• Journal of Periodontal and Implant Science
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• 제50권3호
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• pp.159-170
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• 2020

Purpose: Immunization with Porphyromonas gingivalis heat shock protein 60 (PgHSP60) may have an immunoregulatory effect on atherogenesis. The aim of this study was to determine whether nasal immunization with a PgHSP60 peptide could reduce atherosclerotic plaque formation in apolipoprotein E knockout (ApoE KO) mice. Methods: Seven-week-old male ApoE KO mice were assigned to receive a normal diet, a Western diet, a Western diet and challenge with PgHSP60-derived peptide 14 (Pep14) or peptide 19 (Pep19), or a Western diet and immunization with Pep14 or Pep19 before challenge with Pep14 or Pep19. Results: Atherosclerotic plaques were significantly smaller in mice that received a Western diet with Pep14 nasal immunization than in mice that received a Western diet and no Pep14 immunization with or without Pep14 challenge. An immunoblot profile failed to detect serum reactivity to Pep14 in any of the study groups. Stimulation by either Pep14 or Pep19 strongly promoted the induction of CD4⁺CD25⁺ forkhead box P3 (FoxP3)⁺ human regulatory T cells (Tregs) in vitro. However, the expression of mouse splenic CD4⁺CD25⁺FoxP3⁺ Tregs was lower in the Pep14-immunized mice than in the Pep14-challenged or Pep19-immunized mice. Levels of serum interferon gamma (IFN-γ) and transforming growth factor beta were higher and levels of interleukin (IL) 10 were lower in the Pep14-immunized mice than in the other groups. Induction of CD25^- IL-17⁺ T helper 17 (Th17) cells was attenuated in the Pep14-immunized mice. Conclusions: Nasal immunization with Pep14 may be a mechanism for attenuating atherogenesis by promoting the secretion of IFN-γ and/or suppressing Th17-mediated immunity.

• Journal of Periodontal and Implant Science
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• 제47권3호
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• pp.174-181
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• 2017

Purpose: Epitope spreading is a phenomenon in which distinct subdominant epitopes become major targets of the immune response. Heat shock protein (HSP) 60 from Porphyromonas gingivalis (PgHSP60) and peptide 19 from PgHSP60 (Pep19) are immunodominant epitopes in autoimmune disease patients, including those with periodontitis. It remains unclear whether Pep19 is a dominant epitope in subjects without periodontitis or autoimmune disease. The purpose of this study was to determine the epitope spreading pattern and verify Pep19 as an immunodominant epitope in healthy teenagers using dot immunoblot analysis. The patterns of epitope spreading in age-matched patients with type 1 diabetes mellitus (type 1 DM) and healthy 20- to 29-year old subjects were compared with those of healthy teenagers. Methods: Peptide from PgHSP60, Mycobacterium tuberculosis HSP60 (MtHSP60), and Chlamydia pneumoniae HSP60 (CpHSP60) was synthesized for comparative recognition by sera from healthy subjects and patients with autoimmune disease (type 1 DM). Dot immunoblot analysis against a panel of peptides of PgHSP60 and human HSP60 (HuHSP60) was performed to identify epitope spreading, and a densitometric image analysis was conducted. Results: Of the peptide from PgHSP60, MtHSP60, and CpHSP60, PgHSP60 was the predominant epitope and was most consistently recognized by the serum samples of healthy teenagers. Most sera from healthy subjects and patients with type 1 DM reacted more strongly with PgHSP60 and Pep19 than the other peptides. The relative intensity of antibody reactivity to Pep19 was higher in the type 1 DM group than in the healthy groups. Conclusions: Pep19 is an immunodominant epitope, not only in autoimmune disease patients, but also in healthy young subjects, as evidenced by their robust immunoreactivity. This result suggests that the Pep19-specific immune response may be an initiator that triggers autoimmune diseases.

• 동의생리병리학회지
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• 제17권3호
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• pp.684-692
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• 2003

The herbal extract(YMT_02) is a modified extracts from Yukmijihwang-tang(YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as replenishing yin and tonifying the kidneys herbal medicine for hundred years ia Asian countries. The purpose of this study is to: 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by passive avoidance test, 2) statistical evaluation of candidate gene expression (pentraxin. PEP-19, transthyretin) in rat hippocampus. The hippocampi of YMT_02 and control group were dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Real Time PCR. The results were as follows : 1) passive avoidance test showed enhancing memory retentin by YMT_02 treatment, 2) expression of pentraxin, that accelerate degenerating of neuronal cell, was significantly decreased, 3) the mRNA of genes that has been known to be associated with protecting neuronal cell degeneration, such as PEP-19 and transthyretin, were significantly increased upon YMT_02 treatment. From above results, the administration of YMT_02 which tonify the function of Kidneys could enhance the ability of memory and learning. In addition, the administration of YMT_02 enhance memory retention through modulating particular gene (pentraxin, PEP-19, transthyretin) expressions in hippocampu.

• International Journal of Oral Biology
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• 제39권4호
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• pp.169-176
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• 2014

A positional scanning synthetic peptide combinatorial library (PS-SCL) was screened in order to identify antimicrobial peptides against the cariogenic oral bacteria, Streptococcus mutans. Activity against Streptococcus gordonii and Aggregatibacter actinomycetemcomitans was also examined. The library was comprised of six sub-libraries with the format $O_{(1-6)}XXXXX-NH_2$, where O represents one of 19 amino acids (excluding cysteine) and X represents equimolar mixture of these. Each sub-library was tested for antimicrobial activity against S. mutans and evaluated for antimicrobial activity against S. gordonii and A. actinomycetemcomitans. The effect of peptides was observed using transmission electron microscopy (TEM). Two semi-mixture peptides, RXXXXN-$NH_2$ (pep-1) and WXXXXN-$NH_2$ (pep-2), and one positioned peptide, RRRWRN-$NH_2$ (pep-3), were identified. Pep-1 and pep-2 showed significant antimicrobial activity against Gram positive bacteria (S. mutans and S. gordonii), but not against Gram negative bacteria (A. actinomycetemcomitans). However, pep-3 showed very low antimicrobial activity against all three bacteria. Pep-3 did not form an amphiphilic ${\alpha}$-helix, which is a required structure for most antimicrobial peptides. Pep-1 and pep-2 were able to disrupt the membrane of S. mutans. Small libraries of biochemically-constrained peptides can be used to generate antimicrobial peptides against S. mutans and other oral microbes. Peptides derived from such libraries may be candidate antimicrobial agents for the treatment of oral microorganisms.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1487-1496
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• 2020

Objective: The objective of this study was to look for optimal preparation of hydrolysates of desalted duck egg white powder (DDEWP) by the three different proteases and to investigate their antioxidant and antimicrobial properties. Methods: DDEWP was hydrolyzed by three proteases, including pepsin (PEP), Bacillus spp. (BA) and natokinase (NAT) with three different enzyme concentrations (0.1%, 0.3%, and 0.5%), individually. The important key hydrolysis parameters such as hydrolysis degree, yield, antioxidant and antimicrobial activity were evaluated in this experiment. Results: The results showed that the degree of hydrolysis (DH) of all treatments increased with increasing hydrolysis time and protease concentrations. The antioxidant and antimicrobial activities of the hydrolysates were affected by type and concentration of protease as well as hydrolysis time. Hydrolysis of PEP significantly (p<0.05) obtained the highest yield of hydrolysates, however, both of BA and NAT showed substantially lower DH values and still did not exceed 5% by the end of hydrolysis. Among the different hydrolysates, PEP exhibited significantly higher 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity than BA and NAT. All DDEWP hydrolysates from PEP had low ferrous ion chelating activity (<37%) that was significantly lower than that of NAT (>37% to 92%) and BA (30% to 79%). Besides, DDEWP hydrolysates of PEP presented significantly higher reducing power than BA and NAT. In antimicrobial activities, Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa were not effectively inhibited by any DDEWP hydrolysates of PEP except for Staphylococcus aureus. Especially, the excellent antibacterial activity against S. aureus only was displayed in DDEWP hydrolysates of PEP 0.1%. Conclusion: DDEWP hydrolysates from PEP demonstrated significantly better DH, yield, DPPH radical scavenging activity and reducing power, furthermore, had excellent inhibitory on S. aureus due to large clear zone and moderated inhibitory in bactericidal inhibition.

• The Plant Pathology Journal
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• 제21권3호
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• pp.258-261
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• 2005

We conducted a survey on pepper virus diseases in 31 regions in Korea from November 2001 to December 2004. Using electron microscopy, test plant reaction, rapid immuno-filter paper assay (RIPA), reverse transcription-polymerase chain reaction (RT-PCR) and/or analysis of viral nucleotide sequences, we found a number of viruses from 1,056 samples that we collected. These included Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Pepper mild mottle virus (PMMoV), Broad bean wilt virus 2 (BBWV2), Tobacco mild green mosaic virus (TMGMV), and Tomato spotted wilt virus (TSWV). Of the samples analyzed, $343(32.5\%)$ were infected with CMV, $209(19.8\%)$ with PepMoV, $141(13.4\%)$ with PMMoV, $12(1.1\%)$ with BBWV2, $40(3.8\%)$ with TMGMV, $5(0.5\%)$ with TSWV, $153(14.5\%)$ with CMV and PepMoV, $54 (5.1\%)$ with CMV and PMMoV, $31(2.9\%)$ with PepMoV and PMMoV, $3(0.3\%)$ with CMV and BBWV2, $1(0.1\%)$ with CMV, PepMoV and BBWV2, $8(0.8\%)$ with CMV, PepMoV and PMMoV, and $30 (2.8\%)$ samples were infected with viruses which were not identified. CMV was the most predominant virus in all inspected fields and the number of the samples infected with PMMoV was relatively low as compared PepMoV infection level in pepper. TMGMV was only found in the southern part of Korea, while TSWV was isolated in Anyang and Yesan. However, we did not encounter in this survey the Alfalfa mosaic virus (AMV), Potato virus Y (PVY), Tobacco mosaic virus (TMV), and Pepper vein chlorosis virus (PVCV).

• Journal of Periodontal and Implant Science
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• 제47권5호
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• pp.292-311
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• 2017

Purpose: Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. Methods: Human polyclonal $CD4^+CD25^+CD127^{lo-}$ Tregs (127-Tregs) and $na\ddot{i}ve$ $CD4^+CD25^+CD45RA^+$ Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an $NOD/scid/IL-2R{\gamma}^{-/-}$ mouse model of collagen-induced arthritis. Results: Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of $CD4^+CD25^+$ Tregs at the articular joints in a mechanistic and orchestrated way. Conclusions: We propose human $na\ddot{i}ve$ $CD4^+CD25^+CD45RA^+$ Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis.

• The Plant Pathology Journal
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• 제24권2호
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• pp.152-158
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• 2008

A peculiar virus-like disease of tomato showing yellow mosaic and necrotic spots on leaves and necrosis on veins, petioles and stems was observed at the Tomato Experimental Station (TES), Buyeo, Chungcheongnamdo, Korea. The disease incidence at TES fields ranged from 21 to 35% infecting different tomato cultivars. For this reason, to identify the virus infecting tomato and to characterize the virus based on biology, serology, cytology and at molecular level. Here, leaf samples were randomly collected from different infected tomato cultivars at TES fields and greenhouses and tested by ELISA using Pepper mottle virus (PePMoV) and Tomato mosaic virus (ToMV) antisera. Infected saps were mechanically inoculated in different host plants to test for pathogenicity, symptomatology and host ranges. Infected tissues and ultrathin sections were examined by electron microscopy. Finally, putative coat protein and 3'-untranslated region (CP/3'-UTR) fragment was amplified and cloned for sequence determination and analyzed its genetic relationship to existing PepMoV and PVY sequences at the Genbank. Results showed 69% of the samples were positive with PepMoV, 13% with ToMV and 19 % were doubly infected with PepMoV and ToMV. Symptoms greatly varied from different host plants inoculated with tomato leaf sap infected with PepMoV alone and discussed in detailed in this paper. Electron microscopy from infected tissues showed filamentous particles of 720-750nm in length, a typical morphology and size of PepMoV. In addition, cylindrical inclusion bodies, pinwheels, scrolls and laminates with masses of fibrillar inclusions were also found in ultrathin sections. Alignment of the sequences of the CP/3'-UTR revealed >96% sequence identity with PepMoV and only <61% with PVY. Taken together, all these evidences presented clearly indicated that the causal agent infecting tomato at TES was PepMoV and we designated this PepMoV infecting tomato as Tom-sd2 strain in this study.

• 식물병연구
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• 제24권1호
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• pp.66-74
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• 2018

국내 노지 고추에서의 바이러스 발생률 및 발병현황을 조사하기 위해 2015-2016년까지 포장조사를 수행하였다. 시료 채집은 6월부터 9월까지 월별로 실시하였고 노지 포장에서 바이러스 병징을 보이는 고추에 대해 2015년 424개, 2016년 368개의 시료를 채집하였다. 감염여부는 7종 바이러스(오이모자이크바이러스, 잠두위조바이러스2, 토마토반점위조바이러스, 사탕무황화바이러스, 고추모틀바이러스, 감자바이러스 Y, 고추약한모틀바이러스)에 대한 유전자 진단법을 이용하여 검정하였다. 그 결과, 바이러스 발병률은 2015년에는 91.7%, 2016년에는 98%로 조사되었고, 조사지역에 따라 발생하는 바이러스의 종류는 차이가 있었으나 7종 바이러스가 모두 발생됨을 확인하였다. 2015년 바이러스 종류별 발생률은 CMV, BBWV, BWYV, PMMoV, TSWV, PepMoV 및 PVY가 각각 73.8%, 68.3%, 46.9%, 14.6%, 12.7%, 6.6% 및 3.3%이었으며 2016년 바이러스 발생률은 CMV, BBWV2, BWYV, TSWV, PMMoV, PepMoV 및 PVY가 73.3%, 71.4%, 34.7%, 27.9%, 19.2%, 13.5% 및 3.5%로 조사되었다. 바이러스 발생양상을 분석한 결과, 전체 채집시료의 복합감염 비율이 2015년에는 83%, 2016년에는 86.7%로 단독감염보다 대부분 복합감염의 형태로 발병됨을 알 수 있었다.

• 융합신호처리학회논문지
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• 제19권3호
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• pp.104-109
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• 2018

심전도와 심탄도를 무구속적으로 동시에 측정하는 시스템을 개발하였다. 제안된 시스템은 의자의 보조 등받이에 설치되어 맥파전달시간(PAT), Pre-ejection period (PEP) 등의 심혈관계 역학 정보를 일상생활에서 무구속적으로 측정하는 것을 목적으로 개발되었다. 본 연구에서는, 의자의 등받이에 부착된 용량성 능동전극과 용량성 접지를 이용한 간접접촉 심전도 (IDC-ECG) 측정 방법에 의해 무구속적으로 심전도를 측정하였으며, 등받이에 부착된 EMFi 압력 센서를 사용하여 무구속적으로 심탄도(ballistocardiogram)를 측정하였다. 제안된 시스템으로 심전도와 심탄도가 무구속적으로 동시에 측정됨을 확인하였으며, 측정된 심탄도는 피검자 사이에서 일정한 패턴을 보이는 것을 알 수 있었다. 또한 심전도의 Q파와 심탄도의 첫 번째 첨두간의 시간차가 pre-ejection period와 높은 상관관계를 보이는 것도 확인할 수 있었다. 이 연구에서 개발된 시스템은 심혈관계의 다양한 지표(심전도, 심박수, PAT, PEP 등)를 무구속적으로 일상생활에서 측정할 수 있음을 보였다.