The emergence of antibiotic-resistant bacteria has been increased and become a public health concern worldwide. Many bacterial infections can be sequentially treated with different types of antibiotics. Thus, this study was designed to evaluate the changes in survival, antibiotic susceptibility, mutant frequency, ${\beta}$-lactamase activity, biofilm formation, and gene expression in Klebsiella pneumoniae after exposure to sequential antibiotic treatments of ciprofloxacin and meropenem. Treatments include control (CON; no addition), 1/2 MIC ciprofloxacin addition (1/2CIP), 2 MIC ciprofloxacin addition (2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC ciprofloxacin (16 h-incubation) (1/2CIP-1/2MER-2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC meropenem (16 h-incubation) (1/2CIP-1/2MER-2MER), and initial 1/2 MIC ciprofloxacin addition followed by 2 MIC ciprofloxacin(8 h-incubation) and 2 MIC meropenem(16 h-incubation) (1/2CIP-2CIP-2MER). No growth of K. pneumoniae was observed for the 2CIP throughout the incubation period. The numbers of planktonic cells varied with the treatments (7~10 log CFU/ml), while those of biofilm cells were not significantly different among treatments after 24-h incubation, showing approximately 7 log CFU/ml. Among the sequential treatments, the least mutant frequency was observed at the 1/2CIP-1/2MER-2CIP (14%). Compared to the CON, 1/2CIP-2CIP-2MER decreased the sensitivity of K. pneumoniae to piperacillin, cefotaxime, and nalidixic acid. The highest ${\beta}$-lactamase activity was 22 nmol/min/ml for 1/2CIP-1/2MER-2CIP, while the least ${\beta}$-lactamase activity was 6 nmol/min/ml for 1/2CIP-2CIP-2MER. The relative expression levels of multidrug efflux pump-related genes (acrA, acrB, and ramA) were increased more than 2-fold in K. pneumoniae exposed to 1/2CIP-1/2MER-2MER and 1/2CIP-2CIP-2MER. The results suggest that the sequential antibiotic treatments could change the antibiotic resistance profiles in K. pneumoniae.
Kim, Eun Kyung;Shim, Tae Sun;Lee, Jung Yeon;Oh, Yeon-Mok;Lim, Chae-Man;Lee, Sang Do;Koh, Younsuck;Kim, Dong Soon;Kim, Won Dong;Kim, Woo Sung
Tuberculosis and Respiratory Diseases
/
v.57
no.3
/
pp.226-233
/
2004
Background : Interferon-gamma (IFN-${\gamma}$) is a critical cytokine in the defense against a Mycobacterium tuberculosis infection. Even though IFN-${\gamma}$ has occasionally been used in the treatment of refractory multidrug-resistant tuberculosis (MDR-TB) with some promising results, there is still some controversy regarding the therapeutic efficacy of IFN-${\gamma}$. This study was performed to examine the effect of subcutaneous IFN-${\gamma}$ in the treatment of MDR-TB patients. Methods : Six patients with refractory MDR-TB were enrolled in this study. Two million IU of IFN-${\gamma}$ was administered subcutaneously three times a week with the concomitant administration of antituberculous drugs for at least for 28 weeks. During the IFN-${\gamma}$ therapy, the sputum smear and culture, radiological and clinical evaluations were performed every 4 weeks throughout the study period. Results : The mean age of the 6 patients was 37 years (ranges, 15-61 years). The drug susceptibility test to standard antituberculous drugs revealed resistance to an average of 6.8 (${\pm}1.2$) agents including isoniazid and rifampicin. An average of 10.8 (${\pm}1.3$) antituberculous drugs were prescribed before IFN-${\gamma}$ therapy. The culture became negative in 2 patients (33%) after initiating IFN-${\gamma}$ therapy; one at 8 weeks, and the other at 24 weeks. Finally, after stopping the IFN-${\gamma}$ therapy after 28 weeks, the culture became positive again in the two patients who were culture-negative. The other 4 patients who failed in the culture conversion are still on antituberculous treatment except for one who died of tuberculosis. Conclusion : Even though 28 weeks of subcutaneous IFN-${\gamma}$ therapy in combination with antituberculous drugs was successful in inducing the culture-negative conversion in some patients with refractory MDR-TB, the culture became positive again after stopping the IFN-${\gamma}$ therapy. This suggests that subcutaneous IFN-${\gamma}$ therapy may have suppressive effect on tuberculosis only during the IFN-${\gamma}$ therapy period in some patients. Further studies will be needed to determine the optimum dose, the administration route, the duration of therapy, and the predicting factors of the response to adjuvant IFN-${\gamma}$ therapy.
Kim, Hyoung-Soo;Rho, Kwang-Suk;Kong, Suck-Jun;Sohn, Mal-Hyeun;Kim, Tae-Yoon
Tuberculosis and Respiratory Diseases
/
v.51
no.5
/
pp.409-415
/
2001
Background : Multidrug-resistant tuberculosis(MDR-TB) in patients is mainly caused by acquired drug resistance. However, a small proportion of MDR-TB is caused by initial drug resistance(IDR), which may be somewhat different from acquired drug resistance. This study analyzed the clinical characteristics of IDR in MDR -TB patients to use the results as basic data in managing the disease. Methods : A retrospective study of 30 IDR cases in MDR-TB patients from Jan. 1995 to Dec. 1998 was performed. In order to analyze the clinical characteristics, the age, sex, family history, duration of negative conversion, number of resistant drugs, treatment regimens, duration of treatment, extent of disease and cavitary lesion on the chest X-ray was examined. In order to analyze the level of improvement, the extent of the disease and cavitary lesion on the chest X-ray, tested by Wilcoxon signed rank sum test, and the disease free interval rate of 1-year and 4-year was examined using the Kaplan-Meier method. Results : The mean age of the patients was 46.6 years and the sex ratio 1:1. Six(20%) patients had a family history. The mean negative conversion of the sputum AFB stain was 2.6 months. The number of resistant drugs was 7.6 and the number of used drugs 3.6. Twenty-three(67%) patients were treated for less than 12months and 28(93%) patients were treated with first-line drugs. The extent of the disease and the cavitary lesion on the chest X-ray improved after treatment(p<.05). Among 13 patients who were followed up for 22.6 months, 2(15%) patients relapsed and the disease free interval rate of I-year and 4-year was 85%. Conclusion: It is recommended that the duration of treatment of IDR in MDR-TB with first-line drugs be 9-12 months even if the extent of disease and cavitary lesion on the chest X-ray improves.
Park, Myoung Hee;Song, Eun Young;Kwon, Sung Youn;Park, He Jin;Han, Sung Koo;Shim, Young Soo
Tuberculosis and Respiratory Diseases
/
v.54
no.4
/
pp.367-377
/
2003
Background : It is well known that only 10% of those infected with Mycobacterium tuberculosis actually develop clinical disease, indicating the existence of host genetic factors regulating disease expression. In this study, we investigated HLA-DRB1 and -DQB1 gene polymorphisms in Korean patients with pulmonary tuberculosis (PTB). Methods : HLA-DRB1 and -DQB1 gene polymorphisms were investigated in 67 PTB patients without previous treatment history, 38 drug-sensitive (DS) and 29 multidrug-resistant (MDR) cases, and 200 healthy controls. HLA-DRB1 typing was done using reverse SSO (sequence specific oligonucleotide) and PCR-SSCP (single strand conformational polymorphism) methods and DQB1 typing was done using PCR-RFLP (restriction fragment length polymorphism), PCR-SSCP and PCR-SSP (sequence specific primer) methods. Results : Among the PTB patients, MDR-TB cases showed frequencies of DRB1*0701 and *08032 increased by about two-fold compared to those of normal controls, and likewise for their associated DQB1 alleles, DQB1*0202 and *0601 (15.5% vs. 34.5%, p=0.01). The frequency of HLA-DQB1*0609 was significantly increased in PTB patients (4.0% vs. 14.9%, p=0.004), showing similar increases in both DS and MDR cases. There was also an association of HLA alleles with the clinical severity of the disease according to the extent of lung lesion. Significantly increased frequencies of DRB1*08032 (4.2% vs. 32.6%, p=0.007) and DQB1*0601 (12.5% vs. 34.9%, p=0.047) were observed in more advanced (moderately & far advanced/DS and far advanced/MDR), compared with less advanced (minimal/DS and moderately advanced/MDR) lung lesions. Although DRB1*0701, DQB1*0202 and DQB1*0609 showed significant increases in different subsets of the disease, these HLA alleles did not show consistent association with disease severity. Conclusion : HLA-DRB1*08032 and DQB1*0601 alleles were associated with genetic susceptibility to MDR-TB in Korean patients, and also with disease severity and progression of PTB.
Kim, Su Nam;Won, Chong Bock;Cho, Hye Jung;Eun, Byung Wook;Sim, So Yeon;Choi, Deok Young;Sun, Yong Han;Cho, Kang Ho;Son, Dong Woo;Tchah, Hann;Jeon, In Sang
Pediatric Infection and Vaccine
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v.18
no.2
/
pp.135-142
/
2011
Purpose : Hospital associated infection (HAI) caused by multidrug-resistant (MDR) microorganisms has been recognized as an important issue in the world, especially in critically ill patients such as the patients admitted in the intensive care unit. There are fewer papers about MDR-HAI in pediatric patients compared to adult patients. In this study, we investigated the incidence and associated factors of MDR-HAI in children admitted to the intensive care unit (ICU) of a university hospital. Methods : We retrospectively evaluated 135 children who were admitted in ICU for at least 3 days between January 2009 and December 2010. HAI cases were divided into MDR-HAI group and non-MDR-HAI group. Clinical characteristics and various associated factors were compared between those groups. Results : In 39 patients, 45 cases of ICU-related HAI were developed. ICU-related HAI incidence was 47.7 per 1000 patientdays. Thirty-six cases (80.0%) were MDR-HAI. Acinetobacter baumannii was isolated more commonly in MDR-HAI group. And the followings were found more frequently in MDR-HAI group than non-MDR-HAI group: medical condition as an indication for ICU admission, mechanical ventilation, urinary catheterization and previous use of broad-spectrum antibiotics. Among the risk factors, previous use of broad-spectrum antibiotics was the independent risk factor for MDR-HAI. Conclusion : ICU-related HAI incidence was higher than previously reported. Previous use of broad-spectrum antibiotics was the independent risk factor for MDR-HAI. To investigate the characteristics of MDR-HAI in children admitted in ICU, further studies with a larger sample size over a longer period of time are warranted.
In this study, the hemolysis of Enterococcus faecalis BMSE-HMP strains, isolated from human breast milk, was investigated, and the anti-hemolytic and antimicrobial effects on multidrug-resistant (MDR) bacteria were investigated. The enzyme activity of E. faecalis BMSE-HMP 4 strains was measured, and it was found that the activities of esterase and esterase lipase were the highest. In addition, no hemolytic reaction was observed in any of the isolates. Subsequently, the anti-hemolytic activity against MDR strains causing hemolysis was evaluated. E. faecalis BMSE-HMP002 had the highest anti-hemolytic activity against Staphylococcus aureus CCARM 3855 at 75.71 ± 10.00%. The anti-hemolytic activity against Escherichia coli DC 2 CCARM 0238 and Pseudomonas aeruginosa CCARM 0223 showed that the activity of BMSE-HMP001 was highest at 76.92 ± 2.99% and 87.93 ± 1.93%, respectively. Examination of the antimicrobial effects against the MDR bacteria Staphylococcus spp., Escherichia spp., Pseudomonas spp., Salmonella spp., Klebsiella spp., Enterobacter spp., and E. faecalis BMSE-HMP strains showed antimicrobial effects against both gram-positive and gram-negative strains. Breastfeeding delivers enterococci into the intestinal tract of newborns by lactation, and its usefulness is attracting attention as it has been reported that enterococci have a potential effect on neonatal immune development. In this study, the hemolytic and antimicrobial effects of E. faecalis BMSE-HMP strains on MDR bacteria were investigated, to confirm their potential as useful lactic acid bacteria. Additional studies on the antibiotic resistance and toxicity of the E. faecalis BMSE-HMP strains, isolated in this study, are necessary to prove it safe for use.
Eun Seong Lee;Jeong Woo Park;Ki Hwan Moon;Youngwan Seo
Journal of Life Science
/
v.33
no.12
/
pp.1015-1024
/
2023
Antibiotics have greatly contributed to the treatment and prevention of bacterial diseases in humans, animals, and fish. However, antibiotic misuse has led to the emergence and spread of multidrug-resistant bacteria. In addition to antibiotic discovery research, efforts are being made to combat such multidrug-resistant bacteria using antimicrobial agents, antioxidants, host immune enhancement, probiotics, and bacteriophages, as well as various symptomatic therapies. To discover novel bioactive compounds, it is crucial to adopt approaches that incorporate fresh ideas, new targets, innovative techniques, and untapped resources. Halophytes are plants that grow in high-salt soils and are known to adapt to salt-induced stress through unique metabolic processes that produce secondary metabolites. This study aimed to investigate the effects of extracts of halophytes native to Korea on oxidative stress and to determine whether they exert inhibitory activity against biofilms, which are major pathogenic factors of infectious bacteria. The Acinetobacter baumannii strain ATCC 17978, a representative drug-resistant bacterium, was used to measure anti-biofilm activity. The results showed that Aster spathulifolius, Carex kobomugi, Rosa rugosa, and Asparagus cochinchiensis exerted strong antioxidant and anti-biofilm effects without affecting bacterial growth itself. The halophytes used in this study are promising candidates for the development of pharmaceutical agents with antioxidant and antimicrobial properties.
Purpose: Uptakes of Tc-99m MIBI (MIBI) and Tc-99m tetrofosmin (tetrofosmin) in human non-small cell lung cancer A549, multidrug-resistance associated protein (MRP) expressing cell, were investigated in vitro and in vivo. Materials and Methods: Western blot analysis and immunohistochemistry were used for detection of MRP in A549 cells with anti-MRPr1 antibody. Cellular uptakes of two tracers were evaluated at $100{\mu}M$ of verapamil (Vrp), $50{\mu}M$ of cyclosporin A (CsA) and $25{\mu}M$ of butoxysulfoximide (BSO) after incubation with MIBI and tetrofosmin for 30 and 50 min at $37^{\circ}C$, using single cell suspensions at $1{\times}10^6cells/ml$. Radioactivities in supernatants and pellets were measured with gamma well counter. A549 cells were inoculated in each flanks of 24 nude mice. Group 1 (Gr1) and Gr3 mice were treated with only MIBI or tetrofosmin, and Gr2 and Gr4 mice were treated with 70mg/kg of CsA i.p. for 1 hour before injection of 370KBq of MIBI or tetrofosmin. Mice were sacrificed at 10, 60 and 240 min. Radioactivities of organs and tumors were expressed as percentage injected dose per gram of tissue (%ID/gm). Results: Western blot analysis of the A549 cells detected expression of MRPr1 (190 kDa) and immunohistochemical staining of tumor tissue for MRPr1 revealed brownish staining in cell membrane but not P-gp. Upon incubating A549 cells for 60 min with MIBI and tetrofosmin, cellular uptake of MIBI was higher than that of tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetrofosmin. Percentage increase of MIBI was higher than that of tetrofosmin with Vrp by 623% and 427%, CsA by 753% and 629% and BSO by 219% and 140%, respectively. There was no significant difference in tumoral uptakes of MIBI and tetrofosmin between Gr1 and Gr3. Percentage increases in MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at 240 min) were progressively higher by the time up to 240 min with CsA. Conclusion: These results indicate that MIBI and tetrofosmin are suitable tracers for imaging MRP-mediated drug resistance in A549 tumors. MIBI may be a better tracer than tetrofosmin for evaluating MRP reversal effect of modulators.
Kim, Kyung Hyo;Lee, Jong Eun;Whang, Il Tae;Ryu, Kyung Ha;Hong, Young Mi;Kim, Gyoung Hee;Lee, Keun;Kang, Eun-Suk;Hong, Ki-Sook
Clinical and Experimental Pediatrics
/
v.45
no.3
/
pp.346-353
/
2002
Purpose : Penicillin- and multidrug-resistant S. pneumoniae poses a serious threat to clinicians because the rate of resistance of S. pneumoniae to penicillin in Korea has surged up to the world's highest level. This study was performed to assess the carriage rate, serogroups and antimicrobial susceptibility of S. pneumoniae isolated from oropharynx in children. Methods : From March to July 1998, 209 children under 5 years of age were recruited from five day care centers. The carriage rate for pneumococci was obtained. Antimicrobial susceptibilities were determined with the E-test and agar dilution methods. Serogrouping was performed on 48 of the pneumococcal isolates by the Quellung reaction. Results : The carriage rate of S. pneumoniae was 30.1%. Antimicrobial susceptibility profiles were available for 59 of the isolates. Sixty-six percent of isolates were not susceptible to penicillin, and multidrug-resistance was observed in 76.3% of the isolates. A high proportion of the penicillin-resistant strains showed associated resistance to trimethoprim-sulfamethoxazole, tetracycline, erythromycin, and oxacillin. The most prevalent oropharyngeal serogroups were 19, 6, 3, 23, and 29. Resistance of the pneumococcal isolates to penicillin was different according to the serogroups. All of the strains of serogroup 19, 23, and 29 was resistant to penicillin but 87.5% of serogroup 3 strains were susceptible to penicillin. Conclusion : The resistance rate of S. pneumoniae isolated from oropharynx in children was very high to penicillin and other antimicrobial agents. For the reduction of the drug-resistant rate of S. pneumoniae, clinicians should be required to be more judicious in their use of antimicrobial agents.
Hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA) has been an increasing problem worldwide since the initial reports over 40 years ago. To examine new drug leads with potential antibacterial activities, Various N'-[(-3-substituted-4-oxo-1,3-thiazolidin-2-ylidene]-4-hydroxy benzohydrazide (4a-4.i) and N'-[-(3,4-disubstituted)-1,3-thiazolidin-2ylidene)]-4-hydroxybenzohydrazide from (5.a-5.i) to (10.a-10.i) were synthesized using appropriate synthetic route. The entire test compounds (4.a-4.i) and from (5.a-5.i) to (10.a-10.i) were assayed in vitro against s. aureus strain. The minimum inhibitory concentration (MIC) was determined for test compounds and for reference standards. The test compounds showed significant antibacterial activity against the strains used, when tested in vitro. In general, p-hydroxybenzohydrazide ring and substituted thiazoline ring are essential for antimicrobial activity. Among the compounds tested, compounds 6.f, 7.g, 9.f and 10.f, 10 i were found to be most potent. The test compounds were found nontoxic upto the dose level of 2000 ${\mu}g$/mL. The intact compounds were then subjected for 3D-QSAR studies. 3D-QSAR study based on the principal of alignment of pharmacophoric features by Schrodinger PHASE module. The 3D-QSAR study allowed us to confirm the preferential binding mode of p-hydroxybenzohydrazide inside the active site.
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