1.Production of Antibody against Helicobacter pylori HP0231.
Jongwook LEE ; Soon Youl LEE ; Jae Hag LEE
The Korean Journal of Laboratory Medicine 2006;26(2):98-102
BACKGROUND: Stool antigen detection kits for diagnosis of infection of Helicobacter pylori have been widely used for their convenience, but are mostly imported. Since Helicobacter pylori strains show a distinctive genetic diversity, it is important to find a protein that is a common antigen among various strains and shows a strong immunogenicity for the development of a stool antigen detection kit. HP0231 protein strongly reacts with the sera of patients suffering from gastritis and peptic ulcer. Therefore, HP0231 is an excellent candidate as a target gene for this study. METHODS: Chromosomal DNA from H. pylori was isolated. HP0231 gene was amplified by PCR, cloned into pET28a(+) vector, and overexpressed using isopropyl-beta-D-thiogalactopyranoside in E. coli BL21 (DE3). HP0231 protein was purified by Ni-NTA affinity chromatography followed by electroelution after SDS-PAGE. Rabbits were immunized with the purified HP0231 protein for the production of antibodies. Rabbit anti-HP0231 antibody was partially purified and tested for the sensitivity and specificity using ELISA and Western Blot Analysis. RESULTS: The sequence of the cloned HP0231 gene was identical with the gene sequence from Genbank (AA216016). HP0231 gene was overexpressed and HP0231 protein was purified. Rabbit anti-HP0231 antibody produced after immunization with the purified HP0231 protein reacted with the purified HP0231 protein, cell extracts from cultured H. pylori, and stomach biopsy tissue from patients, but not with cell extracts from cultured E. coli used as a negative control. After 1 million fold dilution, rabbit anti-HP0231 antibody still reacted with 1 microgram of HP0231 protein. CONCLUSIONS: Rabbit anti-HP0231 antibody was produced to detect HP0231 protein of H. pylori and will be tested for the development of a stool antigen detection kit for H. pylori.
Antibodies
;
Biopsy
;
Blotting, Western
;
Cell Extracts
;
Chromatography, Affinity
;
Clone Cells
;
Databases, Nucleic Acid
;
Diagnosis
;
DNA
;
Electrophoresis, Polyacrylamide Gel
;
Enzyme-Linked Immunosorbent Assay
;
Gastritis
;
Genetic Variation
;
Helicobacter pylori*
;
Helicobacter*
;
Humans
;
Immunization
;
Peptic Ulcer
;
Polymerase Chain Reaction
;
Rabbits
;
Sensitivity and Specificity
;
Stomach
2.Clinical Significance of von Willebrand Factor-Cleaving Protease (ADAMTS13) Deficiency in Patients with Sepsis-Induced Disseminated Intravascular Coagulation.
Young Keun KIM ; Jongwook LEE ; Kyung A LEE ; Hee Uk KWON
Infection and Chemotherapy 2009;41(2):78-81
BACKGROUND: Deficiency of von Willebrand factor-cleaving protease, a disintegrin-like and metalloprotease with thrombospondin type 1 motif 13 (ADAMTS13), is thought to be responsible for platelet aggregation and microthrombi formation, which in turn cause typical thrombotic microangiopathies. This deficiency is found in patients with thrombocytopenia-associated multiple organ failure such as thrombocytopenic purpura and disseminated intravascular coagulation (DIC). We evaluated the clinical significance of ADAMTS13 deficiency in patients with sepsis-induced DIC. MATERIALS AND METHODS: Nineteen patients with sepsis-induced DIC were enrolled. ADAMTS13 antigen levels were determined by Enzyme-Linked Immunosorbent Assay (ELISA) and activity levels were measured by fluorescence resonance energy transfer assay. Patients were categorized into two groups according to ADAMTS13 antigen level: less than 350 ng/mL or above. Clinical characteristics and survival were compared between the two groups. RESULTS: ADAMTS13 antigen level was less than 350 ng/mL in 7 patients and was above 350 ng/mL in 12 patients. There were no significant differences between the groups for age, sex, severity of illness, and other clinical characteristics. In patients with ADAMTS13 antigen level less than 350 ng/mL, in-hospital mortality was much higher (100% versus 25%, P=0.003) and 7-day survival was much shorter (P=0.023). CONCLUSION: Deficiency of ADAMTS13 could be thought to be associated with unfavorable outcome in patients with sepsis-induced DIC.
Dacarbazine
;
Disseminated Intravascular Coagulation
;
Enzyme-Linked Immunosorbent Assay
;
Fluorescence Resonance Energy Transfer
;
Hospital Mortality
;
Humans
;
Multiple Organ Failure
;
Platelet Aggregation
;
Prognosis
;
Purpura, Thrombocytopenic
;
Sepsis
;
Thrombospondins
;
Thrombotic Microangiopathies
3.Clinical Significance of von Willebrand Factor-Cleaving Protease (ADAMTS13) Deficiency in Patients with Sepsis-Induced Disseminated Intravascular Coagulation.
Young Keun KIM ; Jongwook LEE ; Kyung A LEE ; Hee Uk KWON
Infection and Chemotherapy 2009;41(2):78-81
BACKGROUND: Deficiency of von Willebrand factor-cleaving protease, a disintegrin-like and metalloprotease with thrombospondin type 1 motif 13 (ADAMTS13), is thought to be responsible for platelet aggregation and microthrombi formation, which in turn cause typical thrombotic microangiopathies. This deficiency is found in patients with thrombocytopenia-associated multiple organ failure such as thrombocytopenic purpura and disseminated intravascular coagulation (DIC). We evaluated the clinical significance of ADAMTS13 deficiency in patients with sepsis-induced DIC. MATERIALS AND METHODS: Nineteen patients with sepsis-induced DIC were enrolled. ADAMTS13 antigen levels were determined by Enzyme-Linked Immunosorbent Assay (ELISA) and activity levels were measured by fluorescence resonance energy transfer assay. Patients were categorized into two groups according to ADAMTS13 antigen level: less than 350 ng/mL or above. Clinical characteristics and survival were compared between the two groups. RESULTS: ADAMTS13 antigen level was less than 350 ng/mL in 7 patients and was above 350 ng/mL in 12 patients. There were no significant differences between the groups for age, sex, severity of illness, and other clinical characteristics. In patients with ADAMTS13 antigen level less than 350 ng/mL, in-hospital mortality was much higher (100% versus 25%, P=0.003) and 7-day survival was much shorter (P=0.023). CONCLUSION: Deficiency of ADAMTS13 could be thought to be associated with unfavorable outcome in patients with sepsis-induced DIC.
Dacarbazine
;
Disseminated Intravascular Coagulation
;
Enzyme-Linked Immunosorbent Assay
;
Fluorescence Resonance Energy Transfer
;
Hospital Mortality
;
Humans
;
Multiple Organ Failure
;
Platelet Aggregation
;
Prognosis
;
Purpura, Thrombocytopenic
;
Sepsis
;
Thrombospondins
;
Thrombotic Microangiopathies
4.Anchorage Dependence and Cancer Metastasis
Dong Ki LEE ; Jongwook OH ; Hyun Woo PARK ; Heon Yung GEE
Journal of Korean Medical Science 2024;39(19):e156-
The process of cancer metastasis is dependent on the cancer cells’ capacity to detach from the primary tumor, endure in a suspended state, and establish colonies in other locations.Anchorage dependence, which refers to the cells’ reliance on attachment to the extracellular matrix (ECM), is a critical determinant of cellular shape, dynamics, behavior, and, ultimately, cell fate in nonmalignant and cancer cells. Anchorage-independent growth is a characteristic feature of cells resistant to anoikis, a programmed cell death process triggered by detachment from the ECM. This ability to grow and survive without attachment to a substrate is a crucial stage in the progression of metastasis. The recently discovered phenomenon named “adherent-to-suspension transition (AST)” alters the requirement for anchoring and enhances survival in a suspended state. AST is controlled by four transcription factors (IKAROS family zinc finger 1, nuclear factor erythroid 2, BTG anti-proliferation factor 2, and interferon regulatory factor 8) and can detach cells without undergoing the typical epithelialmesenchymal transition. Notably, AST factors are highly expressed in circulating tumor cells compared to their attached counterparts, indicating their crucial role in the spread of cancer.Crucially, the suppression of AST substantially reduces metastasis while sparing primary tumors. These findings open up possibilities for developing targeted therapies that inhibit metastasis and emphasize the importance of AST, leading to a fundamental change in our comprehension of how cancer spreads.
5.Treatment Outcomes and Response Pattern of Ustekinumab in Korean Patients with Psoriasis: A Retrospective Single-center Study
Jongwook OH ; TaeGyun KIM ; Min Geol LEE
Korean Journal of Dermatology 2019;57(8):441-447
BACKGROUND: Psoriasis is a chronic immune-mediated inflammatory skin disease affecting 2~3% of the worldwide population. Ustekinumab, an IL-12/23p40 inhibitor, is a biologic reported to be effective and safe in treating psoriasis. However, there are limited data on the treatment outcomes of ustekinumab in patients with psoriasis in Korea. OBJECTIVE: To evaluate the treatment outcomes and response pattern of ustekinumab in patients with psoriasis in Korea. METHODS: This was a retrospective single-center study. Eighty-four patients with psoriasis treated with ustekinumab were analyzed. Each patient's medical records, psoriasis area and severity index (PASI) score, and body surface area were reviewed at baseline and up to week 52. RESULTS: A total of 84 patients were included (male:female=1.8:1). The mean age was 44.5 years. At week 16, 86.7% achieved PASI75, 59.0% achieved PASI90, and 20.5% achieved PASI100. By week 16, 84.8% of subjects had attained PASI75 for the head region, whereas 79.0% had attained it for the lower extremities, indicating a relatively slower treatment response of psoriatic lesions on the lower extremities. Four patients discontinued treatment due to lack of effect. No severe adverse events occurred during the follow-up period. CONCLUSION: Ustekinumab demonstrated highly effective and safe treatment profiles in Korean psoriatic patients, consistent with the previous reports from mainly Western countries. Psoriasis severity and treatment responsiveness may vary with body region.
Body Regions
;
Body Surface Area
;
Follow-Up Studies
;
Head
;
Humans
;
Korea
;
Lower Extremity
;
Medical Records
;
Psoriasis
;
Retrospective Studies
;
Skin Diseases
;
Ustekinumab
6.Evaluation of Asan Helicobacter Test for Diagnosis of Helicobacter pylori Infection.
Jongwook LEE ; Pum Soo KIM ; Kyungwon LEE ; Jae Hag LEE ; Jae Kyoung LEE ; Chang Hoon LEE
Korean Journal of Clinical Microbiology 2003;6(2):156-159
BACKGROUND: Helicobacter pylori is the single most common pathogen that causes chronic bacterial infection in human. The authors designed a new type of urease detection method (Asan Helicobacter test) that can be used for rapid early detection of H. pylori as well as a transport medium. This medium has a strong acidity with a minimal concentration of urea for the purpose of the detection of H. pylori. The current study was to evaluate the bacteriological and clinical usefulness of this medium. METHOD: 252 antral biopsies from patients underwent upper gastrointestinal endoscopies in Inha University Hospital were inserted Asan Helicobacter Test and CLO test. 37 antral biopsies from patients underwent upper gastrointestinal endoscopies in Konyang University Hospital were inserted Asan Helicobacter Test. Biopsies were cultured on nonselective media only. RESULT: The sensitivity and specificity of the Asan Helicobacter test were comparable with the CLO test (88.0% and 94.0%, respectively), and the results agreed in 99.2% of 252 cases with the CLO test. With this transport medium, all 23 specimens that showed positive reaction among 37 patients yielded satisfactory isolation of H. pylori. CONCLUSION: These findings suggest that the reagent in the kit inhibit the growth of microbial contaminant due to low pH and do not suppresses growth of H. pylori due to low concentration of urea. This kit may be used as a transport medium as well as a rapid urease test for H. pylori.
Bacterial Infections
;
Biopsy
;
Chungcheongnam-do*
;
Diagnosis*
;
Endoscopy, Gastrointestinal
;
Helicobacter pylori*
;
Helicobacter*
;
Humans
;
Hydrogen-Ion Concentration
;
Sensitivity and Specificity
;
Urea
;
Urease
7.Prevalence of Helicobacter pylori infection and clarithromycin resistance rate from 2015 to 2018 using the laboratory information system of the Seegene Medical Foundation in Korea: a repeated cross-sectional study
Sunkyung JUNG ; Mi-Na KIM ; Dongeun YONG ; Miae LEE ; Jongwook LEE ; Hae Kyung LEE ; Mi-Kyung LEE ;
Annals of Clinical Microbiology 2024;27(1):19-30
Background:
Numerous studies have examined the prevalence of Helicobacter pylori infection and clarithromycin (CLA) resistance rate of H. pylori. However, in South Korea, there is a lack of research analyzing specimens from local clinics and hospitals using molecular methods. This study aimed to assess the prevalence of H. pylori infection and CLA resistance across sex and age groups, as well as to explore regional variations in CLA resistance and its characteristics.
Methods:
Data from a laboratory information system from 2015 to 2018 were retrospectively analyzed to determine the prevalence of H. pylori infection and CLA resistance rate. The 23S ribosomal RNA genes of H. pylori were analyzed using a dual priming oligonucleotide-based multiplex polymerase chain reaction method.
Results:
The overall prevalence of H. pylori infection was 50.5%(12,000/23,773), with a significantly higher prevalence among males (53.5%) than females (47.0%). The CLA resistance rate was 28.3%, with a significantly higher rate among females (34.9%) than males (23.8%). Age group analysis revealed that the highest prevalence of H. pylori infection was among individuals in their 40s, whereas the highest CLA resistance rate was observed among those in their 60s. The CLA resistance rate exhibited an upward trend and varied among patients based on their place of residence, and A2143G mutation was the most prevalent across all regions.
Conclusion
The prevalence of H. pylori infection and CLA resistance rate in Korea remain high and vary according to sex, age, and region. To effectively eradicate H. pylori, it is crucial to periodically monitor regional CLA resistance patterns and conduct CLA susceptibility testing before prescription.
8.Emergence of Plasmid-Mediated CMY-2 beta -Lactamase Produced by Clinical Isolates of Escherichia coli in Korea.
Chang Hoon LEE ; Jae Seok KIM ; Young UH ; Jongwook LEE ; Kyungwon LEE ; Wonkeun SONG
The Korean Journal of Laboratory Medicine 2005;25(2):98-103
BACKGROUND: Of the plasmid-mediated AmpC beta-lactamases (ABLs), CMY-2 is the most prevalent and is distributed in many countries. However, little is known about the emergence and characteristics of CMY-2 among Escherichia coli isolates in Korea. The aims of this study were to detect the emergence of the CMY-2 beta-lactamase in clinical isolates of E. coli from various regions in Korea. METHODS: Eighteen cefoxitin non-susceptible isolates of 1, 130 consecutive, nonrepeat isolates of E. coli at five university hospitals were tested for antimicrobial susceptibility by the broth microdilution method. The cefoxitin non-susceptible isolates were further investigated by AmpC disk tests, double disk synergy (DDS) tests, isoelectric focusing, CMY-2-specific PCR, DNA sequencing, and plasmid analysis. RESULTS: Seven (0.6%) isolates of plasmid-mediated ABL-producing E. coli were found at three of the five hospitals; all seven isolates produced CMY-2 beta-lactamase and one of the isolates was also tested positive by the DDS test. All isolates demonstrated different plasmid patterns by plasmid analysis. CONCLUSIONS: Our data indicate that CMY-2-producing E. coli has emerged and is prevalent in the medical institution in Korea. Therefore, constant surveillance is needed to prevent its further spread.
beta-Lactamases
;
Cefoxitin
;
Escherichia coli*
;
Hospitals, University
;
Isoelectric Focusing
;
Korea
;
Plasmids
;
Polymerase Chain Reaction
;
Sequence Analysis, DNA
9.Evaluation of Urea Breath Test for the Detection of Helicobacter pylori Infection.
Jongwook LEE ; Nam Keum LEE ; Soo Hwan PAI ; Pum Soo KIM ; Won CHOI ; Don Hang LEE ; Hyung GIL ; Young Soo KIM
Korean Journal of Clinical Microbiology 2000;3(2):111-115
BACKGROUND: Helicobacter pylori (H. pylori) is closely associated with gastritis, peptic ulcer and gastric carcinoma. We evaluated the reliability and usefulness of 73C-urea breath test (13C-UBT) for the detection of H. pylori infection and searched for the cut-off value of the test. METHOD : We investigated 45 patients, who underwent esophagoduodenoscopy with multiple biopsy specimens taken for culture, histology and rapid urease test, and 13C-UBT. Sensitivity and specificity of UBT were calculated against the combined biopsy-based test results. RESULT: Of 45 patients, 26 were found to be H. pylori-positive according to combined biopsy-based test-results. Sensitivity and specificity of the 13C-UBT were 100.0% and 89.5 %, respectively. CONCLUSION: The urea breath test provides a simple and reliable and noninvasive method of assessing HL pylori infection status.
Biopsy
;
Breath Tests*
;
Gastritis
;
Helicobacter pylori*
;
Helicobacter*
;
Humans
;
Peptic Ulcer
;
Sensitivity and Specificity
;
Urea*
;
Urease
10.Evaluation of Urea Breath Test for the Detection of Helicobacter pylori Infection.
Jongwook LEE ; Nam Keum LEE ; Soo Hwan PAI ; Pum Soo KIM ; Won CHOI ; Don Hang LEE ; Hyung GIL ; Young Soo KIM
Korean Journal of Clinical Microbiology 2000;3(2):111-115
BACKGROUND: Helicobacter pylori (H. pylori) is closely associated with gastritis, peptic ulcer and gastric carcinoma. We evaluated the reliability and usefulness of 73C-urea breath test (13C-UBT) for the detection of H. pylori infection and searched for the cut-off value of the test. METHOD : We investigated 45 patients, who underwent esophagoduodenoscopy with multiple biopsy specimens taken for culture, histology and rapid urease test, and 13C-UBT. Sensitivity and specificity of UBT were calculated against the combined biopsy-based test results. RESULT: Of 45 patients, 26 were found to be H. pylori-positive according to combined biopsy-based test-results. Sensitivity and specificity of the 13C-UBT were 100.0% and 89.5 %, respectively. CONCLUSION: The urea breath test provides a simple and reliable and noninvasive method of assessing HL pylori infection status.
Biopsy
;
Breath Tests*
;
Gastritis
;
Helicobacter pylori*
;
Helicobacter*
;
Humans
;
Peptic Ulcer
;
Sensitivity and Specificity
;
Urea*
;
Urease