CCR3 is a chemokine receptor that mediates the accumulation of allergic inflammatory cells, including eosinophils and Th2 cells, at inflamed sites. The regulatory sequence of the CCR3 gene, contains two Runt-related transcription factor (RUNX) 1 sites and two PU.1 sites, in addition to a functional GATA site for transactivation of the CCR3 gene. In the present study, we examined the effects of the cis-acting elements of RUNX1 and PU.1 on transcription of the gene in EoL-1 eosinophilic cells and Jurkat T cells, both of which expressed functional surface CCR3 and these two transcription factors. Introduction of RUNX1 siRNA or PU.1 siRNA resulted in a modest decrease in CCR3 reporter activity in both cell types, compared with transfection of GATA-1 siRNA. Cotransfection of the two siRNAs led to inhibition in an additive manner. EMSA analysis showed that RUNX1, in particular, bound to its binding motifs. Mutagenesis analysis revealed that all point mutants lacking RUNX1- and PU.1-binding sites exhibited reduced reporter activities. These results suggest that RUNX1 and PU.1 participate in transcriptional regulation of the CCR3 gene.
Eosinophils ; Mutagenesis ; RNA, Small Interfering ; T-Lymphocytes ; Th2 Cells ; Transcription Factors ; Transcriptional Activation ; Transfection

Neutrophils and eosinophils, 2 prominent granulocytes, are commonly derived from myelocytic progenitors through successive stages in the bone marrow. Our previous genome-wide transcriptomic data unexpectedly showed that genes encoding a multitude of neutrophil primary granule proteins (NPGPs) were markedly downregulated during the end period of eosinophilic terminal differentiation when cord blood (CB) cluster of differentiation (CD) 34+ cells were induced to differentiate toward the eosinophil lineage during a 24-day culture period. Accordingly, this study aimed to examine whether NPGP genes were expressed on the way to eosinophil terminal differentiation stage and to compare their expression kinetics with that of genes encoding eosinophil-specific granule proteins (ESGPs). Transcripts of all NPGP genes examined, including proteinase 3, myeloperoxidase, cathepsin G (CTSG), and neutrophil elastase, reached a peak at day 12 and sharply declined thereafter, while transcript of ESGP genes including major basic protein 1 (MBP1) attained maximum expression at days 18 or 24. Growth factor independent 1 (GFI1) and CCAAT/enhancer-binding protein α (C/EBPA), transactivators for the NPGP genes, were expressed immediately before the NPGP genes, whereas expression of C/EBPA, GATA1, and GATA2 kinetically paralleled that of eosinophil granule protein genes. The expression kinetics of NPGPs and ESGPs were duplicated upon differentiation of the eosinophilic leukemia cell line (EoL-1) immature eosinophilic cells. Importantly, confocal image analysis showed that CTSG was strongly coexpressed with MBP1 in differentiating CB eosinophils at days 12 and 18 and became barely detectable at day 24 and beyond. Our results suggest for the first time the presence of an immature stage where eosinophils coexpress NPGPs and ESGPs before final maturation.
Bone Marrow ; Cathepsin G ; Cell Line ; Eosinophils* ; Fetal Blood ; Granulocytes ; Hypereosinophilic Syndrome ; Kinetics ; Leukocyte Elastase ; Myeloblastin ; Neutrophils* ; Peroxidase ; Trans-Activators

PURPOSE: Epigenetic alterations of specific genes have recently been identified as diagnostic biomarkers for human cancers. However, there are currently no standardized epigenetic biomarkers for drug sensitivity in human gastrointestinal cancer. Therefore, the aim of this study is to identify a novel epigenetic biomarker in gastrointestinal cancer. MATERIALS AND METHODS: Using bisulfite sequencing and pyrosequencing analysis, DNA methylation patterns of gastric, colon primary tissues and their cancer cells were analyzed, and histone modifications were analyzed using chromatin immunoprecipitation assay. In addition, cancer cells were exposed to cisplatin and treated with a DNA methyltransferase inhibitor. RESULTS: We report that in human gastric and colon cancers, latrophilin 2 (LPHN2) is silenced by epigenetic modifications, including CpG island methylation and aberrant histone modifications. We also confirmed that LPHN2 was silenced by DNA hypermethylation in primary gastric and colon tumor tissues compared to their normal counterparts. Interestingly, we found that cancer cells with methylated LPHN2 showed higher sensitivity to cisplatin. Also, 5-aza- 2′-deoxycytidine combined with cisplatin decreased the cytotoxicity of cisplatin in cancer cells with methylated LPHN2. In addition, LPHN2 knockdown in cancer cells with high LPHN2 expression sensitized these cells to the anti-proliferative effects of cisplatin. CONCLUSION: In human gastrointestinal cancer, we found that LPHN2 is regulated by epigenetic modifications, and that cancer cells with lower LPHN2 expression show higher sensitivity to cisplatin. Therefore, the methylation status of LPHN2 is a potential novel epigenetic biomarker for cisplatin treatment in human gastric and colon cancers.
Biological Markers ; Chromatin Immunoprecipitation ; Cisplatin ; Colon ; Colonic Neoplasms ; CpG Islands ; DNA ; DNA Methylation ; Epigenomics* ; Gastrointestinal Neoplasms* ; Histones ; Humans* ; Methylation

BACKGROUND: Acute renal failure is one of the leading causes of postoperative morbidity and mortality. The purpose of this study was to determine the risk factors that are associated with acute renal failure after colorectal surgery. MATERIALS AND METHODS: Five hundred seventy patients who operated colorectal surgery at the Yeungnam University Medical Center over three years from 2004 to 2006 were enrolled in this study. The effects of gender, age, ASA classification, concomitant disease, surgery type and duration, reoperation, urogenital manipulation, medication, hypotension, hypovolemia, transfusion, and postoperative ventilatory care on the occurrence of acute renal failure after colorectal surgery were studied. RESULTS: The major risk factors of acute renal failure after colorectal surgery were age of patients (P=0.003), ASA classification (P<0.001), concomitant disease (P<0.001), duration of the time surgery (P=0.034), reoperation (P=0.001), use of intraoperative diuretics (P=0.005), use of postoperative diuretics (P<0.001), intraoperative hypotension (P=0.018), intraoperative transfusion (P<0.001), postoperative transfusion (P<0.001), and postoperative ventilatory care (P=0.001). CONCLUSION: Multiple factors cause synergistic effects on the development of acute renal failure after colorectal surgery. Therefore, efforts to reduce the risk factors associated with acute renal failure are needed. In addition, intensive postoperative care should be provided to all patients.
Academic Medical Centers ; Acute Kidney Injury* ; Classification ; Colorectal Surgery* ; Diuretics ; Humans ; Hypotension ; Hypovolemia ; Mortality ; Postoperative Care ; Reoperation ; Risk Factors*

BACKGROUND: The purpose of this study was to determine the adequate loading and maintenance doses of N-acetylcyseteine (NAC) for patients suffering from acute ROS-induced injury. METHODS: Concentrations of extra cellular NAC, cysteine (Cys), cystine (Cyst2), and methionine (Met) were measured in vitro, at which more than 50% of the intracellular ROS raised by paraquat were suppressed using Swiss 3T3 fibroblasts. An in vivo pharmacokinetic study followed on a healthy subject to determine the proper loading and maintenance doses of reduced NAC following intravenous administration of 25 mg/kg NAC. RESULTS: In vivo, NAC suppressed ROS in a dose dependant manner. 10 mM of NAC suppressed about 50% of ROS, and was comparable to 10 micro M of Cys and Met and 400 micro M of Cys2. In vitro, the elimination of half-life was achieved at 2.88+/-1.14 h for NAC and at 3.68+/-1.84 h for total NAC. The body clearances were 1.23+/-0.77 L h (-1) kg (-1) and 0.56+/-0.27 L h (-1) kg (-1) and the volumes of distribution were 3.07+/-0.10 L kg (-1) and 3.00+/-0.11 L kg (-1), respectively. The loading and maintenance NAC doses used to reach the target concentration of 10 mM, were 5010 mg. kg (-1) and 2250 mg min (-1) kg (-1), respectively. CONCLUSION: NAC provides an antioxidant effect on ROS produced by paraquat in vivo. However, in vitro, our results showed that the intravenous NAC dose could not be estimated from NAC plasma concentration or its metabolites.
Sulfur/*blood ; Reactive Oxygen Species ; Humans ; Glutathione/blood ; Amino Acids/*blood/chemistry ; Acetylcysteine/*administration & dosage/pharmacokinetics/pharmacology

Objective: Here, we investigated whether cytokines in the cervicovaginal fluid (CVF) can be predictive markers of preterm birth (PTB). Methods: A multi-center prospective cohort study was conducted on 59 singleton pregnant women hospitalized for preterm labor (PTL) and/or preterm premature rupture of membranes (pPROM) between 22 weeks and 36 weeks 6 days of gestation from 2014 to 2015. The levels of 13 inflammatory cytokines (macrophage inflammatory protein [MIP]-1α, MIP-1β, tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, IL-8, IL-17α, granulocyte colony stimulating factor [G-CSF], IL-7, IL-4, IL-5, IL-10, and IL-13) were measured using a multiplex bead-based immunoassay and that of fetal fibronectin (fFN) was measured using enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed using Student’s t-test, Mann-Whitney U test, Pearson’s correlation, and receiver operating characteristic (ROC) curve analysis in SPSS version 20.0. Results: Among the 13 cytokines assessed, the levels of 3 cytokines (MIP-1α, IL-6, and IL-7) were negatively correlated with gestational age at delivery (P=0.028, P=0.002, and P=0.018, respectively). Sensitivities of MIP-1α, IL-6, and IL-17α were 70%, 80%, and 75%, respectively, and their specificities were 57%, 65%, and 69%, respectively. The sensitivity and specificity of fFN were 33% and 95%, respectively. Conclusion In symptomatic women diagnosed with PTL and/or pPROM, cytokines from cervicovaginal fluid, especially IL-6 and IL-17α, could be better predictive markers of PTB than fFN.

Objective: Here, we investigated whether cytokines in the cervicovaginal fluid (CVF) can be predictive markers of preterm birth (PTB). Methods: A multi-center prospective cohort study was conducted on 59 singleton pregnant women hospitalized for preterm labor (PTL) and/or preterm premature rupture of membranes (pPROM) between 22 weeks and 36 weeks 6 days of gestation from 2014 to 2015. The levels of 13 inflammatory cytokines (macrophage inflammatory protein [MIP]-1α, MIP-1β, tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, IL-8, IL-17α, granulocyte colony stimulating factor [G-CSF], IL-7, IL-4, IL-5, IL-10, and IL-13) were measured using a multiplex bead-based immunoassay and that of fetal fibronectin (fFN) was measured using enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed using Student’s t-test, Mann-Whitney U test, Pearson’s correlation, and receiver operating characteristic (ROC) curve analysis in SPSS version 20.0. Results: Among the 13 cytokines assessed, the levels of 3 cytokines (MIP-1α, IL-6, and IL-7) were negatively correlated with gestational age at delivery (P=0.028, P=0.002, and P=0.018, respectively). Sensitivities of MIP-1α, IL-6, and IL-17α were 70%, 80%, and 75%, respectively, and their specificities were 57%, 65%, and 69%, respectively. The sensitivity and specificity of fFN were 33% and 95%, respectively. Conclusion In symptomatic women diagnosed with PTL and/or pPROM, cytokines from cervicovaginal fluid, especially IL-6 and IL-17α, could be better predictive markers of PTB than fFN.

BACKGROUND/AIMS: Data are lacking regarding the management of chronic hepatitis B (CHB) with resistance to clevudine (CLV). This study evaluated the efficacy of different rescue therapies for CLV-resistant CHB. METHODS: Patients with CLV-resistant CHB were enrolled in the cohort, and all patients developed virologic breakthrough during CLV therapy and had confirmed-genotypic resistance to CLV (rtM204I mutation) before enrollment. RESULTS: Of the 107 patients, 12 received adefovir (ADV), 21 received a CLV plus ADV combination (CLV+ADV), 34 received a lamivudine plus ADV combination (LAM+ADV), and 40 received entecavir (ETV) therapy for 48 weeks. The CLV+ADV group had the lowest hepatitis B virus (HBV) DNA level (p<0.0001) and showed the greatest reduction of HBV DNA levels from baseline compared to all other groups (p=0.004) at week 48. HBV DNA was undetectable (<70 IU/mL) in 0%, 57.1%, 21.2%, and 27.5% (p=0.003) of the patients in each group, respectively, at week 48. At the end of the study, the mean alanine transaminase (ALT) level, rate of ALT normalization, and rate of hepatitis B envelope antigen loss or seroconversion did not differ between groups. CONCLUSIONS: CLV+ADV combination therapy in patients with CLV-resistant CHB more effectively suppresses HBV replication than ETV, ADV, or LAM+ADV therapy.
Alanine Transaminase ; Cohort Studies* ; DNA ; Hepatitis B ; Hepatitis B virus ; Hepatitis B, Chronic* ; Hepatitis, Chronic* ; Humans ; Lamivudine ; Seroconversion

BACKGROUND: Cefditoren is a third generation orally administered cephalosporin with excellent activity against respiratory pathogens. This study was performed to determine the comparative antibacterial activity of cefditoren against clinical isolates of respiratory tract infection. MATERIALS AND METHODS: According to the NCCLS guideline, in vitro activities of cefditoren and other antibiotics were tested against respiratory pathogens including 117 isolates of Streptococcus pneumoniae, 60 isolates of Haemophilus influenzae, and 31 isolates of Moraxella catarrhalis. RESULTS: The level of cefditoren activity against S. pneumoniae (MIC50/90, 0.5/1 microgram/mL) was superior to amoxicillin+/-clavulanate (MIC50/90, 4/16 microgram/mL), clarithromycin (MIC50/90, >32/>32 microgram/mL), and most of the marketed cephalosporins (MIC50/90, 8-64/16-128 microgram/mL). Although the MIC of cefditoren was relatively higher than those of new fluoroquinolone agents (MIC50/90, 0.03-1/0.06-1 microgram/mL), it was comparable to ceftriaxone (MIC50/90, 0.5/1 microgram/mL). In addition, cefditoren was active against two quinolone resistant pneumococci strains with MIC of 0.5 microgram/mL. In detail, cefditoren was active against pneumococci strains with MIC50 and MIC90 values of 0.015/0.12, 0.12/0.5, and 0.5/1 microgram/mL for penicillin-susceptible, -intermediate, and -resistant pneumococci, respectively. Cefditoren was also active against all respiratory isolates of H. influenzae (MIC50/90, 0.03/0.06 microgram/mL) and M. catarrhalis (MIC50/90, 0.03/0.05 microgram/mL) irrespective of beta-lactamase production or ampicillin resistance. CONCLUSION: Cefditoren is considered to be a good option for outpatient treatment of respiratory infections, particulary if there is concern about S. pneumoniae infection with decreased susceptibility to penicillin or beta-lactamase producing organisms.
Ampicillin Resistance ; Anti-Bacterial Agents ; beta-Lactamases ; Ceftriaxone ; Cephalosporins ; Clarithromycin ; Haemophilus influenzae ; Humans ; Influenza, Human ; Moraxella (Branhamella) catarrhalis ; Outpatients ; Penicillins ; Pneumonia ; Respiratory Tract Infections ; Streptococcus pneumoniae

BACKGROUND: Due to advances in various diagnostic methods, recent studies reported changes in the pattern of etiology of fever of unknown origin (FUO). To identify the current pattern of the causes of FUO, we analyzed the etiology of recently diagnosed FUO at a university hospital in Korea. MATERIALS AND METHODS: We reviewed 69 cases that fulfilled the criteria of classic FUO and retrospectively analyzed the etiology and decisive methods of diagnosis. RESULTS: The etioloies of FUO were infectious disease, non-infectious inflammatory disease, malignancy and miscellaneous cases in 22 (31.9%), 8 (11.6%), 4 (2.3%) and 21 (30.4%) patients, respectively. In 15 (21.7%) cases the cause could not be identified. Among infectious diseases, tuberculosis and suspected typhoid fever were the most common causes of infection (8 case, 11.7%) with tuberculosis being the most common confirmed infection. Adult onset Still's disease (13 cases, 4.4%) and drug-related fever (13 cases, 18.8%) were the most common cause of non-infectious inflammatory disease and miscellaneous causes, respectively. Decisive methods of final diagnosis were by observation of clinical course in 35 (64.8%), radiologic examination in 10 (18.5%), serologic or biochemical test in 5 (9.3%) and tissue biopsy in 4 (7.4%); none were diagnosed by culture. CONCLUSION: Infection remains the most common etiology of classic FUO in Korea and observing the clinical course is the most commonly used method for decisive diagnosis and its importance should be emphasized in approaching patients with FUO.
Adult* ; Biopsy ; Communicable Diseases ; Diagnosis ; Fever of Unknown Origin* ; Fever* ; Humans ; Korea* ; Retrospective Studies ; Still's Disease, Adult-Onset ; Tuberculosis ; Typhoid Fever