• Title/Summary/Keyword: in vivo and in vitro

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Comparison of lymphocyte DNA damage levels and total antioxidant capacity in Korean and American diet

  • Lee, Min Young;Kim, Hyun A;Kang, Myung Hee
    • Nutrition Research and Practice
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    • v.11 no.1
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    • pp.33-42
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    • 2017
  • BACKGROUND/OBJECTIVE: This study aims to measure the in vitro antioxidant capacity of Korean diet (KD) with American diet (AD) as a control group and to examine the ex vivo DNA damage reduction effect on human lymphocytes. MATERIALS/METHODS: The KD applied in this study is the standard one-week meals for Koreans (2,000 kcal/day) suggested by 2010 Dietary Reference Intakes for Koreans. The AD, which is the control group, is a one-week menu (2,000 kcal/day) that consists of foods that Americans would commonly take in according to the National Health and Nutrition Examination Survey. The antioxidant capacity of each menu was measured by means of the total phenolic assay and 3 in vitro antioxidant activity assays (2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC), Oxygen radical absorbance capacity ($ORAC_{ROO{\cdot}}$)), while the extent of ex vivo lymphocyte DNA damage was measured by means of the comet assay. RESULTS: When measured by means of TEAC assay, the in vitro antioxidant capacity of the KD of the day was higher than that of the AD (P < 0.05) while there was no significant difference in total phenolic contents and DPPH and ORAC assays. The ex vivo lymphocyte DNA damage protective effect of the KD was significantly higher than that of the AD (P < 0.01). As for the one-week menu combining the menus for 7 days, the total phenolic assay (P < 0.05) and in vitro antioxidant capacity (P < 0.001, DPPH; P < 0.01, TEAC) of the KD menu were significantly higher than those of the AD menu. Likewise, the ex vivo DNA damage reduction rate of the Korean seven-day menu was significantly higher than that of the American menu (P < 0.01). CONCLUSION: This study demonstrates that the high antioxidant capacity and DNA damage protective effect of KD, which consists generally of various plant foods, are higher than those of typical AD.

Effect of resveratrol on the metastasis of 4T1 mouse breast cancer cells in vitro and in vivo

  • Lee, Hyun-Sook;Ha, Ae-Wha;Kim, Woo-Kyoung
    • Nutrition Research and Practice
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    • v.6 no.4
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    • pp.294-300
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    • 2012
  • We investigated the effects of resveratrol on metastasis in in vitro and in vivo systems. 4T1 cells were cultured in the presence of various concentrations (0-30 ${\mu}mol/L$) of resveratrol. For experimental metastasis, BALB/c mice were injected intravenously with 4T1 cells in the tail vein, and were orally administered various concentrations (0, 100, or 200 mg/kg Body weight) of resveratrol for 21 days. After resveratrol treatment, cell adhesion, wound migration, invasion, and MMP-9 activity were significantly decreased in a dose-dependent manner in 4T1 cells (P < 0.05). The numbers of pulmonary nodules were significantly decreased in mice fed the resveratrol (P < 0.05). The plasma MMP-9 activity was decreased in response to treatment with resveratrol in mice (P < 0.05). We conclude that resveratrol inhibits cancer metastasis both in vitro and in vivo, and this inhibition is likely due to the decrease in MMP-9 activity caused by resveratrol.

Inhibitory Effect of Salvia miltiorrhiza Extract on Growth of Some Cancer Cells (단삼(Salvia Miltiorrhiza) 추출물의 암세포 증식 억제 효과에 관한 연구)

  • 정국찬;이지영;김동청;서성옥;황우익
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.726-731
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    • 2000
  • This study was performed to evaluate the antitumor activities of water and ethanol (EtOH) extract of Salvia miltiorrhiza in vitro and in vivo. The proliferation of the human hepatoma (HepG2), rectum cancer (HRT-18) and colon cancer (HT-29) cells was inhibited by administration of extracts in a dose-dependent manner. Particularly, EtOH extract inhibited proliferation of the cells more effectively than water extract did. The morphology of cells induced by EtOH extract was characterized by reduction of cell size and deformatin. Oral administration of the EtOH extract (3 mg/head) to tumor-bearing mice inhibited the tumor (sarcoma-180) growth by 35% and prolonged their survival rate by 61%. The EtOH extract was shown to be nontoxic at 37.5% mg/head/day on the acute toxicity test. These studies suggest that the EtOH extract of Salvia miltiorrhiza may have antitumor activity in vitro and in vivo.

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Production of Bovine Embryos Using Follicular Oocytes Matured In Vitro (체외성숙 난포란을 이용한 소배의 생산)

  • 박수봉
    • Journal of Embryo Transfer
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    • v.5 no.1
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    • pp.21-27
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    • 1990
  • The technique for maturation of follicular oocyte has been devised to provide such a low cost and in ptentifut number supply of bovine embryo. Some of problems concerning production of bovine embtyo in vitro were discussed in this paper. Bovine follicular oocytes cultured in vitro achieved normal fertilization but cleavage rates to blastocyst were low compared to the oocyte matured in vivo. It has been concluded that a deficient cytoplasmic maturation occurs in the oocytes matured in vitro. These results indicate that the studies for maturation of bovine follicular oocytes in vitro need improvement of culture conditions and to define the characteristics that might be indicative of healthy oocyte.

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Comparison of Gene Coding Clones Content in In vivo and In vitro Methyl-Filtration Libraries of Maize(Zea may)

  • Lee, Myung-Chul;Wing, Rod A;Suh, Seok-Cheol;Eun, Moo-Young
    • Korean Journal of Plant Resources
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    • v.20 no.6
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    • pp.491-498
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    • 2007
  • It has been hypothesized that efficient exclusion of methylated retrotransposons and repeated DNA region is one of the rapid and cost-effective approaches for comprehensive gene discovery in large genome size of maize. Three kinds of methylation-sensitive restriction enzymes, HapII, MspI and McrBC, were used to identify the restriction frequency of cytosine methylation sites in maize genome. Roughly 60% of total maize genomic DNA was restricted less than 500bp by McrBC, and the most of restricted small size fraction was composed retrotransposon. In order to validate the efficient construction of gene-rich shotgun library, we compare two gene-rich methyl-filtration shotgun libraries using in vivo and in vitro methyl-filtration system. The size selected DNA fraction by Sau3A-McrBC enzyme treated was very stable and has not appeared modification in E. coli, but most insert DNA size of partially digested with Sau3A were decrease less than 500bp by bacterial methylation-modification system. In compare of retroelements portion, A 44.6% of the sequences were retroelement in unmethyl-filtered library, and the most of them was Copia type, such as Prem, Opie and Ji. The portion of retroelement was drastically decreased to 25% and 20% by in vivo and in vitro filtration system, respectively.

In vitro and In vivo Evaluation of Novel Gel Formulations of Testosterone for Transdermal Delivery

  • Heo, Sung-Koun;Cho, Young-Seok;Han, Sang-Dae;Chang, Jin-Kang;Yoon, Eun-Ju;Ko, Dae-Woong;Lim, Chang-Baeg;Chung, Suk-Jae;Shim, Chang-Koo;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.35 no.5
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    • pp.329-332
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    • 2005
  • HPMC-based novel gel formulations for the transdermal delivery of testosterone (TS) were developed, and the effect of various skin permeation enhancers was studied in vitro and in vivo. In vitro hairless mouse skin permeation of TS from the gel was investigated using Keshary-Chien diffusion cells for 8 hours at $37^{\circ}C$. In vivo plasma concentration profiles of TS after applying the gel on the abdominal skin of rat were determined using a commercial radioimmunoassay kit. Hairless mouse skin permeation of TS increased with the addition of permeation enhancers both in vitro and in vivo. Combination of diethanolamine (2%) and N-methylpyrrolidone (NMP, 6%) was the most effective among tested. Plasma concentration of TS significantly increased for at least 24 hours with the addition of diethanolamine and NMP. These results suggest the feasibility of the development of a HPMC-based gel formulation for the transdermal delivery of TS.

Metabolism and excretion of novel pulmonary-targeting docetaxel liposome in rabbits

  • Wang, Jie;Zhang, Li;Wang, Lijuan;Liu, Zhonghong;Yu, Yu
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.1
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    • pp.45-54
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    • 2017
  • Our study aims to determine the metabolism and excretion of novel pulmonary-targeting docetaxel liposome (DTX-LP) using the in vitro and in vivo animal experimental models. The metabolism and excretion of DTX-LP and intravenous DTX (DTX-IN) in New Zealand rabbits were determined with ultra-performance liquid chromatography tandem mass spectrometry. We found DTX-LP and DTX-IN were similarly degraded in vitro by liver homogenates and microsomes, but not metabolized by lung homogenates. Ultra-performance liquid chromatography tandem mass spectrometry identified two shared DTX metabolites. The unconfirmed metabolite $M_{un}$ differed structurally from all DTX metabolites identified to date. DTX-LP likewise had a similar in vivo metabolism to DTX-IN. Conversely, DTX-LP showed significantly diminished excretion in rabbit feces or urine, approximately halving the cumulative excretion rates compared to DTX-IN. Liposomal delivery of DTX did not alter the in vitro or in vivo drug metabolism. Delayed excretion of pulmonary-targeting DTX-LP may greatly enhance the therapeutic efficacy and reduce the systemic toxicity in the chemotherapy of non-small cell lung cancer. The identification of $M_{un}$ may further suggest an alternative species-specific metabolic pathway.

Evaluation of a Sample-Pooling Technique in Estimating Bioavailability of a Compound for High-Throughput Lead Optimazation (혈장 시료 풀링을 통한 신약 후보물질의 흡수율 고효율 검색기법의 평가)

  • Yi, In-Kyong;Kuh, Hyo-Jeong;Chung, Suk-Jae;Lee, Min-Haw;Shim, Chang-Koo
    • Journal of Pharmaceutical Investigation
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    • v.30 no.3
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    • pp.191-199
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    • 2000
  • Genomics is providing targets faster than we can validate them and combinatorial chemistry is providing new chemical entities faster than we can screen them. Historically, the drug discovery cascade has been established as a sequential process initiated with a potency screening against a selected biological target. In this sequential process, pharmacokinetics was often regarded as a low-throughput activity. Typically, limited pharmacokinetics studies would be conducted prior to acceptance of a compound for safety evaluation and, as a result, compounds often failed to reach a clinical testing due to unfavorable pharmacokinetic characteristics. A new paradigm in drug discovery has emerged in which the entire sample collection is rapidly screened using robotized high-throughput assays at the outset of the program. Higher-throughput pharmacokinetics (HTPK) is being achieved through introduction of new techniques, including automation for sample preparation and new experimental approaches. A number of in vitro and in vivo methods are being developed for the HTPK. In vitro studies, in which many cell lines are used to screen absorption and metabolism, are generally faster than in vivo screening, and, in this sense, in vitro screening is often considered as a real HTPK. Despite the elegance of the in vitro models, however, in vivo screenings are always essential for the final confirmation. Among these in vivo methods, cassette dosing technique, is believed the methods that is applicable in the screening of pharmacokinetics of many compounds at a time. The widespread use of liquid chromatography (LC) interfaced to mass spectrometry (MS) or tandem mass spectrometry (MS/MS) allowed the feasibility of the cassette dosing technique. Another approach to increase the throughput of in vivo screening of pharmacokinetics is to reduce the number of sample analysis. Two common approaches are used for this purpose. First, samples from identical study designs but that contain different drug candidate can be pooled to produce single set of samples, thus, reducing sample to be analyzed. Second, for a single test compound, serial plasma samples can be pooled to produce a single composite sample for analysis. In this review, we validated the issue whether the second method can be applied to practical screening of in vivo pharmacokinetics using data from seven of our previous bioequivalence studies. For a given drug, equally spaced serial plasma samples were pooled to achieve a 'Pooled Concentration' for the drug. An area under the plasma drug concentration-time curve (AUC) was then calculated theoretically using the pooled concentration and the predicted AUC value was statistically compared with the traditionally calculated AUC value. The comparison revealed that the sample pooling method generated reasonably accurate AUC values when compared with those obtained by the traditional approach. It is especially noteworthy that the accuracy was obtained by the analysis of only one sample instead of analyses of a number of samples that necessitates a significant man-power and time. Thus, we propose the sample pooling method as an alternative to in vivo pharmacokinetic approach in the selection potential lead(s) from combinatorial libraries.

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Real-Time Monitoring of Catheter-Related Biofilm Infection in Mice

  • Liu, Xu;Yin, Hong;Xu, Xianxing;Cheng, Yuanguo;Cai, Yun;Wang, Rui
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1728-1733
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    • 2015
  • This study was done to establish a mouse model for catheter-related biofilm infection suitable to bioluminescence imaging (BLI). Biofilm formation of Pseudomonas aeruginosa (P. aeruginosa) Xen5 grown on catheter disks in vitro and in an implanted mouse model was real-time monitored during a 7-day study period using BLI. The numbers of integrated brightness (IB) and viable bacterial count (VBC) in the biofilm disks in vitro were highest at 24 h after inoculation; the IB of biofilm in vivo was increased until 24 h after implantation. A statistical correlation was observed between IB and VBC in vitro by linear regression analysis. The actual VBC value in vivo can be estimated accurately by IB without sacrifice. In addition, we monitored the change in white blood cells (WBCs) during infection. The number of WBCs on day 7 was significantly higher in the infection group than in the control group. This study indicates that BLI is a simple, fast, and sensitive method to measure catheter biofilm infection in mice.

Protective Effect of Ginseng Polysaccharide Fraction on CCl4-induced Hepatotoxicity in vitvo ana in vivo (인삼 다당분획의 in vitro, in vivo에서 사염화탄소 유발 간독성에 대한 보호효과)

  • Kim, Young-Sook
    • Journal of Ginseng Research
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    • v.19 no.2
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    • pp.108-113
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    • 1995
  • Effect of ginseng polysaccharide fraction was examined for $CCl_4$-induced hepatotoxicity in vitro and in vivo. In $CCl_4$-injured primary cultured rat hepatocytes, treatment of the polysaccharide fraction (0.1, 0.3, 1.0 mg/ml) significantly Inhibited the release of LDH and GOT into the culture medium in a dose-dependent manner. Oral administration of the polysaccharide fraction (100, 200 mg/kg) inhibited the decrease of body weight and the increase of the ratio of liver to body weight in $CCl_4$-intoxicated rats. Elevation of GOT, GPT and ALP activity in the serum by $CCl_4$-induced hepatotoxicity was suppressed by administration of ginseng polysaccharide fraction. MDA levels increased in the serum as well as in the liver tissue by treatment with $CCl_4$ showed a tendency to be 연w in the rats given to the polysaccharide fraction. These results suggest that the polysaccharide fraction may be active substance responsible for antihepatotoxic effect of Panax ginseng.

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