• 제목/요약/키워드: microplate

검색결과 151건 처리시간 0.027초

작약(Paeoniae radix) 추출물의 식후 과혈당 억제작용 (Inhibitory Effect of Extracts from Paeoniae radix on Postprandial Hyperglycemia)

  • 지승택;이성진;이강은;손용태;정요경
    • 한국식품영양과학회지
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    • 제31권1호
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    • pp.131-135
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    • 2002
  • 본 연구에서는 작약 추출물의 식후 과혈당 억제작용을 조사하였다. 작약 유기용매(hexane, ethyl acetate, butanol, water) 추출물들을 high performance liquid chromatogrphy를 이용하여 분획한 분획물(fraction)들으 $\alpha$-glucosidase(EC 3.2.1.20) 저해제를 탐색하였다. 작약 ethyl acetate 추출물의 11, 12, 18, 19번 분획물들은 20 $\mu\textrm{g}$/mL에서 각각 85%, 84%, 77%, 77%의 강력한 저해 활성을 나타내었으며, 작약 ethyl acetate 추출물의 10~19번 분획을 경구투여한 in vivo당 (maltose) 부하 실험에서도 양성대조군과 비교하여 유의성 있게 22%의 혈당을 낮추었다. 따라서 본 연구의 in vitro와 in vivo 실험을 통하여 확인 된 $\alpha$-glucosidase 저해 활성을 갖는 작약 분획물들은 식후의 혈당상승 작용을 억제하는 새로운 nutraceutical 소재와 항당뇨 신약개발을 위한 탐색자원으로서 매우 가치있는 자원으로 기대된다.

Optimization of SNP Genotyping Assay with Fluorescence Polarization Detection

  • Cai Chun Mei;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • 한국작물학회지
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    • 제50권5호
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    • pp.361-367
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    • 2005
  • Single nucleotide polymorphisms (SNPs) are valuable DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed. In this report, one of effective and high throughput SNP genotyping assays, which was named the template-directed dye-terminator incorporation with fluorescence polarization detection (FP-TDI) was described. Although the most of this assay succeed, the objective of this work was to deter­mine the reasons for the failures, find ways to improve the assay and reduce the running cost. Ninety $F_2$-derived soybean, Glycine max (L.) Merr., RILs from a cross between 'Pureunkong' and 'Jinpumkong 2' were genotyped at four SNPs. FP measurement was done on $Victot^3$ microplate reader (perkinelmer Inc., Boston, MA, USA). Increasing the number of thermal cycles in the single-base extension step increased the separation of the FP values between the products corresponding to different genotypes. But in some assays, excess of heterozygous genotypes was observed with increase of PCR cycles. We discovered that the excess heterozygous was due to misincorporation of one of the dye­terminators during the primer extension reaction. After pyrophosphatase incubation and thermal cycle control, misincoporation can be effectively prevented. Using long amplicons instead of short amplicons for SNP genotyping and decreasing the amount of dye terminator and Acyclopol Taq polymerase to 1/2 or 1/3 decreased the cost of the assay. With these minor adjustments, the FP-TDI assay can be used more accurately and cost-effectively.

Field efficacy of a combined vaccine supplemented with recombinant Pasteurella multocida toxin subunits against atrophic rhinitis

  • Kang, Mi Lan;Shin, Seung Won;Rayamahji, Nabin;Seo, Yeon Soo;Lee, Su In;Lee, Won Hyung;Yoo, Han Sang
    • 대한수의학회지
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    • 제48권1호
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    • pp.53-60
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    • 2008
  • We have investigated efficiency of a recombinant subunit Pasteurella multocida toxin (PMT) that was mixed with a vaccine consisted of inactivated whole cells of Bordetella bronchiseptica, P. multocida (types A and D). For verification of the efficacy of the vaccine, all experimental pigs (suckling piglets, sow and gilts) in the three farms were vaccinated. Antibody titers against B. bronchiseptica and P. multocida type A of the vaccinated pigs by microplate agglutination were significantly higher than those of the control pigs (p < 0.05). Similar patterns were observed in the analysis of anti- PMT neutralizing antibody by serum neutralizing method using Vero cell (p < 0.05). Anti- P. multocida type D antibody titer of the vaccinated sows and gilts by ELISA showed significant differences with those of the non-vaccinated pigs (p < 0.05). Although antibody titers increased, it was unable to find out the difference in the clinical signs between the vaccinated and non-vaccinated pigs. However, the increase in body weight of the vaccinated piglets was observed in comparison with the non-vaccinated piglets on a farm. At slaughtering of the pigs, pathological lesions in the turbinate bones of the vaccinated pigs were significantly lower than those of the non-vaccinated pigs (p < 0.001). These results suggested that efficacy of the vaccine in pigs demonstrated to protect against atrophic rhinitis in Korea.

강아지풀 및 강아지풀모사로봇의 이동특성에 관한 연구 (Locomotion Characteristics of a Foxtail and a Foxtail-like Robot)

  • 이민수;김영혁;임상혁;김병규
    • 대한기계학회논문집A
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    • 제34권12호
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    • pp.1893-1899
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    • 2010
  • 평평한 바닥 위에서 강아지풀을 수직으로 누르면 강아지풀은 앞으로 이동한다. 이 현상을 통해서 강아지풀의 이동거리는 강아지풀 처짐량에 의존하는 것을 알아내었다. 그러므로 강아지풀을 누르는 힘, 면적, 속도 변화에 따른 강아지풀의 이동거리에 영향을 주는 요인을 실험을 통해 알아보았다. 이 결과를 바탕으로 강아지풀의 섬모를 모사한 나일론 섬모를 제작하여, 나일론 섬모에 작용하는 힘과 처짐량에 따른 이동거리를 이론적, 실험적으로 알아내었다. 또한 로봇과 관의 내벽 사이 간극 차이에 따라 발생하는 로봇 다리의 각도변화에 따른 이동거리에 대해서 연구하였다. 그리고 소화기와 혈관의 치료를 위한 이동로봇의 섬모 제작에 관한 설계 변수를 제공하기 위해, 로봇 다리의 직경과 탄성계수의 관계에 대해서 연구하였다.

이정환(二精丸)이 노화과정에 미치는 영향 (Effects of Ichungwhan on the Aging Process)

  • 정지천;현민경
    • 대한한방내과학회지
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    • 제26권2호
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    • pp.379-389
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    • 2005
  • Objectives: It is well known that aging and aging-related diseases are linked to the increased level of oxidative stress caused by reactive oxygen species(ROS) and reactive nitrogen species(RNS). Nonprotein-SH decreases during aging, while substances such as ROS, nitric oxide(NO), peroxynitrite($ONOO^-$), myeloperoxidase(MPO), and dityrosine show a significant increase. This study investigated the effect of Ichungwhan on the aging process by examining its effect on the generation of the above-mentioned substances. Methods: Four comparison groups of SD rats were used. They were 6 month-old rats, 24 month-old rats, and 24 month-old rats fed with food containing 0.1% and 0.3% of Ichungwhan extract. The amount of NO, $ONOO^-$, and ROS in the rats' kidneys were examined using a fluorescence microplate reader. The reagents used for this purpose include: dihydrorhodamine 123 (DHR 123), 2',7' -dichlorodihydrofluorescein, diacetate(DCFDA), and 4,5-diaminofluorescein(DAF-2). A spectrophotometer was used to investigate the reactivity of nonprotein-SH and myeioperoxidase(MPO), using reagents such as trichloroacetic acid(TCA) and tetramethylbenzidine(TMB). The amounts of MPO protein and dityrosine were measued by western blot. Results: The observed effects of Ichungwhan on rats were as follows: increase of nonprotein-SH; effective decrease of RNS level by suppression of the generation system of $ONOO^-$ and NO; decrease of ROS level; decrease of the MPO reactivity and the subsequent reduction of amount of MPO protein; retardation of dityrosine formation. It can be hypothesized, therefore, that Ichungwhan affects both the earlier and later phases of the molecular inflammatory process, and retards the aging process. Conclusions: Empirical evidence in this study supports a role for Ichungwhan in generation mechanisms of aging process-linked substances ROS, NO, $ONOO^-$, nonprotein-SH, MPO and dityrosine. Affects contrary to the aging process observed in rats beg further empiricism to investigate potential application of Ichungwhan as a medication for age-related diseases in humans.

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Glutathione is the Major Defensive Mechanism against Oxidative Stress in Human Embryonic Stem Cell

  • 이건섭;이영재;김은영;박세필;임진호
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.78-78
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    • 2003
  • Embryonic stem (ES) cells, derived from preimplantation embryo, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In contrast, terminally differentiated cells do not usually alter their nature but frequently die or transform if they are exposed to inappropriate external stimulations. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES cells (MB03) and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is $H_2O$$_2$. Approximately 1$\times$10$^4$ cells were plated in 96 well plate and serum starved for overnight. The conditioned cells were exposed to a various concentration of $H_2O$$_2$ fur 24 hrs and loaded with neutral red (50$\mu\textrm{g}$/ml) for 4 hrs, washed with PBS for 2 min three times, and entrapped dye was dissolved out using acetic ethanol. Cytotoxicity was determined by reading the amount of dye in the medium using microplate reader. equipped with 575 nm filter. Relative amount of the dye entrapped within MB03 or HeLa were not significantly different when cells were exposed up to 0.4 mM $H_2O$$_2$. However, this sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM $H_2O$$_2$, while it was approximately 54% in MB03 suggesting that this concentration of $H_2O$$_2$ is the defensive threshold for HeLa cells. The resistance to oxidative stimulation reversed, however, when cells were co-treated with BSO (L-buthionine- 〔S, R〕-sulfoximine) which chelates intracellular GSH. This result suggests that cellular GSH is the major defensive mechanism of human ES cells. Induction of enzymes involved in GSH metabolism and type of cell death is currently being studied.

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Phenylpropanoids of Plant Origin as Inhibitors of Biofilm Formation by Candida albicans

  • Raut, Jayant Shankar;Shinde, Ravikumar Bapurao;Chauhan, Nitin Mahendra;Karuppayil, Sankunny Mohan
    • Journal of Microbiology and Biotechnology
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    • 제24권9호
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    • pp.1216-1225
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    • 2014
  • Biofilm-related infections of Candida albicans are a frequent cause of morbidity and mortality in hospitalized patients, especially those with immunocompromised status. Options of the antifungal drugs available for successful treatment of drug-resistant biofilms are very few, and as such, new strategies need to be explored against them. The aim of this study was to evaluate the efficacy of phenylpropanoids of plant origin against planktonic cells, important virulence factors, and biofilm forms of C. albicans. Standard susceptibility testing protocol was used to evaluate the activities of 13 phenylpropanoids against planktonic growth. Their effects on adhesion and yeast-to-hyphae morphogenesis were studied in microplate-based methodologies. An in vitro biofilm model analyzed the phenylpropanoid-mediated prevention of biofilm development and mature biofilms using XTT-metabolic assay, crystal violet assay, and light microscopy. Six molecules exhibited fungistatic activity at ${\leq}0.5mg/ml$, of which four were fungicidal at low concentrations. Seven phenylpropanoids inhibited yeast-to-hyphae transition at low concentrations (0.031-0.5 mg/ml), whereas adhesion to the solid substrate was prevented in the range of 0.5-2 mg/ml. Treatment with ${\leq}0.5mg/ml$ concentrations of at least six small molecules resulted in significant (p < 0.05) inhibition of biofilm formation by C. albicans. Mature biofilms that are highly resistant to antifungal drugs were susceptible to low concentrations of 4 of the 13 molecules. This study revealed phenylpropanoids of plant origin as promising candidates to devise preventive strategies against drug-resistant biofilms of C. albicans.

하악구치부 보철공간을 위한 상악구치부의 분절골절단 및 상방 정위 (SURGICAL REPOSITIONING OF THE EXTRUDED DENTO-ALVEOLAR SEGMENTS BY THE SINGLE-STAGE POSTERIOR MAXILLARY SEGMENTAL OSTEOTOMY)

  • 김명래;김충;김형섭
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제23권4호
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    • pp.338-347
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    • 2001
  • Purpose: This is to review the cases of posterior maxillary segmental osteotomies to regain the interarch spaces for dental implants in the posterior mandible. Materials & Methods: Seven patients who presented with alveolar extrusion of upper posterior molars underwent segmental osteotomies by single-stage Kufner's buccal approach under the intravenous sedation and local anesthesia. The posterior maxillary cento-alveolar segments were repositioned upward using pre-fabricated palato-occlusal resin splints and immobilized with osteosynthesis microplates and screws. Dental implants were installated simultaneously. The regained spaces, tooth vitality, periodontal healing, relapse, tenderness on function, and complications including maxillary sinus involvements were evaluated periodically for over one year after the surgeries. Results: The single-tage procedures were completed within 80 minutes without any surgical complications. The posterior maxillary segments were repositioned upward to regain the interarch spaces ranging from 2.5 to 5.5mm. All teeth involved in the procedures keep their vitalities. The repositioned segments were maintained showing neither evidence of periodontal break-down nor tenderness to function. One patient whose segments had not been immobilized by osteosynthesis plate resulted in 2mm down-ward relapse in post-operative 8 months. A case of postoperative nasal bleeding from the posterior-lateral wall resulted in oroantral fistula and chronic maxillary sinusitis later. Conclusion: The extruded dento-alveolar segments of the posterior maxilla were repositioned properly by Kufner's one-stage segmental osteotomies. One microplate can be of help to keep the position until the osseous healing enough to support the masticatory force.

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건칠의 항주름 효과와 미백효과 연구 (The effect of wrinkle care and whitening by Rhus verniciflua Stokes)

  • 안재형;김경준
    • 한방안이비인후피부과학회지
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    • 제29권3호
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    • pp.124-133
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    • 2016
  • Objectives : Recently, the demands for the effective and safe depigmentating and anti-aging agents of the skin have increased due to the medical, pharmaceutical and cosmetic reasons. The aim of this study is to search new materials from the traditional herbal medicines which inhibit the aging process of skin in vitro.Methods : Human dermis cell (HS68) was used to test the effect of Rhus verniciflua Stokes (RVS). 80% ethanol or water extracts were screened for their inhibitory activities against elastase. Elastase inhibition effect was tested by microplate reader instrument. And MMP-1 suppression effect of RVS was tested by western blot. These cells were investigated the viability by MTS assay. And also the inhibition effect of tyrosinase by RVS was tested.Results : RVS (final concentrstion 1 ㎎/㎖ appeared over 30% of inhibition of elastase activity. So we are investigated anti wrinkle effects of Rhus verniciflua Stokes look through MMP-1 inhibition activity, also Extracts of RVS showed higher anti-tyrosinase activity than arbutin as final concentration 1 mg/ml. These results suggest that herbal medicines could be strong potential sources of inhibition of anti-aging and whitening effects for the skin.Colclusions : RVS was the best suppressor candidate of elastase activity among other control oriental drugs. It was found that RVS did not have toxicity to cells. We found proper concentration of RVS to treat in HS68 culture and investigated not only wrinkle care effect but also whitening effect of RVS.

ELISA를 이용한 돼지 톡소플라스마병의 조기 진단에 관한 연구 (Use of the enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine)

  • 서명득;장동화;주후돈
    • 대한수의학회지
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    • 제29권4호
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    • pp.567-575
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    • 1989
  • This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

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