• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.495-501
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• 2009

A new spore display method is presented that enables recombinant proteins to be displayed on the surface of Bacillus spores via fusion with InhA, an exosporium component of Bacillus thuringiensis. The green fluorescent protein and $\beta$-galactosidase as model proteins were fused to the C-terminal region of InhA, respectively. The surface expression of the proteins on the spores was confirmed by flow cytometry, confocal laser scanning microscopy, measurement of the enzyme activity, and an immunogold electron microscopy analysis. InhA-mediated anchoring of foreign proteins in the exosporium of Bacillus spores can provide a new method of microbial display, thereby broadening the potential for novel applications of microbial display.

• Biomedical Science Letters
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• v.28 no.4
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• pp.237-246
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• 2022

Ischemia-reperfusion results in excess reactive oxygen species (ROS) that affect myocardial cell damage. ROS production inhibition is effectively proposed in treating cardiovascular diseases including myocardial hypertrophy. Studies have shown that oxidizing cultured cells in in vitro experiments gradually decreases the permeability of mitochondrial membranes time- and concentration-dependent, resulting in increased mitochondrial membrane damage due to secondary ROS production and cardiolipin loss. However, recent studies have shown that 5-iodo-6-amino-1,2-benzopyrone (INH₂BP), an anticancer and antiviral drug, inhibited peroxynitrite-induced cell damage in in vitro and alleviated partial or overall inflammation in animal experiments. Therefore, in this paper, we studied the preventive effect of INH₂BP on H9c2 cells derived from mouse heart damaged by oxidative stress using 700 μM of hydrogen peroxide. As a result of oxidative stress to H9c2 cells by hydrogen peroxide whether the treatment of INH₂BP or not, hydrogen peroxide caused serious damage in H9c2 cells. These results were confirmed with cell viability and Hoechst 33342 assays. And this damage was through cell death. However, it was confirmed that H9c2 cells pretreated with INH₂BP significantly reduced cell death by hydrogen peroxide. In addition, measurements with DCF-DA assay to determine whether ROS is produced in H9c2 cells treated with only hydrogen peroxide produced ROS significantly, but H9c2 cells pretreated with INH₂BP significantly reduced ROS production by hydrogen peroxide. Taken together, it is believed that INH₂BP can be useful for the prevention and treatment of cardiovascular diseases induced through oxidative stress such as heart damage caused by ischemia/reperfusion.

• Journal of Yeungnam Medical Science
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• v.41 no.2
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• pp.113-119
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• 2024

Background: Missing isoniazid (INH) resistance during tuberculosis (TB) diagnosis can worsen the outcomes of INH-resistant TB. The BD MAX MDR-TB assay (BD MAX) facilitates the rapid detection of TB and INH and rifampin (RIF) resistance; however, data related to its performance in clinical setting remain limited. Moreover, its effect on treatment outcomes has not yet been studied. Methods: We compared the performance of BD MAX for the detection of INH/RIF resistances to that of the line probe assay (LPA) in patients with pulmonary TB (PTB), using the results of a phenotypic drug sensitivity test as a reference standard. The treatment outcomes of patients who used BD MAX were compared with those of patients who did not. Results: Of the 83 patients included in the study, the BD MAX was used for an initial PTB diagnosis in 39 patients. The sensitivity of BD MAX for detecting PTB was 79.5%. The sensitivity and specificity of BD MAX for INH resistance were both 100%, whereas these were 50.0% and 95.8%, respectively, for RIF resistance. The sensitivity and specificity of BD MAX were comparable to those of LPA. The BD MAX group had a shorter time interval from specimen request to the initiation of anti-TB drugs (2.0 days vs. 5.5 days, p=0.001). Conclusion: BD MAX showed comparable performance to conventional tests for detecting PTB and INH/RIF resistances. The implementation of BD MAX as a diagnostic tool for PTB resulted in a shorter turnaround time for the initiation of PTB treatment.

• Tuberculosis and Respiratory Diseases
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• v.60 no.1
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• pp.38-43
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• 2006

Background : Even though two-month rifampicin (RMP, R) and pyrazinamide (PZA, P) treatment has some advantages over isoniazid (INH, H) treatment for latent tuberculosis infection (LTBI), it was withdrawn from the list of treatment regimens for LTBI because of reported cases of severe hepatotoxicity. The purpose of this study was to estimate the frequency of hepatotoxicity of RMP and PZA treatment excluding INH in a Korean population. Method : TIn order to recruit patients who were prescribed RMP and PZA excluding INH, 256 INH-resistant tuberculosis patients were investigated through retrospective medical record analysis. A standard four-drug regimen was changed to a RMP/PZA-containing regimen excluding INH in 64 patients (RZ+ group). In the same study period, 146 patients who were prescribed an INH/RMP/PZA-containing standard regimen were randomly selected as a control (HRZ+ group). Clinical characteristics including liver diseases and the frequency of drug-induced hepatitis were compared between the RZ+ and HRZ+ groups. Result : The mean age of patients in the RZ+ group was 50.2 (${\pm}16.2$) and the male-to-female ratio was 36:28. The frequency of underlying liver diseases was 10.9% (7/64), which was not significantly different from that of the HRZ+ group (4.1%, 6/146). Even though the treatment duration of RZ+ ($5.5{\pm}4.8months$) was longer that than that of HRZ+ ($2.7{\pm}2.3months$), the frequency of toxic hepatitis was not significantly different between RZ+ and HRZ+ groups, 3.5% (2/57) and 7.1% (10/140), respectively. Conclusion : Hepatotoxicity was mild and occurred in a minor proportion of patients in a Korean population prescribed an RMP/PZA-containing regimen. A future prospective study including more patients is needed.

• Journal of the Korean Chemical Society
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• v.52 no.3
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• pp.281-294
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• 2008

synergistic action caused by iodide ions on the corrosion inhibition of aluminium (Al) in 0.5 M HCl in the presence of Azadirachta Indica (AZI) plant extract has been investigated using potintiodynamic polarization and impedance techniques. It is found that AZI extract inhibits the corrosion of aluminium in 0.5 M HCl. The inhibition efficiency increases with the increase in AZI extract concentration, until 24% v/v of AZI extract, then Inh.% is decreased with father increase in AZI extract concentration. The adsorption of this extract in the studied concentration is found to obey Frewendlish adsorption isotherm. The addition of iodide ions enhances the inhibition efficiency to a considerable extent. The increase in Inh.% values in presence of fixed concentration of iodide ions indicates that AZI extract forms an insoluble complex at lower AZI extract concentrations by undergoing a joint adsorption. But at higher concentrations of AZI extract, competitive adsorption is found between iodide ions and the formed complex leading to less Inh.%.The Inh.% decreased in presence of iodide ions with AZI extract than in presence of AZI extract alone at all studied iodide concentrations. The synergism parameter Sq is defined and calculated from surface coverage values. This parameter in the case of AZI extract is found to be more than unity, indicating that the enhanced inhibition efficiency caused by the addition of iodide ions.

• Tuberculosis and Respiratory Diseases
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• v.44 no.5
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• pp.992-1000
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• 1997

Background : Since up to 90% of a theophylline dose is biotransformed, probably by interaction with one or more the variants of the cytochrome P-450 drug metabolism system, anti-tuberculosis agents including drugs influencing microsomal enzyme systems, such as isoniazid and rifampicin. may be affect the elimination of theophylline. Method : The effect of combination therapy with isoniazid(INH), rifampicin(RFP), ethambutol(EMB) and pyrazinamide(PZA) on the pharmacokinetics of theophylline was evaluated by a computer program using Bayesian method. Three group were divided as follows. Group I is control, Group II is treated with INH. RFP, EMB and PZA and Group III is treated with INH, RFP and EMB. All of them were ilon-smoker who were normal in liver and renal functions, and not administered drugs affecting on the clearance of theophylline with exception of anti-tuberculous agents. Results : When it compared control with test groups, the clearance of theophylline in Group II and Group III was significantly decreased(p<0.001), and half life in Group II and Group III showed significant elevation(p<0.001). However there were no significant differences in clearance and half life between the Group II and Group III. Conclusion : These results suggest that theophylline dose may be need of readjustment in concurrent medication of anti-tuberculous agents including INH, RFP, and EMB.

• Tuberculosis and Respiratory Diseases
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• v.51 no.5
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• pp.416-425
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• 2001

Background : The incidence of drug-resistant tuberculosis has recently decreased in Korea. However, it is still one of the major obstacles in treating pulmonary tuberculosis. This study was performed to determine the prevalence and clinical characteristics associated with drug-resistance in pulmonary tuberculosis at the tertiary referral hospital in Pusan, Korea. Methods : The medical records of 138 patients, who had been diagnosed as active pulmonary tuberculosis were retrospectively reviewed, and results of drug susceptibility from May 1997 to June 2000. The relationships among those with a history of previous tuberculosis treatment, the extent of lung involvement, the presence of cavities on the initial chest X-ray films and patterns of drug resistance were analyzed. Results : The total number of patients who had drug resistance to at least one drug was 55(39.9%). Among them 34(24.6%) had resistance to isoniazid(INH) and rifampin(RFP). There was drug resistance in 20(22%) of 91 patients without previous tuberculosis therapy, and among them 9(9.9%) were multi-drug resistant. Thirty-two(74.5%) out of 47 patients with previous therapy were drug-resistant and 25(53.2%) were multidrug resistant. For all 138 patients, resistance to INH was the most common(34.1%), followed by RFP(26.1%) and ethambutol(EMB)(14.5%). Drug resistance to INH, RFP, PZA and streptomycin(SM) were independently associated with a history of previous treatment(odds ratio; 9.43, 9.09, 8.93 and 21.6 respectively, p<0.01). The extent of lung involvement on the chest films was significantly associated with the drug resistance to INH and RFP(odds ratio; 2.12 and 2.40 respectively, p<0.01). The prevalence of drug resistance to RFP, INH and RFP was significantly more common in patients with a cavitary lesion on the chest films by multivariate analysis(odds ratio; 4.17 and 4.81 respectively, p<0.05). Conclusion : This study revealed that patients with a prior treatment history for pulmonary tuberculosis, and the presence of a cavitary lesion on chest films had a higher prevalence of anti-tuberculosis drug resistance. A very careful clinical and microbiological examination is needed for patients with such characteristics.

• Tuberculosis and Respiratory Diseases
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• v.65 no.3
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• pp.177-182
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• 2008

Background: Isoniazid (INH, H) is a key drug of the standard first-line regimen for the treatment of tuberculosis (TB), yet some reports have suggested that treatment efficacy was maintained even though INH was omitted from the treatment regimen. Methods: One hundred forty C57BL/6 mice were infected with the H37Rv strain of M. tuberculosis with using a Glas-Col aerosol generation device, and this resulted in depositing about 100 bacilli in the lung. Four weeks after infection, anti-TB treatment was initiated with varying regimens for 4-8 weeks; Group 1: no treatment (control), Group 2 (4HREZ): 4 weeks of INH, rifampicin (R), pyrazinamide (Z) and ethambutol (E), Group 3: 1HREZ/3REZ, Group 4: 4REZ, Group 5: 4HREZ/4HRE, Group 6: 1HREZ/3REZ/4RE, and Group 7: 4REZ/4RE. The lungs and spleens were harvested at several time points until 28 weeks after infection, and the colony-forming unit (CFU) counts were determined. Results: The CFU counts increased steadily after infection in the control group. In the 4-week treatment groups (Group 2-4), even though the culture was negative at treatment completion, the bacilli grew again at the 12-week and 20-week time points after completion of treatment. In the 8-week treatment groups (Groups 5-7), the bacilli did not grow in the lung at 4 weeks after treatment initiation and thereafter. In the spleens of Group 7 in which INH was omitted from the treatment regimen, the culture was negative at 4-weeks after treatment initiation and thereafter. However, in Groups 5 and 6 in which INH was taken continuously or intermittently, the bacilli grew in the spleen at some time points after completion of treatment. Conclusion: TThe exclusion of INH from the standard first-line regimen did not affect the treatment outcome in a murine model of TB in the early stage of disease. Further studies using a murine model of chronic TB are necessary to clarify the role of INH in the standard first-line regimen for treating TB.

• Korean Journal Plant Pathology
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• v.10 no.1
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• pp.39-46
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• 1994

Pseudomonas fluorescens Biovar III strains S-2 antagonistic to Rhizoctonia solani was subjected to Tn5 mutagenesis by the transposon vector pGS9. Ampicillin and kanamycin resistant (Ampr, Kmr) transconjugants were recovered at a frequency of 1.3$\times$10-7 per initial recipient cell, when recipient cells were washed twice in TE buffer before conjugation. Of the ca. 3000 transconjugants, a frequency of noninhibitory (Inh-), nonfluorescent (Flu-) and auxotorphic (Pro-) mutants were 0.27%, 0.47% and 0.40%, respectively. In these mutants, all Inh- mutants showed the same colony morphology as wild type, whereas all Flu- and Pro- mutants inhibited the growth of R. solani. These mutants were also susceptible to chloramphenicol, indicating only the Tn5 element, except for parts of pGS9, was integrated into the recipient genome. In a Southern blot analysis, the Tn5 element inserted into one site on the chromosome for each of the chosen mutants. However, Tn5 insertion sites of Inh-, and Pro- mutants were differed in each other. These indicate that the genes essential for R. solani inhibition, fluorescent production and auxotrophic are chromosomally located, but not linked to each other.

• Annals of Laboratory Medicine
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• v.38 no.6
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• pp.569-577
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• 2018

Background: The increasing prevalence of drug-resistant tuberculosis (TB) infection represents a global public health emergency. We evaluated the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform [QMAP], QuantaMatrix, Seoul, Korea) to rapidly identify the Mycobacterium tuberculosis complex (MTBC) and detect rifampicin (RIF) and isoniazid (INH) resistance-associated mutations. Methods: A total of 200 clinical isolates from respiratory samples were used. Phenotypic anti-TB drug susceptibility testing (DST) results were compared with those of the QMAP system, reverse blot hybridization (REBA) MTB-MDR assay, and gene sequencing analysis. Results: Compared with the phenotypic DST results, the sensitivity and specificity of the QMAP system were 96.4% (106/110; 95% confidence interval [CI] 0.9072-0.9888) and 80.0% (72/90; 95% CI 0.7052-0.8705), respectively, for RIF resistance and 75.0% (108/144; 95% CI 0.6731-0.8139) and 96.4% (54/56; 95% CI 0.8718-0.9972), respectively, for INH resistance. The agreement rates between the QMAP system and REBA MTB-MDR assay for RIF and INH resistance detection were 97.6% (121/124; 95% CI 0.9282-0.9949) and 99.1% (109/110; 95% CI 0.9453-1.0000), respectively. Comparison between the QMAP system and gene sequencing analysis showed an overall agreement of 100% for RIF resistance (110/110; 95% CI 0.9711-1.0000) and INH resistance (124/124; 95% CI 0.9743-1.0000). Conclusions: The QMAP system may serve as a useful screening method for identifying and accurately discriminating MTBC from non-tuberculous mycobacteria, as well as determining RIF- and INH-resistant MTB strains.