hausdorff distance for each sam-pled mesh pair by shooting the rays fired from the extracted skeleton. As we use a mouse picking interface for analyzing a local shape inter-actively, we provide an interaction and multiresolution based analysis for the local shape changes. In this paper, our experiment shows that our approach is robust to the rotation and the scale, especially effective to discriminate the changes between local shapes of hippocampus and more-over to increase the speed of analysis without degrading accuracy by using a hierarchical level-of-detail approach.

Comparison of Behavioral Response between Intranasal and Submucosal Midazolam Adminstration (소아 진정 치료 시 구강 점막 하와 비점막 Midazolam 투여의 행동 반응 비교)

  • Kim, Yun-Hee;Jung, Sang-Hyuk;Baek, Kwang-Woo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.3
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    • pp.427-436
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    • 2008
  • Purpose. The objective of this study was to evaluate the behavioral response and assess the effectiveness of additional intranasal (IN) and submucosal (SM) administration of midazolam during pediatric sedation for dental procedure. Material and methods. Thirty-three cases of healthy (ASAⅠ), uncooperative children aged from 24 to 72 month old at pediatric dental clinic of Ewha Womans University Hospital were selected for this study. Children received oral chloral hydrate 50 mg/kg with hydroxyzine 1.0 mg/kg. After waiting for 45 minutes, midazolam 0.2 mg/kg was administrated via IN route and via SM route randomly maintaining 50% of $N_2O$. A pulse oximeter and a capnograph were used for measuring vital signs ($SpO_2$, PR, RR, $EtCO_2$) throughout the sedation. Behavioral response was evaluated as Quiet (Q), Crying (C), Movement (M) or Struggling (S) in every 2 minutes for 40 minutes. Results. There were also no statistically significant differences in vital signs of the two groups. The behavioral response for the first ten minutes during sedation was a statistically significant difference (P < 0.05) between the two groups. After the first ten minutes, it was revealed that there was no significant difference. Conclusion. This study demonstrated that the addition of IN midazolam to the combination of oral chloral hydrate with hydroxyzine and nitrous oxide/oxygen inhalation is as safe and effective as that of SM midazolam in pediatric sedation for dental procedure.

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Characterization of Bacteriocin Produced from Isolated Strain of Bacillus sp. (Bacillus 속 분리주가 생산하는 박테리오신의 특성 조사)

  • Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.202-210
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    • 2017
  • As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

Changes in isoflavone content and quality characteristics of Cheonggukjang prepared with Bacillus subtilis HJ18-3 and KACC 15935 (Bacillus subtilis HJ18-3과 KACC 15935를 이용하여 제조한 청국장의 품질특성과 isoflavone 함량의 변화)

  • Lee, Kyung Ha;Choi, Hye Sun;Choi, Yoon Hee;Park, Shin Young;Song, Jin
    • Food Science and Preservation
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    • v.21 no.1
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    • pp.121-128
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    • 2014
  • This study was conducted in order to investigate the change of isoflavone composition (glycoside and bio-active aglycone), and to evaluate the quality characteristics of Cheonggukjang, which was prepared by different bacillus strains. After the 48-hour fermentation, the contents of daidzein, genistein, and glycitein in the Bacillus subtilis HJ18-3 have significantly increased up to approximately $89.06{\pm}3.59$, $10.36{\pm}0.28$, and $101.37{\pm}3.67ug/g$, respectively. The contents of daidzein, genistein, and glycitein in the Bacillus subtilis KACC 15935 were $38.88{\pm}5.39$, $12.58{\pm}2.14$, and $80.13{\pm}0.71ug/g$, respectively. The original content of daidzein was 3.96 ug/g, while genistein and glycitein were not measured. However, the contents of daidzen and genistein in HJ18-3 and in KACC 15935 were decreased. The ${\alpha}$-Amylase and cellulase activities of Chungkookjang in HJ18-3 were higher than in the KACC 15935. The contents of Chungkookjang in HJ18-3 were $29.70{\pm}11.66$ and $4861.3{\pm}388.07unit/g$, respectively. The amino type nitrogen contents and ammonia type nitrogen contents of Chungkookjang in KACC 15935 were higher than in the HJ18-3. These results suggested that it could be used to increase the bioactivity via fermentation with the Bacillus subtilis possessing a ${\beta}$-glucosidase activity with a view towards the development of functional foods.

A Study on the Antidotal Effects of Dietary Garlic Juice on Lead Poisoning Rats (납중독 흰쥐에서 식이 마늘 즙의 해독효과에 관한 연구)

  • Sheo, Hwa-Jung;Seo, Yu-Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.3
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    • pp.342-350
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    • 2005
  • In the present work antidotal effect of dietary garlic was studied on lead-intoxicated rat. One of 5 groups of young Wistar sp. male rat, aged 4 weeks for control were fed only normal diet. Lead (25 ㎎/㎏.bw/week) was administered to other four groups for plumbism model over 4 weeks, of which three groups were supplemented with one of the following raw garlic juice: 1.10 (1% diet), 2.21 (2%) and 3.31 (3%) ㎎/㎏.bw/day respectively. Body weight gain rates in all garlic group significantly increased, especially in 2% garlic group that showed 9.8% net gain, as compared with only-lead treated group but lower values than control. The fecal and urinary lead excretion in all garlic groups significantly increased in a dose dependent fashion with highest value of 9.59% net gain in 3% garlic group as compared to lead treated control group. In comparison with lead treated control group, all garlic groups showed significantly increased hemoglobin contents, hematocrit values (Hct), red blood cell (RBC) count, mean corpuscular volume (MCV), and δ-amino levulinic acid dehydratase (δ-ALAD) activities. The values of 2% and 3% garlic groups remarkably increased while no significant difference between the values of 2% and 3% garlic groups was observed. The ALT activities, blood urea nitrogen (BUN) and creatinine (CR) in all garlic groups significantly decreased as compared with lead-treated control group. The values of 2% garlic group were the lowest and significantly different from the values of 1% and 3% garlic groups. The results showed that 2%-3% garlic juice in diet had obviously antidotal effects in lead-poisoned rats by promoting lead excretion. However, mega dose garlic such as in 3% garlic group might have some adverse effects on hepatic and renal functions in rats. In conclusion, the dietary habit to take ordinary garlic sauce in appropriate amount, may be helpful for preventing lead or other heavy metal intoxication.