• 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.

• 한국동물위생학회지
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• 제39권2호
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• pp.117-124
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• 2016

In this study, the virucidal efficacy of a fumigant containing 20% ortho-phenylphenol against classical swine fever virus (CSFV) and porcine reproductive and respiratory syndrome virus (PRRSV) was examined. After each carrier deposited with CSFV and PRRSV suspensions was exposed to the fumigant in a $25-m^3$ test room for 15 h, all carriers were neutralized and diluted, and each diluted suspension was inoculated into each proper cell line. After incubation, CSFV and PRRSV viability in each cell line was examined and 50% tissue culture infectious dose $(TCID_{50})/mL$ was calculated. In the results, the concentration of viable virus in all of pathogen control-carriers was more than $2{\times}10^5TCID_{50}/mL$, and there were no cytotoxicity in all of toxicity control-carriers. In addition, the fumigant inactivated ${\geq}4.8{\log}_{10}(TCID_{50}/mL)$ of both CSFV and PRRSV. These findings will be useful for preventing the spread of CSFV and PRRSV infection.

• 대한수의학회지
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• 제36권1호
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• pp.169-179
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• 1996

This study was carried out to examine the role of small cells and oval cells in cholangiocarcinogenesis in the hamsters infected with Clonorchis(C) sinensis. Forty two female Syrian golden hamsters were divided into two groups. Group I was for the induction of the cholangiocarcinoma, which was infected orally with C sinensis and given dimethylnitrosamine(15ppm) in drinking water for 4 weeks. Group II was served as control. More than 5 heads of hamsters in each group were sacrificed at 4, 7, 11 and 15 weeks after the beginning of the experiment. The livers were examined histopathologically, electron microscopically and immunohistochemically. The results obtained were as follows; 1. Cholangiocarcinomas were occurred in 1 of 6 animals at 11 weeks and in 4 of 6 animals at 15weeks after the beginning of the experiment. 2. Small cells and oval cells were proliferated around the portal triads from 4 weeks and peaked at 11 weeks, and slightly decreased after then. 3. The strong positive reaction to the $\alpha$-fetoprotein was shown in many of small cells and oval cells. But ductlike oval cells, which were arranged rosette form, showed week positive reaction to the $\alpha$-fetoprotein. 4. Most of small cells and oval cells showed negative reaction to the cytokeratin. But weak positive reaction in ductlike oval cells, and moderate positive reaction in cholangiocarcinoma cells were observed. These results suggested that cholangiocarcinoma induced by infection of C sinensis was believed to originate from the proliferated small cells around the portal triads which would be able to differentiate to the oval cells, ductlike oval cells, and cholangiocarcinoma cells gradually.

• 구강회복응용과학지
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• 제23권4호
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• pp.303-312
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• 2007

Recently several studies have been developed not only to apply bone materials to bony defect, but also to use osteogenic and osteoinductive materials to form bone more effectively. In 1998 Mark et al applied gel formation of PRP(platelet-rich plasma) in bony transplantation for mandibular reconstruction as one of the method of stimulating bone formation in maxillofacial area, which is contain of varies growth factors. After he reported that PRP accelerate bone formation, which is used in varies bone transplantation and augmentation with a good result. Especially there are amount of growth factors in PRP, and PRP increase angiogenesis, cell division, and mesenchymal cell growth. Moreover it is capable of osteoconduction, hemostatitis, anti-infection, forming the shape at transplantation, ease of handling, and recipient site stability. So it is known that success rate is high in bone transplantation. However PRP need tissue adhesive to make plasma to solid form. Thrombin and calcium chloride, component of PRP, is extracted from autogenic donor. So it is expensive to extract and there is possibility of hepatitis, AIDS, and hematogenous metastasis. After all, tissue adhesive have the limitation and danger of use. So we are willing to introduce that we had get some idea after using PRF(platelet-rich fibrin) in the various hard and soft tissue bony defect, which is self extracted simply and contain growth factors.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.26-26
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• 2015

Phytophthora infestans is the causal agent of potato and tomato late blight, one of the most devastating plant diseases. P. infestans secretes effector proteins that are both modulators and targets of host plant immunity. Among these are the so-called RXLR effectors that function inside plant cells and are characterized by a conserved motif following the N-terminal signal peptide. In contrast, the effector activity is encoded by the C terminal region that follows the RXLR domain. Recently, I performed in planta functional profiling of different RXLR effector alleles. These genes were amplified from a variety of P. infestans isolates and cloned into a Potato virus X (PVX) vector for transient in planta expression. I assayed for R-gene specific induction of hypersensitive cell death. The findings included the discovery of new effector with avirulence activity towards the Solanum bulbocastanum Rpi-blb2 resistance gene. The Rpi-blb2 encodes a protein with a putative CC-NBS-LRR (a coiled-coil-nucleotide binding site and leucine-rich repeat) motif that confers Phytophthora late blight disease resistance. We examined the components required for Rpi-blb2-mediated resistance to P. infestans in Nicotiana benthamiana. Virus-induced gene silencing was used to repress candidate genes in N. benthamiana and to assay against P. infestans infections. NbSGT1 was required for disease resistance to P. infestans and hypersensitive responses (HRs) triggered by co-expression of AVRblb2 and Rpi-blb2 in N. benthamiana. RAR1 and HSP90 did not affect disease resistance or HRs in Rpi-blb2-transgenic plants. To elucidate the role of salicylic acid (SA) in Rpi-blb2-mediated resistance, we analyzed the response of NahG-transgenic plants following P. infestans infection. The increased susceptibility of Rpi-blb2-transgenic plants in the NahG background correlated with reduced SA and SA glucoside levels. Furthermore, Rpi-blb2-mediated HR cell death was associated with $H_2O_2$, but not SA, accumulation. SA affects basal defense and Rpi-blb2-mediated resistance against P. infestans. These findings provide evidence about the roles of SGT1 and SA signaling in Rpi-blb2-mediated resistance against P. infestans.

• 한국임상수의학회지
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• 제23권1호
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• pp.9-13
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• 2006

Anaplasma phagocytophilum의 주요 표면단백질인 Msp2 (p44)는 세균의 표면에 발현되는 주요한 항원 단백질이다. 본 실험에서는 A. phagocytophilum이 주요 숙주세포인 호중구나 HL-60 세포에 침입하는 데 있어, 세균의 주요 표면 단백질인 Msp2와 숙주세포의 표면에 발현되는 PSGL-1과의 반응 여부를 알아보았다. 그 결과, 재조합 단백질인 Msp2나 순수하게 분리된 A. phagocytophilum이 PSGL-1/FucT IV 유전자가 형질전환되어 PSGL-1이 발현되는 BJAB 세포와는 결합하지만, 순수한 BJAB 세포와는 전현 반응하지 않는 것을 관찰할 수 있었다(p<0.01 & p<0.01). 또한, 순수하게 분리된 A. phagocytophilum과 형질전환된(PSGL-1/FucT IV) BJAB 세포와의 결합이 Msp2의 단-클론항체나 Msp2 재조합 단백질의 농도에 따라 억제됨을 관찰할 수 있었다(p<0.05 & p<0.01). 따라서 A. phagocytophilum의 Msp2가 숙주세포인 호중구의 표면에 발현되는 PSGL-1과 직접적으로 결합하는 부착물질임을 알 수 있었다.

• 대한수의학회지
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• 제38권1호
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• pp.71-76
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• 1998

The effects of chitosan on mastitis in lactating holstein cows were evaluated. Fifty six cows with intramammary infection(IMI) from nine farms were selected and the cows were fed with diets which contained 15~20g chitosan per day for 5~7 days. The milk samples were obtained from cows at 7 days and 14 days after administration to determine effect of the curing of mastitis and the reduction of somatic cell counts(SCC). The average value of SCC levels in quarter milk from the cows administrated with chitosan significantly decreased up to 31.8% and 47.7% at 7 and 14 days, respectively(P<0.05). The cure rates of chitosan for Stapylococcus aureus, coagulase-negative Staphylococci, Streptococci spp, other gram positive bacteria and coliforms were 30.4, 42.8, 33.3, 66.6 and 54.5 % respectively. Twenty three out of 64 cases were cured by feeding with chitosan. The results showed that administration of chitosan could reduce SCC in milk and improve cure rates of bovine mastitis caused by microorganisms. The further studies will be pursued to study on the mechanism of chitosan in the immune responses of cows with mastitis.

• 대한수의학회지
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• 제32권1호
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• pp.77-82
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• 1992

세포 비병원성 소 설사병 바이러스가 감염된 우태아 폐세포를 이용 바이러스를 순수분리하여 이화학적 성상을 검사하였던 바 다음의 결과를 얻었다. 바이러스의 비중은 $1.090{\sim}1,114gm/cm^3$로 검출되었으며 $1,098gm/cm^3$에서 최대 감염가를 나타내었다. 면역 전자현미경을 이용한 형태학적 분석결과 30~80nm의 바이러스성 입자를 관찰할 수 있었으며 유전인자의 추출 분석결과 병원성 바이러스와의 전기영동상 유전자 규모에 있어서 차이는 인정되지 않았다. 그러나 비세포 병원성 소 설사병 바이러스는 병원성 바이러스에 비하여 세포외 배출이 적었으며 세포내에서 보다 많이 존재하는 것으로 관찰되었다.

• 대한한의학회지
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• 제19권1호
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• pp.419-429
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• 1998

Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) has been used for the treatment of allergic disease. IgE is normally one of the least abundant immunoglobulin (Ig) isotypes in the serum of both humans and several species of experimental animals: however a number of different stimuli can result in profound increases in IgE levels relative to other isotypes. In rodents, infection with many parasitic helminths can cause approximately 100-fold elevation in IgE within 2 wks. Immunization of mice with small amounts of protein antigens on alum also results in 10-fold to fold increase in total serum IgE, much of it specific for the immunizing antigen. In this experiment, I investigated the effect of an aqueous extract of Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) on a in vivo and in vitro IgE production. PTFE dose-dependently inhibited the serum levels of IgE induced by antigens. The regulation of IgE synthesis is influenced by T cells and T cell derived factors. IL -4, a T cell-derived cytokine, has been shown to stimulate murine IgE synthesis both in vitro and in vivo. Current evidence suggests that IL-4 induces IgE synthesis in the mouse by stimulating H chain isotype switch. Lipopolysaccharide (LPS) plus IL-4 cause about l00-fold increase in IgE secretion by murine B cells. The effects of PTFE on the IL-4-dependent IgE response by mouse whole spleen cells were studied. Whole spleen cells were cultured for 7 days in the presence of LPS plus IL-4 and PTFE and the supernatants were assayed for IgE. IL-4 dependent IgE production of LPS-stimulated whole spleen cells was inhibited by PTFE. Moreover, in the present study using U266Bl human IgE-bearing B cells, I found that PTFE inhibited the production of IgE activated by LPS plus IL-4. These results indicate that PTFE have antiallergic activity by inhibition of IgE production from B cells.

• The Korean Journal of Physiology and Pharmacology
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• 제14권4호
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• pp.235-240
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• 2010

Toll-like receptors (TLRs) functionally expressed in salivary epithelial cells, but their roles remain elusive. Among TLRs family, TLR3 is activated by dsRNA, a byproduct of viral infection. The aim of this study was to investigate the role of TLR3 in the inflammatory immune responses using HSG cells. Reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR and ELISA were performed to identify expression of TLRs and TLR3-mediated chemokine inductions. The chemotaxis assay of activated T lymphocytes was also performed. Treatment of HSG cells with polyinosinic: polycytidylic acid (poly(I:C)) significantly increased interferon-$\gamma$-inducible protein 10 (IP-10), interferoninducible T-cell $\alpha$ chemoattractant (I-TAC), and regulated on activation, normal T-cells expressed and secreted (RANTES) gene expressions in a concentration-dependent manner. Anti-TLR3 antibody blocked the increases of IP-10 and I-TAC genes. Poly(I:C)-induced increases of IP-10 and I-TAC were also confirmed at protein levels from cell lysates, but their release into extracellular medium was detected only in IP-10. We found that the culture media from HSG cells stimulated with poly(I:C) significantly increases T lymphocyte migration. Our results suggest that TLR3 plays an important role in chemokine induction, particularly IP-10, in salivary epithelial cells.