{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T14:19:56Z","timestamp":1782915596222,"version":"3.54.5"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2002,8,1]],"date-time":"2002-08-01T00:00:00Z","timestamp":1028160000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2002,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            In this study, we propose a simple and reproducible host-cell-based assay for the screening of antimycobacterial drugs that is suitable for drug discovery. The method evaluates both antimycobacterial activity of the drugs and their cytotoxicity to host cells. The basis of this simple fibroblast-based assay (SFA) is that cells of human lung fibroblast cell line MRC-5, which are highly sensitive to mycobacterial cytotoxicity, are killed by virulent\n            <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n            strain H\n            <jats:sub>37<\/jats:sub>\n            Rv bacilli in response to the viability of bacilli. Clinically used antimycobacterial drugs inhibited the mycobacterial cytotoxicity to MRC-5 cells in a dose-dependent manner. MICs of isoniazid, streptomycin, rifampin, and ethambutol determined by this SFA (0.428, 1.816, 0.013, and 3.465 \u03bcg\/ml, respectively) were within 1 log of MICs determined by the broth dilution test (BDT) using Middlebrook 7H9 medium. The MIC of pyrazinamide, which exhibits bactericidal activity only at a high dose by BDT (1,231 \u03bcg\/ml at pH 6.6 and 492 \u03bcg\/ml at pH 5.8), was 3.847 \u03bcg\/ml in the modified method of SFA. On the other hand, sodium azide, a toxic agent for both mammalian cells and bacteria, exhibited cytotoxicity to fibroblasts at a dose lower than that required to inhibit mycobacterial growth. Thus, this fibroblast-based method enabled us to evaluate both antibacterial activity of drugs and their cytotoxicity to human cells within a short period of time.\n          <\/jats:p>","DOI":"10.1128\/aac.46.8.2533-2539.2002","type":"journal-article","created":{"date-parts":[[2002,7,28]],"date-time":"2002-07-28T23:07:23Z","timestamp":1027897643000},"page":"2533-2539","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["Simple Fibroblast-Based Assay for Screening of New Antimicrobial Drugs against\n            <i>Mycobacterium tuberculosis<\/i>"],"prefix":"10.1128","volume":"46","author":[{"given":"Takemasa","family":"Takii","sequence":"first","affiliation":[{"name":"Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya"},{"name":"Department of Microbiology, Colorado State University, Fort Collins, Colorado"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshifumi","family":"Yamamoto","sequence":"additional","affiliation":[{"name":"Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taku","family":"Chiba","sequence":"additional","affiliation":[{"name":"Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiyoji","family":"Abe","sequence":"additional","affiliation":[{"name":"Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Kiyose, Tokyo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John T.","family":"Belisle","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Colorado State University, Fort Collins, Colorado"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick J.","family":"Brennan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Colorado State University, Fort Collins, Colorado"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kikuo","family":"Onozaki","sequence":"additional","affiliation":[{"name":"Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.6.1542"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.6.1536"},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1093\/jac\/31.2.289","volume":"31","year":"1993","unstructured":"Banerjee, D. K., and B. R. Patel. 1993. Evaluation of the activity of a number of antimicrobial agents against mycobacteria within mouse macrophages by a radiometric method. J. Antimicrob. Chemother.31:289-302.","journal-title":"J. Antimicrob. Chemother."},{"key":"e_1_3_2_5_2","first-page":"1274","volume":"132","year":"1985","unstructured":"Carlone, N. A., G. Acocella, A. M. Cuffini, and M. Forno-Pizzoglio. 1985. Killing of macrophage-ingested mycobacteria by rifampicin, pyrazinamide, and pyrazinoic acid alone and in combination. Am. Rev. Respir. Dis.132:1274-1277.","journal-title":"Am. Rev. Respir. Dis."},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.12.3232-3239.1997"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.5.1004"},{"key":"e_1_3_2_8_2","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1164\/ajrccm\/137.5.1141","volume":"137","year":"1988","unstructured":"Crowle, A. J., N. Elkins, and M. H. May. 1988. Effectiveness of ofloxacin against Mycobacterium tuberculosis and Mycobacterium avium, and rifampin against M. tuberculosis in cultured human macrophages. Am. Rev. Respir. Dis.137:1141-1146.","journal-title":"Am. Rev. Respir. Dis."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/0041-3879(88)90036-0","volume":"69","year":"1988","unstructured":"Crowle, A. J., J. A. Sbarbaro, and M. H. May. 1988. Effects of isoniazid and of ceforanide against virulent tubercle bacilli in cultured human macrophages. Tubercle69:15-25.","journal-title":"Tubercle"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1164\/arrd.1986.134.5.1052"},{"key":"e_1_3_2_11_2","first-page":"839","volume":"130","year":"1984","unstructured":"Crowle, A. J., J. A. Sbarbaro, F. N. Judson, G. S. Douvas, and M. H. May. 1984. Inhibition by streptomycin of tubercle bacilli within cultured human macrophages. Am. Rev. Respir. Dis.130:839-844.","journal-title":"Am. Rev. Respir. Dis."},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(84)90147-9"},{"key":"e_1_3_2_13_2","first-page":"491","volume":"4","year":"2000","unstructured":"Heifets, L., M. Higgins, and B. Simon. 2000. Pyrazinamide is not active against Mycobacterium tuberculosis residing in cultured human monocyte-derived macrophages. Int. J. Tuber. Lung Dis.4:491-495.","journal-title":"Int. J. Tuber. Lung Dis."},{"key":"e_1_3_2_14_2","unstructured":"Heifets L. B. 1991. Drug susceptibility tests in the management of chemotherapy of tuberculosis p. 89-122. In L. B. Heifets (ed.) Drug susceptibility in the chemotherapy of mycobacterial infection. CRC Press Inc. Boca Raton Fla."},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.2.400"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1522-7243(199909\/10)14:5<255::AID-BIO543>3.0.CO;2-W"},{"key":"e_1_3_2_17_2","unstructured":"Inderieid C. B. and M. Salfinger. 1995. Antimicrobial agents and susceptibility tests: mycobacteria p. 1385-1404. In P. R. Murray E. J. Baron M. A. Pfaller F. C. Tenover and R. H. Yolken (ed.) Manual of clinical microbiology 6th ed. ASM Press Washington D.C."},{"key":"e_1_3_2_18_2","unstructured":"Maher D. and M. C. Raviglion. 1999. The global epidemic of tuberculosis: a World Health Organization prospective p. 104-115. In D. Schlossberg (ed.) Tuberculosis and nontuberculous mycobacterial infections 4th ed. W. B. Saunders Company Philadelphia Pa."},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/82139"},{"key":"e_1_3_2_20_2","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1179\/joc.1997.9.4.247","volume":"9","year":"1997","unstructured":"Marone, P., L. Bono, E. Carretto, D. Barbarini, and S. Telecco. 1997. Rapid drug susceptibility of Mycobacterium avium complex using a fluorescence quenching method. J. Chemother.9:247-250.","journal-title":"J. Chemother."},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/82142"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002849900095"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01588176"},{"key":"e_1_3_2_24_2","first-page":"127","volume":"75","year":"1994","unstructured":"Reddy, M. V., S. Srinivasan, B. Andersen, and P. R. Gangadharam. 1994. Rapid assessment of mycobacterial growth inside macrophages and mice, using the radiometric (BACTEC) method. Tuber. Lung Dis.75:127-131.","journal-title":"Tuber. Lung Dis."},{"key":"e_1_3_2_25_2","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/0732-8893(95)00147-7","volume":"22","year":"1995","unstructured":"Reisner, B. S., A. M. Gatson, and G. L. Woods. 1995. Evaluation of mycobacteria growth indicator tubes for susceptibility testing of Mycobacterium tuberculosis to isoniazid and rifampin. Diagn. Microbiol. Infect. Dis.22:325-329.","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.4.1144-1149.1999"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.12.3127-3131.1997"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/162.1.201"},{"key":"e_1_3_2_29_2","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1016\/S0962-8479(96)90045-3","volume":"77","year":"1996","unstructured":"Sbarbaro, J. A., M. D. Iseman, and A. J. Crowle. 1996. Combined effect of pyrazinamide and ofloxacin within the human macrophage. Tuber. Lung Dis.77:491-495.","journal-title":"Tuber. Lung Dis."},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1998.9147"},{"key":"e_1_3_2_31_2","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1006\/bbrc.1997.7689","volume":"240","year":"1997","unstructured":"Srivastava, R., D. Kumar, and B. S. Srivastava. 1997. Recombinant Mycobacterium aurum expressing Escherichia coli \u03b2-galactosidase in high throughput screening of antituberculosis drugs. Biochem. Biophys. Res. Commun.240:536-539.","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1997.6837"},{"key":"e_1_3_2_33_2","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1089\/107999001750133258","volume":"21","year":"2001","unstructured":"Takii, T., C. Abe, A. Tamura, S. Ramayah, J. T. Belisle, P. J. Brennan, and K. Onozaki. 2001. Interleukin 1 or tumor necrosis factor \u03b1 augmented cytotoxic effect of mycobacteria on human fibroblasts: application to evaluation of pathogenesis of clinical isolates of M. tuberculosis and M. avium complex. J. Interferon Cytokine Res.21:187-196.","journal-title":"J. Interferon Cytokine Res."},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.24.6.976-981.1986"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.1.121-124.1997"},{"key":"e_1_3_2_36_2","unstructured":"World Health Organization. Anti-tuberculosis drug resistance in the world (World Health Organization Geneva 2000). [Online.] http:\/\/www.who.int\/gtb\/publications\/dritw\/contents.htm\n."},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.67.3.1239-1245.2001"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.181.7.2044-2049.1999"}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.46.8.2533-2539.2002","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.46.8.2533-2539.2002","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,21]],"date-time":"2022-02-21T17:47:20Z","timestamp":1645465640000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AAC.46.8.2533-2539.2002"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,8]]},"references-count":37,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2002,8]]}},"alternative-id":["10.1128\/AAC.46.8.2533-2539.2002"],"URL":"https:\/\/doi.org\/10.1128\/aac.46.8.2533-2539.2002","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"value":"0066-4804","type":"print"},{"value":"1098-6596","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,8]]},"assertion":[{"value":"2001-08-30","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2002-05-15","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2002-08-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}