• Tuberculosis and Respiratory Diseases
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• v.56 no.3
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• pp.261-268
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• 2004

Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.3
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• pp.225-232
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• 2021

This study sought to investigate the distribution, antimicrobial resistance rate, and bacterial co-infection frequency of non-tuberculous mycobacteria (NTM) in a single center in Incheon, South Korea. A total of 8,258 specimens submitted for tuberculosis (TB)/NTM real-time PCR tests during the years 2015 to 2020 were retrospectively reviewed. In total, 296 specimens (3.6%) were NTM positive, and the positivity increased from 2.5% (30/1,209) in 2015 to 3.8% (66/1,740) in 2020. Of 296 NTM specimens, 54.7% (162/296) were identified as the Mycobacterium avium complex (MAC) followed by the Mycobacterium abscessus complex (MABC) 20.9% (62/296), M. fortuitum 6.4% (19/296) and M. flavescens 3.4% (10/296). Of the NTM-positive specimens, 76.7% (227/296) were tested for drug resistance. The results showed multidrug-resistant NTM in 40.1% (91/227) and extensively drug-resistant NTM in 59.9% (136/227) of these specimens. Of the 145 isolates taken for bacterial culture, bacteria/fungi co-infection with NTM accounted for 43.4% (63/145), in which the most common bacterial species was Klebsiella pneumonia (23.8%, 15/63). This study is the first report on the distribution and antimicrobial resistance of NTM in Incheon. As the proportion of NTM infections increases, active treatment and thorough infection control are required for effective management.

• The Korean Journal of Nuclear Medicine
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• v.38 no.1
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• pp.85-98
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• 2004

Purpose: Cellular uptake of $^{99}mTc$-sestamibi (MIBI) and $^{99}mTc$-tetrofosmin (TF) is low in cancer cells expressing multidrug resistance(MDR) by p-glycoprotein(Pgp) or multidrug related protein(MRP). Verapamil is known to increase cellular uptake of MIBI in MDR cancer cells, but is recently reported to have different effects on tracer uptake in certain cancer cells. This study was prepared to evaluate effects of verapamil on cellular uptake of MIBI and TF in several cancer cells. Materials and Methods: Celluar uptakes of Tc-99m MIBI and TF were measured in erythroleukermia K562 cell, breast cancer MCF7 cell, and human ovarian cancer SK-OV-3 cells, and data were compared with those of doxorubicin-resistant K562(Ad) cells. RT-PCR and Western blot analysis were used for the detection of mdr1 mRNA and Pgp expression, and to observe changes in isotypes of PKC enzyme. Effects of verapamil on MIBI and TF uptake were evaluated at different concentrations upto $200{\mu}M\;at\;1{\times}10^6\;cells/ml\;at\;37^{\circ}C$. Radioactivity in supernatant and pellet was measured with gamma counter to calculate cellular uptake ratio. Toxicity of verapamil was measured with MTT assay. Results: Cellular uptakes of MIBI and TF were increased by time in four cancer cells studied. Co-incubation with verapamil resulted in an increase in uptake of MIBI and TF in K562(Adr) cell at a concentration of $100{\mu}M$ and the maximal increase at $50{\mu}M$ was 10-times to baseline. In contrast, uptakes of MIBI and TF in K562, MCF7, SK-OV3 cells were decreased with verapamil treatment at a concentration over $1{\mu}M$. With a concentration of $200{\mu}M$ verapamil, MIBI and TF uptakes un K562 cells were decreased to 1.5 % and 2.7% of those without verapamil, respectively. Cellular uptakes of MIBI and TF in MCF7 and SK-OV-3 cells were not changed with $10{\mu}M$, but were also decreased with verapamil higher than $10{\mu}M$, resulting 40% and 5% of baseline at $50{\mu}M$. MTT assay of four cells revealed that K562, MCF7, SK-OV3 were not damaged with verapamil at $200{\mu}M$. Conclusion: Although verapamil increases uptake of MIBI and TF in MDR cancer cells, cellular uptakes were further decreased with verapamil in certain cancer cells, which is not related to cytotoxicity of drug. These results suggest that cellular uptakes of both tracers might differ among different cells, and interpretation of changes in tracer uptake with verapamil in vitro should be different when different cell lines are used.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.428-436
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• 2016

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.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.3
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• pp.256-262
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• 2017

Essential oil from herb is known to exert pharmacological effects on the human body. In this study we investigated the antibacterial activity of 4 essential oils (teetree, rosemary, melisa, and lavender), as well as the blended mixture oil of teetree, rosemary, and melisa (TRM) on three bacteria, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Antibacterial analysis was performed using the standard disk diffusion method, and minimum inhibition concentration was determined by the broth microdilution method with different concentrations of essential oils (0.5, 1, 2 and 3 mg/mL). After incubation at $37^{\circ}C$ for 24 h, the antibacterial activity was assessed by measuring the zone of growth inhibition surrounding the disks. Herb oil with the inhibition zones showed varied values ranging from6 to 25 mm. However, the components of herb oil of TRM are as highly active as the teetree oil against pathogens, generating large inhibition zones for both gram negative and positive bacteria (13~22 mm and 8 mm inhibition zones). In the analysis for MIC, TRM showed growth-inhibitory effects at 0.0625% for S. aureus and E. coli, and 1.25% for P. aeruginosa. This result demonstrated that the anti-microbial activity of TRM was greater than a single herb oil, including oxacillin, rosemary, and teetrea. As a single herb oil, both rosemary and teetrea also had an anti-microbial effect by itself, and we can expect that the blended oil mixture may exert a synergistic effect against multidrug resistant bacteria, suggesting its future application in natural preservative agents for health food and cosmetics.

• Journal of Food Hygiene and Safety
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• v.26 no.3
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• pp.222-226
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• 2011

Salmonellosis is a major bacterial zoonosis that causes self-limited enteritis to fatal infection in animals and food-borne infection and typhoid fever in humans. Multidrug-resistant strains of Salmonella spp. has increased over the last several decades and recently causes more serious problems in public health. The present study was investigated bacteriocidal effects of sodium chlorate, sodium azide, sodium cyanide, and sodium salts mixture containing sodium chlorate, sodium azide, and sodium cyanide on infection with S. typhimurium in macrophage RAW 264.7 cells, and antibacterial effects of sodium salts mixture for murine salmonellosis. In infection assay of S. typhimurium in RAW 264.7 cells, bacterial survival rates within macrophage in all treated groups was significantly reduced comparing to that of the control group with the passage of incubation time. Administration of sodium salts mixture showed a therapeutic effect for S. typhimurium infected ICR mice. The mortality of mice treated with sodium salts mixture was 70% until 12 days, while that of control mice was 100% until 9 days after S. typhimurium infection. The results of this study strongly indicate that sodium salts mixture has a potency treatment for murine salmonellosis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.5
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• pp.436-446
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• 2006

The purpose of this study was to isolate and identify the bacteria in chronic maxillary sinusitis (CMS) lesions from 3 patients and to determine the antimicrobial susceptibility of them against 10 antibiotics. One of them was odontogenic origin and the others were non-odontogenic origin. Pus samples were collected by needle aspiration from the lesions and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, CO2, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene (16S rDNA) nucleotide sequencing method. To test the sensitivity of the bacteria isolated from the maxillary sinusitis lesions against seven antibiotics, penicillin G, amoxicillin, tetracycline, ciprofloxacin, cefuroxime, erythromycin, clindamycin, and vancomycin, minimum inhibitory concentration (MIC) was performed using broth dilution assay. Our data showed that enterobacteria such as Enterobacter aerogenes (30%), Klebsiella pneumoniae (25%), and Serratia marcescens (15%) were predominately isolated from the lesion of non-odontogenic CMS of senile patient (70 year old). Streptococcus spp. (40.3%), Actinomyces spp. (27.4%), P. nigrescens, M. micros, and P. anaerobius strains were isolated in the lesion of odontogenic CMS. In the lesion of non-odontogenic CMS, Streptococcus spp. (68.4%), Rothia spp. (13.2%), and Actinomyces sp. (10.5%) were isolated. The susceptibility pattern of 10 antibiotics was determined according to the host of the bacteria strains ratter than the kinds of bacterial species. Even though the number of CMS was limited as three, these results indicate that antibiotic susceptibility test must be accompanied with treatment of CMS. The combined treatment of two or more antibiotics is better than single antibiotic treatment in the presence of multidrug-resistant bacteria in the CMS lesions.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4353-4358
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• 2014

Background: PI3/AKT and NF-kB signaling pathways are constitutively active in acute myeloid leukemia and cross-talk between the two has been shown in various cancers. However, their role in acute myeloid leukemia has not been completely explored. We therefore used cell penetrating inhibitor peptides to define the contributions of AKT and NF-kB to survival and multi drug resistance (MDR) in HL-60 cells. Materials and Methods: Inhibition of AKT and NF-kB activity by AKT inhibitor peptide and NBD inhibitor peptide, respectively, resulted in decreased expression of mRNA for the MDR1 gene as assessed by real time PCR. In addition, treatment of HL-60 cells with AKT and NBD inhibitor peptides led to inhibition of cell viability and induction of apoptosis in a dose dependent manner as detected by flow cytometer. Results: Finally, co-treatment of HL-60 cells with sub-optimal doses of AKT and NBD inhibitor peptides led to synergistic apoptotic responses in AML cells. Conclusions: These data support a strong biological link between NF-kB and PI3-kinase/AKT pathways in the modulation of antiapoptotic and multi drug resistant effects in AML cells. Synergistic targeting of these pathways using NF-kB and PI3-kinase/AK inhibitor peptides may have a therapeutic potential for AML and possibly other malignancies with constitutive activation of these pathways.

• Korean Journal of Clinical Pharmacy
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• v.27 no.2
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• pp.92-98
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• 2017

Background: Nebulized colistimethate is increasingly used, because there are problems such as renal dysfunction and low distribution within the lungs when colistimethate is administered intravenously. This study was designed to compare and analyze the changes in renal function by of nebulized colistimethate treatment for its safe administration. Methods: This study retrospectively reviewed the electronic medical records of adult patients above 19 years old, receiving only the nebulized colistimethate at least 4 days in Yonsei university health system from Nov 2014 to Aug 2015. Acute kidney injury (AKI) was determined by using the RIFLE criteria (Risk, Injury, Failure, Loss and End-stage renal disease) according to serum creatinine (SCr) levels before and after use of nebulized colistimethate. Results: 48 patients were included our study and their SCr increased significantly after nebulized colistimethate treatment ($SCr_0$ vs. $SCr_1$; $0.85{\pm}0.80$ vs. $1.00{\pm}0.82mg/dL$, n=48, p<0.001), but the changes were in normal range according to the standards at Yonsei university health $system^a$. Among 48 patients, 38 patients were in the non-AKI group (79.2%), and 10 patients developed AKI (20.8%). Within the AKI group, 2 patients were in the Injury group (20%) and the other 8 in the Risk group (80%). Conclusion: There was no significant difference in age, dosage and duration of treatment between AKI group and non-AKI group (p>0.05). The study has a significance in that it reviewed the safety of nebulized colistimethate only treatment to national patients, analyzing its nephrotoxicity. It has confirmed that nebulized colistimethate is a safer method than intravenous injection, and requires to establish a guideline for the use of nebulized colistimethate in further studies with broader patient groups. $^a$ : SCr Male 0.68-1.19 mg/dL, Female 0.49-0.91 mg/dL.

• Korean Journal of Environmental Biology
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• v.25 no.4
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• pp.342-348
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• 2007

In a previous study, a new antibacterial activity was found from Paenibacillus polymyxa DY1 isolated from Korean soil, which showed a good growth inhibition against various multidrug resistant enteric bacteria. Thus the effect of nutritional factors and physicochemical conditions on the growth of P. polymyxa DY1 cells and the production of antibacterial activity were evaluated in this study. For the growth of P. polymyxa DY1 cells the optimum condition reaches by incubating at $35^{\circ}C$ for 48 hours by shaking at 180 rpm in the basal medium containing 0.6% dextrose, 1.4% ammonium phosphate dibasic, 0.9% peptone, 2.4% glutamine, and 2 mM sodium molybdate with initial pH 8.0. A maximum level of antibacterial activity was obtained when P. polymyxa DY1 was incubated at $35^{\circ}C$ for 48 hours by shaking at 180 rpm in the basal medium with initial pH 8.0 containing 1% dextrose, 1.5% ammonium sulfate, 1.1% casamino acid, 1.9% aspartic acid, and 2 mM ferrous sulfate.