• 제목/요약/키워드: in vitro lab

검색결과 269건 처리시간 0.024초

경피흡수제형의 in vitro 약물방출실험을 위한 연속확산 장치의 개발 (Development of Automated Diffusion Cell for Determining In Vitro Drug Release from Transdermal Device)

  • 변영로;최영권;정서영;김영하
    • 약학회지
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    • 제34권3호
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    • pp.161-165
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    • 1990
  • An automated, simple, and reliable method was developed for determining in vitro drug release rate from transdermal delivery dosage forms. The patch is held in position in the heating block by sandwiching it between the middle plate and the bottom plate of diffusion cell. The dissolution profile of the commercially available transdermal scopolamine patch was determined over a 72-h period, and the results were compared with those obtained with other methods; paddle-over-disk method, reciprocating method, and diffusion cell method. It was demonstrated that the flow-through method is equivalent in terms of release rate profile and accumulated released drug amount over the lifetime of the dosage form tested. Also this method is simple, reliable and reproducible. Therefore, this technique can be used in a quality control for assuring product uniformity.

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저장기간, 배지종류 및 viscin이 멸종위기종 꼬리겨우살이의 기내 종자발아에 미치는 영향 (Effects of Storage Duration, Medium and Viscin on in vitro seed Germination in Endangered Species, Loranthus tanakae)

  • 이수광;이송희;강호덕
    • 한국산림과학회지
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    • 제99권4호
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    • pp.618-624
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    • 2010
  • 본 연구는 멸종위기에 처한 꼬리겨우살이 종자로부터 기내배양체계 기반확립을 위해 기내 종자발아 실험을 실시하였다. 꼬리겨우살이 종자는 배지에 치상 1주 후 원기근이 형성되어 발아가 진행되었다. 배지종류와 viscin 유 무에 따른 꼬리겨우살이의 기내 종자발아율은 viscin이 제거된 종자를 SH 배지에 치상하였을 때 가장 높은 발아율(69%)을 나타냈고, 고형지지물 종류와 농도에 따른 발아율은 0.35% gelrite 첨가 시 가장 높은 발아율(75%)을 보였다. 저장 기간과 저장 상태에 따른 발아율은 채집 직후(3일 후) 조건에서 가장 높은 발아율을 나타냈고, 저장기간이 8주, 16주로 길어짐에 따라 발아율은 감소하였다. 흡착판과 흡기근은 White 기본배지에 치상하였을 때 각각 95%, 8%의 형성율을 나타내었다. 꼬리겨우살이의 기내 종자발아과정은 원기근 형성, 흡착판 형성, 흡기근 형성 순으로 진행되었다.

Relationship between Molecular Structure Characteristics of Feed Proteins and Protein In vitro Digestibility and Solubility

  • Bai, Mingmei;Qin, Guixin;Sun, Zewei;Long, Guohui
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권8호
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    • pp.1159-1165
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    • 2016
  • The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as ${\alpha}$-helices and ${\beta}$-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility ($p{\leq}0.003$); moreover, the relatively quantitative amounts of ${\alpha}$-helices, random coils, and ${\alpha}$-helix to ${\beta}$-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility ($p{\leq}0.004$). On the other hand, the percentage of ${\beta}$-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001) and solubility (p = 0.002). These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the ${\alpha}$-helix-to-${\beta}$-sheet ratio can be used to predict the nutritional value of feed proteins.

Cytotoxicity of Two Novel Cisplatin Analogues, $(CPA)_2Pt[DOLYM]$ and (DACH)Pt[DOLYM], to Human Cancer Cells In Vitro

  • Choi, Sang-Un;Kim, Kwang-Hee;Choi, Eun-Jung;Park, Sung-Hee;Kim, Kwan-Mook;Shon, Youn-Soo;Lee, Chong-Ock
    • Archives of Pharmacal Research
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    • 제22권2호
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    • pp.151-156
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    • 1999
  • Despite the impressive antitumor activity of cisplatin, two major limitations of the drug, that is severe side effects and drug-resistance of cancer cells, make its use difficult of r cancer therapy. These limitations have resulted in a greate deal of effort having been expended into structural modifications of cisplatin. In this study, we tested two novel cisplatin analogues, (CPA)2 Pt[DOLYM] (COMP-I) and (DACH)Pt[DOLYM] (CoMP-II), for the mode of cytotoxic action against human tumor cells comparing with cisplatin and carboplatin in vitro. These two novel analogues had considerable cytotoxic activities against five kinds of human solid tumor cells, and especially COMP-II was more effective on HCT15 colon cancer cells than other compounds. In addition, COMP-II had cytostatic activity at low concentrations (10~0.3${\mu}g/ml$), but other compounds revealed little effect on tumor growth at the low concentration.

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Current approaches for assisted oocyte maturation in camels

  • Saadeldin, Islam M.;Cho, Jongki
    • 한국동물생명공학회지
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    • 제36권3호
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    • pp.162-167
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    • 2021
  • Camel (camelus dromedarius) is a unique large mammalian species that can survive harsh environmental conditions and produce milk, meat, and wool. Camel reproduction is inferior when compared to other farm animal species such as cattle and sheep. Several trials have been reported to increase camel reproduction and production through assisted reproductive techniques (ARTs) such as in vitro fertilization and cloning. For these reasons, obtaining enough mature oocytes is a cornerstone for ARTs. This demand would be improved by the oocyte in vitro maturation (IVM) systems. In this review, the current approaches and views from different laboratories using ARTs and the IVM to produce embryos in vitro in camel species. For the last two decades, conventional IVM system was the common approach, however, recently the bi-phasic IVM system has been introduced and showed promising improvement in IVM of camel oocytes. Detailed studies are needed to understand camel meiosis and IVM to efficiently increase the production of this species.

3D Bioprinted GelMA/PEGDA Hybrid Scaffold for Establishing an In Vitro Model of Melanoma

  • Duan, Jiahui;Cao, Yanyan;Shen, Zhizhong;Cheng, Yongqiang;Ma, Zhuwei;Wang, Lijing;Zhang, Yating;An, Yuchuan;Sang, Shengbo
    • Journal of Microbiology and Biotechnology
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    • 제32권4호
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    • pp.531-540
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    • 2022
  • Due to the high incidence of malignant melanoma, the establishment of in vitro models that recapitulate the tumor microenvironment is of great biological and clinical importance for tumor treatment and drug research. In this study, 3D printing technology was used to prepare GelMA/PEGDA composite scaffolds that mimic the microenvironment of human malignant melanoma cell (A375) growth and construct in vitro melanoma micro-models. The GelMA/PEGDA hybrid scaffold was tested by the mechanical property, cell live/dead assay, cell proliferation assay, cytoskeleton staining and drug loading assay. The growth of tumor cells in two- and three-dimensional culture systems and the anti-cancer effect of luteolin were evaluated using the live/dead staining method and the Cell Counting Kit-8 (CCK-8) method. The results showed a high aggregation of tumor cells on the 3D scaffold, which was suitable for long-term culture. Cytoskeleton staining and immunofluorescent protein staining were used to evaluate the degree of differentiation of tumor cells under 2D and 3D culture systems. The results indicated that 3D bioprinted scaffolds were more suitable for tumor cell expansion and differentiation, and the tumor cells were more aggressive. In addition, luteolin was time- and dose-dependent on tumor cells, and tumor cells in the 3D culture system were more resistant to the drug.

생기액(生肌液)의 세포독성 및 자궁경부암 바이러스 (HPV 16 type) 암 유발인자 E6와 E7의 작용에 미치는 효과 (The Effects of Somatid on the Cytotoxicity of Cancer Cells and Human Papillomavirus Type 16 E6 and E7 Oncogenes)

  • 정옥;조영식;조정원;이경애;심정현;조민철;이홍수;염영일;김상범;박순희;윤도영
    • 약학회지
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    • 제44권4호
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    • pp.340-346
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    • 2000
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, CaSki) and HaCaT keratinocytes in a dose response manner, In vitro binding assay and ELISA showed that Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by Somatid. Our data suggested that Somatid inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas caused by HPV.

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Heat-Shock Protein 70 as a Tumor Antigen for in vitro Dendritic Cell Pulsing in Renal Cell Carcinoma Cases

  • Meng, Fan-Dong;Sui, Cheng-Guang;Tian, Xin;Li, Yan;Yang, Chun-Ming;Ma, Ping;Liu, Yun-Peng;Jiang, You-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권20호
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    • pp.8947-8950
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    • 2014
  • Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

Lactic acid bacteria strains selected from fermented total mixed rations improve ensiling and in vitro rumen fermentation characteristics of corn stover silage

  • Huang, Kailang;Chen, Hongwei;Liu, Yalu;Hong, Qihua;Yang, Bin;Wang, Jiakun
    • Animal Bioscience
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    • 제35권9호
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    • pp.1379-1389
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    • 2022
  • Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×106 cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.