Hyung Hoi Kim

Increase of NKG2D ligands and sensitivity to NK cell-mediated cytotoxicity of tumor cells by heat shock and ionizing radiation

In this study, we have investigated if current cancer therapeutic modalities including hyperthermia and ionizing radiation can increase the expression of NKG2D ligands in human cancer cell lines. The expressions of NKG2D ligands were induced by both heat shock and ionizing radiation in various cell lines including KM12, NCI-H23, HeLa and A375 cells with peaks at 2 h and 9 h after treatment, respectively, although inducibility of each NKG2D ligand was various depending on cell lines. During the induction of NKG2D ligands, heat shock protein 70 was induced by heat shock but not by ionizing radiation. These results were followed by increased susceptibilities to NK cell-mediated cytolysis after treatment with heat shock and ionizing radiation. These results suggest that heat shock and ionizing radiation induce NKG2D ligands and consequently might lead to increased NK cell-mediated cytotoxicity in various cancer cells.

High-resolution melting curve analysis for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis clinical isolates.

ABSTRACT We evaluated high-resolution melting (HRM) curve analysis as a tool for detecting rifampin (RIF) and isoniazid (INH) resistance in Mycobacterium tuberculosis in an accurate, affordable, and rapid manner. Two hundred seventeen M. tuberculosis clinical isolates of known resistance phenotype were used. Twenty-nine known rpoB mutant DNAs, including rare mutations, were also included. Four pairs of primers were designed: rpoB-F/R (for codons 516 to 539 of rpoB), rpoB-516F/R (for codons 508 to 536 of rpoB), katG-F/R (for the codon 315 region of katG), and inhA-F/R (for the nucleotide substitution of C to T at position −15 of inhA). An HRM curve was generated for each isolate after real-time PCR differentiated the mutant from the wild-type strains. DNA sequencing of the target regions was performed to confirm the results of the HRM curve analysis. All but one of the 73 RIF-resistant (RIF-R) strains and all 124 RIF-susceptible (RIF-S) isolates were correctly identified by HRM curve analysis of rpoB. Twenty-seven of 29 known rpoB mutants were detected. In HRM curve analysis of katG and inhA, 90 INH-R strains that harbored katG or inhA mutations, or both, and all INH-S strains were correctly identified. Ten phenotypically INH-R strains not harboring katG or inhA mutations were not detected. The HRM curve analysis will be a useful method for detection of RIF and INH resistance in M. tuberculosis in a rapid, accurate, simple, and cost-effective manner.

Evaluation of the Sysmex UF-100 Urine Cell Analyzer as a Screening Test to Reduce the Need for Urine Cultures for Community-Acquired Urinary Tract Infection

We evaluated the UF-100 flow cytometer (TOA Medical Electronics, Kobe, Japan) as a screening test for community-acquired urinary tract infection (UTI) to reduce the need for bacterial cultures. By comparing the test results for 330 urine samples with quantitative urine cultures, we established cutoff criteria for the UF-100. To rule out hospital-acquired UTI, all urine samples were from new patients who had not been admitted to a hospital within the previous month. Abacterial cutoff value of 3,000/microL provided the best discrimination for community-acquired UTI, with a sensitivity of 94.4% and a specificity of 73.4%compared with urine culture. It was possible to forgo 58.2% of cultures with only 4 false-negative results. With a bacterial cutoff value of 1,500/microL, the sensitivity improved to 100%, but the specificity declined to 49.8%, and only 38.5% of cultures could be avoided without any false-negative results. Screening with the UF-100 for community-acquired UTI is acceptable for routine use. It would improve the efficiency of the routine microbiology laboratory, and unnecessary antibiotic prescriptions could be reduced.

Detection of YMDD Motif Mutants by Oligonucleotide Chips in Lamivudine-Untreated Patients with Chronic Hepatitis B Virus Infection

Lamivudine, a nucleoside analogue, has been used widely as an effective antiviral agent for the treatment of patients with chronic hepatitis B virus (HBV) infection. However, the YMDD motif mutation of HBV polymerase resistant to lamivudine occurs very frequently after long term therapy. We developed an oligonucleotide chip for the detection of YMDD motif mutants resistant to lamivudine and investigated the prevalence of the mutants in patients with chronic HBV infection who had not been treated by lamivudine before. Forty patients who had not been treated with lamivudine were included in this study. Serum samples were tested by the oligonucleotide chips designed for detection of wild-type YMDD motif, M552V and M552I. Samples were confirmed by restriction fragment length polymorphism (RFLP) and direct sequencing. M552I mutants were detected by the oligonucleotide chips in 7.5% (3/40) of chronic HBV infected patients (2 chronic hepatitis and 1 cirrhosis). The results were in accordance with those of RFLP. YMDD motif mutants occur as natural genome variabilities in patients with chronic HBV infection who had not been treated with lamivudine before. Oligonucleotide chip technology is a reliable and useful diagnostic tool for the detection of mutants resistant to antiviral therapy in chronic HBV infection.

Evaluation of the Performances of AdvanSure TB/NTM Real Time PCR Kit for Detection of Mycobacteria in Respiratory Specimens

BACKGROUND PCR is a widely used method for rapid and accurate diagnosis of mycobacteriosis. The sensitivity and specificity of a real time PCR kit newly developed in Korea were evaluated for detecting mycobacteria in respiratory specimens. METHODS One hundred twenty nine Mycobacterium tuberculosis (TB) culture positive respiratory specimens (82 AFB stain positive and 47 stain negative specimens) were used for evaluation of the sensitivity. Nine non-tuberculous mycobacteria (NTM) culture positive specimens were also included. For evaluation of the specificity, 48 AFB stain and culture negative respiratory specimens from patients who were initially not fully excluded from mycobacterial diseases (specificity group 1) were used. Other 51 respiratory specimens from patients who were not suspected of mycobacterial diseases were also included (specificity group 2). Real time PCR was performed by using AdvanSure TB/NTM real time PCR Kit (LG Lifescience, Korea) and SLAN real time PCR detection system (LG Lifescience). The target genes of TB and NTM were IS6110 and rpoB, respectively. RESULTS Among 129 TB culture positive specimens, 82 of 82 AFB stain positive specimens (100%) and 35 of 47 (74.5%) stain negative specimens revealed real time PCR positivity for TB, resulting in sensitivity of 90.7%. Five of nine NTM culture positive specimens resulted in real time PCR positivity for NTM (55.6%). Forty seven of 48 specimens (97.9%) and all 51 specimens (100%) of the specificity group 1 and 2, respectively, were real time PCR negative for TB and NTM. CONCLUSIONS AdvanSure TB/NTM real time PCR Kit should be useful for detecting TB in respiratory specimens with high sensitivity and specificity.

Performance of an Enzyme-Linked Immunosorbent Assay for Detection of Clonorchis sinensis Infestation in High- and Low-Risk Groups

ABSTRACT Clonorchis sinensis is still a common parasite in South Korea. An enzyme-linked immunosorbent assay (ELISA) was developed to replace the usual intradermal test, and its performance in an area of endemicity was evaluated. From 2004 to 2006, 182 adults were included. The patient group consisted of 51 adults; 43 patients showed fluke eggs by microscopy, and 8 had their disease diagnosed clinically. The negative-control group included 131 adults: 98 at low risk and 33 at high risk of infestation. Both intradermal testing and ELISA were performed for all patients, and the sensitivity and specificity were evaluated. Statistical analysis for specificity looked at two groups: those at high risk and those at low risk. The sensitivity of the C. sinensis ELISA was 80.4%, while that of the intradermal test was 56.9% (P = 0.019). The specificities of the C. sinensis ELISA in the low-risk and high-risk groups were 93.9% and 33.3%, respectively, while those of the intradermal test were 85.7% and 30.3%, respectively (P = 0.0968, P = 0.9979). The C. sinensis ELISA may be more useful than the intradermal test; however, because of its low specificity, it may not be used independently for the diagnosis of C. sinensis infestation or a prevalence survey in a high-risk group.

Decreased emergency department length of stay by application of a computerized consultation management system.

OBJECTIVES Consultation difficulty occurs in hospitals located in many countries, and it is understood that strategies to improve the emergency department (ED) consultation process are needed. The authors constructed a computerized consultation management system in the ED of a tertiary care teaching hospital to improve the consultation process and evaluate the influence of the consultation management system on ED length of stay (LOS) and the throughput process. METHODS Consultation management system software was developed and integrated into the hospital information system. The development process took place between June 2008 and May 2009. Before the development of the consultation system, ED personnel contacted on-call physicians of the specialty department, who are usually residents, by cellular phones. After the system had been developed, ED personnel selected the department and on-call physician in the specialty department using the consultation management software and activated the automatic consultation process when specialty consultation was necessary. If the treatment plan had not been registered for 3 hours, all of the residents in the specific department are notified of the delay in the treatment plan with a short message service (SMS) message. If an admission or discharge order had not been made in 6 hours, all of the residents and faculty staff in the specific department receive SMS messages stating the delay in disposition. ED patient data were collected from the hospital information system for 40 days before the system was developed (June 1, 2008, to July 10, 2008) and 40 days after the system was implemented (June 1, 2009, to July 10, 2009). RESULTS The median ED LOS decreased significantly, from 417.5 minutes (interquartile range [IQR] = 178.8-1,247.5 minutes) in the presystem period to 311.0 minutes (IQR = 128.0-817.3 minutes) in the postsystem period (p < 0.001). Also, the median time to disposition order decreased significantly, from 336.0 minutes (IQR = 145.0-943.0 minutes) to 235.0 minutes (IQR = 103.0-21.5; p = 0.001). No significant reduction was observed in the interval between the time of disposition decision and the time when the patients left the ED. Significant reductions of ED LOS were observed after implementing the system (p < 0.001) regardless of whether the visit occurred during the weekday daytime (09:00-17:00 hours), holiday and weekend daytime (09:00-17:00 hours), or nighttime (17:00-09:00 hours next day). CONCLUSIONS This study found decreased ED LOS by implementation of a computerized consultation management system in a tertiary care teaching hospital. The automated consultation and monitoring process formalized communication between physicians providing ED patient care in the academic ED with high consultation and admission rates.

Optimization of cryopreservation condition for hematopoietic stem cells from umbilical cord blood.

The conditions for cryopreservation of CD34(+) hematopoietic stem cells (HSC) from umbilical cord blood (UCB) were optimized with a new cryo-medium containing 10% ethylene glycol (EG) and 2% dimethyl sulfoxide (Me(2)SO) using a controlled-rate freezing (CRF) method. After the cryopreservation of mononuclear cells (MNC) from UCB, recoveries of MNC, CD34(+) cells, and total colony-forming units (CFU) were significantly improved compared to those in the control cryo-medium containing 10% Me(2)SO and 2% Dextran-40 (P<0.05). This study shows that the new cryo-medium and CRF method provide better recoveries of MNC, HSC and total CFU than the control cryo-medium and isopropylalcohol freezing (IPA) method. Therefore, this cryo-medium, combined with the CRF method, is valuable for optimizing cryopreservation conditions for HSC from UCB to obtain satisfactory HSC recovery.

Association of chronic viral hepatitis B with insulin resistance

AIM To investigate the relationship between chronic viral hepatitis B (CVHB) and insulin resistance (IR) in Korean adults. METHODS A total of 7880 adults (3851 men, 4029 women) who underwent a comprehensive medical examination were enrolled in this study. Subjects diagnosed with either diabetes mellitus, or any other disorder that could influence their insulin sensitivity, were rejected. Anthropometry, metabolic risk factors, hepatitis B surface antigen, hepatitis B surface antibody, hepatitis B core antibody, fasting plasma glucose and insulin were measured for all subjects. Homeostasis model assessment (HOMA), quantitative insulin check index (QUICKI), and Mf(fm) index were used for determining insulin sensitivity. Each participant was categorized into a negative, recovery, or CVHB group. To compare variables between groups, a t-test and/or one-way analysis of variance were used. Partial correlation coefficients were computed to present the association between insulin resistance and other variables. Multiple logistic regression analysis was used to assess the independent association between CVHB and IR. RESULTS The mean age of men and women were 48.9 and 48.6 years, respectively. Subjects in the CVHB group had significantly higher waist circumference [(86.0 ± 7.7 cm vs 87.3 ± 7.8 cm, P = 0.004 in men), (78.3 ± 8.6 cm vs 80.5 ± 8.5 cm, P < 0.001 in women)], cystatin C [(0.96 ± 0.15 mg/dL vs 1.02 ± 0.22 mg/dL, P < 0.001 in men), (0.84 ± 0.15 mg/dL vs 0.90 ± 0.16 mg/dL, P < 0.001 in women)], fasting insulin [(5.47 ± 3.38 μU/mL vs 6.12 ± 4.62 μU/mL, P < 0.001 in men), (4.57 ± 2.82 μU/mL vs 5.06 ± 3.10 μU/mL, P < 0.001 in women)] and HOMA index [(1.24 ± 0.86 vs 1.43 ± 1.24, P < 0.001 in men), (1.02 ± 0.76 vs 1.13 ± 0.87, P = 0.033 in women)] compared to control group. The HOMA index revealed a positive correlation with body mass index (BMI) (r = 0.378, P < 0.001), waist circumference (r =0.356, P < 0.001), percent body fat (r = 0.296, P < 0.001), systolic blood pressure (r = 0.202, P < 0.001), total cholesterol (r = 0.134, P < 0.001), triglycerides (r = 0.292, P < 0.001), cystatin C (r = 0.069, P < 0.001) and uric acid (r = 0.142, P < 0.001). The QUICKI index revealed a negative correlation with BMI (r = -0.254, P < 0.001), waist circumference (r = 0-0.243, P < 0.001), percent body fat (r = -0.217, P < 0.001), systolic blood pressure (r = -0.132, P < 0.001), total cholesterol (r = -0.106, P < 0.001), triglycerides (r = -0.205, P < 0.001), cystatin C (r = -0.044, P < 0.001) and uric acid (r = -0.096, P < 0.001). For subjects identified with IR, the odds ratio of an accompanying diagnosis of chronic hepatitis B was 1.534 (95% CI: 1.158-2.031, HOMA index criteria) or 1.566 (95% CI: 1.124-2.182, QUICKI criteria) after adjustment for age, gender, BMI, and amount of alcohol consumption. CONCLUSION Our study demonstrates that CVHB is associated with IR. CVHB may need to be monitored for occurrence of IR and diabetes mellitus.

Usefulness of Plasma Procalcitonin to Predict Severity in Elderly Patients with Community-Acquired Pneumonia

Background Community-acquired pneumonia (CAP) is one of the leading causes of death among the elderly. Several studies have reported the clinical usefulness of serum procalcitonin, a biomarker of bacterial infection. However, the association between the levels of procalcitonin and the severity in the elderly with CAP has not yet been reported. The aim of this study was to evaluate usefulness of procalcitonin as a predictor of severity and mortality in the elderly with CAP. Methods This study covers 155 CAP cases admitted to Pusan National University Hospital between January 2010 and December 2010. Patients were divided into two groups (≥65 years, n=99; <65 years, n=56) and were measured for procalcitonin, C-reactive protein (CRP), white blood cell, confusion, uremia, respiratory rate, blood pressure, 65 years or older (CURB-65) and pneumonia severity of index (PSI). Results The levels of procalcitonin were significantly correlated with the CURB-65, PSI in totals. Especially stronger correlation was observed between the levels of procalcitonin and CURB-65 in the elderly (procalcitonin and CURB-65, ρ=0.408 with p<0.001; procalcitonin and PSI, ρ=0.293 with p=0.003; procalcitonin and mortality, ρ=0.229 with p=0.023). The correlation between the levels of CRP or WBC and CAP severity was low. The existing cut-off value of procalcitonin was correlated with mortality rate, however, it was not correlated with mortality within the elderly. Conclusion The levels of procalcitonin are more useful than the levels of CRP or WBC to predict the severity of CAP. However, there was no association between the levels of procalcitonin and mortality in the elderly.