• Journal of the Korean Academy of Child and Adolescent Psychiatry
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• v.17 no.1
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• pp.27-31
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• 2006

Objectives : We examined whether D8/17 expression in Tourette syndrome children who suggested PANDAS were higher than comparison group, and there was my clinical difference by D8/l7 expression. Methods : Nine Tourette's syndrome children suggested PANDAS and two ADHD children without tic disorder were evaluated far percentage of D8/17 expression-positive B cells by immunofluorescence flow cytometric assay and anti-streptolysin O titer. Results : The frequency of D8/17 positive B lymphocyte rate was significantly higher in Tourette's syndrome than ADHD, whose average rate were 77.9 and 24.8, respectively. Among 9 TD patients,4 patients showed above 90% D8/l7 expression. There was high concordance expression rate between mother (98.4%) and daughter (99.0%) The significant relation between percentage of D8/17 expression and tic severity were not detected. The significant relation between percentage of D8/17 expression and anti-streptolysin O titer were not detected, however in 66.7% TD patients showed above 100IU/ml. Conclusion : We concluded that subgroup of TD children are streptococcal infected tic disorder, so called PANDAS.

• Parasites, Hosts and Diseases
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• v.54 no.4
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• pp.385-391
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• 2016

The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.

• Journal of Yeungnam Medical Science
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• v.15 no.2
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• pp.286-296
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• 1998

Different techniques for culturing respiratory epithelial cells have been developed to overcome the limitations of studies on in vivo and on bioptic material. Traditionally, culture systems are divided into organ cultures, explant cultures and dissociated cell cultures. The first two contain both epithelial and non-epithelial cells. However, in monolayer cultures of dissociated cells only epithelial cells are present, the effects observed are caused by a pure epithelial responses. The purpose of this study is to establish primary culture method of human nasal epithelium (HNEC) by monolayer culture of dissociated cells to evaluate the role of the epithelial cells in the allergic and non-allergic nasal inflammatory reactions. HNEC was prepared by primary culture method of monolayer culture of dissociated cells from human inferior nasal turbinate mucosa of septal deviation patients. Primary cultured cells were characterized by indirect immunofluorescence assay and transmission electron microscopy. The immunoreactivities of cytokeratin-pan and cytokeratin No. 8 were observed in cultured HNEC. However, the immnoreactivities of vimentin and von Willebrand factor were not observed in cultured HNEC. The tonofilaments and desmosome were observed in cultured HNEC. The cultured epithelial cells were identified to be pure nasal epithelial cells. The monolayer culture of dissociated cells could successfully be employed for further study to investigate the role of the epithelial cells in allergic or non-allergic nasal inflammatory diseases.

• Parasites, Hosts and Diseases
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• v.36 no.4
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• pp.269-275
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• 1998

Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7％) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG 1), and 22 kDa （SAG2) bands. With N. caninum antigen, the number of reactive bands was reduced, however a 43 kDa band reacted in three cases in Toxoplasma-positive sera in addition to one in Toxoplasma-negative control sera. All sera of the Toxoplasma-positive group labeled surface membrane of T. gondii, but reacted differently with N. caninum. Fluorescence was detected in surface membrane, subcellular organelles, or both in N. caninum. And one case in the Toxoplasma-negative group also reacted with N. caninum strongly in subcellular organelles. This suggested that the antibody against N. caninum may be present in human sera although the positive rate was very low in this study. The possibility of human infection with N. caninum remains to be evaluated further.

• Parasites, Hosts and Diseases
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• v.48 no.4
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• pp.319-324
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• 2010

A family of calcium-dependent protein kinases (CDPKs) is a unique enzyme which plays crucial roles in intracellular calcium signaling in plants, algae, and protozoa. CDPKs of malaria parasites are known to be key regulators for stage-specific cellular responses to calcium, a widespread secondary messenger that controls the progression of the parasite. In our study, we identified a gene encoding Plasmodium vivax CDPK4 (PvCDPK4) and characterized its molecular property and cellular localization. PvCDPK4 was a typical CDPK which had well-conserved N-terminal kinase domain and C-terminal calmodulin-like structure with 4-EF hand motifs for calcium-binding. The recombinant protein of EF hand domain of PvCDPK4 was expressed in Echerichia coli and a 34 kDa product was obtained. Immunofluorescence assay by confocal laser microscopy revealed that the protein was expressed at the mature schizont of P. vivax. The expression of PvCDPK4-EF in schizont suggests that it may participate in the proliferation or egress process in the life cycle of this parasite.

• Parasites, Hosts and Diseases
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• v.55 no.5
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• pp.491-503
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• 2017

The effects of tyrosine kinase inhibitors (TKIs) were evaluated on growth inhibition of intracellular Toxoplasma gondii in host ARPE-19 cells. The number of tachyzoites per parasitophorous vacuolar membrane (PVM) was counted after treatment with TKIs. T. gondii protein expression was assessed by western blot. Immunofluorescence assay was performed using Programmed Cell Death 4 (PDCD4) and T. gondii GRA3 antibodies. The TKIs were divided into 3 groups; non-epidermal growth factor receptor (non-EGFR), anti-human EGFR 2 (anti-HER2), and anti-HER2/4 TKIs, respectively. Group I TKIs (nintedanib, AZD9291, and sunitinib) were unable to inhibit proliferation without destroying host cells. Group II TKIs (lapatinib, gefitinib, erlotinib, and AG1478) inhibited proliferation up to 98% equivalent to control pyrimethamine ($5{\mu}M$) at $20{\mu}M$ and higher, without affecting host cells. Group III TKIs (neratinib, dacomitinib, afatinib, and pelitinib) inhibited proliferation up to 98% equivalent to pyrimethamine at $1-5{\mu}M$, but host cells were destroyed at $10-20{\mu}M$. In Group I, TgHSP90 and SAG1 inhibitions were weak, and GRA3 expression was moderately inhibited. In Group II, TgHSP90 and SAG1 expressions seemed to be slightly enhanced, while GRA3 showed none to mild inhibition; however, AG1478 inhibited all proteins moderately. Protein expression was blocked in Group III, comparable to pyrimethamine. PDCD4 and GRA3 were well localized inside the nuclei in Group I, mildly disrupted in Group II, and were completely disrupted in Group III. This study suggests the possibility of a vital T. gondii TK having potential HER2/4 properties, thus anti-HER2/4 TKIs may inhibit intracellular parasite proliferation with minimal adverse effects on host cells.

• The Journal of Korean Society of Virology
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• v.28 no.3
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• pp.247-265
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• 1998

To determine the sequence and expression of the VP7 gene of Korean isolates (CAU-9), viral RNA was purified and used for cDNA amplification by RT-PCR. The VP7 cDNA was cloned, sequenced, and expressed using baculovirus expression system. The result showed that the sequence homologies CAU-9 compared with foreign isolated strains Wa, 417, TMC-II, 95B and SA11 were ranged from 74.0% to 95.1 % of nucleotide sequence and 35% to 43% of amino acid sequence, respectively. High homology of CAU-9 was observed in Japanease isolates 417 (nucleotide sequence homology was 95.1% and amino acid sequence homology was 43%). To express VP7 gene, the VP7 cDNA was cloned into pCR-Bac vector and inserted into the genome of baculovirus adjacent to the polyhedrin promoter by cotransfection of Spodoptera frugiperda (Sf9) insect cells with wild type baculovirus DNA. In antigenic analysis of Sf9 cells inoculated with the recombinant VP7, immunofluorescence assay revealed positive for viral antigens. In metabolic labeling of Sf9 cell lysates infected with recombinant baculoviruses, it was revealed that the protein of 34 kDa was expressed. The limited study of expressed VP7 protein inoculated with guinea pigs failed to elicit neutalizing antibody. As a results, the sequence analysis and expression of VP7 protein of rotavirus CAU-9 isolated from an infant in Korea could permit the conformation and development of virus like particles which may be useful in designing vaccine strategy.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.441-446
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• 1986

Previous studies have demonstrated that phagocytosis of encapsulated bacteria needs the opsonization of bacteria with immunoglobulin and complement. Several investigators have studied the role of specific antibody to the bacteria. The purpose of this study is to investigate the role of specific anti-Actinobacillus actinomycetemcomitans Y4($A{\alpha}Y4$) antibody, which was obtained from the immunized rabbit serum for phagocytosis of $A{\alpha}Y4$ by PMNL. For this study, specific and nonspecific IgG were separated from the sera of the rabbits and PMNL were isolated from 15 healthy adults. By an enzyme-linked immunosorbent assay, the results showed that the binding capacity of anti-$A{\alpha}Y4$ IgG to $A{\alpha}Y4$ was much higher than that of nonspecific IgG; 0.75 and 0.14(O.D. at 400nm), respectively. The oxygen consumption of PMNL, phagocytizing $A{\alpha}Y4$ which was opsonized with specific $A{\alpha}Y4$ IgG(37.13 nmol/min/$1{\times}10^7$ PMNL), was significantly higher than that with nonspecific IgG(27.95 nmol/min/$1{\times}10^7$ PMNL, p<0.01). In immunofluorescence microscopic examination, the difference between the numbers of the ingested $A{\alpha}Y4$ opsonized with specific anti-$A{\alpha}Y4$ IgG and nonspecific IgG reached to statistically significant level; $184{\pm}11.4$ and $133.2{\pm}8.3$ per 100 PMNL, p<0.05. These results suggest that specific anti-$A{\alpha}Y4$ IgG has a significant role in PMNL phagocytosis of encapsulated $A{\alpha}Y4$ and also it can be available to adopt this system to develop anti-capsular antibody to $A{\alpha}Y4$ for enhancing and emphasizing the phagocytic activity against this bacterium.

• Journal of Periodontal and Implant Science
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• v.45 no.3
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• pp.120-126
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• 2015

Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.

• Polymer(Korea)
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• v.38 no.3
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• pp.364-370
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• 2014

The retinal pigment epithelium (RPE) plays an important role in maintaining a healthy retina and the degeneration of RPE caused a number of retinal diseases. The transplantation of RPE has recently become a possible therapeutic modality for retinal degeneration. To transplant RPE cells securely, substrates are essential, and then as a substrate, we fabricated films using silk that has unique mechanical properties and biocompatibility. After the FTIR spectra, contact angle and biodegradation of silk films were confirmed, RPE cells were seeded and the influence of RPE cells on silk films was examined. We measured the cell adhesion, cell viability, morphology and specific mRNA expression by MTT assay, SEM, immunofluorescence and RT-PCR. In this study, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells cultured on silk films were great, and thereby we were able to confirm the potential applications of silk films as tissue engineering carrier for regeneration of retina.