• Title/Summary/Keyword: pMB4

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Molecular Identification of Pseudanabaena Strains and Analysis of 2-MIB Production Potential in the North Han River System (북한강 수역에 분포하는 Pseudanabaena 균주의 동정 및 2-MIB 생산 잠재성 분석)

  • Kim, Keonhee;Lee, Sejin;Seo, Kyunghwa;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.53 no.4
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    • pp.344-354
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    • 2020
  • Identification of the target species of 2-MIB (2-methyllisoborneol) production is crucial in the management of off-flavor problem in the freshwater system. This study was conducted to identify 2-MIB-producing Pseudanabaena strains occurring in the North Han River system using molecular genetic method. Eleven phenotypes of Pseudanabaena were isolated from several mainstream sites of the North Han River, including Sambong-ri, Joam-myun, and Lake Uiam areas. Despite of morphological similarity of the strains, the phylogenetic analysis using 16S rDNA classified them into different species with low genetic similarity (40~55%). Isolated Pseudanabaena strains were converged to four species; Pseudanabaena cinerea, P. yagii, P. mucicola, and P. redekei. Among them, the 2-MIB synthesizing gene (mibC) was detected in P. cinerea, P. yagii, and P. redekei. However, actual 2-MIB production was detected only in P. cinerea and P. redekei based on gas chromatography analysis. This study is the first report of the molecular identification of Pseudanabaena strains and their 2-MIB production potential in Korea. The results of this study provides an evidence of species diversity of Pseudanabaena occurring in the North Han River.

Tumor Cell Clone Expressing the Membrane-bound Form of IL-12p35 Subunit Stimulates Antitumor Immune Responses Dominated by $CD8^+$ T Cells

  • Lim, Hoyong;Do, Seon Ah;Park, Sang Min;Kim, Young Sang
    • IMMUNE NETWORK
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    • v.13 no.2
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    • pp.63-69
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    • 2013
  • IL-12 is a secretory heterodimeric cytokine composed of p35 and p40 subunits. IL-12 p35 and p40 subunits are sometimes produced as monomers or homodimers. IL-12 is also produced as a membrane-bound form in some cases. In this study, we hypothesized that the membrane-bound form of IL-12 subunits may function as a costimulatory signal for selective activation of TAA-specific CTL through direct priming without involving antigen presenting cells and helper T cells. MethA fibrosarcoma cells were transfected with expression vectors of membrane-bound form of IL-12p35 (mbIL-12p35) or IL-12p40 subunit (mbIL-12p40) and were selected under G418-containing medium. The tumor cell clones were analyzed for the expression of mbIL-12p35 or p40 subunit and for their stimulatory effects on macrophages. The responsible T-cell subpopulation for antitumor activity of mbIL-12p35 expressing tumor clone was also analyzed in T cell subset-depleted mice. Expression of transfected membranebound form of IL-12 subunits was stable during more than 3 months of in vitro culture, and the chimeric molecules were not released into culture supernatants. Neither the mbIL-12p35-expressing tumor clones nor mbIL-12p40-expressing tumor clones activated macrophages to secrete TNF-${\alpha}$. Growth of mbIL-12p35-expressing tumor clones was more accelerated in the $CD8^+$ T cell-depleted mice than in $CD4^+$ T cell-depleted or normal mice. These results suggest that $CD8^+$ T cells could be responsible for the rejection of mbIL-12p35-expressing tumor clone, which may bypass activation of antigen presenting cells and $CD4^+$ helper T cells.

Cloning of the MLS Antibiotics Inducible Resistance Gene and Its Control Mechanism -Inducible Resistance to MLS Antibiotics of pMB4 Plasmid Isolated from Streptococcus sp. TR-1- (MLS계 항생물질 유도 내성 유전자의 크로닝과 유전자의 조절기전 -Streptococcus sp. TR-1에서 분리한 pMB 4 Plasmid의 MLS계 항생물질 유도내성-)

  • Jeong, Soon-Hak;Kwak, Jin-Hwan;Kim, Hee-Sun;Shim, Mi-Ja;Choi, Eung-Chil;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.34 no.2
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    • pp.139-146
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    • 1990
  • Streptococcus sp. TR-1 which has inducible resistance to MLS antibiotics was isolated from soil samples in Korea. Streptococcus sp. TR-1 was cultured in Lysis broth, then a plasmid was isolated by modified Elliker method. Bacillus subtilis UOTO277 was transformed with that plasmid. This result showed that the plasmid has the gene relating with inducible resistance to MLS antibiotics. It was named pMB4 and its size was determined about 2.4 Kb by results of digestion with various restriction enzymes. Restriction endonuclease cleavage site map of pMB4 plasmid was made by double digestion of the plasmid. pMB4 plasmid has different restriction endonuclease site map from the other plasmids that have been discovered in Streptococcus sp. so far. And it could be identified that pMB4 plasmid does not have homology with ermK of Bacillus licheniformis EMR but has homology with ermC of Staphylococcus aureus from the results of Southern hybridization.

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Application of array comparative genomic hybridization in Korean children under 6 years old with global developmental delay

  • Lee, Kyung Yeon;Shin, Eunsim
    • Clinical and Experimental Pediatrics
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    • v.60 no.9
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    • pp.282-289
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    • 2017
  • Purpose: Recent advancements in molecular techniques have greatly contributed to the discovery of genetic causes of unexplained developmental delay. Here, we describe the results of array comparative genomic hybridization (CGH) and the clinical features of 27 patients with global developmental delay. Methods: We included 27 children who fulfilled the following criteria: Korean children under 6 years with global developmental delay; children who had at least one or more physical or neurological problem other than global developmental delay; and patients in whom both array CGH and G-banded karyotyping tests were performed. Results: Fifteen male and 12 female patients with a mean age of $29.3{\pm}17.6months$ were included. The most common physical and neurological abnormalities were facial dysmorphism (n=16), epilepsy (n=7), and hypotonia (n=7). Pathogenic copy number variation results were observed in 4 patients (14.8%): 18.73 Mb dup(2)(p24.2p25.3) and 1.62 Mb del(20p13) (patient 1); 22.31 Mb dup(2) (p22.3p25.1) and 4.01 Mb dup(2)(p21p22.1) (patient 2); 12.08 Mb del(4)(q22.1q24) (patient 3); and 1.19 Mb del(1)(q21.1) (patient 4). One patient (3.7%) displayed a variant of uncertain significance. Four patients (14.8%) displayed discordance between G-banded karyotyping and array CGH results. Among patients with normal array CGH results, 4 (16%) revealed brain anomalies such as schizencephaly and hydranencephaly. One patient was diagnosed with Rett syndrome and one with $M{\ddot{o}}bius$ syndrome. Conclusion: As chromosomal microarray can elucidate the cause of previously unexplained developmental delay, it should be considered as a first-tier cytogenetic diagnostic test for children with unexplained developmental delay.

Biosorption of Methylene Blue from Aqueous Solution using Dried Rhodotorula glutinis Biomass

  • Dae Haeng Cho;Jaesung Lee;Eui Yong Kim
    • Korean Chemical Engineering Research
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    • v.61 no.2
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    • pp.273-277
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    • 2023
  • The biosorption of methylene blue (MB) from aqueous solution was investigated using dried Rhodotorula glutinis as the biosorbent. The effects of pH, initial dye concentration, biosorbent dosage, and kinetic studies were determined to obtain valuable information for biosorption. Results indicated that most of the adsorbed MB bound within 30 minutes of contact and the MB adsorption capacity increased from 21.1 to 101.8 mg/g with the initial MB concentration increased from 50 to 300 mg/L. Additionally, the MB adsorption capacity gradually increased from pH 4.0 to 9.0, reaching its peak at an initial pH of 9.0. As the biomass load was increased from 0.25 to 4.0 g/L, the MB removal efficiency increased from 14.1 to 84.5%. The Langmuir model provided the best fit throughout the concentration range, and the maximum adsorption capacity (qmax) and Langmuir constant (b) were determined to be 135.14 mg/g and 0.026 l/mg, respectively. Furthermore, the biosorbent process of R. glutinis was found to follow pseudo-second-order kinetics and the calculated qeq,cal value showed good agreement with the experimental qeq value. Overall, the biosorption of MB by R. glutinis can be characterized as a monolayer, single site type phenomenon, and the rate-limiting step was determined to be the chemical reaction between the adsorbent and the adsorbate.

A cone-beam computed tomography study of the prevalence and location of the second mesiobuccal root canal in maxillary molars

  • Seong-Ju Lee ;Eun-Hye Lee ;Se-Hee Park ;Kyung-Mo Cho ;Jin-Woo Kim
    • Restorative Dentistry and Endodontics
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    • v.45 no.4
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    • pp.46.1-46.8
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    • 2020
  • Objectives: This study aimed to investigate the incidence and location of the second mesiobuccal root (MB2) canal in maxillary molars with the aid of various measuring points and lines using cone-beam computed tomography (CT). Materials and Methods: A total of 205 images of patients who underwent cone-beam CT examinations between 2011 and 2015 as part of their dental diagnosis and treatment were included. There were 76 images of the maxillary first molar and 135 images of the maxillary second molar. Canal orifices were detected at -1 mm from the top of the pulpal floor on cone-beam CT images. Image assessment was performed by 2 observers in reformatted image planes using software. Assessments included measurement of the distance between the MB1 and MB2 canals, and the angles between the lines connecting the MB1-MB2 and distobuccal (DB)-palatal (P) canals. The data were analyzed using the student's t-test. Results: The prevalence of the MB2 canal was 86.8% in the first molar and 28.9% in the second molar. The angle between the lines connecting the MB1-MB2 and DB-P canals was 2.3° ± 5.7° in the first molar and -3.95° ± 7.73° in the second molar. The distance between the MB1 and MB2 canals was 2.1 ± 0.44 mm in the first molar and 1.98 ± 0.42 mm in the second molar. Conclusions: The angles between the lines connecting the MB1-MB2 and DB-P canals was almost parallel. These findings may aid in the prediction of the location of the MB2 canal orifice.

Improvement of Quality in Treated Water by the pH Adjustment of Raw Water (원수 pH 조정에 의한 정수 수질 개선)

  • Jeong, Gwan-Jo;Lee, Kyeong-Woo;Kim, Hyun-Hee;Jeong, Eui-Sun;Park, Hyeon;Han, Sun-Hee
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.5
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    • pp.469-476
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    • 2010
  • The purpose of the study is to find ways to decrease turbidity and residual aluminum by improving the efficiency of coagulation process through controlling the pH of the source water with $CO_2$ when the pH increases by algal bloom or by the characteristics of the source water. Water quality parameters were monitored before and after $CO_2$ addition in February, March, April, and December, when the pH of the source water is over 8.0 and constant regardless of day and night. Water quality parameters closely related with evaluation of treated drinking water quality were monitored in detail, e.g. aluminum, turbidity, particle counts, TOC, THMs, 2-MIB, and geosmin. According to the results, inorganic water quality parameters such as turbidity, particle counts, and aluminum were decreased due to improved efficiency of the coagulation process. It was concluded that the pH of the water in the arrival basin must be controlled below 7.4 by adding $CO_2$ when the pH of the source water increasing. By controlling pH with $CO_2$, the water quality could be maintained within the drinking water quality goal of Seoul City (<30 particle/mL of particle count, <0.05 NTU of turbidity and <0.02 mg/L of aluminum). The change of the pH could not affect the concentrations of DBP's (e.g., THMs, CH, and HAAs) and taste/odor causing compounds (e.g., 2-MIB and geosmin). 2-MIB and geosmin were affected more by their initial concentrations in the source water.

Prognostic Implications of the MIB-1 Labeling Index in Astrocytic Tumors (성상세포성 종양에서 MIB-1증식지수와 예후의 연관성)

  • Kim, Choong Hyun;Bak, Koang Hum;Kim, Jae Min;Ko, Yong;Oh, Suck Jun;Hong, Eun Kyung
    • Journal of Korean Neurosurgical Society
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    • v.30 no.4
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    • pp.430-436
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    • 2001
  • Objective : The proliferative potential of intracranial glioma affects the histological malignancy and prognosis of patients with these tumors. In this study, we present the relationship between MIB-1 labeling index(LI) and clinical variables which might play the major role in determining the prognosis of patient with astrocytic tumors. Patients and Methods : Excised tumor specimens from a total of 52 patients were stained to detect monoclonal MIB-1-Ki-67 antibody by avidin-biotin complex immunohistochemistry. The MIB-1 LI was evaluated with histological grades, demograpghic data, and survival time. The statistical significance of their correlation was analyzed by Pearson correlation test. Results : The 52 patients included 30 male patients and 22 female patients. The tumors according to the criteria of the World Health Organization(WHO) classification were verified as pleomorphic xanthoastrocytoma in one, pilocytic astrocytomas 4, astrocytomas 1, anaplastic astrocytomas 3, and glioblastomas 31. MIB-1 LI in astrocytic ttumors showed no correlation with age and gender. However, the patients under 10 years had the longest survival time, whereas short survival time was observed in the older patients. The mean MIB-1 LI of different tumor grades were as follows : pleomorphic xanthoastrocytoma, $4.40{\pm}0.00$ ; pilocytic astrocytoma, $4.53{\pm}3.09$ ; astrocytoma, $5.50{\pm}6.03$ ; anaplastic astrocytoma, $12.68{\pm}12.50$ ; Glioblastoma, $21.31{\pm}19.63$. Although the levels of MIB-1 LI were varied in individual tumors, the MIB-1 LI was increased in parallel with the histological grades. Glioblstomas showed significantly higher MIB-1 LI compared with that of anaplastic astrocytomas and low grade astrocytomas (p = 0.001). The mean survival time of entire group of patients was also well correlated with MIB-1 LI in astrocytic tumors(p = 0.015). Moreover, the mean survival time of the entire group of patients with Lis < 10 was $125.33{\pm}113.57weeks$, and the mean survival of those with $Lis{\geq}10$ was $60.71{\pm}62.58weeks$. This difference was also statistically significant(p = 0.004). Conclusion : The results of this study suggest that MIB-1 LI correlates with histological grades and might play a significant role in predicting the survival of patients with astrocytic tumors.

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Cloning of Inducible MLS Antibiotics Resistance Genes and their Expression Control Mechanism - ermC-4, a macrolide-lincosamide-streptogramin B resistance determinant on pMB4 from Staphylococcus aureus TR-1 (MLS계 항생물질 유도내성 유전자의 크로닝과 유전자의 발현조절 기전 - Staphylococus aureus TR-1균주의 프라스미드 pMB4에 존재하는 MLS 내성 유전자 ermC-4)

  • 김수환;최응칠;김병각;심미자
    • YAKHAK HOEJI
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    • v.35 no.1
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    • pp.22-29
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    • 1991
  • pMB4 is a 2.4-kilobase plasmid of Staphylococcus aureus TR-1 that confers inducible resistance to the macrolide-lincosamide-streptogramin B(MLS) antibiotics. By subcloning studies, it was found that the MLS resistance determinant was located at 1.0Kb fragment between Sau3AI and TaqI sites. DNA sequence of the MLS resistant determinant, named ermC-4 was determined, and found to be highly homologous with that of ermC. Because the leader peptide sequence of ermC-4 was identical with that of ermC, the expression of the resistance gene is thought to be controlled by posttranscriptional attenuation in S. aureus TR-1.

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The Significance of $p27^{KIP1}$, MIB-1, bcl-2 and p53 Expression in the Differential Diagnosis of Follicular Adenoma and Carcinoma of the Thyroid Gland (갑상선 여포상 선종과 암종의 감별진단에서 $p27^{KIP1}$, MIB-1, bcl-2, p53 발현의 유용성에 대한 연구)

  • Kang Mi-Seon;Yoon Hye-Kyoung;Kim Sang-Hyo;Yoon Ki-Young;Lee Choong-Han;Choi Kyung-Hyun;Hur Bang;Roh Mi-Sook;Hong Sook-Hee
    • Korean Journal of Head & Neck Oncology
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    • v.17 no.2
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    • pp.139-147
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    • 2001
  • Objectives: The most important differential point of follicular carcinoma from adenoma is capsular invasion or angioinvasion of follicular cells. Serial sections for examination of levels of tumor margins are necessary to watch the invasion. However, the interpretation of capsular invasion or angioinvasion is sometimes not feasible on the routine staining of tumor tissue. The aim of this study is to evaluate the clinical significance of expressions of $p27^{KIP1}$, MIB-1, bcl-2 and p53 in differential diagnosis of follicular adenoma and carcinoma. Materials and Methods: 16 cases of follicular carcinoma and 26 cases of follicular adenoma were entered on study of immunohistochemical stains for $p27^{KIP1}$, MIB-1, bcl-2 and p53. In carcinoma cases, correlation between the above markers, patient's age, tumor size, infiltration pattern and metastasis was studied. Results: $p27^{KIP1}$ labelling index (LI) of follicular carcinoma and adenoma was $4.89{\pm}6.92$ and $14.52{\pm}9.17$, respectively, but there was no significant difference between adenoma and carcinoma (p=0.2560). MIB-1 LI of carcinoma and adenoma was $4.11{\pm}3.89$ and $0.80{\pm}0.75$, respectively, and MIB-1 LI was significantly higher in carcinoma (p=0.0000). bcl-2 expression was seen in 2(12.5%) of 16 carcinoma cases and 130(50.0%) of 26 adenoma cases, and bcl-2 expression rate was higher in adenoma than in carcinoma(p=0.014). In one adenoma and one carcinoma case, p53 expression was noted. In follicular adenoma with atypia compared to adenoma without atypia, lower $p27^{KIP1}$ LI, higher MIB-1 LI and lower bcl-2 expression rate were seen. In follicular carcinoma, MIB-1 LI was significantly higher in invasive carcinoma(p=0.045) and was relatively increased in tumors larger than 3.0cm, showing angioinvasion and distant metastasis. But $p27^{KIP1}$ LI was higher in cases over 40 years old(p=0.008) and with conspicuous capsular invasion. There were no positive correlations between expressions of MIB-1, bcl-2 and p53. Conclusion: MIB-1 labelling index and bcl-2 expression could be helpful for differential diagnosis of follicular adenoma and carcinoma, but p53 showed very low expression rate and no significance in differential diagnosis. $p27^{KIP1}$ labelling index reveals decreasing tendency in carcinoma compared with adenoma, MIB-1 LI was considered as a poor prognostic marker in follicular carcinoma, but $p27^{KIP1}$ LI was higher in carcinoma cases over 40 years old with showing conspicuous capsular invasion. Further study for the significance of $p27^{KIP1}$ labelling index in follicular neoplasms is necessary to evaluate diagnostic value of follicular carcinoma.

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