• Parasites, Hosts and Diseases
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• 제52권4호
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• pp.367-376
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• 2014

Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with $GRA2_{25-105}$, $GRA3_{39-138}$, $ROP2_{324-561}$, and $MIC2_{1-284}$ domains had respectively higher value of IgG avidity. The $rGST-GRA2_{25-105}$ and $rGST-GRA3_{39-138}$ were soluble, while $rGST-ROP2_{324-561}$ and $rGST-MIC2_{1-284}$ were not. $GRA2_{31-71}$, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The $rGST-GRA2_{31-71}-ROP2_{324-561}$, a chimeric protein, appeared to be soluble. Moreover, $rGST-GRA2_{31-71}-MIC2_{1-284}$ was also soluble and had higher IgG avidity comparing to $rGST-MIC2_{1-284}$. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

• 한국기계가공학회지
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• 제15권6호
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• pp.115-121
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• 2016

The aircraft flight control system controls an aircraft's direction and flying attitude, and actuators are key components of control systems. Actuators can be classified as Geared Rotary Actuator (GRA) and Ball Screw Actuator (BSA). GRA is used in mid-sized aircraft, and BSA is used in larger aircraft. A two-stage differential GRA model was suggested in this paper, and structural analysis and performance tests were performed. According to the analysis and experiment, the stiffness of the two-stage differential GRA was 17.57% higher than that of the conventional GRA, and the structural safety was improved.

• Parasites, Hosts and Diseases
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• 제46권4호
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• pp.209-216
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• 2008

A monoclonal antibody against Toxoplasma gondii of Tg556 clone (Tg556) blotted a 29 kDa protein, which was localized in the dense granules of tachyzoites and secreted into the parasitophorous vacuolar membrane (PVM) after infection to host cells. A cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg556, and the full-length was completed by 5'-RACE of 2,086 bp containing an open reading frame (ORF) of 669 bp. The ORF encoded a polypeptide of 222 amino acids homologous to the revised GRA3 but not to the first reported one. The polypeptide has 3 hydrophobic moieties of an N-terminal stop transfer sequence and 2 transmembrane domains (TMD) in posterior half of the sequence, a cytoplasmic localization motif after the second TMD and an endoplasmic reticulum (ER) retrival motif in the C-terminal end, which suggests GRA3 as a type III transmembrane protein. With the ORF of GRA3, yeast two-hybrid assay was performed in HeLa cDNA expression library, which resulted in the interaction of GRA3 with calcium modulating ligand (CAMLG), a type II transmembrane protein of ER. The specific binding of GRA3 and CAMLG was confirmed by glutathione S-transferase (GST) pull-down and immunoprecipitation assays. The localities of fluorescence transfectionally expressed from GRA3 and CAMLG plasmids were overlapped completely in HeLa cell cytoplasm. In immunofluorescence assay, GRA3 and CAMLG were shown to be co-localized in the PVM of host cells. Structural binding of PVM-inserted GRA3 to CAMLG of ER suggested the receptor-ligand of ER recruitment to PVM during the parasitism of T. gondii.

• 약학회지
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• 제46권6호
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• pp.411-415
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• 2002

We compared the hypoglycemic effects of Ginseng Radix Alba (GRA) and Mori Folium (MF) in multiple low dose (MLD) streptozotocin(STZ)-induced diabetic rats. In order to induce hyperglycemic state 25 mg/kg of STZ was injected intraperitoneally for 5 consecutive days. SD rats were randomly divided into diabetic control and treatment groups. Treatment groups were administered with either 500 mg/kg of GRA (G500), 500 mg/kg of MF (M500), or 250 mg/kg of GRA mixed with same dose of MF (GM250) for 3 weeks. Blood glucose level and body weight were measured every 5th day. G500 and M500 both significantly reduced blood glucose levels as compared to the diabetic control group (diabetic control, 458.3$\pm$25.4 mg/dl; G500, 275.0$\pm$12.0; M500, 278.0$\pm$15.4; GM250, 324.0$\pm$18.4). While body weight in diabetic control group was decreased slightly after 3 weeks, treatment groups showed gradual increases of body weight during 3 week-period. Plasma insulin level was increased by treatment with GRA, but those levels in M500 and GM250 groups were similar to the diabetic control (normal control, 32.0$\pm$13.9 $\mu$IU/mι; diabetic control, 12.4$\pm$1.9; G500, 17.5$\pm$3.4; M500, 11.1$\pm$3.2; GM250, 10.5$\pm$t3.7). Urine glucose levels were also remarkably reduced in all treatment groups (normal control, 0.0$\pm$0.0 g/day; diabetic control, 11.4$\pm$2.5; G500, 4.9$\pm$0.2; M500, 5.7$\pm$1.6 ; GM250, 8.2$\pm$0.2). At the second and third week of the treatment, food and water intakes were determined. At the third week, food and water intakes were significantly decreased in all treatment groups. Taken together, we may conclude that GRA and MF alone may prevent or delay the development of hyperglycemia, however, synergistic hypoglycemic activity was not be seen in group treated with mixed formula of GRA and MF when compared to GRA or MF alone.

• Parasites, Hosts and Diseases
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• 제51권2호
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• pp.247-253
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• 2013

Neospora caninum is the etiologic agent of bovine neosporosis, which affects the reproductive performance of cattle worldwide. The transmembrane protein, NcSRS2, and dense-granule protein, NcGRA7, were identified as protective antigens based on their ability to induce significant protective immune responses in murine neosporosis models. In the current study, NcSRS2 and NcGRA7 genes were spliced by overlap-extension PCR in a recombinant adenovirus termed Ad5-NcSRS2-NcGRA 7, expressing the NcSRS2-NcGRA7 gene, and the efficacy was evaluated in mice. The results showed that the titer of the recombinant adenovirus was $10^9TCID_{50}/ml$. Three weeks post-boost immunization (w.p.b.i.), the IgG antibody titer in sera was as high as 1:4,096. IFN-${\gamma}$ and IL-4 levels were significantly different from the control group (P<0.01). This research established a solid foundation for the development of a recombinant adenovirus vaccine against bovine N. caninum.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.261.2-261.2
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• 2002

Hypoglycemic effect of GRA was examined in multiple low dose(MLD) streptozotocin(STZ)-induced diabetic rats with regard to time of administration. Experimental methods: 20 mg/kg of STZ in 100 mM citrate buffer(pH 4.5) was injected intraperitoneally for 5 consecutive days. In co-treatment groups, GRA was administered intraperitoneally for 3 weeks at dose of 150 or 300 kg/kg. (omitted)

• Parasites, Hosts and Diseases
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• 제53권3호
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• pp.345-348
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• 2015

Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa, which infects all warm-blood animals, including humans. In the present study, we examined sequence variation in dense granule 20 (GRA20) genes among T. gondii isolates collected from different hosts and geographical regions worldwide. The complete GRA20 genes were amplified from 16 T. gondii isolates using PCR, sequence were analyzed, and phylogenetic reconstruction was analyzed by maximum parsimony (MP) and maximum likelihood (ML) methods. The results showed that the complete GRA20 gene sequence was 1,586 bp in length among all the isolates used in this study, and the sequence variations in nucleotides were 0-7.9% among all strains. However, removing the type III strains (CTG, VEG), the sequence variations became very low, only 0-0.7%. These results indicated that the GRA20 sequence in type III was more divergence. Phylogenetic analysis of GRA20 sequences using MP and ML methods can differentiate 2 major clonal lineage types (type I and type III) into their respective clusters, indicating the GRA20 gene may represent a novel genetic marker for intraspecific phylogenetic analyses of T. gondii.

• Parasites, Hosts and Diseases
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• 제51권5호
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• pp.503-510
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• 2013

Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.

• Parasites, Hosts and Diseases
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• 제39권3호
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• pp.233-240
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• 2001

Although some reports have been published on the protective effect of antibodies to Toxoplasma gondii surface membrane proteins, few address the inhibitory activity of antibodies to dense granular proteins (GRA proteins) . Therefore, we performed a series of experiments to evaluate the inhibitory effects of monoclonal antibodies (mAbs) to GRA proteins (GRA2, 28 kDa; GRA6, 32 kDa) and surface membrane protein (SAGI, 30 kDa) on the invasion of T. gondii tachyzoites. Passive immunization of mice with one of three mAbs following challenge with a lethal dose of tachyzoites significantly increased survival compared with results for mice treated with control ascites. The survival times of mice challenged with tachyzoties pretreated with anti-GRA6 or anti-SAG 1 mAb were significantly increased. Mice that received tachyzoties pretreated with both mAb and complement had longer survival times than those that received tachyzoites pretreated with mAb alone. Invasion of tachyzoites into fibroblasts and macrophages was significantly inhibited in the anti-GRA2, anti-GRA6 or anti-SAG 1 mAb pretreated group. Pretreatment with mAb and complement inhibited invasion of tachyzoites in both fibroblasts and macrophages. These results suggest that specific antibodies to dense-granule molecules may be useful for controlling infection with T. gondii.

• 한국건설관리학회논문집
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• 제24권2호
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• pp.37-49
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• 2023

본 연구에서는 제조업 분야에서 보편적으로 활용되어 온 제약이론을 바탕으로 강교 제작공장의 생산성을 향상시키기 위한 자원할당 방법론을 제안한다. 이를 위해 도장 공정을 병목공정으로 정의한 후, 공정 전용 자원할당(OSRA), 제품 전용 자원할당(PSRA), 범용 자원할당(GRA) 등 3가지의 자원할당 방법론을 개발하였다. 강교 공급사슬 시뮬레이션 모델을 활용한 성능평가 실험 결과, GRA 방법론이 재공재고수(NWIP)와 대기시간(WT) 측면에서 가장 우수한 성능을 보였다. 특히, 강교 제작공장의 부하와 부하 변동성이 증가할수록 다른 두 전용 자원할당 방식과의 성능 격차는 더욱 커졌다. 평균적으로 GRA는 NWIP과 WT를 OSRA 대비 36.2%, 34.6%, PSRA 대비 71.0%, 70.4% 감소시켰다. 재공재고수와 대기시간의 감소는 병목현상의 완화를 의미하며, 이는 결국 강교 제작공장의 생산성이 향상되었음을 의미한다.