• 제목/요약/키워드: Cell-based assay

검색결과 663건 처리시간 0.024초

Sexing of Sheep Embryos Produced In vitro by Polymerase Chain Reaction and Sex-specific Polymorphism

  • Saravanan, T.;Nainar, A. Mahalinga;Kumanan, K.;Kumaresan, A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권5호
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    • pp.650-654
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    • 2003
  • The accuracy of Polymerase chain reaction (PCR) assay in sexing of sheep embryos was assessed in this study. A total of 174 ovine embryos produced in vitro at different stages of development (2, 4-8 cell stages, morula and blastocyst) were sexed. The universal primers (P1-5EZ and P2-3EZ) used in this assay amplified ZFY/ZFX-specific sequences and yielded a 445 bp fragment in both sexes. Restriction enzyme analysis of ZFY/ZFX-amplified fragments with Sac I exhibited polymorphism between sexes, three and two fragments in males and in females, respectively. For verification of accuracy, blood samples of known sex were utilized as positive controls in each test. The mean percentages of sex identification by this method at 2 cell, 4-8 cell, morula and blastocyst were $73.00{\pm}5.72$, $89.77{\pm}3.79$, $3.33{\pm}8.08$ and $79.6{\pm}9.09$, espectively with the over all male to female ratio of 1:0.87. It is concluded that the ZFY/ZFX based method is highly reliable for the sexing of sheep embryos.

Biological Effects of Vinca minor extract; Tyrosinase inhibition, stimulation of ROS generation and increasement of cell migration activity in keratinocytes

  • Kim, Jun-Sub;Yu, Il-Hwan;Joo, Ji-Hye;Nam, Gyeong Hoe;Jung, Kyung-Hwan;Chung, Young Soo;Lee, Hyang-Yeol
    • 한국응용과학기술학회지
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    • 제33권4호
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    • pp.788-794
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    • 2016
  • Vinca alkaloids from plant Vinca minor have been investigated for their effects of tyrosinase inhibition, stimulation of ROS generation and increasement of cell migration activity. The methanolic crude extract and the water-soluble fraction exhibited $IC_{50}$ value of 3.1 mg/mL and 2.1 mg/mL. Vinca minor extract treatment significantly increased ROS levels in HaCaT cells, in a concentration-dependent manner. Treatments of Vinca minor extract led to increase wound closure when compared with non-treatment. Low dose (0.1% or 0.3%) of extracts have not significantly affected, compared with that in controls. By contrast, 0.5% extract have dramatic effect on wound healing activity of keratinocytes. Effects of Vinca minor extract in a filter-based cell mobility assay appear similar to that of wound closure assay, which suggests that the Vinca minor extract have wound healing effects on skin.

Non-B, Non-T Acute Lymphoblastic Leukemia in a Cat

  • Sumin Cha;Hyunwoo Kim;Hyeona Bae;Minjeong Kang;Rankyung Jung;Minji Kim;DoHyeon Yu
    • 한국임상수의학회지
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    • 제40권4호
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    • pp.298-302
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    • 2023
  • A 7-year-old neutered male, domestic shorthair cat presented anorexia and lethargy. The complete blood cell count revealed severe non-regenerative anemia, lymphocytic leukocytosis, neutropenia, and thrombocytopenia. On the peripheral blood smear examination, medium to large lymphoblastic cells with moderate amounts of basophilic cytoplasm were observed in up to 70% of peripheral leukocytes. Feline leukemia and immunodeficiency viruses were not detected using a commercial diagnostic kit. While splenomegaly and blunt margins of the caudoventral liver were observed in abdominal radiography, changes in the intra-abdominal lymph nodes were not remarkable. Ultimately, flow cytometric immunophenotyping from the peripheral blood revealed a negative for B-cell markers (CD21-/CD79a-) and T-cell markers (CD3-/CD4-/CD5-/CD8-). Based on the hematological examination and the immunophenotyping assay, the cat was diagnosed with non-B, non-T acute lymphoblastic leukemia. Here, we report a rare case of non-B, non-T acute lymphoblastic leukemia to raise awareness and provide information on clinical symptoms and laboratory test and immunophenotyping analysis results.

Effect of Oral Administration of Korean Red Ginseng on Influenza A (H1N1) Virus Infection

  • Kim, Jin-Young;Kim, Hyoung-Jin;Kim, Hong-Jin
    • Journal of Ginseng Research
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    • 제35권1호
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    • pp.104-110
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    • 2011
  • Korean red ginseng (RG), which is a ginseng treated by heating and steaming, has biological activity similar to Panax ginseng. The effect of ginseng on influenza infection has not been studied although it is known to have a broad range of biological activities. The aim of the study is to investigate the effect of RG extract on influenza A (H1N1) virus infection. We investigated the inhibitory effect of RG extract on plaque formation by influenza A virus in a cell-based plaque assay, and the effect of orally administered RG on influenza A virus infection in mice. RG extract, which was applied at a non-cytotoxic concentration, inhibited plaque formation by influenza A virus in the cell-based plaque assay. The orally administered RG extract ameliorated body weight loss and significantly increased survival in mice infected with influenza A virus. Our results suggest that RG extract has components that reduce the severity of infection by influenza A virus and could potentially be used as a complement to treatment of influenza A virus infections.

Human sebocyte-based assay system for the screening of compounds to lower the lipid synthesis in sebaceous gland

  • Mun, Yeun-Ja;Lee, Seung-Yon;Im, Sook-Jung;Ahn, Sung-Hun;Lee, Jason;Woo, Won-Hong
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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    • pp.508-518
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    • 2003
  • SZ95 cell is an immortalized human sebaceous gland cell line that shows the morphologic, phenotypic and functional characteristics of normal human sebocytes. Sebocytes may play crucial parts in the pathophysiologic processes and disorders of the pilosebaceous unit. The secretory activity of the sebaceous gland is remarkably species-specific and acne is an exclusively human disease. Thus, this SZ95 cells offer possibilities for investigations on the physiology of the sebaceous gland and its role in sebum-associated skin disease such as acne. In this study, we investigated the effects of 13-cis-retinoic acid (13-cis-RA) and spironolactone, frequently used as therapeutic agents of acne, on the lipid synthesis and proliferation of human sebocytes. Cell proliferation was determined by MTT assay and cytoplasmic lipid droplets was shown by Oil-red a staining. Total lipid levels were biochemically estimated by the sulfo-phospho-vanilline reagent. 13-cis-RA and spironolactone significantly inhibited proliferation and lipid levels in a dose-dependent manner. Combined treatment with testosterone and 13-cis-RA or spironolactone resulted in a lower total lipid levels than that with androgen alone. These observations indicate that 13-cis-RA and spironolactone are potent inhibitors of both cell proliferation and lipid synthesis in human sebocytes. We will provide experimental evidence that this human sebocyte cell line serves as an adequate tool for evaluating the anti-lipogenic activity of various compounds potentially useful for the bioactive cosmeceutical ingredients on acne skin, and studying the intracellular biochemical markers depending on the types of compounds from various sources.

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한약처방제의 인체 위암 세포주에 대한 세포독성 효과에 관한 연구 (Cytotoxic Effect of Korean Traditional Prescriptions on the Human Gastric Cancer Cell Lines)

  • 김은해;은영아;강봉주;성현제;박갑주
    • 생약학회지
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    • 제28권4호
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    • pp.233-238
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    • 1997
  • ln order to search for antigastric cancer agents from Korean traditional prescriptions. We selected 41 traditional prescriptions, based on a review of the Korean traditional medicine books. Both boiling water and methanol extracts were tested, by means of the Sulforhodamine B (SRB) protein assay. Six of the 41 water extracts; #3, #34, #35, #38, #40, #41 showed efficacy against gastric cancer cell (AGS: Human gastric carcinoma, ATCC HTB 103). #3 inhibited 50% cancer cell growth1 at the concentration of $152\;{\mu}g/ml$, #34, #35, #38, #40 and #41 inhibited 50% cancer cell growth at the concentration of $145\;{\mu}g/ml$, $129\;{\mu}g/ml$, $173\;{\mu}g/ml$, $10\;{\mu}g/ml$ and $19\;{\mu}g/ml$ respectively. Ten of the 41 methanol extracts; #1, #3, #32, #33, #35, #36, #37, #38, #41 were active. #1 inhibited 50% cancer cell growth at the concentration of $206\;{\mu}g/ml$, #3, #32, #33, #35, #36, #37, 738, #40, #41 inhibited 50% cancer cell growth at the concentration of $133\;{\mu}g/ml$, $159\;{\mu}g/ml$, $199\;{\mu}g/ml$, $147\;{\mu}g/ml$, $113\;{\mu}g/ml$, $187\;{\mu}g/ml$, $130\;{\mu}g/ml$, $9\;{\mu}g/ml$, $15\;{\mu}g/ml$ respectively. Prescription #3, #35, #38, #40, #41 were also interesting because both methanol and water extracts were active.

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MHC class I 분자들에 의해 제시되는 Epitope을 인지하는 CD8+ T 림프구의 결핵균 감염에 대한 면역반응의 연구: 결핵 환자와 PPD+ 건강개체에 존재하는 결핵균 항원에 특정한 CD8+ T세포 (The Study of MHC class I Restricted CD8+ T Cell Mediated Immune Responses against Mycobacterium tuberculosis Infection: Evidence of M. tuberculosis S pecific CD8+ T Cells in TB Patients and PPD+ Healthy Individuals)

  • 조장은;이경화;박승규;천선희;조상래;조성애
    • IMMUNE NETWORK
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    • 제3권3호
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    • pp.235-241
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    • 2003
  • Background: The protective immunity against tuberculosis (TB) involves both CD4+ T cells and CD8+ T cells. In our previous study, we defined four Mycobacterium tuberculosis derived peptide epitopes specific for HLA-$A^*0201$ restricted CD8+ T cells ($ThyA_{30-38}$, $RpoB_{127-135}$, $85B_{15-23}$, $PstA1_{75-83}$). In this study, we investigated the immune responses induced by these peptide specific CD8+ T cells in latently and chronically infected people with TB. Methods: We characterized these peptide specific CD8+ T cell population present in PBMC of both TB patients and PPD+healthy people using IFN-${\gamma}$elispot assay, intracellular staining and HLA-A2 dimer staining. Results: The frequency of peptide specific CD8+ T cell was in the range of 1 to 25 in $1.7{\times}10^5$ PBMC based on ex vivo IFN-${\gamma}$ elispot assay, demonstrating that these peptide specific CD8+ T cell responses are induced in both TB patients and PPD+ people. Short term cell lines (STCL) specific for these peptides proliferated in vitro and secreted IFN-${\gamma}$ upon antigenic stimulation in PPD+ donors. Lastly, HLA-$A^*0201$ dimer assays indicated that $PstA1_{75-83}$ specific CD8+ T cell population in PPD+ healthy donors is heterogeneous since approximately 25~33% of $PstA1_{75-83}$ specific CD8+ T cell population in PPD+ healthy donors produced IFN-${\gamma}$ upon peptide stimulation. Conclusion: Our results suggest that MHC class I restricted CD8+ T cell mediated immune responses to M. tuberculosis infection are induced in both TB patients and PPD + people; however, the CD8+ T cell population is functionally heterogeneous.

저근백피의 Virus-Cell Fusion 저해활성 성분 (Virus-Cell Fusion Inhibitory Compounds from Ailanthus altissima Swingle)

  • 장영수;문영희;우은란
    • 생약학회지
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    • 제34권1호통권132호
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    • pp.28-32
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    • 2003
  • In order to search for the anti-HIV agents from natural products, eighty MeOH extracts of medicinal plants were applied to a syncytia formation inhibition assay which is based on the interaction between the HIV-1 envelope glycoprotein gp120/gp41 and the cellular membrane protein CD4 of T lymphocytes. Among them, Ailanthus altissima showed a potent virus-cell fusion inhibitory activity. Repeated column chromatoghaphy of the methylene chloride fraction of A. altissima afforded compounds 1$({\beta}-sitosterol-3-O-{\beta}-D-glucoside)$, 2(tetramethoxycoumarin), and 3(ocotillone). Virus-cell fusion inhibitory activity of compound 3(ocotillone) was $70.76{\pm}4.09%$ at the concentration of $100\;{\mu}g/ml$.

Virtual Screening and Testing of GSK-3 Inhibitors Using Human SH-SY5Y Cells Expressing Tau Folding Reporter and Mouse Hippocampal Primary Culture under Tau Cytotoxicity

  • Chih-Hsin Lin;Yu-Shao Hsieh;Ying-Chieh Sun;Wun-Han Huang;Shu-Ling Chen;Zheng-Kui Weng;Te-Hsien Lin;Yih-Ru Wu;Kuo-Hsuan Chang;Hei-Jen Huang;Guan-Chiun Lee;Hsiu Mei Hsieh-Li;Guey-Jen Lee-Chen
    • Biomolecules & Therapeutics
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    • 제31권1호
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    • pp.127-138
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    • 2023
  • Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer's disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine's screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (TauRD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC50 of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 TauRD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 TauRD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 TauRD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.

Rapid determination of baculovirus titers an antibody-based assay

  • Kwon, M.S.;Dojimal, T.;Park, Enoch-Y.
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.315-319
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    • 2003
  • A novel method is developed to yield virus titers in 10 h, is easy to .perform using 96-well plates, and applicable to both any Autographa californica nucleopolyhyderovirus (AcNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV)-based recombinant baculovirus. This assay uses an antibody to a DNA-binding protein to detect the infected cells via immune-staining. The titer is determined by counting foci produced due to infection of virus under a fluorescent microscopy. The required incubation period was shortened considerably because infected cells expressed viral antigens at the post infection time of 4 h. Therefore, 10 hours were enough to estimate the virus titer including virus infection time, insect cell culture, and estimation of virus titer.

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