• 제목/요약/키워드: biochemical

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H2AX Directly Interacts with BRCA1 and BARD1 via its NLS and BRCT Domain Respectively in vitro (H2AX의 BRCA1 NLS domain과 BARD1 BRCT domain 각각과의 in vitro 상호 결합)

  • Bae, Seung-Hee;Lee, Sun-Mi;Kim, Su-Mi;Choe, Tae-Boo;Kim, Cha-Soon;Seong, Ki-Moon;Jin, Young-Woo;An, Sung-Kwan
    • KSBB Journal
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    • v.24 no.4
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    • pp.403-409
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    • 2009
  • H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.

Effect of Tremella fuciformis Berk on Anti-stress activities during Long-Term and Short-Term in Mice (마우스에서 흰목이버섯 (Tremella fuciformis Berk)의 장.단기적 항스트레스 효과)

  • Ko, Min-Seok;Lee, Seung-Jin;Kang, Sang-Mo
    • KSBB Journal
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    • v.24 no.2
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    • pp.131-139
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    • 2009
  • The objective of this study was to evaluate the protective effect of aqueous extracts from Tremella fuciformis Berk(Tf AE) against stress during long-term and short-term in ICR mice. All the animals were randomly divided into two groups which had been bred for 5 months that were treated by immobilization stress for 8 weeks (total 7 months breeding, equivalent to human beings aged 20) with or without Tf AE, and one out of two groups was continuously bred until they become 18 months old (equivalent to human beings aged 60) without Tf AE. Afterwards, the changes of serum and hepatic metabolites were investigated on the basis of the index of stress-related in vivo oxidative damage. As a result, it was found that stress increases serum triglyceride (TG) and aspartate aminotransferase (AST) and decreases serum HDL-cholesterol in the long-term (total 18 months breeding) and short-term (total 7 months breeding). In addition, stress concerned the decrease of total antioxidant status (TAS) and superoxide dismutase (SOD) as well as the increase of malondialdehyde (MDA) in liver. On the other hand, Tf AE-fed groups reversed all these biochemical indices. These results suggest that stress in one's youth causes negative results in TG, HDL-cholesterol, LDL-cholesterol, AST, TAS, SOD and MDA measured in one's senescent. The administration of Tf AE in the stressed mice decreases serum TG and AST that are increased by stress, and exerts influence on the increase of serum HDL-cholesterol. Also Tf AE recovered the values of liver TAS, SOD and MDA in the stressed mice. In conclusion, Tf AE represented protective effect in the stressed mice to some degree.

Assessment of Microbiological Quality for Raw Materials and Cooked Foods in Elementary School Food Establishment (초등학교에 공급되는 급식용 식재료 및 조리식품의 미생물학적 품질평가)

  • Shin, Weon-Sun;Hong, Wan-Soo;Lee, Kyung-Eun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.3
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    • pp.379-389
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    • 2008
  • This study was conducted to assess the microbiological quality of raw and cooked foods served in the elementary school food service. Raw and cooked food samples were collected from 11 selected elementary schools in both June to July and September to October of 2005. Petrifilm plates were used to determine (in duplicate) total aerobic colony counts (PAC), Enterobacteriaceae (PE), coliform counts (PCC), and E. coli counts (PEC). Heavy contamination of Enterobacteriaceae (from 0.08 to 7.40 log CFU/g) and total coliform (0.50 to 6.52 log CFU/g) were observed in raw materials and cooked foods. Escherichia coli (E. coli) were detected in the sample of currant tomato (3.70 log CFU/g), sesame leaf (3.59 log CFU/g), dropwort (0.20 log CFU/g), crown daisy (3.15 log CFU/g), parsley (3.00 log CFU/g), peeled green onion (1.74 log CFU/g), frozen pork (0.65 log CFU/g), frozen beef (0.20 or 1.50 log CFU/g), chicken (1.78 log CFU/g), and young radish leaf seasoned with soybean paste (1.24 log CFU/g). Multiplex PCR system was used to determine the food-borne pathogens: Salmonella spp., Bacillus cereus (B. cereus), E. coli O157:H7, Staphylococcus aureus, Listeria monocytogenes (L. monocytogenes), Vibrio parahaemolyticus, Campylobacter jejuni (C. jejuni), Shigella spp., B. cereus was detected in 19 samples of raw materials and 8 samples of cooked foods. With regard to quantitative analysis, B. cereus counts exceeded 5.46, 3.48 and 1.79 log CFU/g in sesame leaf, peeled green onion and seasoned mungbean jelly, respectively. E. coli O157:H7 was detected on 2 samples of frozen beefs, and its biochemical characteristics of one beef sample was confirmed with API 20E kit (93.7%). L. monocytogenes was detected in fried rice paper dumpling, but the presumptive colonies were not detected onto the conventional plate. C. jejuni was detected in peeled & washed onion.

Effect of Cryopreservation Day on Pregnancy Outcomes in Frozen-thawed Blastocyst Transfer (동결 해동한 포배 이식에 있어서 동결시기가 임신결과에 미치는 영향)

  • Kim, Hyun-Jung;Kim, Chung-Hyon;Lee, Joong-Yeup;Kwon, Jae-Hee;Hwang, Do-Yeong;Kim, Ki-Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.57-64
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    • 2010
  • Objectives: Likewise fresh cycle, it is also important to select right blastocysts for transfer in purpose of improving the pregnancy and implantation rates in frozen-thawed embryo transfer (ET) cycles. To investigate the relationship between the developmental velocity at the time of cryopreservation and pregnancy rates, we compared pregnancy rates between the day 5 cryopreservation group and the day 6 cryopreservation group. Methods: Transfers of frozen-thawed blastocysts which had been cryopreserved by vitrification on day 5 or day 6 were performed between January 2006 and June 2007. Ethylene glycol, DMSO, and pull and cut straws were used for vitrification and artificial shrinkage was done in expanded blastocysts. Thawing was performed on the day before transfer and thawed blastocysts were cultured in for 15~18 hrs in Quinn's blastocyct media. Blastocyst survival was assessed before transfer and post-thaw survival was defined as >50% of cells remaining intact and blastocoele re-expansion by the time of transfer. Results: Transfers of thawed blastocyst had been cryopreserved on day 5 were 52 cycles and 41 transfer cycles were cryopreserved on day 6. Patient characteristics, the number of transferred embryos and the survival rate of thawed blastocysts were not different in each cryopreservation day. But the biochemical pregnancy, clinical pregnancy, ongoing pregnancy, and implantation rate were significantly high in transfer of frozen-thawed blastocyst which were cryopreserved on day 5. Conclusions: The clinical pregnancy and implantation rate of day-5 blastocyst showed significantly higher than those of day-6 blastocyst in frozen-ET cycles. This result indicated that developmental rate of blastocyst at cryopreservation time in frozen-thawed cycle is discriminative marker of pregnancy outcome as like in fresh cycle.

Current Status of the Research on the Postharvest Technology of Melon(Cucumis melo L.) (멜론(Cucumis melo L.) 수확 후 관리기술 최근 연구 동향)

  • Oh, Su-Hwan;Bae, Ro-Na;Lee, Seung-Koo
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.442-458
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    • 2011
  • Among Cucubitaceae, melon (Cucumis melo) is one of the most diversified fruits, with various forms, sizes, pulps, and peel colors, In addition, it is a commercially important crop because of its high sweetness, deep flavor, and abundant juice. In the species, there are both climacteric and non-climacteric melons depending on the respiration and ethylene production patterns after harvest. Ethylene is also considered a crucial hormone for determining sex expression, Phytohormones other than ethylene interact and regulate ripening, There are some indices that can be used to evaluate the optimum harvest maturity. The harvest time can be estimated after the pollination time, which is the most commonly used method of determining the harvest maturity of the fruit. Besides the physiological aspects, the biochemical alterations, including those of sweetness, firmness, flavor, color, and rind, contribute to the overall fruit quality. These changes can be categorized based on the ethylene-dependent and ethylene-independent phenomena due to the ethylene-suppressed transgenic melon. After harvest, the fruits are precooled to $10^{\circ}C$ to reduce the field heat, after which they are sized and packed. The fruits can be treated with hot water ($60^{\circ}C$ for 60 min) to prevent the softening of the enzyme activity and microorganisms, and with calcium to maintain their firmness. 1-methylenecyclopropene (1-MCP) treatment also maintains their storability by inhibiting respiration and ethylene production. The shelf life of melon is very short even under cold storage, like other cucurbits, and it is prone to obtaining chilling injury under $10^{\circ}C$. In South Korea, low-temperature ($10^{\circ}C$) storage is known to be the best storage condition for the fruit. For long-time transport, CA storage is a good method of maintaining the quality of the fruit by reducing the respiration and ethylene. For fresh-cut processing, washing with a sanitizing agent and packing with plastic-film processing are needed, and low-temperature storage is necessary. The consumer need and demand for fresh-cut melon are growing, but preserving the quality of fresh-cut melon is more challenging than preserving the quality of the whole fruit.

The effects of pregnancy on alveolar bone turnover during experimental tooth movement in rats (백서에서 실험적 치아이동시 임신이 치조골 교체(Turnover)에 미치는 영향에 대한 연구)

  • Kim, Young-Sun;Lee, Ki-Soo
    • The korean journal of orthodontics
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    • v.30 no.4 s.81
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    • pp.413-421
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    • 2000
  • The purpose of this study was to observe the effects of pregnancy on the experimental tooth movement and alveolar bone turnover process of Sprague-Dawley female rat. Sixty rats were divided into pregnant-tooth movement group(P-Tm), normal-tooth movement group(N-Tm) and normal group(N). Maxillary first molar appliances were inserted bilaterally and activated to 40grams. To measure the amount of tooth movement, x-ray was taken 2 times after appliance insertion and before sacrifice. Animals were sacrificed at 1,3,7,14 days(N=5). Just after sacrifice, alveolar bones were collected and frozen immediately for biochemical analysis. Tooth movement was assessed cephalometrically and tartrate-resistant acid(TRAP) and alkaline phosphatase (ALP) activities were measured in extracts of paradental alveolar bone. The results were as follows: 1. The amount of tooth movement in P-Tm group was greater than that of N-Tm group(p<0.01). 2. Alveolar bone ALP of normal tooth movement group was not significantly different from the control, TRAP was significantly different from the control(p<0.01). In normal tooth movement group, alveolar bone ALP was increased gradually and peak(day 7) fell off significantly at day 14(p<0.05). The Peak of alveolar bone TRAP(day 7) fell off slightly, sustained day 14(p<0.01). 3. Alveolar bone ALP and TRAP of pregnant tooth movement group were not significantly different from that of normal tooth movement group. In pregnant tooth movement group, alveolar bone ALP was increased at day 3(p<0.01) and fell off significantly at day 7-14, alveolar bone TRAP were increased at day 3 and sustained day 14. 4. The peak of alveolar bone phosphatases in pregnant tooth movement group(day3) preceded the peak in normal tooth movement group(day7) (p<0.01). According to the above results, we suggested that bone resorption activity was increased in alveolar bone of pregnant rat, and the degree of tooth movement in pregnancy may be greater than that of normal group because of high bone turnover of alveolar bone in pregnant rat.

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The Predictors of Effectiveness on Urokinase Instillation Therapy into Loculated Pleural Effusion. (소방이 형성된 흉막질환에서 유로키나제 주입치료의 예후인자)

  • Song, Kee-San;Bang, Jei-So;Kwak, Seung-Min;Cho, Chul-Ho;Park, Chan-Sup
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.621-628
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    • 1997
  • Background : As the pleural inflammation progresses, exudative pleural fluid becomes loculated rapidly with pleural thickening. Complete drainage is important to prevent pleural fibrosis, entrapment and depression of lung function. Intrapleural urokinase instillation therapy has been advocated as a method to facilitate drainage of gelatinous pleural fluid and to allow enzymatic debriment of pleural surface. This study was designed to investigate the predictors of effectiveness of intrapleural urokinase in the treatment of loculated pleural effusion. Method : Thirty-five patients received a single radiographically guided pig-tail catheter ranging in size from 10 to 12 French. Twenty-two patients had tuberculous pleural effusions, and 13 had non-tuberculous postpneumonic empyemas. A total of 240,000 units of urokinase was dissolved in 240 ml of normal saline and the aliquots of 80mL was instilled into the pleural cavity via pig-tail catheter per every 8hr. Effectiveness of intrapleural urokinase instillation therapy was assessed by biochemical markers, ultrasonography, and technical details. A greater than 50% improvement on follow-up chest radiographs was defined as success group. Result : Twenty-seven of 35 (77.1%) patients had successful outcome to urokinase instillation therapy. Duration of symptoms before admission was shorter in success group ($11.8{\pm}6.9day$) than in failure group ($26.62{\pm}16.5day$) (P<0.05). Amount of drained fluid during urokinase therapy was larger in success group ($917.1{\pm}392.7ml$) than in failure group ($613.8{\pm}259.7ml$) (P<0.05). Pleural fluid glucose was higher in success group ($89.7{\pm}35.9mg/dl$) than in failure group ($41.2{\pm}47.1mg/dl$) (P<0.05). Pleural fluid LDH was lower in success group ($878.4{\pm}654.3IU/L$) than in failure group ($2711.1{\pm}973.1IU/L$) (P<0.05). Honeycomb septated pattern on chest ultrasonography was observed in six of eight failure group, but none of success group (P<0.05). Conclusion : Longer duration of symptoms before admission, smaller amount of drained fluid during urokinase therapy, lower glucose value, higher LDH value in pleural fluid examination, and honeycomb septation pattern on chest ultrasonograph were predictors for failure group of intrapleural urokinase instillation therapy.

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The Usefulness of Pleural IFN-$\gamma$ Level in Differential Diagnosis of Tuberculous Pleural Effusion and Malignant Pleural Effusion (결핵성 흉막염과 악성 흉수의 감별진단에 있어서 흉수 IFN-$\gamma$ 측정의 유용성)

  • Kim, Myung-Sun;Yang, Sung-Eun;Chi, Hyun-Sook;Kim, Woo-Sung;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.280-289
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    • 1998
  • Background: It is sometimes difficult to differentiate tuberculous pleural effusion from malignant pleural effusion by clinical symptoms, signs, by routine tests of pleural fluid, and by pathologic studies. And recently, it was discovered that cytokines such as IL-2, IFN-$\gamma$, TNF-$\alpha$ are elevated in tuberculous pleural fluid, and there have been several attempts to diagnose tuberculous pleural effusion by using these immunological mediators. There are several studies regarding the diagnostic value of IFN-$\gamma$, and there are two studies in Korea. But the diagnostic values of IFN-$\gamma$ in these studies were slightly lower than those in other countries. To compare the diagnostic value of IFN-$\gamma$ with those of CEA and ADA, and to determine the sensitivity and specificity of IFN-$\gamma$ in Korean, we mesured IFN-$\gamma$, CEA level and ADA activity in pleural effusions. Methods: ADA activity, IFN-$\gamma$ level and CEA level as well as cell count, differential count, and biochemical assays such as protein content and lactate dehydrogenase were measured in 40 cases of tuberculous pleuritis and 42 cases of malignant pleural effusion. Results: Tuberculous pleural fluid showed higher levels of IFN-$\gamma$ and ADA ($832.6{\pm}357.2$ pg/ml and $82.5{\pm}25.9$ U/L, respectively) than those of malignant pleural effusion ($2.6{\pm}8.0$ pg/ml and $19.2{\pm}10.9$ U/L, respectively) (p<0.01). Malignant pleural effusions showed higher median value (102.2 ng/ml) than tubercalous pleural effusions (1.8 ng/ml) (p<0.01). The sensitivities of IFN-$\gamma$, ADA, CEA were 0.97, 0.87, 0.67 and the specificities of IFN-$\gamma$, ADA, CEA were 1.0, 0.97, 1.0, respectively. There was no significant correlation between ADA activity and IFN-$\gamma$ level. Conclusion: This study showed that IFN-$\gamma$ test would be a very useful clinical test for differential diagnosis of tuberculous pleuritis and malignant pleural effusion because it is very sensitive and specific, although it is an expensive test.

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Histomorphometry and Stability Analysis of Loaded Implants with two Different Surface Conditions in Beagle Dogs (하중을 가한 두 가지 표면의 임플란트에 관한 조직형태학적 분석 및 안정성 분석 (비글견을 이용한 연구))

  • Kim, Sang-Mi;Kim, Dae-Gon;Cho, Lee-Ra;Park, Chan-Jin
    • Journal of Dental Rehabilitation and Applied Science
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    • v.24 no.4
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    • pp.337-349
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    • 2008
  • Despite an improved bone reactions of Mg-incorporated implants in the animals, little yet has been carried out by the experimental investigations in functional loading conditions. This study investigated the clinical and histologic parameters of osseointegrated Mg-incorporated implants in delayed loading conditions. A total of 36 solid screw implants (diameter 3.75 mm, length 10mm) were placed in the mandibles of 6 beagle dogs. Test groups included 18 Mg-incorporated implants. Turned titanium Implants served as control. Gold crowns were inserted 3 months. Radiographic assessments and stabilitytests were performed at the time of fixture installation, $2^{nd}$ stage surgery, 1 and 3 months after loading. Histological observations and morphometrical measurements were also performed. Of 36 implants, 32 displayed no discernible mobility, corresponding to successful clinical function. There was no statistically significant difference between test implants and controls in marginal bone levels (p=0.413) and RFA values. The mean BIC % in the Mg-implants was $54.4{\pm}20.2%$. The mean BIC % in the turned implant was $48.9{\pm}8.0%$. These differences between the Mg-implant and control implant were not statistically significant (P=0.264). In the limitation of this study, bone-to-implant contact (BIC) and bone area of Mg-incorporated oxidized implant were similar to machine-turned implant. The stability analysis showed no significantly different ISQ values and marginal bone loss between two groups. Considering time-dependent bone responses of Mg-implant, it seems that Mg-implants enhanced bone responses in early loading conditions and osseointegrated similarly to cp Ti implants in delayed loading conditions. However, further investigations are necessary to obtain long-term bone response of the Mg-implant in human.

Production and fermentation characteristics of seafood kimchi started with Leuconostoc mesenteriodes SK-1 isolated from octopus baechu kimchi (문어 배추김치에서 분리한 Leuconostoc mesenteroides SK-1을 이용한 수산물김치의 제조 및 발효 특성)

  • Jang, Mi-Soon;Jung, Ko-Eun;Yun, Jae-Ung;Nam, Ki-Ho
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1050-1057
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    • 2016
  • This study was carried out to investigate the effectiveness of Leuconostoc mesenteroides isolated from octopus baechu kimchi as a potential starter for seafood kimchi. L. mesenteroides is lactic acid bacterium currently used as a starter for kimchi production. We selected the most effective L. mesenteroides strain from the 7 strains isolated from octopus baechu kimchi and, based on biochemical properties and 16S rRNA sequencing, identified the selected strain as L. mesenteroides SK-1. The SK-1 strain exhibited acid-tolerance, good survival capacity, and excellent dextran productivity. We investigated the effects the SK-1 of starter on seafood kimchi fermentation. Octopus baechu kimchi was fermented with L. mesenteroides SK-1 at $4^{\circ}C$ for 35 d. The decrease in pH and increase in acidity in octopus baechu kimchi fermented with the SK-1 starter occurred more quickly than that in the control kimchi indicating that. Octopus baechu kimchi with SK-1 starter has a relatively slow rate of increase in lactic acid production. As a result, octopus baechu kimchi prepared with L. mesenteroides SK-1 can be maintained at a suitable ripening degree over an extended period of time compared to that of the control kimchi, Moreover, the octopus baechu kimchi started with L. mesenteroides SK-1 has excellent sensory properties, including a refreshing taste, and a weak sour odor.