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In Vitro Magnetometric Evaluation far Toxicity to Alverolar Macrophage of Arsenic Compounds (In Vitro 자계(磁界) 측정에 의한 비소화합물의 폐포 Macrophage 독성 평가)

  • Cho, Young-Chae
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.4
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    • pp.467-472
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    • 1999
  • Objectives: This study was conducted to evaluate the cytotoxicity of gallium arsenide(GaAs), indium phosphide(InP) and indium arsenide(InAs) all of which are used a$ the semiconductor eletments in semiconductor industry. Methods: Cytotoxicity id the alveolar macrophage was evaluated by the measurement of in vitro magnetometry, LDH release assay and histological examination. Results: The relaxation curves by the in vitro magnetometry showed that GaAs has the cytotoxicity for the alveolar macrophage which is more significant in the higher dosages, while this cytotoxicity is not appeared in the groups added with InP or InAs or PBS. In the decay constant for two minutes after magnetization, GaAs-added groups showed a significant decrease with increasing doses, but both InP- and InAs-added groups did not show any significance. The LDH release assay showed a dose-dependent increasing tendency in the GaAs-, InP- and InAs-added groups. In terms of cellular morphological changes, GaAs-added groups revealed such severe cellular damages as prominent destructions in cell membranes and their morphological changes of nucleus, while InP- and InAs-added groups remained intact in intracellular structures, except for cytoplasmic degenerations. Conclusions: It is suggested that GaAs is more influential to cytotoxicity of alveolar macrophages than InP and InAs.

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A Study of Correcting Technology based POI for Pedestrian Location-information Detecting in Traffic Connective Transferring System (교통 연계 환승 시스템의 보행자 위치정보 수집을 위한 POI 기반 위치 보정 기술 연구)

  • Jung, Jong-In;Lee, Sang-Sun
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.10 no.2
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    • pp.84-93
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    • 2011
  • In order to provide the real time and proper information to the pedestrian who is using the transport connection and transfer center through data collecting and processing process, the design of the test-bed (Gimpo airport)'s communication construction and the technology of the pedestrian location tracking has been researched. The design of the communication construction should make sure that it can provide believable data to the user of the transfer center. At the same time, the location tracking should also be considered, so that the require of the communication efficiency and the location tracking efficiency can be met together. In order to make the efficient location tracking technology, the problems related to the commercial technology based real time location identification will be resolved and the new approach method was proposed and be applied and analysed to the test-bed. The wireless access points can be located in the most real-world situation which has added the characteristics of the real building to the electronic map, and through the analysis of theirs location, they can be set as the mainly necessary points for the communication construction design and the location tracking and the method to locate that points has been proposed. How to set, how to apply it to the test-bed and the examination result will be introduced in this paper.

Identification of Potential Substrates of N-acteylglucosamine Kinase by a Proteomic Approach (프로테오믹스를 이용한 N-아세틸글루코사민 인산화효소 기질단백질의 동정)

  • Lee, HyunSook;Moon, Il Soo
    • Journal of Life Science
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    • v.23 no.4
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    • pp.586-594
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    • 2013
  • Post-translational O-GlcNAc modification (O-GlcNAcylation) of serine or threonine is a new protein modulation mechanism. In contrast to the classical glycosylation, O-GlcNAcylation occurs in a one-step transfer of O-GlcNAc on both nuclear and cytoplasmic proteins. In contrast to the general consensus that O-GlcNAc is a final modification, a recent paper (J Proteome Res. 2011 10:2725-2733) showed the presence of O-GlcNAc-P on a synaptic assembly protein AP180. This finding raises a fundamental question about its prevalence. To address this question, we used proteomics to identify those proteins that were phospho-signal enriched by GlcNAc kinase (NAGK). Comparison of pDsRed2-$NAGK_{WT}$-transfected HEK293T cell extract with pDsRed2-$NAGK_{D107A}$-transfected control culture revealed 15 phospho-signal increased spots. Excluding those spots that had no detectable amount of protein expression yielded 7 spots, which were selected for ID determination. Among these, two duplicate spots (two $HSP90{\beta}$ and two ENO1 spots) were shown to be O-GlcNAcylated, two (dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P) were not known to be involved in O-GlcNAcylation, and one (heat shock protein gp96 precursor or grp94) was a glycoprotein. The increase in the phospho-levels of O-GlcNAc by NAGK strongly indicates that these proteins are phosphorylated on O-GlcNAc. Our present data support the idea that O-GlcNAc is not a terminal modification.

Molecular Cloning and Expression of a Cellulolytic Xylanase Gene from Bacillus circulans in Escherichia coli (Bacillus circulans 기원의 Cellulolytic Xylanase 유전자의 대장균에서의 클로닝 및 발현)

  • 이동석;김지연;김한복
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.196-202
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    • 2000
  • A gene for cellulolytic xylanase of Bacillus circulnns ATCC21365 was cloned on pUC 19 in Eschwichia coli. The recombinant plasniid pXLI80 contained an 1.8 id, inselt composed of0.5 kb and 1.3 kb PslI fragments derived from B, circulans. The 0.5 kh fragment in the upstream region of 1.3 kb one was confirmed lo be indispensable for not only expression but also hyperexpression of the cloned gene. The transformant overproduced the xylanase 135 times greater than that produced by the orlginal B circulnns. The optimum pH and temperature of the cloned enzyme we]-e pH 5.2 and $60^{\circ}C$, respectively. Heal pretl-eatment at TEX>$55^{\circ}C$C for 1 Indid not cause inhibition of the activity of this enzyme. The elm.ynie could hydl-olyre CMC and lichenan as well as xylan to produce xylose(or GI), xylohiose(or G2) and xylolnose(or G3) as inah products. Hence We defined the cloned enzyme as a cellulolytic xylanase. The SDS-PAG electrophoretic mobility and zyiiogram of this enzyme derived from whole cell extracts or c~~lture supematants or E. coli(pXL180) indicated a molecular weight of 45,000 and nonprocessing of the enzyme in the peilplasln of E. coli.

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Evaluation of 99mTc-MAG3-2-nitroimidazole for hypoxic tumor imaging

  • Lee, Yun-Sang;Kim, Young Joo;Jeong, Jae Min
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.5 no.1
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    • pp.18-25
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    • 2019
  • 2-Nitroimidazole derivatives have been reported to accumulate in hypoxic tissue. We prepared a novel $^{99m}Tc-MAG_3$-2-nitroimidazole and evaluated the feasibility for hypoxia imaging agent. $Bz-MAG_3$-2-nitroimidazole was synthesized by direct coupling of $Bz-MAG_3$ and 2-nitroimidazole using dicyclohexylcarbodiimide. $Bz-MAG_3$-2-nitroimidazole was labeled with $^{99m}Tc$ in the presence of tartaric acid and $SnCl_2-2H_2O$ at $100^{\circ}C$ for 30 min. And the reaction mixture was purified by $C_{18}$ Sep-pak cartridge. The labeling efficiency and the radiochemical purity were checked by ITLC-SG/acetonitrile. The tumor was grown in balb/c mice for 8~13 days after the subcutaneous injection of tumor cells, CT-26 (murine colon adenocarcinoma cell). Biodistribution study and tumor autoradiography were performed in the xenografted mice after i.v injection of 74 kBq/0.1 mL and 19 MBq/0.1 mL of $^{99m}Tc-MAG_3$-2-nitroimidazole, respectively. In vivo images of $^{99m}Tc-MAG_3$-2-nitroimidazole in tumor bearing mice were obtained 1.5 hr post injection. The labeling efficiency was $45{\pm}20%$ and the radiochemical purity after purification was over 95%. Paper electrophoresis confirmed negative charge of $^{99m}Tc-MAG_3$-2-nitroimidazole. $^{99m}Tc-MAG_3$-2-nitroimidazole was very stable at room temperature and its protein binding was 53%. The $^{99m}Tc-MAG_3$-2-nitroimidazole exhibited high uptake in the liver, stomach and intestine. In biodistribution study using tumor bearing mice, the uptakes (% ID/g) of the tumor were $0.5{\pm}0.1$, $0.4{\pm}0.0$, $0.2{\pm}0.1$ and $0.1{\pm}0.1$ at 5, 15, 30 min and 4 hrs. Tumor/muscle ratio were $1.4{\pm}0.1$, $2.2{\pm}0.83$, $3.0{\pm}0.9$, and 3.7 (n=2) for 5, 15, 30 min and 4 hrs. The uptake in hypoxic area was found higher than in non-hypoxic area of tumor tissue by autoradiography. In vivo images showed the relatively faint uptake to the hypoxic tumor region. $^{99m}Tc-MAG_3$-2-nitroimidazole was successfully synthesized and found feasible for imaging hypoxia.

Synthesis and biological evaluation of diagnostic reagent for prostate cancer using copper-64 radioisotope

  • Ahn, Heesu;Kim, Mi Hyun;Han, Sang Jin;Woo, Sang Keun;Kim, Jung Young;Lee, Kyu Chul;Lim, Il Han;Lee, Yong Jin
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.4 no.2
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    • pp.65-72
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    • 2018
  • Prostate specific membrane antigen (PSMA) is a cell surface membrane protein, which is overexpressed in most prostate cancer. Recently, PET imaging with $[^{68}Ga]$PSMA-HBED-CC has been widely used for the diagnosis of recurrent prostate cancer and the studies on the diagnostic potential of $^{64}Cu$-labeled PSMA ligands reported actively. In this study, we monitored with biological evaluation in vivo and PET imaging of $^{64}Cu$-labeled PSMA ligand ($[^{64}Cu]$PSMA-617). The radiolabelling efficiency and stability of $[^{64}Cu]$PSMA-617 were confirmed by radio-thin layer chromatography. The radiolabeling efficiency of $[^{64}Cu]$PSMA-617 showed over 95%, and stabilities of intact remained over 98% in both human and mouse serum for 48 h. In normal male mice, in vivo uptake of $[^{64}Cu]$PSMA-617 in several organs was measured at 2, 4, 6, 24, 48 h after injection. Rapid blood clearance was observed for $[^{64}Cu]$PSMA-617. The high uptake was observed in the lung, liver, intestines and kidneys at 2 h postinjection, but was low in the other organs (1-2 %ID/g) at 4 h. The dynamic PET/CT images of 22RV1 tumor-bearing nude mice were acquired during 60 min and additionally acquired 24 h and 48 h after injection. In dynamic PET images, $[^{64}Cu]$PSMA-617 uptake ratio in tumors versus muscle was increased as time elaplsed until 60 minutes and remained in tumors at 48 h. In these results, the PET/CT imaging using $[^{64}Cu]$PSMA-617 in prostate cancer is expected to be useful for the diagnosis and treatment of prostate cancer patients.

Comparative Genomic Analysis of Pathogenic Factors of Pectobacterium Species Isolated in South Korea Using Whole-Genome Sequencing

  • Jee, Samnyu;Kang, In-Jeong;Bak, Gyeryeong;Kang, Sera;Lee, Jeongtae;Heu, Sunggi;Hwang, Ingyu
    • The Plant Pathology Journal
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    • v.38 no.1
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    • pp.12-24
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    • 2022
  • In this study, we conducted whole-genome sequencing with six species of Pectobacterium composed of seven strains, JR1.1, BP201601.1, JK2.1, HNP201719, MYP201603, PZ1, and HC, for the analysis of pathogenic factors associated with the genome of Pectobacterium. The genome sizes ranged from 4,724,337 bp to 5,208,618 bp, with the GC content ranging from 50.4% to 52.3%. The average nucleotide identity was 98% among the two Pectobacterium species and ranged from 88% to 96% among the remaining six species. A similar distribution was observed in the carbohydrate-active enzymes (CAZymes) class and extracellular plant cell wall degrading enzymes (PCWDEs). HC showed the highest number of enzymes in CAZymes and the lowest number in the extracellular PCWDEs. Six strains showed four subsets, and HC demonstrated three subsets, except hasDEF, in type I secretion system, while the type II secretion system of the seven strains was conserved. Components of human pathogens, such as Salmonella pathogenicity island 1 type type III secretion system (T3SS) and effectors, were identified in PZ1; T3SSa was not identified in HC. Two putative effectors, including hrpK, were identified in seven strains along with dspEF. We also identified 13 structural genes, six regulator genes, and five accessory genes in the type VI secretion system (T6SS) gene cluster of six Pectobacterium species, along with the loss of T6SS in PZ1. HC had two subsets, and JK2.1 had three subsets of T6SS. With the GxSxG motif, the phospholipase A gene did locate among tssID and duf4123 genes in the T6SSa cluster of all strains. Important domains were identified in the vgrG/paar islands, including duf4123, duf2235, vrr-nuc, and duf3396.

Cell Surface Antigenic Relationship of Pathogenic Mycobacteria (병원성 Mycobacteria의 세포표면항원간의 항원적 상관 관계)

  • Kwon, Hyuk-Han;Saito, Hajime;Kim, Sang-Jae
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.5
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    • pp.483-494
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    • 1993
  • Cell surface antigenic relationships between pathogenic mycobacteria have been investigated by the enzyme-linked immunosorbent assay using phenolkilled cells and their rabbits antisera. Homologous and heterologous reactions of Mycobacterium avium-intracellulare antisera before and after homologous and heterologous absorption revealed a close antigenic relationship between strains of the same species and between species if they were members of M. avium(MA)-intracellulare(MI)-scrofulaceum(MG) complex. MAI sera showed a considerable reaction with M. kansasii(MK) and tuberculosis(MTB), but not with the other species. MA(K40004) antiserum reacted with other mycobacteria except few strains of MI and 50~89% of homologous reaction was reduced by heterologous absorption with cells of MI or MS. Intraspecific reaction of MI antisera was natural1y stronger than interspecific reaction and different in extent due to a magnitude of antigenic sharing. Antigenic relationships between N-260D, N-260R, N-260T, and K41014 was somewhat closer than that with N-242D, N-257T, N-28ID, and N-275T. M. nonchromogenicum(MNC) antisera showed a strong interspecific reaction with exception of M. chelonei(MC) and triviale(MTV) to which they reacted weakly or none. Antigenic sharing with M. terrae(MTR) and MG(K30003) was next to intraspecific sharing. NC-3 shared antigens considerably with MA, MC, and M. fortuitum(MF) while NC-11 did not. MTR antisera showed a strong cross-reaction with MI but their homologous reaction was not reduced by MI absorption indicating a paucity of shared antigen of MTR surface. Intraspecific antigenic sharing of course was large with on exception between T-8 and T-13. A considerable amount of antigenic sharing was also found with MNC, MC and MF. Unlike T-8 serum, T-13 antiserum strongly cross-reacted with MA, MG, MK, and MTB. In general, antigenic relationships of mycobacteria, that have been elucidated in this study, well conformed to taxons delineated by the various biological and biochemical means.

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Animal Experiments on an Antithrombogenic Small-Caliber Vascular Prostheses and Vascualr Patch : Observation in Canine Models (항혈전성 소구경 인조 혈관 및 봉합편에 대한 동물 실험)

  • 김수철;김원곤;유세영
    • Journal of Chest Surgery
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    • v.36 no.2
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    • pp.63-72
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    • 2003
  • Although a variety of synthetic vascular grafts are available in modern vascular surgery, no ideal prosthesis ha,4 yet been developed. Small-caliber vascular grafts with low flow, as used in the lower extremity, continue to become thrombosed at unacceptable rates. We have developed and evaluated the new antithrombogenic blood contacting surfaces in canine model. Material and Method: Two now antithrombogenic blood contacting surfaces(Polyvinylalcohol -Polyurethane(PVA-PU) blend and natural Graphite-polyurethane(G-PU) blend) have been developed and evaluated in canine model, using vascular grafts and patches. The luminal surfaces of the test vascular grafts(5 mm ID) were fabricated by dipping a glass rod in PVA-PU blend solution(50 % PVA) using phase separation method. Mongrel dogs of either sex weighing 18-22 kg were anesthetized by endotracheal intubation using halothane and their lungs were ventilated with a volume-cycled ventilator, Maintenance anesthesia with 0.5-1.0% halothane and supplemental oxygen was used. Two pairs were used for comparison in the bilateral femoral arteries for both vascular grafts(PVA-PU vs. PU) and vascular patches(G-PU vs. PU). Bilateral groin incisions were made and the arteries were exposed and clamped. After an excision of 1 cm of the artery between clamps, a grail of 2.5 cm in length was implanted end-to-end using 6-0 polypropylene suture. The vascular patch was implanted as a form of on-lay patch. Animals were sacrificed at 1, 2, 4, 6, 8 and 16 weeks for vascular grafts and 1, 2. 4 and 6 weeks for vascular patches. Result The vascular grafts of PVA-PU blends showed patent lumina in the 2 and 16 weeks animals, while those of PU showed a patent lumen in 2 weeks animal. PVA-PU graft of 16 weeks showed a fairly clean luminal surface. A light microscopic finding of this graft demonstrated good tissue infiltration through porosity, The animals with vascular patches showed patent arteries in both groups except 2 weeks animal. Scanning electron microscopy of the luminal surfaces of G-PU patches in 4 and 6 weeks animals showed endothelial cell covering with microvilli. PU patches showed qualitatively less endothelial cell covering. Conclusion: In conclusion, PVA-PU and G-PU blends can be a promising blood contacting surfaces for application in a synthetic vascualr graft. However, further animal study is needed to determine the real long-term effects of these methods of surface modifications.

Effect of Thyroid Hormone on the Electrical Activity of Rabbit Heart (토끼심장의 전기적 활동에 대한 갑상선 호르몬의 영향)

  • Hong, Seong-Geun;Kwun, Jong-Kuk;Chung, Soon-Il
    • The Korean Journal of Physiology
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    • v.20 no.1
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    • pp.17-29
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    • 1986
  • The present study was carried out to observe the effect of triiodothyronine on heart, one of the target organ of thyroid hormone. There are many reports that tachycardia, arrythmia, and agumentation of sodium, potassium pump activity are caused in hyperthyroid animal. To examine these cardiac positive chronotropic effects on sinoatrial (SA) node and atrial muscle, hyperthyroid state was induced experimentally by the injecion of 3,3',5-1-triiodothyronine $(T_3)$ in $3{\sim}6$ month-old rabbits. Then intracellular recordings by inserting glass microelectrode into cell were obtained in SA node and atrial muscle. The results can be summarized as follows : 1) Heartbeat was increased from $169.6{\pm}28.0\;to\;264.2{\pm}18.0$ beats per minute, while body weight was decreased to 68f of the initial body weight (Day 1). 2) In experimental group, the duration of action potential at 80% repolarization was decreased from $148.0{\pm}29.1\;to\;107{\pm}13.6msec$. This suggested the increase heartbeat. 3) The firing rate in hyperthyroid group markedly reduced under the 15 mM potassium Tyrode (p<0.005). 4) In hyperthyroid group, depolarization of atrial muscle cell was lowered significantly in 15 mM (p<0.05), 20 mM (p<0.05) potassium Tyrode solution. 5) Sodium-potassium pump activities in experimental group were higher than those in control group in both SA node (p<0. 1) and atrial muscle (p<0.025). 6) In lower concentration of $MnCl_2$, the excitability of SA node in hyperthyroid group was decreased more than that in control group. Effective inhibitory dose $(ID_{50})$ as 0.6 mM in hyperthyroid statd and 1.1 mM in control group.

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