• 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.

• 대한수의학회지
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• 제36권1호
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• pp.1-9
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• 1996

Functional and morphological characteristics of the exocrine pancreas in genetic model BB rat of insulin dependent diabetes medllitus(IDDM) were carried out. Wistar rat was used as control animal. Flow rate of pancreatic juice, output of amylase and protein, and plasma glucose and insulin levess were examined. Also light and ultrastructural characteristics of the exocrine pancreas were observed. Pancreatic flow rate, output of amylase and protein, and insulin level were lower;glucose level was higher comparing with those of the control Wistar rat. In Wistar rat, exocrine pancreas was typical light microscopically. Zymogen granules and cell organelles were well developed in fine structure. Cell size of the periinsular acini was larger, and number of zymogen granules were more than those of the teleinsular acini. Most acinar cells were dark cells which containe well-developed RER in their cytoplasm. On the other hand, some light cells which have the dilated RER cisterns were found. In BB rat exocrine pancreas, cell size of per-and tele-insular acini similar to that of Wistar rat. The number of light cells occupied 40-50% compairing with that of Wistar rat. Zymogen granules were lower in number than that of Wistar rat and divied into three types in morphological characteristics ; type I showing normal structure, type II showing the wide hallo and small electron dense core in center of the zymogen granule and type III not having the electron dense core in the zymogen granule. The present ratio of type I, type II and type III are less than 5%, 30-40% and more than 50%, respectively.

• Journal of Plant Biology
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• 제39권2호
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• pp.123-126
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• 1996

The pattern of seed storage protein, vicilin, deposition and site of intracellular localization was examined in cotyledon cells of pea (Pisum sativum) seed using the immunocytochemical methods. The vicilin was confined to the cisternae fo the rough endoplasmic reticulum and dictyosome as well as protein granules newly formed in rough endoplasmic reticulum. Vacuolar protein deposites and protein bodies were also labelled by gold particles. After small protein bodies were formed in the rough endoplasmic reticulum, they were transported to large protein bodies and then fused together. Electron dense protein granule, elaborated in the dictyosome, appears to be transported from dictyosome to protein body. A few unlabelled protein granules seem to be accumulated in other type of proteins than vicilin.

• Bulletin of the Korean Chemical Society
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• 제35권10호
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• pp.3071-3076
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• 2014

Proteomic analyses of the excretory/secretory proteins from the RH strain of Toxoplasma gondii have been performed to understand their functions in the host-parasite interaction. A total of 34 proteins were identified from LC/MS/MS analysis and their abundance was estimated by spectral counting methods. Among them, 8 species of micronemal proteins (MICs), 2 species of rhoptry proteins (ROPs), and 6 species of dense granular proteins (GRAs) were confirmed. Besides these, 18 species of protein were newly identified, and their cellular functions were estimated from sequence analysis. The three most abundant of the 34 identified extractor/secretory proteins-GRA1, GRA7 and GRA2-were confirmed to be highly expressed in T. gondii using the spectral count method. This phenomenon is another demonstration of the importance of GRA proteins for the penetration and survival of T. gondii.

• 대한수의학회지
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• 제33권4호
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• pp.573-577
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• 1993

청설모(Sciurus vulgaris corea)의 췌장에 분포하는 glucagon 및 somatostatin 세포의 분포와 그들 분비과립의 특징을 밝히기 위해 면역조직화학적 및 면역전자현미경적 방법으로 관찰하였다. Glucagon 면역반응세포는 주로 췌도의 주변부에 분포하였고, 때로는 외분비부에서 관찰되었다. 이 세포의 분비과립은 직경이 240~320nm였으며, 전자밀도가 높은 core를 전자밀도가 낮은 halo가 둘러싸고 특히 gold particle들은 전자밀도가 높은 core에 강한 반응을 보였다. Somatostatin면역반응세포는 췌도의 주변부를 따를 면역반응이 아주 약한 세포들로 산재하였으며 또한 외분비에서 단독 혹은 소집단으로 관찰되었다. 이 세포의 분비과립은 전자밀도가 낮고 직경이 250~275nm였으며, gold particle는 분비과립에 중등도로 고른 반응을 보였다.

• Journal of Ginseng Research
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• 제46권3호
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• pp.387-395
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• 2022

Background: Fermentation may alter the bioavailability of certain compounds, which may affect their efficacy and pharmacological responses. This study investigated the antiplatelet effects of red ginseng extract (RGE) and fermented red ginseng extract (FRG). Methods: A rodent model was used to evaluate the antiplatelet and antithrombotic effects of the extracts. Rats were orally fed with human equivalent doses of the extracts for 1 week and examined for various signaling pathways using standard in vivo and ex vivo techniques. Light transmission aggregometry was performed, and calcium mobilization, dense granule secretion, integrin α_IIbβ₃-mediated signaling molecules, cyclic nucleotide signaling events, and various protein molecules were evaluated ex vivo in collagen-stimulated washed platelets. Furthermore, antithrombotic properties were evaluated using a standard acute pulmonary thromboembolism model, and the effects on hemostasis were investigated using rat and mice models. Results: Both RGE and FRG significantly inhibited platelet aggregation, calcium mobilization, and dense granule secretion along with integrin-mediated fibrinogen binding and fibrinogen adhesion. cAMP levels were found to be elevated in RGE-treated rat platelets. Ginseng extracts did not exert any effect on prothrombin time and activated partial thromboplastin time. RGE-treated mice showed significantly better survival under thrombosis than FRG-treated mice, with no effects on hemostasis, whereas FRG-treated mice exhibited a slight increment in bleeding time. Conclusion: Both extracts, especially RGE, are remarkable supplements to maintain cardiovascular health and are potential candidates for the treatment and prevention of platelet-related cardiovascular disorders.

• 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.

• Applied Biological Chemistry
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• 제39권5호
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• pp.379-383
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• 1996

조생종 쌀인 오대벼 품종을 대상으로 종자의 배유세포에 있는 전분복합체와 단백질체의 외부형태 및 미세 구조의 특징을 광학현미경과 전자현미경으로 관찰하였다. 배유세포는 긴 막대모양으로 세로 단면이 장방형 또는 마름모형을 하고 있고, 세포벽은 $0.5\;{\mu}m$의 두께로 균질한 막상 구조물로 이루어져 있다. 또한, 세포내에는 구형 또는 타원형의 전분복합체가 치밀하게 채워져 있으며, 직경이 $18{\sim}25\;{\mu}m$로 다양한 크기로 존재하였다. 전분복합체는 12면체 또는 14면체의 중앙 전분과립을 중심으로 약 $5\;{\mu}m$ 크기의 전분과립들이 동심원상으로 2 내지 3층으로 형성되어 있다. 배유세포에서 단백질체는 호분층에서 보다 아주 드물게 관찰되었지만, 전분복합체의 주변에 인립하여 산재하고 있다. 단백질체는 직경이 약 $3\;{\mu}m$로 구형 또는 난원형의 형태를 하고 있으며 미세구조적으로 중심부는 전자밀도가 높게, 한계막의 주변부는 비교적 전자밀도가 낮게 관찰되었다.

• 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.

• Parasites, Hosts and Diseases
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• 제48권3호
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• pp.195-201
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• 2010

We studied on the proteomic characteristics of Toxoplasma gondii KI-1 tachyzoites which were originally isolated from a Korean patient, and compared with those of the well-known virulent RH strain using 2-dimensional electrophoresis (2-DE), mass spectrometry, and quantitative real-time PCR. Two-dimensional separation of the total proteins isolated from KI-1 tachyzoites revealed up to 150 spots, of which 121 were consistent with those of RH tachyzoites. Of the remaining 29 spots, 14 showed greater than 5-fold difference in density between the KI-1 and RH tachyzoites at a pH of 5.0-8.0. Among the 14 spots, 5 from the KI-1 isolate and 7 from the RH strain were identified using MALDI-TOF mass spectrometry and database searches. The spots from the KI-1 tachyzoties were dense granule proteins (GRA 2,3,6, and 7), hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGRPTase), and uracil phosphoribosyltransferase (UPRTase). The spots from the RH strain were surface antigen 1 (SAG 1), L-lactate dehydrogenase (LDH), actin, chorismate synthase, peroximal catalase, hexokinase, bifunctional dihydrofolate reductase-thymidylate synthase (DHTR-TS), and nucleosidetriphosphatases (NTPases). Quantitative real-time PCR supported our mass spectrometric results by showing the elevated expression of the genes encoding GRA 2,3, and 6 and UPRTase in the KI-1 tachyzoites and those encoding GRA 7, SAG 1, NTPase, and chorismate synthase in the RH tachyzoites. These observations demonstrate that the protein compositions of KI-1 and RH tachyzoites are similar but differential protein expression is involved in virulence.