• Title/Summary/Keyword: Drug screening

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Detection of Point Mutations in the rpoB Gene Related to Drug Susceptibility in Mycobacterium Tuberculosis using an Oligonucleotide Chip (올리고뉴클레오티드 칩(Oligonucleotide Chip)을 이용한 항결핵제 감수성과 관련된 Mycobacterium tuberculosis rpoB 유전자의 점돌연변이 판별 방법)

  • Kim, Hyun-Jung;Kim, Seong-Keun;Shim, Tae-Sun;Park, Yong-Doo;Park, Mi-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.29-41
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    • 2001
  • Background : The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance. Methods : In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type. Results : Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made. Conlcusion : An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drugsusceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.

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Factors affecting Diabetic Eye disease and Kidney disease Screening in Diabetic Patients (당뇨병 환자의 당뇨성 안질환 및 신장질환 합병증 검사 수검 여부에 영향을 주는 요인)

  • Kang, Jeong-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.4
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    • pp.226-235
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    • 2020
  • This study was undertaken to investigate factors that affect the assessment of complications in diabetic eye and kidney diseases. Data was obtained from the National Community Health Survey, 2017. The subjects included were 25,829 respondents who had been diagnosed with diabetes. Logistic regression analysis was applied to determine the factors affecting associated diabetic eye disease (fundus examination) and kidney disease (microalbuminuria examination) complications. The diabetic eye disease complication rate was 35.6%, and diabetic kidney disease complication rate was 39.8%. Complications arising due to diabetes were determined to be 35.6% for eye diseases and 39.8% for kidney related diseases. Ed. Notes: The original sentence is not very lucid. I have suggested an alternate edit. I leave it to the author's discretion to accept or reject the same. Please delete whichever sentence is not suitable. Walking activity (OR=1.03, OR=1.02), hemoglobin A1c (HbA1c) recognition (OR=2.33, OR=2.33), blood glucose level recognition (OR=1.61, OR=1.71), diabetes drug therapy (OR=2.67, OR=3.05), and diabetic management education (OR=1.45, OR=1.47) were more likely to be evaluated for eye and kidney disease complications. Our results indicate that to increase the rate of screening for diabetic complications, it is necessary to develop a diabetes management system that includes the type and timing of diabetic complications, as well as different promotional methods that recognize HbA1C and blood glucose levels. Ed. Notes: Do you mean 'screening' methods? Please revise appropriately, if required. In addition, it is essential to develop a guideline for the management of diabetes mellitus, and to incorporate a screening test for diabetic complications in the national screening system.

Screening for Various Herb Medicines Extracts HSV 1-2 (수종 한약 처방 전탕액(煎湯液)의 허피즈바이러스에 대한 효능 검색)

  • Lim, Seong-Woo
    • The Journal of Internal Korean Medicine
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    • v.21 no.2
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    • pp.291-297
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    • 2000
  • Objective : In order to find antiviral compounds against Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, a convenient virus-induced cytopathic effect(CPE) inhibition assay was introduced. Methods : Fourteen purchased herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Results : Cytotoxic concentration (CC) of the $H_2O$ extracts of Hyongbangpaedoksan against HSV-1 and HSV-2 was 181.12. This is high level cytotoxic concentration compared with the contrast. Therefore, we assumed that the high level cytotoxic concentration of herbal medicine play a major role in improvement of antiviral activity at the first infective cell. But antiviral effect was unable to figure out for selective index(Sl)=CC50/EC50. The other herbal medicines were unable to showed potent anti-HSV activity. Conclusions : The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.

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Review of measurement of prostate specific antigen: in the aspect of insurance medicine (전립선암 선별을 위한 PSA 측정의 보험의학적 의미)

  • Park, Kwang-Il
    • The Journal of the Korean life insurance medical association
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    • v.29 no.1
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    • pp.16-21
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    • 2010
  • The measurement of prostate specific antigen (PSA) in screening for prostate cancer is recently performed as a routine check-up in clinical medicine and insurance medicine. Several factors may affect serum PSA levels. As prostate size increases with increasing age, the PSA concentration also rises. Increasing body mass index (BMI) is associated with a lower mean PSA concentration. Inhibitors of 5-alpha-reductase such as finasteride and dutasteride produce a 50 percent or greater decrease in serum PSA during the first three months of therapy, which persists as long as the drug is continued. Men who are regularly taking non-steroidal antiinflammatory drugs (NSAIDs) or acetaminophen have lower PSA levels. Emerging concepts regarding PSA testing that may help refine the interpretation of an elevated concentration include: PSA density, PSA velocity, and Free versus complexed or bound PSA. With many insurance companies, PSA level has become part of a standard battery of blood tests, along with HIV, cholesterol, liver enzymes, and other predictors of premature death. But, there is no clear proof of benefit, so we have to monitor the value of PSA test as a prostate cancer screening test in insurance medicine.

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A tendency of herbal medicines on anti-viral effect (항바이러스 효과에 응용되는 한약의 연구 동향)

  • Lim, Sung-Woo
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.199-209
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    • 1998
  • In order to find antiviral effect against Human immunodeficiency virus(HIV), Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, publicated 29 paters on anti-viral effect of herbal medicines and a convenient virus-induced cytopathic effect (CEP) inhibition assay was introduced. The major virus on experiment are HIV, Hepatitis B virus and HSV-1,2. Those of other studies showed inhibition of infected virus DNA replication and screening test of herbal medicines. More than 15 extractions were prepared by pure water boiling from herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Cytotoxic concentration (CC) of the $H_2O$ extracts of Padoo against HIV was <4.0, Hyungbangpaedoksan against HIV was 9.3, Whangyonhaedoktang against HIV-1 and HSV-2 was 15.3. These are high level cytotoxic concentration compared with the contrast. But antiviral effect was unable to figure out for selective $index(SI)=CC_{50}/EC_{50}$. The other herbal medicines were unable to showed potent anti-HIV and anti-HSV activity. The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.

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Screening for Chemosensitizers from Natural Plant Extracts through the Inhibition Mechanism of P-glycoprotein

  • Ahn, Hee-Jeong;Song, Im-Sook
    • Journal of Pharmaceutical Investigation
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    • v.40 no.5
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    • pp.269-275
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    • 2010
  • P-gp plays a critical role in drug disposition and represents a mechanism for the development of multidrug resistance. Flavonoids, a major class of natural compounds widely present in foods and herbal products, have been shown to inhibit P-gp. Therefore, the aim of this study was to identify new candidate chemosensitizers by screening various plant extracts. The ability of natural plant extracts to inhibit P-gp activity was assessed by measuring cellular accumulation of calcein AM, daunorubicin and vincristine in P-gp overexpressing MDCKII-MDR1 cells. Among more than 800 plant extracts, eight were found to inhibit P-gp activity. Curcuma aromatica extract produced greatest inhibition, followed by Curcuma longa and Dalbergia odorifera extracts. Extracts of Aloe ferox, Curcuma zedoariae rhizome, Zanthoxylum planispinum, and Ageratum conyzoides showed moderate inhibitory effects. Curcumin and quercetin exhibited similar inhibition of P-gpmediated efflux of daunorubicin and vincristine, and flavones had a lesser effect. When chemosensitizing effect was evaluated by measuring daunorubicin sensitivity to MDCKII-MDR1 cells in the presence of natural plant extracts, Curcuma aromatica showed the most potent chemosensitizing effect based on daunorubicin cytotoxicity. In conclusion, natural plant extracts such as Curcuma aromatica can potently inhibit P-gp activity and may have potential as a novel chemosensitizers.

Methanol Extract of Cinnamomum cassia Represses Cellular Proliferation and Gli-mediated Transcription in PANC-1 Human Pancreatic Cancer Cells

  • Lee, Hwa Jin
    • Natural Product Sciences
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    • v.20 no.3
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    • pp.170-175
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    • 2014
  • Twenty five methanolic plant extracts were investigated to determine the anticancer activity against sonic hedgehog (shh)/Gli signaling pathway dependent cancer, PANC-1 human pancreatic cancer cells, through three screening programs. All extracts were inspected their inhibitory properties on sonic hedgehog-conditioned medium (shh-CM) induced alkaline phosphatase (ALP) activity in C3H10T1/2 mouse mesenchymal stem cells to examine whether the plant extracts affect the shh/Gli signaling pathway. Next, plant extracts were screened the ability to suppress the cell proliferation of PANC-1 human pancreatic cancer cells. Finally, active plant extracts from the two screening systems were evaluated for the suppressive effect on Gli-mediated transcriptional activity in PANC-1 cells. Among active plants, Cinnamomum cassia suppressed Gli-mediated transcriptional activity leading to the down-regulated expression of Gli-target genes such as Gli-1 and Patched-1 (Ptch-1). This study provides the consideration for the important role of natural products in drug discovery process as well as the basis for the further analysis of active plant and potential identification of novel bioactive compounds as inhibitors of Gli and therapeutic candidates against shh/Gli signaling pathway dependent cancers.

Innate Color Preference of Zebrafish and Its Use in Behavioral Analyses

  • Park, Jong-Su;Ryu, Jae-Ho;Choi, Tae-Ik;Bae, Young-Ki;Lee, Suman;Kang, Hae Jin;Kim, Cheol-Hee
    • Molecules and Cells
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    • v.39 no.10
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    • pp.750-755
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    • 2016
  • Although innate color preference of motile organisms may provide clues to behavioral biases, it has remained a longstanding question. In this study, we investigated innate color preference of zebrafish larvae. A cross maze with different color sleeves around each arm was used for the color preference test (R; red, G; green, B; blue, Y; yellow). The findings showed that 5 dpf zebrafish larvae preferred blue over other colors (B > R > G > Y). To study innate color recognition further, tyrosinase mutants were generated using CRISPR/Cas9 system. As a model for oculocutaneous albinism (OCA) and color vision impairment, tyrosinase mutants demonstrated diminished color sensation, indicated mainly by hypopigmentation of the retinal pigment epithelium (RPE). Due to its relative simplicity and ease, color preference screening using zebrafish larvae is suitable for high-throughput screening applications. This system may potentially be applied to the analysis of drug effects on larval behavior or the detection of sensory deficits in neurological disorder models, such as autism-related disorders, using mutant larvae generated by the CRISPR/Cas9 technique.

Use of DNA-Specific Anthraquinone Dyes to Directly Reveal Cytoplasmic and Nuclear Boundaries in Live and Fixed Cells

  • Edward, Roy
    • Molecules and Cells
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    • v.27 no.4
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    • pp.391-396
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    • 2009
  • Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

Q-omics: Smart Software for Assisting Oncology and Cancer Research

  • Lee, Jieun;Kim, Youngju;Jin, Seonghee;Yoo, Heeseung;Jeong, Sumin;Jeong, Euna;Yoon, Sukjoon
    • Molecules and Cells
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    • v.44 no.11
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    • pp.843-850
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    • 2021
  • The rapid increase in collateral omics and phenotypic data has enabled data-driven studies for the fast discovery of cancer targets and biomarkers. Thus, it is necessary to develop convenient tools for general oncologists and cancer scientists to carry out customized data mining without computational expertise. For this purpose, we developed innovative software that enables user-driven analyses assisted by knowledge-based smart systems. Publicly available data on mutations, gene expression, patient survival, immune score, drug screening and RNAi screening were integrated from the TCGA, GDSC, CCLE, NCI, and DepMap databases. The optimal selection of samples and other filtering options were guided by the smart function of the software for data mining and visualization on Kaplan-Meier plots, box plots and scatter plots of publication quality. We implemented unique algorithms for both data mining and visualization, thus simplifying and accelerating user-driven discovery activities on large multiomics datasets. The present Q-omics software program (v0.95) is available at http://qomics.sookmyung.ac.kr.