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Jung Min Lee

Publications and source records attributed to Jung Min Lee.

At least 19 recordsLinked to original sources
PubMed2006

Isolation and characterization of promoters from Lactococcus lactis ssp. cremoris LM0230.

DNA fragments showing promoter activity were obtained from the chromosomal DNA of Lactococcus lactis ssp. cremoris LM0230 by using a promoter-screening vector pBV5030, which contains a promoterless chloramphenicol acetyltransferase gene. Ten fragments were identified based on their ability to confer resistance against chloramphenicol in Escherichia coli. DNA sequencing revealed that all the fragments had a consensus region recognized by the sigma factor and only the nucleotide sequence of fragment 15C had the identical consensus region with the promoter P2 from L. lactis ssp. lactis MG1614. To compare their promoter strengths, an E. coli-lactococcal shuttle vector pWL1 containing a luciferase gene as the reporter gene was constructed based on lactococcal plasmid pMG36e. The putative promoter regions of 10 fragments exhibiting promoter activity were characterized in E. coli and L. lactis by measuring the luciferase activity, among which the putative promoter P6C exhibited the highest promoter activity both in E. coli JM109 and L. lactis ssp. cremoris MG1363. The luciferase system endowed significantly different expression levels enough to compare promoter strengths in E. coli and lactococcal host. The transcription-initiation sites of P6C and P13C were mapped by primer extension, which showed that they corresponded to a purine residue. The characterized promoters could be useful for the industrial production of heterologous proteins in L. lactis in case the proteins require a high safety level.

Base Sequence↗
PubMed2006

[Natural history of HBeAg negative chronic hepatitis B virus infection: a cohort study].

BACKGROUND/AIMS: The long-term virologic and biochemical changes in patients with HBeAg negative HBV infection, especially in Asia, remain unclear. To address this issue, we conducted a 3 year- retrospective, cohort study. METHODS: A total of 157 patients with HBeAg negative HBV infection who were monitored without treatment were reviewed between January 1999 and March 2004. Those patients were followed up every 3 months with liver function tests and serologic tests. All patients were stratified into 3 groups; inactive carrier (IC), viremic carrier (VC) and chronic hepatitis (CH). Serum HBV DNA was measured by a hybridization assay (sensitivity: 1.4 x 10(5)) genomes/mL, Digene Diagnostics, Silver Spring, USA). RESULTS: The median age of enrolled patients was 42.7 years (M:F=2.3:1). By single time-point observations, the 3 year-cohort prevalence of HBeAg negative CH varied from 12.7 to 35.8% (median 20.7%) HBeAg negative CH was accumulated over time (P=0.002) and transition rates among three groups after 3 years of follow-up are as follows: IC to CH, 6.0%; IC to VC, 4.1%; VC to CH, 23.2%. VC seems to be a disease state in the middle of transition from IC to CH. CONCLUSIONS: We demonstrated the dynamic changing patterns of HBeAg negative CH with time, of which the change from IC or VC to CH was dominant.

Adult↗
PubMed2005

In vivo bone formation by human marrow stromal cells in biodegradable scaffolds that release dexamethasone and ascorbate-2-phosphate.

An unsolved problem with stem cell-based engineering of bone tissue is how to provide a microenvironment that promotes the osteogenic differentiation of multipotent stem cells. Previously, we fabricated porous poly(D,L-lactide-co-glycolide) (PLGA) scaffolds that released biologically active dexamethasone (Dex) and ascorbate-2-phosphate (AsP), and that acted as osteogenic scaffolds. To determine whether these osteogenic scaffolds can be used for bone formation in vivo, we seeded multipotent human marrow stromal cells (hMSCs) onto the scaffolds and implanted them subcutaneously into athymic mice. Higher alkaline phosphatase expression was observed in hMSCs in the osteogenic scaffolds compared with that of hMSCs in control scaffolds. Furthermore, there was more calcium deposition and stronger von Kossa staining in the osteogenic scaffolds, which suggested that there was enhanced mineralized bone formation. We failed to detect cartilage in the osteogenic scaffolds (negative Safranin O staining), which implied that there was intramembranous ossification. This is the first study to demonstrate the successful formation of mineralized bone tissue in vivo by hMSCs in PLGA scaffolds that release Dex and AsP.

Alkaline Phosphatase↗
PubMed2005

Cloning and characterization of the bifunctional alcohol/acetaldehyde dehydrogenase gene (adhE) in Leuconostoc mesenteroides isolated from kimchi.

A bifunctional alcohol/acetaldehyde dehydrogenase (AdhE) gene (adhE) was cloned from Leuconostoc mesenteroides C7 (LMC7), which is the dominant lactic acid bacterium produced during heterofermentation of kimchi. The nucleotide sequence of the DNA fragment containing putative adhE, which is 2685 bp long and encodes an 886 amino acid polypeptide, exhibits 99% homology with Leu. mesenteroides sp. cremoris. The deduced AdhE comprises two conserved domains: alcohol dehydrogenase (Adh) and acetaldehyde dehydrogenase (Aldh). Moreover, two NAD-binding sites were observed, based on the presence of the GXGXXG motif. A pADHE containing the adhE gene expressed AdhE at the translational level in Escherichia coli BL21, which was at a higher level than in E. coli DH5alpha and E. coli JM109. The AdhE of LMC7 showed Adh and Aldh activities that, when expressed in E. coli. BL21, were 7.5 and 5.7 U mg(-1) , respectively.

Alcohol Dehydrogenase↗
PubMed2005

Cloning and characterization of the gene encoding phosphoketolase in Leuconostoc mesenteroides isolated from kimchi.

The gene encoding phosphoketolase, which is 2749 bp long and contains 814 amino acid polypeptides with a total molecular mass of 91.9 kDa, was cloned from Leuconostoc mesenteroides C7 (LMC7) and expressed in Escherichia coli. It exhibited a homology of >58% with phosphoketolases from other lactic acid bacteria. The phosphoketolase of LMC7 belongs to the xylulose 5-phosphate (X5P)/fructose 6-phosphate (F6P) phosphoketolase (Xfp) family, which is an enzyme with dual specificity for X5P and F6P. The members of this family contain typical thiamin pyrophosphate (TPP) binding sites as reported for other TPP-dependent enzymes, and several highly conserved regions as signature patterns for phosphoketolases. The plasmid pGPK containing the Xfp gene (xfp) exhibits phosphoketolase activity in E. coli. The specific activities of the enzyme from E. coli BL21 and E. coli EC101 harboring xfp were 0.28 and 0.14 units/mg, respectively. They both exhibited a 1.5-fold increase in the production of acetic acid from acetyl phosphate compared with their corresponding original strain.

Aldehyde-Lyases↗
PubMed2005

Determination of catechin in aqueous solution by chemiluminescence method.

A method to determine catechin in aqueous solution by measuring chemiluminescence intensities using a stopped flow system has been studied. The lucigenin-hydrogen peroxide chemiluminescence reaction was chosen for the determination of catechin. Fe(II) ion was added to the chemiluminescence system to increase the sensitivity. The chemiluminescence intensity from the lucigenin system was increased by the addition of catechin. Effects of flow rates of reagent and sample and concentrations of lucigenin, hydrogen peroxide, Fe(II) ion and KOH were investigated. The calibration curve for catechin was linear over the range from 1.0x10(-6) to 1.0x10(-3) M and the detection limit was 3.0x10(-7) M under the optimal experimental conditions.

Acridines↗
PubMed2005

Efficacy of a traditional Korean medicine, Chung-Sang-Bo-Ha-Tang, in a murine model of chronic asthma.

Traditional herbal medicines may be viable alternatives to corticosteroid therapy for treatment of asthma. However, the therapeutic mechanisms of herbal compounds remain a matter of considerable debate. This study was performed to evaluate the effects of Chung-Sang-Bo-Ha-Tang (CSBHT), a herbal compound administrated therapeutically to asthma patients for centuries, on airway inflammation and remodeling in a murine model of chronic asthma. BALB/c mice sensitized to ovalbumin (OVA) were chronically challenged with aerosolized OVA for 6 weeks. During the last 2 weeks, some mice were treated daily with CSBHT by intragastric feeding. Dexamethasone (Dex)-treated, phosphate-buffered saline (PBS)-treated, and naive mice served as controls. The effects of CSBHT on airway inflammation, lung pathology, and cytokine production were evaluated. Mice exposed to recurrent airway challenge with OVA had chronic inflammation and characteristics of airway remodeling, including subepithelial fibrosis, epithelial hypertrophy, and goblet cell hyperplasia. CSBHT was as effective as Dex at moderately reducing these changes compared to the PBS-treated mice. In addition, IL-5 and IFN-gamma levels in supernatants of Concanavalin A (Con A)-activated splenocyte cultures were reduced in mice treated with CSBHT. Treatment with CSBHT during the last 2 weeks of challenge modulated airway inflammation and remodeling in a murine model of chronic asthma. Thus, CSBHT may effectively delay the progression of airway inflammation and remodeling.

Administration, Oral↗
PubMed2005

Long-term clinical and histological outcomes in patients with spontaneous hepatitis B surface antigen seroclearance.

BACKGROUND/AIMS: During the natural course of hepatitis B virus (HBV) infection, the long-term clinical and histological outcomes following spontaneous hepatitis B surface antigen (HBsAg) seroclearance remain unclear. METHODS: Between 1984 and 2003, 49 (9.5%) out of 432 inactive HBsAg carriers had no detectable level of circulating HBsAg. Fifteen of 49 patients had undergone paired peritoneoscopic liver biopsies. RESULTS: During a mean follow-up period of 19.6 months after HBsAg seroclearance, 5 of 49 (10.2%) patients were noted to have HCC. Liver cirrhosis (P=0.040), a history of perinatal infection (P=0.005) and long-standing duration (at least 30 years) of HBsAg positivity (P=0.002) were associated with a significantly higher risk of developing HCC. Despite HBsAg seroclearance, HBV DNA was detected in the liver tissues from all 15 patients who underwent paired liver biopsies. Necroinflammation was significantly ameliorated (P<0.0001). On the other hand, amelioration of the fibrosis score did not reach a statistically significant level (P=0.072). Interestingly, aggravation of liver fibrosis was evident in 2 patients (13.3%) including one who had rapidly progressed to overt cirrhosis. CONCLUSIONS: In patients with spontaneous HBsAg seroclearance, necroinflammation was markedly improved and liver fibrosis was unchanged or regressed despite occult HBV infection. However, HCC developed in a minority of cases.

Adult↗
PubMed2005

The effect of simvastatin on the proliferation and differentiation of human bone marrow stromal cells.

Statins have been postulated to affect the bone metabolism. Recent experimental and epidemiologic studies have suggested that statins may also have bone protective effects. This study assessed the effects of simvastatin on the proliferation and differentiation of human bone marrow stromal cells (BMSCs) in an ex vivo culture. The bone marrow was obtained from healthy donors. Mononuclear cells were isolated and cultured to osteoblastic lineage. In the primary culture, 10(-6) M simvastatin diminished the mean size of the colony forming units-fibroblastic (CFU-Fs) and enhanced matrix calcification. At near confluence, the cells were sub-cultured. Thereafter, the alkaline phosphatase (ALP) activities of each group were measured by the time course of the secondary culture. Simvastatin increased the ALP activity in a dose dependent manner, and this stimulatory effect was more evident during the early period of culture. A 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay was performed during the secondary culture in order to estimate the effect of simvastatin on the proliferation of human BMSCs. When compared to the control group, simvastatin significantly decreased the proliferation of cells of each culture well. 10(-6) M of simvastatin also significantly enhanced the osteocalcin mRNA expression level. This study shows that simvastatin has a stimulatory effect on bone formation through osteoblastic differentiation, and has an inhibitory effect on the proliferative potential of human BMSCs.

Alkaline Phosphatase↗
PubMed2005

Prevalence and clinical characteristics of the metabolic syndrome in middle-aged Korean adults.

BACKGROUND: The aim of this study was to analyze the prevalence and clinical characteristic of the metabolic syndrome of adults, over 40 years old, living in Korea. METHODS: This study was carried out for 2 years, 2003-2004, on total 5,330 individuals (2,197 men and 3,133 women) selected by the stratified random cluster sampling among adults over 40 years old. Metabolic syndrome was defined based on both the NCEP-ATP III criteria and Modified ATP III criteria applying the WHO-APR (Asian Pacific Region)'s abdominal obesity criteria (waist circumference > 90 cm in men, 80 cm in women) instead of NCEP-ATP III criteria. RESULTS: Using NCEP-ATP III criteria, the age-adjusted overall prevalence of metabolic syndrome was 24.8% (17.6% in men, 30.0% in women). Age-adjusted overall prevalence of metabolic syndrome as defined by modified-ATP III criteria was 34.3% (26.3% in men, 40.1% in women). The prevalence of metabolic syndrome for each age group (40-49, 50-59, 60-69, > or = 70) in men was as follows: 18.8%, 17.4%, 18.3%, 14.5%. In women: 22.3%, 32.7%, 39.9%, 39.3%. The prevalence of hypertriglyceridemia (triglycerides > or = 1.7 mmol/l) was well correlated with the changing pattern of the prevalence of metabolic syndrome both in men and women. CONCLUSIONS: The peak age of metabolic syndrome in men was age 40 through 49, and the prevalence decreased with aging. Therefore, early intervention for risk factors of metabolic syndrome might be required in men. On the other hand, prevention for cardiovascular disease will be needed for perimenopausal women due to considerably increased prevalence in the age 50 through 59.

Adult↗
PubMed2004

Dispersion polymerization of methyl methacrylate with a novel bifunctional polyurethane macromonomer as a reactive stabilizer.

A novel macromonomer of vinyl-terminated bifunctional polyurethane was synthesized and applied to the dispersion polymerization of MMA in ethanol. The existence of the vinyl terminal groups and the grafted macromonomer with PMMA was verified using 1H NMR and 13C NMR. The stable and monodisperse PMMA microspheres having a weight-average diameter of 5.09 microm and a good uniformity of 1.01 were obtained with 20 wt% polyurethane macromonomer. The molecular weight increased, but the size of the synthesized PMMA microspheres decreased with the macromonomer concentration since the macromonomer acts as a reactive stabilizer in dispersion polymerization. Furthermore, the molecular weight of the PMMA prepared by the use of the macromonomer was approximately twofold higher than that prepared by a conventional stabilizer, poly(N-vinylpyrrolidone). The higher molecular weight is thought to originate from the grafting and a possibly slight crosslinking of PMMA molecules due to the bifunctional reactive groups at the ends of macromonomer chains.

Magnetic Resonance Spectroscopy↗
PubMed2004

Quantitative profiling of plasma peptides in asthmatic mice using liquid chromatography and mass spectrometry.

Asthma is increasing in prevalence worldwide as a result of factors associated with a Western lifestyle. However, simple and reliable diagnostic and prognostic markers are yet to be found. In an attempt to identify protein biomarker profiles among small molecular weight ranges, we employed an approach combining liquid chromatography with mass spectrometry, instead of two-dimensional gel electrophoresis (2-DE), which has previously been used to analyze protein expression patterns. Here we described its application to compare plasma peptides from control and chronic asthma mice. Peptides were quantitatively profiled as a multidimensional peptide mass fingerprint by a combination of reverse-phase high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. They were identified by peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight tandem mass spectrometry. In this study, we quantitatively identified the fragment f of complement 3 (C3f), which is important in inflammation. C3f was significantly higher in controls than chronic asthma mice. Our strategy allowed the detection and identification of different plasma peptides between control and chronic asthma mice on a proteomic scale. Therefore, these results suggest that native small peptides detected by non-2-DE techniques may be useful and specific biomarkers of disease.

Animals↗
PubMed2004

Construction of a pTOC-T vector using GST-ParE toxin for direct cloning and selection of PCR products.

Using linker insertion mutations, we determined the most stable region of the parE gene which encodes a toxic protein (ParE) that inhibits growth of Escherichia coli. The toxicity of ParE was sustained until a 144 bp linker was inserted into this region. We have developed a 3' T-overhang vector based on these characteristics of the GST-ParE toxin, and named pTOC-T. Because pTOC-T uses a post-segregational killing system, all transformants grown up on the plates can be considered as recombinants containing foreign DNA. pTOC-T not require X-Gal, IPTG or other substrates for selection. This T-vector using a positive selection system can be applied to various E. coli strains such as XL1-Blue, BL21, DH5alpha, JM109, and JM110.

Bacterial Toxins↗
PubMed2004

Characterization of pC7 from Lactobacillus paraplantarum C7 derived from Kimchi and development of lactic acid bacteria--Escherichia coli shuttle vector.

A cryptic plasmid pC7 was characterized, which was isolated from Lactobacillus paraplantarum C7 derived from Kimchi, a traditional korean vegetable food. The plasmid pC7 is a circular molecule of 2,134 base-pairs in length with a G + C content of 38.5%. The nucleotide sequence analysis revealed the presence of an open reading frame encoding a putative 317 amino acids protein homologous to replication proteins RepA. Furthermore, a putative double and a single-strand origin were identified. Together with features of nucleotide sequences, the detection of single-stranded intermediate DNA in Lb. paraplantarum demonstrated that pC7 replicates via a rolling circle mechanism. A cloning vector for lactic acid bacteria was developed on the basis of the pC7 replicon, into which were inserted an erythromycin resistance gene as a marker, multiple cloning sites, and Escherichia coli ColE1 replication origin. E. coli and several species of Lactobacillus and Leuconostoc can be transformed with the resultant vector. Therefore, pC7 derivatives may be useful LAB-E. coli shuttle vectors, which are essential in engineering important strains in food fermentation.

Amino Acid Sequence↗
PubMed2004

[Reappraisal of HBV genotypes and clinical significance in Koreans using MALDI-TOF mass spectrometry].

BACKGROUND/AIMS: Recent studies have shown that the genotype of hepatitis B virus (HBV) may correlate with the disease natural history and treatment outcome. However, several of these studies used low sensitivity assays in a small number of patients, and this has precluded an accurate evaluation of Korean HBV genotypes. We analyzed Korean HBV genotypes in a large population by employing a new technology, restriction fragment mass polymorphism (RFMP) using MALDI-TOF mass spectrometry, in a sensitive and specific manner. METHODS: Between February 1995 and December 2003, a total of 475 patients with chronic HBV infection were enrolled. The assay is based on the mass measurement of oligonucleotides having genotypic variations of the S gene. Clinical features including the virologic status and disease progression were also evaluated. RESULTS: The median age of the total patients was 35.5 years. Out of 475 patients, there were 162 (34.1%) inactive carriers, 172 (36.2%) had chronic hepatitis, 77 (16.2%) had liver cirrhosis and 64 (13.5%) had hepatocellular carcinoma (HCC). There were 454 patients (95.6%) with genotype C, 4 patients (0.8%) with genotype A, 16 patients (3.4%) with the mixed A and C genotype [7 patients (1.4%) with A A], and 1 patient (0.2%) with B genotype. Comparing genotype A and C, genotype A patients were all inactive carriers without HCC, whereas genotype C patients included those with chronic hepatitis, liver cirrhosis and HCC. CONCLUSIONS: HBV genotype C is highly prevalent in Korea. Although it is a small percentage, genotype A also exists and it seems to take a more benign clinical course than genotype C. Further studies are necessitated to assess the relationship between HBV genotypes and the various aspects of the diseases' clinical course.

Adolescent↗
PubMed2003

New integration vector using a cellulase gene as a screening marker for Lactobacillus.

The new integration vector for Lactobacillus, pJC4, was developed using the extracellular endoglucanase A gene (celA) of Clostridium thermocellum as a screening marker. pJC4 was transformed into four Lactobacillus species, Lb. johnsonii, Lb. gasseri, Lb. bulgaricus, and Lb. plantarum. In each species, the pJC4 integrants were easily and accurately detected by the appearance of a clear halo on a cellulase screening plate without any false transformants. Polymerase chain reaction and Southern hybridization indicated that all transformants with clear halos contained pJC4 in their chromosomal DNAs. The celA gene could be a useful screening marker for other lactic acid bacteria.

Cellulase↗
PubMed2003

Selective beta-cell loss and alpha-cell expansion in patients with type 2 diabetes mellitus in Korea.

In the presence of obesity, beta-cell mass needs to be increased to compensate for the accompanying demands and maintain euglycemia. However, in Korea, the majority of type 2 diabetic patients are nonobese. We determined the absolute masses, relative volumes, and ratio of alpha- and beta-cell in the pancreas and islets in normal and diabetic Korean subjects to correlate these findings with the clinical characteristics. Whole pancreases procured from organ donors were divided into 24 parts (control 1, n = 9). Tissue was also obtained by surgical resection after 35 partial pancreatectomies: in 25 diabetic patients, 10 age- and body mass index (BMI)-matched patients of benign or malignant pancreatic tumor without diabetes mellitus (DM) (control 2). Morphometric quantifications were performed. In control 1, the relative volume of beta-cells was 2.1 +/- 0.9%, and the total beta-cell mass was 1.3 +/- 0.3 g. The relative volume of beta-cells was found to be variable (control 1, 2.1 +/- 0.9%; control 2, 1.9 +/- 0.7%; DM, 1.4 +/- 1.0%; P < 0.05 DM vs. control 1 and 2) and showed good correlation with BMI (control 1, r(2) = 0.64; DM, r(2) = 0.55; all subjects, r(2) = 0.38; P < 0.05). Notably, in type 2 diabetic patients, the ratio of alpha-cell area to beta-cell area in the islet was higher than in control 1 and 2 (0.81 +/- 0.4 vs. 0.29 +/- 0.2, 0.20 +/- 0.1, P < 0.05). Additionally, significant alpha-cell expansion and a decreased beta-cell fraction were predominantly observed in larger islets (islet area, >6415 micro m(2); P < 0.05) in control 1 and diabetic patients. The relative volume of beta-cell was found to be correlated with BMI in diabetic patients and normal organ donors. Moreover, decreased beta-cell but increased alpha-cell proportion in the islets suggests for a selective beta-cell loss in the pathogenesis of Korean type 2 diabetes.

Adult↗