• Title/Summary/Keyword: In vivo

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Silencing of Mutant p53 Leads to Suppression of Human Breast Xenograft Tumor Growth in vivo (돌연변이 p53 단백질의 Silencing에 의한 사람유방암세포의 in vivo 항 종양 효과)

  • Park, Won Ick;Park, Se-Ra;Park, Hyun-Joo;Bae, Yun-Hee;Ryu, Hyun Su;Jang, Hye-Ock;Bae, Moon-Kyoung;Bae, Soo-Kyung
    • KSBB Journal
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    • v.31 no.1
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    • pp.52-57
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    • 2016
  • Mutant p53 (R280K) is highly expressed in MDA-MB-231 triple-negative human breast cancer cells. Currently, we reported the role of mutant p53-R280K in mediating the survival of MDA-MB-231 cells in vitro. The present study was undertaken to determine whether mutant p53-R280K affects breast cancer cell growth in vivo. To this end, we used small interfering RNA to knockdown the level of mutant p53-R280K in MDA-MB-231 cells. Silencing of mutant p53-R280K in MDA-MB-231 cells causes substantial tumor regression of established xenografts in vivo. In xenograft model for breast cancer, silencing of mutant p53-R280K in MDA-MB-231 cells significantly inhibited the tumor growth. Moreover, TUNEL assay showed more occurrence of apoptotic cells in mutant p53-R280K silenced tumors compared to control. Our data indicate that mutant p53-R280K has an important role in mediating tumor growth of MDA-MB-231 cells in vivo. Taken together, this study suggests that endogenous mutant p53-R280K could be used as a therapeutic target for breast cancer cells harboring this TP53 missense mutation.

Effects of Aqueous Extract of Diospyros Kaki Calyx on Anti-thrombotic Activity in vitro and in vivo (시체(柿蒂)의 in vitro와 in vivo 항혈전 효능 연구)

  • Baek, Kyung-Min;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.139-147
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    • 2011
  • Objectives : The aim of this study is to research an anti-thrombus effect by Diospyros Kaki Calyx. Methods : The healthy human plasma were gained and used in vitro study such as factor X activity (FXa) inhibition, prothrombinase inhibition, prothrombin time (PT) and activated partial thromboplastin time. Fifteen SD rats were divided into three groups ; intact control group (orally administrated with distilled water 5ml/kg) and two experimental group treated with extract of diospyros kaki calyx (EKC). Experimental rats were orally 600 mg/kg concentration of EKC and 200 mg/kg concentration of EKC. After an hour from administration, we anesthetized rats and made arteriovenous (AV) shunt rat models to study weight of thrombus, took whole blood to study content of thromboxane B2 and blood clotting time. Results : In vitro, EKC significantly increased inhibitory activity of FXa, prothrombinase compared with intact control group ($^*P$ <0.05). PT and aPTT were increased in EKC treated (600 mg/kg) group compared with intact control group ($^*P$ <0.05). In vivo, blood clotting time of experiment group treated with EKC 600 mg/kg were significantly increased compare with that of intact control group (p<0.05) and content of thromboxane B2 was significantly decreased in group treated with EKC 600 mg/kg in serum. The weight of thrombus were significantly reduced in group treated with EKC 600 mg/kg compared with intact control group (p<0.05). But in vivo experiment study, those parameters of group treated with EKC 200 mg/kg were relatively decreased compared with those of intact control group without statistical significance. Conclusions : EKC has an antithrombic activity because of inhibition internal course such as FXa and prothrombin. And EKC inhibited a hole blood clotting in vivo experiment by low content of thromboxane B2.

Seeing is Believing: Illuminating the Source of In Vivo Interleukin-7

  • Kim, Grace Yoon-Hee;Hong, Chang-Wan;Park, Jung-Hyun
    • IMMUNE NETWORK
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    • v.11 no.1
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    • pp.1-10
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    • 2011
  • Interleukin-7 (IL-7) is an essential cytokine for T cells. However, IL-7 is not produced by T cells themselves such that T cells are dependent on extrinsic IL-7. In fact, in the absence of IL-7, T cell development in the thymus as well as survival of naive T cells in the periphery is severely impaired. Furthermore, modulating IL-7 availability in vivo either by genetic means or other experimental approaches determines the size, composition and function of the T cell pool. Consequently, understanding IL-7 expression is critical for understanding T cell immunity. Until most recently, however, the spatiotemporal expression of in vivo IL-7 has remained obscured. Shortage of such information was partly due to scarce expression of IL-7 itself but mainly due to the lack of adequate reagents to monitor IL-7 expression in vivo. This situation dramatically changed with a recent rush of four independent studies that describe the generation and characterization of IL-7 reporter mice, all utilizing bacterial artificial chromosome transgene technology. The emerging consensus of these studies confirmed thymic stromal cells as the major producers of IL-7 but also identified IL-7 reporter activities in various peripheral tissues including skin, intestine and lymph nodes. Strikingly, developmental and environmental cues actively modulated IL-7 reporter activities in vivo suggesting that IL-7 regulation might be a new mechanism of shaping T cell development and homeostasis. Collectively, the availability of these new tools opens up new venues to assess unanswered questions in IL-7 biology in T cells and beyond.

Enhanced visualization of the root canal morphology using a chitosan-based endo-radiopaque solution

  • Shashirekha Govind;Amit Jena;Satabdi Pattanaik;Mahaprasad Anarasi ;Satyajit Mohapatra;Vinay Shivagange
    • Restorative Dentistry and Endodontics
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    • v.46 no.3
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    • pp.33.1-33.12
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    • 2021
  • Objectives: This study aimed to investigate the efficacy of ionic and non-ionic-based contrast media (in vitro study) and the combinatorial effect of chitosan-based endo-radiopaque solution (CERS) (in vivo study) for visualization of the root canal anatomy. Materials and Methods: In vitro study (120 teeth): The root canal of maxillary premolars and molars (in vitro group 1 and 2 respectively, n = 60 each) were analyzed using 4 different contrast media (subgroups: Omnipaque 350, Iopamidol, Xenetix 350, and Urografin 76; n = 15 each) in combination with 5.25% sodium hypochlorite (NaOCl). Based on the results of the in vitro study, in vivo study (80 teeth) was done to compare Xenetix 350 + 5.25% NaOCl with CERS (in vivo group 1 and 2 respectively, n = 40 each) on maxillary and mandibular premolars and molars. Two endodontists used radiovisiography to assess the depth of ingress and identify the aberrant root anatomy after access cavity preparation, and after initial cleaning and shaping of canals. Kruskal-Wallis test was used for in vitro comparison (p < 0.05), and Wilcoxon signed-rank test and Mann-Whitney U test for in vivo analysis (p < 0.01). Results: In vitro study, Xenetix 350 + 5.25% NaOCl facilitated a significant higher visualization (p < 0.05). For in vivo study, CERS had a statistically significant depth of ingress (p < 0.01), and was efficient in identifying the aberrant root canal anatomy of premolars and molars. Conclusions: CERS facilitates better visualization of the root canal anatomy of human premolars and molars.

Genotoxicological Safety of Gamma-Irradiated Salted and Fermented Anchovy Sauce (감마선 조사된 멸치액젓의 유전독성학적 안전성 평가)

  • 육홍선;차보숙;김동호;이주운;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.7
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    • pp.1192-1200
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    • 2004
  • Gamma irradiations at 5 or 10 kGy were applied to salted and fermented anchovy sauce, for improving the hygiene Quality and evaluating the genotoxicological safety. In vitro genotoxicological safety of irradiated sauces was evaluated by Salmonella Typhimurium (TA98, TA100, TAI535 and TAI537) and E. coli WP2 uvrA, reversion assay, SOS chromotest (Escherichia coli PQ37), and chromosome aberration test (Chinese hamster lung fibroblast cells) in the absence or presence of an exogenous metabolizing system (S9 mix). The gamma-irradiated samples were not significantly different from nonirradiated-control for three in vitro tests (p<0.05). :In vivo micronucleus test using ICR mice (male) was not significantly different from the control at p<0.05. The salted and fermented anchovy sauce exposed to 5 or 10 kGy-gamma ray revealed negative results in these three in vitro mutagenetic tests and in vivo micronucleus test upto 50,000 $\mu$g/plate, respectively. The results indicated that 5 or 10 kGy gamma-irradiated salted and fermented anchovy sauces did not show any mutagenicity.

Pharmacokinetic Modelling and Simulation of the Counter-transport in the Hepatic Transport of Organic Anions (음이온계 약물의 간수송과정에 있어서 대향수송의 약물동력학적 모델링 및 시뮬레이션)

  • Song, Suk-Gil;Lee, Jun-Seup;Chung, Youn-Bok
    • YAKHAK HOEJI
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    • v.49 no.4
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    • pp.275-283
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    • 2005
  • The purpose of the present study was to kinetically investigate the carrier-mediated uptake in the hepatic transport of organic anions, and to simulate the 'in vivo counter-transport' phenomena, using kinetic model which was developed in this study. The condition that the mobility of carrier-ligand complex is greater than that of free carrier is not essential for the occurrence of 'counter-transport' phenomenon. To examine the inhibitory effects on the initial uptake of organic anions by the liver, it is necessary to judge whether the true counter-transport mechanism (trans-stimulation) is working or not. Effects of bromophenol blue (BPB) or bromosulfophthalein (BSP) on the plasma disappearance curves of a 1-anilino-8-naphthalene sulfonate (ANS) were then kinetically analyzed based on a flow model, in which the ligand is eliminated only from the peripheral compartment (liver compartment). Moreover, 'in vivo counter-transport' phenomena were simulated based on the perfusion model which incorporated the carrier-mediated transport and the saturable intracellular binding. The 'in vivo counter-transport' phenomena in the hepatic transport of a organic anions were well demonstrated by incorporating the carrier-mediated process. However, the 'in vivo counter-transport' phenomena may be also explained by the enhancement of back diffusion due to the displacement of intracellular binding. In conclusion, one should be more cautious in interpreting data obtained from so-called 'in vivo counter-transport' experiments.

The Statistical Technique for Dosimetric Meta-Analysis at in-vivo and in-vitro Papers about Bioeffects of RF Electromagnetic Field (RF 전자기장 생체 영향 연구 문헌에서 노출량 메타분석을 위한 통계적 기법)

  • Choi, Sung-Ho;Kim, Nam
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.14 no.12
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    • pp.1311-1320
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    • 2003
  • This paper proposed the estimating method of electromagnetic dosimetric reliability at in-vivo and in-vitro experiments. For more accurate consequences of these researches, we have tried to find out any correlations among output power, power density and specific absorption rate(SAR) with the results of in-vivo, in-vitro tests and SAR reports of cellular phone and PDA. In the case of in-vivo tests, the power density has close statistical correlations with SAR value and in the event of in-vitro tests, the output power has considerable statistical correlations with SAR containing duty factor. We analysed the coefficient of determination to estimate the dosimeoic uncertainty. If we use this method before evaluating techniques of measurement and analysis at both in-vivo and in-vitro experiments, we will conduct more accurate reliability test.

siRNA-mediated Inhibition of hTERC Enhances Radiosensitivity of Cervical Cancer

  • Chen, Min;Xing, Li-Na
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.5975-5979
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    • 2012
  • Background: To investigate the influence of telomerase activity, apoptosis, radiosensitivity of cervical cancer after siRNA-mediated knockdown of telomerase RNA and evaluate in vivo growth with gene interference. Methods: We studied siRNA-targeting-telomerase RNA transfection into the Hela cell line. Expression of hTERC mRNA was detected by RT-PCR and telomerase activity was measured by the TRAP assay. Growth inhibition was determined by MTT assay and radiosensitivity of the cervical cancer cells was examined by colony formation assay. In addtion, effects of hTERC inhibition in vivo were studied by injection of siRNA-transfected Hela cells into nude mice. Results: The hTERC siRNA effectively downregulated the expression of hTERC mRNA and also reduced the telomerase activity to 30% of the untreated control vlaue. The viability of hTERC siRNA transfected Hela cells was reduced by 44.7% after transfection. After radiation treatment, the radiosensitivity of Hela cells with hTERC knockdown was increased. In vivo, the tumors developing from the hTERC siRNA-transfected cells were of reduced size, indicating that the hTERT siRNA also depressed the tumorigenic potential of the Hela cells. Conclusions: Our results supported the concept of siRNA-mediated inhibition of telomerase mRNA which could inhibit the expression of hTERC and telomerase activity. Furthermore, radiosensitivity was upregulated after knockdown the hTERC in vivo and in vitro.

In Vivo Blood Compatibility of PU-PEO-SO3 as Coating Material for Blood Sac of Left Ventricular Assist Device ( LVAD) (좌심실보조장치의 혈액주머니용 코팅재료로서 PU-PEO-SO3의 in vivo 혈액적합성에 관한 연구)

  • 한동근;김종원
    • Journal of Biomedical Engineering Research
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    • v.15 no.1
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    • pp.19-26
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    • 1994
  • Sulfonated poly (ethyleneoxide)-grafted polyurethane (PU-PEO-$SO_3$) prepared by bulk modification was coated on a blood sac for electrohydraulic left ventricular assist device (ELVAD) implanted in dogs and its in vivo blood compatibility on shear stress was studied as compared with untreated Po. The effect of the wall shear stress on the protein adsorption unlike platelet adhesion is dependent on the surface characteristics of the material, although less proteins seem to be adsorbed in the region of the high shear stress. The thickness of total proteins adsorbed on PU-PEO-SOJ (400 ${\AA}$) by trans¬mission electron microscopy(TEM) was considerably lower than that of untreated PU(l,000~1,600 ${\AA}$), but PU-PEO-$SO_3$ showed high albumin adsorption, low fibrinogen and IgG adsorption, and low platelet adhesion as compared with untreated PU, suggesting that PU-PEO-$SO_3$ is more in vivo blood compatible. Therefore, it appears that such a blood compatible PU-PEO-$SO_3$ is useful for blood contacting biomaterials including artificial organs.

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Effect of Fusion Condition on In Vitro Development of Caprine Cloned Oocytes with Nuclear Transfer (재래산양에 있어서 핵이식란의 융합조건이 융합 및 체외발달에 미치는 영향)

  • 박희성;김태숙;이윤희;정수영;이명열;홍승표;박준규;김충희;정장용
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.127-132
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    • 2004
  • This study was conducted to examine the effects of electric stimulation conditions on in vitro developmental ability of caprine embryos after somatic cell nuclear transfer. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean native goats. The caprine ear cells were cultured in vitro in serum-starvation condition (TCM-l99 + 0.5% FBS) for 3 to 5 days of cell confluence. The zona pellucida of in vivo and in vitro matured oocytes were partially drilled using laser system. Single somatic cell was individually transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol. After the electofusion, embryos were activated by electric stimulation or Ionomycin + 6-DMAP. Nuclear transfer embryos were cultured in mSOF medium supplemented with 0.8% BSA 6∼7 days at 39 , 5% $CO_2$, 5% $O_2$, 90% $N_2$. The fusion rate of donor cells was 60.4% and 40.3 % in ear cell and fetal fibroblast, and cleavage rate were 40.6% and 48.2%, respectively. No significant difference was found in the fusion and cleavage rate in different donor cells. Nuclear transferred oocytes were fused by electric pulses of 1.30∼1.40, 2.30∼2.39 and 2.40∼2.46 ㎸/cm. There was no significant difference among different electric pulses in fusion rates (26.7, 34.8 and 43.8%). The cleavage rate was higher (p<0.05) in 1.30∼1.40 ㎸/cm (82.9%) than 2.30∼2.39 ㎸/cm (43.8%) and 2.40∼2.46 ㎸/cm. (51.8%). The fusion rates of recipient oocyte source were 1st (43.5% and 23.6%), 2nd (55.7% and 39.2%) and 3rd (66.1% and 52.8%) in in vivo and in vitro oocytes. However, fusion ratee were significantly higher (p<0.05) in in vivo than in vitro oocyte. The cleavage rate of fused oocytes from in vivo and in vitro sources were 52.6% and 54.4%, respectively. No significant difference was found in the cleavage rate according to the recipient oocyte source. These results suggest that factors such as field pulse of electric stimulation and oocyte source could affect in vitro developmental ability of nuclear transplanted caprine oocytes.