• IMMUNE NETWORK
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• 제10권1호
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• pp.5-14
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• 2010

Background: There have been several reports describing the capability of ginseng extracts as an adjuvant. In this study, we tested if ginsan, a polysaccharide extracted from Panax ginseng, was effective in enhancing antibody response to orally delivered Salmonella antigen. Methods: Ginsan was treated before oral salmonella antigen administration. Salmonella specific antibody was determined by ELISA. mRNA expression was determined by RT-PCR. Cell migration was determined by confocal microscopy and flow cytometry. COX expression was detected by western blot. Results: Ginsan treatment before oral Salmonella antigen delivery significantly increased both secretory and serum antibody production. Ginsan increased the expression of COX in the Peyer's patches. Various genes were screened and we found that CCL3 mRNA expression was increased in the Peyer's patch. Ginsan increased dendritic cells in the Peyer's patch and newly migrated dendritic cells were mostly found in the subepithelial dome region. When COX inhibitors were treated, the expression of CCL3 was reduced. COX inhibitor also antagonized both the migration of dendritic cells and the humoral immune response against oral Salmonella antigen. Conclusion: Ginsan effectively enhances the humoral immune response to orally delivered antigen, mediated by CCL3 via COX. Ginsan may serve as a potent vaccine suppliment for oral immunization.

• BMB Reports
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• 제31권5호
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• pp.453-458
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• 1998

Human thrombopoietin (hTPO) was expressed to high levels in insect cells using the baculovirus expression system. Full-length hTPO cDNA containing a native signal peptide sequence was amplified by PCR from a human fetal liver cDNA library and cloned into the Autographa californica nuclear polyhedrosis virus (AcNPV) expression vector. Immunoblot analysis with antiserum against hTPO indicated that an approximately 55 kDa protein was produced in recombinant AcNPV infected insect cells. Recombinant hTPO was produced 4-fold higher in Trichoplusia ni (Tn5) cells than in Spodoptera frugiperda (Sf9) cells. with most of the hTPO produced in Tn5 cells secreted into the culture medium. Addition of tunicamycin in the culture medium resulted in the reduction of the size of hTPO to 35-38 kDa, and most of the protein remained within the cell. These results suggest that N-glycosylation of hTPO is required for the secretion of the protein into the culture medium in insect cells. hTPO produced in insect cells induced proliferation and maturation of megakaryocyte progenitors, indicating that it is in a biologically active form.

• BMB Reports
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• 제48권8호
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• pp.445-453
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• 2015

Endoplasmic Reticulum (ER) is an organelle where most secretory and membrane proteins are synthesized, folded, and undergo further maturation. As numerous conditions can perturb such ER function, eukaryotic cells are equipped with responsive signaling pathways, widely referred to as the Unfolded Protein Response (UPR). Chronic conditions of ER stress that cannot be fully resolved by UPR, or conditions that impair UPR signaling itself, are associated with many metabolic and degenerative diseases. In recent years, Drosophila has been actively employed to study such connections between UPR and disease. Notably, the UPR pathways are largely conserved between Drosophila and humans, and the mediating genes are essential for development in both organisms, indicating their requirement to resolve inherent stress. By now, many Drosophila mutations are known to impose stress in the ER, and a number of these appear similar to those that underlie human diseases. In addition, studies have employed the strategy of overexpressing human mutations in Drosophila tissues to perform genetic modifier screens. The fact that the basic UPR pathways are conserved, together with the availability of many human disease models in this organism, makes Drosophila a powerful tool for studying human disease mechanisms. [BMB Reports 2015; 48(8): 445-453]

• Parasites, Hosts and Diseases
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• 제44권3호
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• pp.251-254
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• 2006

Monoclonal antibody (mAb) Tg786 against Toxoplasma gondii has been found to detect a 42-kDa rhoptry protein (ROP6) which showed protease activity and host cell binding characteristics after secretion. Using the mAb, a colony containing a 3'-UTR was probed in a T. gondii cDNA expression library. A full length cDNA sequence of the rhoptry protein was completed after 5'-RACE, which consisted of 1,908 bp with a 1,443 bp ORF. The deduced amino acid sequence of ROP6 consisted of a polypeptide of 480 amino acids without significant homology to any other known proteins. This sequence contains an amino terminal stop transfer sequence downstream of a short neutral sequence, hydrophilic middle sequence, and hydrophobic carboxy terminus. It is suggested that the ROP6 is inserted into the rhoptry membrane with both N- and C-termini.

• Asian-Australasian Journal of Animal Sciences
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• 제13권4호
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• pp.440-445
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• 2000

Thirty lactating Javanese thin-tail ewes (12 ewes had been injected, prior to mating, with 700 IU pregnant mare serum gonadotropin, and 18 ewes with saline as a control) were used to evaluate the effect of superovulation on milk production during lactation and mammary chemical indices at the end of lactation. Thirteen ewes (9 control and 4 superovulated ewes) were fed at low and the other 17 ewes (9 control and 8 superovulated ewes) were fed at high quality ration. Superovulated ewes, either fed at low or high quality ration, had dramatically higher milk yields (57%). At the end of lactation, superovulated ewes had higher mammary dry fat-free tissue, mammary DNA concentration, total mammary DNA and RNA contents than nonsuperovulated ewes. Superovulation did not affect mammary RNA and collagen concentrations, and total collagen content. Ration quality did not significantly increase milk production during lactation and mammary chemical indices at the end of lactation. The observed increase in milk production in the superovulated ewes was probably due to the increased mammary secretory cell number and their synthetic activities during lactation as a result of the increased endogenous hormonal stimulation of mammary growth and development during pregnancy.

• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.430-434
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• 2003

In vivo antagonistic effect of Lactobacillus helveticus CU 631 and Lactobacillus spp. against typical enteritis causing pathogen Salmonella enteritidis KU 101 have been determined, which showed an increase in survival rate and the decline in viable cell numbers of pathogen in liver and spleen at sacrifice. A signifcant difference in the antagonistic effect against KU 101 were observed, which was species and/or strain dependent of Lactobacillus (p<0.01), the survival rate of the mice in the Salmonella infection by feeding L. helveticus CU 631 has been shown to be 157%, whereas those of L. rhamnosus GG ATCC 53103, L. acidophilus ATCC 4356, L. johnsonii C-4 were 137%, 132%, 119% respectively on the basis of lactobacilli non-associated control KU101 fed mice to be 100%. Viable cells of S. enteritidis KU101 in the liver and in the spleen at sacrifice were decreased in Lactobacillus spp. fed group with no significant difference. The higher level of total secretory IgA concentration in the intestinal fluid of lactobacilli fed mice than control mice have been observed. In vitro antagonistic activity of Lactobacillus spp. against KU101 have been determined, a prominent antagonistic activity of CU 631 against KU 101 were demonstrated.

• Applied Microscopy
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• 제9권1호
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• pp.35-56
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• 1979

To investigate ultrastructural characteristics of cancer cells of the nervous system, 25 cases; i.e. astrocytoma(9), oligodendroglioma(1), medulloblastoma(1), meningioma(5), pinealoma(2) and pituitary adenomas(7). The common findings were marked irregularity of nuclear membrane with pronounced infoldings, clumping of heterochromatin along inner nuclear membrane, enlargement of nucleolus, and frequent observations of nuclear bodies and nuclear inclusions. But these findings are also the signs that can be observed in hyperactive cells. Thus, ultrastructural characteristics of cancerous nucleus are the great variability of nuclear size, shape and composition. but none of them appear to be specific. Among cytoplasmic organelles, massive fibrils are characteristic of astrocytoma and meningiomas, cytoplasmic protofibrils such as glial process and microvesicles in oligodendroglioma, secretory granules are characteristic in pituitary adenomas, and fine filamentous fibrils and desmosomes are characteristic of fibroblastic type of meningioma. Intercellular relationships and cell membrane specialization are important features in the differential diagnosis of various undifferentiated tumors. The frequent resolution of difficult diagnosis problems by electron microscopy outweighs the disadvantages of this technique, such as the expense and time required.

• Applied Microscopy
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• 제22권2호
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• pp.127-140
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• 1992

This research was undertaken to determine the effects of the cyclophosphamide (CP) on the epididymal corpus of the male rat in terms of ultrastructural alteration and protein analysis by SDS-PAGE at different groups; control group, 1 week group, 3 weeks group and 5 weeks group were treated with saline (control group) or CP at doses of 20mg/kg/week, 1 time a week, respectively. In the cytoplasm of the principal cells on the epididymal corpus, the mitochondria were significantly swollen or disrupted. The lumens of rough endoplasmic reticulum (rER) were also dilated and the number of secretory vesicles and lysosomes were increased respectively. CP caused changes in protein concentrations in the corpus of epididymis after CP treatment. Total proteins of 31 to 36 species were expressed in the corpus fluid. Then the more CP was increased, the more concentration of proteins caused to decrease, synthesize or increase in epididymal corpus. In contrast to the control group, in particular 88KD and the other 8 proteins in the corpus fluid, were decreased or disappeared respectively, whereas acid phosphatase and the other 9 proteins in the corpus fluid, were increased or synthesized respectively. The other proteins are not showed distinctive difference. It is suggested that treatment with CP alters the specific cell organelles and proteins in segment of the epididymal corpus.

• 한국동물학회지
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• 제37권2호
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• pp.281-288
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• 1994

한국산 거머리(Erpobdella lineate)의 전, 후 흠반을 광학현미경과 투과전자현미경을 사용하여 조직화학적 및 미세구조적 연구를 수행한 결과 다음과 같았다. 거머리 전, 후 홍안에서 관찰된 상피세포는 불규칙한 단층원주형 상피로 되어 있으며, 상피세포의 상단에는 큐티를층이 있고 측면원형질 막은 거치상을 이루면서 여러개의 desmosome이 관찰되 었다. 큐티클층은 projection eplcuticularis, amorphorous stratum 및 fibrous stratum 등 3부분으로 구분되었다. 전, 후 출반의 상피조직 사이에서 공히 a형 분비과립과 b혐 분비과립 등이 관찰되었는데, 이 세포에서 분비된 과립들은 중성점액다당류로 확인되었다. 전 흠반의 횡단면 상피조직 밑에서 많은 근육세포들조 형성된 원형의 집단들이 다수 관찰되었는데, 이는 흠반의 흡수기능과 밀접한 관계가 있었다 비교적 통근형태의 근육세포들은 세포의 원형질막 내측에 많은 근섬유 다발을 지니고 있고 그 중앙에는 cristae가 발달된 많은 수의 사립체들이 모여 있었다. 전, 후 출반의 결합조직 내에서 5종류(A, B, C, D 및 I 등)의 분비과립들이 관찰되었다. 그 중 C, D 과립은 전, 후 흡반에서 공통으로 관찰되고, A, B 과립은 전 흠반에서, E 과립은 후 흠반에서만 각각 관찰되었는데, 이들 역시 모두 중성점액다당류로 확인되었다.

• 대한수의학회지
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• 제57권1호
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• pp.9-15
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• 2017

To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.