• IMMUNE NETWORK
• /
• 제8권1호
• /
• pp.29-37
• /
• 2008

Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.

• IMMUNE NETWORK
• /
• 제3권1호
• /
• pp.38-46
• /
• 2003

Background: Platelet-activating factor (PAF) induces nuclear factor $(NF)-{\kappa}B$ activation and angiogenesis and increases tumor growth and pulmonary tumor metastasis in vivo. The role of $NF-{\kappa}B$ activation in PAF-induced angiogenesis in a mouse model of Matrigel implantation, and in PAF-mediated pulmonary tumor metastasis were investigated. Methods: Angiogenesis using Matrigel and experimental pulmonary tumor metastasis were tested in a mouse model. Electrophoretic mobility shift assay was done for the assessment of $NF-{\kappa}B$ translocation to the nucleus. Expression of angiogenic factors, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\alpha}$, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were tested by RT-PCR and ELISA. Results: PAF induced a dose- and time-dependent angiogenic response. PAF-induced angiogenesis was significantly blocked by PAF antagonist, CV6209, and inhibitors of $NF-{\kappa}B$ expression or action, including antisense oligonucleotides to p65 subunit of $NF-{\kappa}B$ (p65 AS) and antioxidants such as ${\alpha}$-tocopherol and N-acetyl-L-cysteine. In vitro, PAF activated the transcription factor, $NF-{\kappa}B$ and induced mRNA expression of $TNF-{\alpha}$, $IL-1{\alpha}$, bFGF, VEGF, and its receptor, KDR. The PAF-induced expression of the above mentioned factors was inhibited by p65 AS or antioxidants. Also, protein synthesis of VEGF was increased by PAF and inhibited by p65 AS or antioxidants. The angiogenic effect of PAF was blocked when anti-VEGF antibodies was treated or antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF was co-administrated, but not by antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF each alone. PAF-augmented pulmonary tumor metastasis was inhibited by p65 AS or antioxidants. Conclusion: These data indicate that PAF increases angiogenesis and pulmonary tumor metastasis through $NF-{\kappa}B$ activation and expression of $NF-{\kappa}B$-dependent angiogenic factors.

• IMMUNE NETWORK
• /
• 제13권4호
• /
• pp.116-122
• /
• 2013

This study was conducted to determine whether CD4 T cell responses to citrullinated fibrinogen occur in patients with rheumatoid arthritis (RA), especially in HLA-DR4-positive subjects. Whole peripheral blood mononuclear cells (PBMCs) of RA patients and control subjects were stimulated with citrullinated fibrinogen peptides, and T-cell production of proliferation and proinflammatory cytokines, such as interferon-${\gamma}$(IFN-${\gamma}$) and interleukin-17A (IL-17A), were measured. In addition, CD4 T cells from RA patients were stimulated with the citrullinated fibrinogen peptide, $Fib-{\alpha}$ R84Cit, identified as a DRB1*0401-restricted T cell epitope in HLA-DR4 transgenic mice, and the degree of T cell activation was examined similarly. No proliferative responses to the citrullinated fibrinogen peptides were observed in whole PBMCs or CD4 T cells from RA patients. Furthermore, no increased production of IFN-${\gamma}$ or IL-17A was found in whole PBMCs or CD4 T cells stimulated with the citrullinated fibrinogen peptides, although these cells responded to recall antigen, a mixture of tetanus toxoid, purified protein derivative (PPD) from Mycobacterium tuberculosis, and Candida albicans. The results of this study indicate that anti-citrulline immunity in RA patients may be mediated by fibrinogen because there is no evidence of CD4 T cell-mediated immune responses to citrullinated fibrinogen peptides.

• 한국재료학회:학술대회논문집
• /
• 한국재료학회 2011년도 추계학술발표대회
• /
• pp.7-7
• /
• 2011

Quantum beam technology has been expected to develop breakthroughs for nanotechnology during the third basic plan of science and technology (2006~2010). Recently, Green- or Life Innovations has taken over the national interests in the fourth basic science and technology plan (2011~2015). The NIMS (National Institute for Materials Science) has been conducting the corresponding mid-term research plans, as well as other national projects, such as nano-Green project (Global Research for Environment and Energy based on Nanomaterials science). In this lecture, the research trends in Japan and NIMS are firstly reviewed, and the typical achievements are highlighted over key nanotechnology fields. As one of the key nanotechnologies, the quantum beam research in NIMS focused on synchrotron radiation, neutron beams and ion/atom beams, having complementary attributes. The facilities used are SPring-8, nuclear reactor JRR-3, pulsed neutron source J-PARC and ion-laser-combined beams as well as excited atomic beams. Materials studied are typically fuel cell materials, superconducting/magnetic/multi-ferroic materials, quasicrystals, thermoelectric materials, precipitation-hardened steels, nanoparticle-dispersed materials. Here, we introduce a few topics of neutron scattering and ion beam nanofabrication. For neutron powder diffraction, the NIMS has developed multi-purpose pattern fitting software, post RIETAN2000. An ionic conductor, doped Pr2NiO4, which is a candidate for fuel-cell material, was analyzed by neutron powder diffraction with the software developed. The nuclear-density distribution derived revealed the two-dimensional network of the diffusion paths of oxygen ions at high temperatures. Using the high sensitivity of neutron beams for light elements, hydrogen states in a precipitation-strengthened steel were successfully evaluated. The small-angle neutron scattering (SANS) demonstrated the sensitive detection of hydrogen atoms trapped at the interfaces of nano-sized NbC. This result provides evidence for hydrogen embrittlement due to trapped hydrogen at precipitates. The ion beam technology can give novel functionality on a nano-scale and is targeting applications in plasmonics, ultra-fast optical communications, high-density recording and bio-patterning. The technologies developed are an ion-and-laser combined irradiation method for spatial control of nanoparticles, and a nano-masked ion irradiation method for patterning. Furthermore, we succeeded in implanting a wide-area nanopattern using nano-masks of anodic porous alumina. The patterning of ion implantation will be further applied for controlling protein adhesivity of biopolymers. It has thus been demonstrated that the quantum beam-based nanotechnology will lead the innovations both for nano-characterization and nano-fabrication.

• 한국균학회지
• /
• 제41권1호
• /
• pp.1-8
• /
• 2013

Cop9 signalosome(CSN)은 최초 식물 발달 과정에서의 빛에 의한 전사 조절 과정에서의 억제 유전자로 처음 분리된 이후 이들이 다양한 진핵 생물 에서 매우 잘 보존되어 있음이 알려지게 되었다. 이들은 대부분 8개의 subunit으로 구성되며 26S proteasome lid와 eIF3와 구조적으로는 물론 기능적으로도 유사성을 보인다고 알려져 있다. 이들은 특히 Cullin-Ring ubiquitin ligases(CRL)의 구성 요소인 Cullin의 deneddylation을 매개하여 ubiquitin ligase의 활성을 조절한다고 알려져 있으며, 또한 세포 주기 및 checkpoint 조절에 관여한다고 보고되었다. 분열효모의 경우 CSN1 및 CSN2 결손 세포에서 S-phase로서의 진행이 지연됨이 관찰되었고 감마선 혹은 UV에 좀더 민감해지는 현상이 관찰되어 CSN이 checkpoint 조절에 관여한다는 것을 보여주었다. 곰팡이의 CSN 경우 구조적으로 더욱 상위 개체들의 그것과 더욱 유사한데, CSN이 생체 시계 리듬, 빛과 연관한 호르몬 생산, 곰팡이의 발달 과정 및 생식 주기를 조절함이 보고되었다. 또한 Aspergillus nidulans의 경우 상위개체에서 보여준 DNA 합성 및 손상, 세포 주기 조절에서의 기능이 알려지면서 CSN은 곰팡이 생활사에 필수적인 여러 과정들을 조절하는 중요한 인자임을 알 수 있다. 이로써 식물이나 포유동물 등에서 보고되었던 CSN의 주요 기능을 미생물에서도 대부분 공유하고 있음을 알 수 있고 이들이 CRL을 통한 주요 세포 활성 조절 연구에 좋은 툴로서 활용할 수 있음을 시사하고 있다.

• 한국근적외분광분석학회:학술대회논문집
• /
• 한국근적외분광분석학회 2001년도 NIR-2001
• /
• pp.1062-1062
• /
• 2001

The concept of “precision agriculture” or “site-specific farming” is usually confined to the fields of soil science, crop science and agronomy. However, because plants grow in soil, animals eat plants, and humans eat animal products, it could be argued (perhaps with some poetic licence) that the fields of feed quality, animal nutrition and animal production should also be considered in this context. NIR spectroscopy has proved over the last 20 years that it can provide a firm foundation for quality measurement across all of these fields, and with the continuing developments in instrumentation, computer capacity and software, is now a major cog in the wheel of precision agriculture. There have been a few giant leaps and a lot of small steps in the impact of NIR on the animal world. These have not been confined to the amazing advances in hardware and software, although would not have occurred without them. Rapid testing of forages, grains and mixed feeds by NIR for nutritional value to livestock is now commonplace in commercial laboratories world-wide. This would never have been possible without the pioneering work done by the USDA NIR Forage Research Network in the 1980's, following the landmark paper of Norris et al. in 1976. The advent of calibration transfer between instruments, algorithms which utilize huge databases for calibration and prediction, and the ability to directly scan whole grains and fresh forages can also be considered as major steps, if not leaps. More adventurous NIR applications have emerged in animal nutrition, with emphasis on estimating the functional properties of feeds, such as in vivo digestibility, voluntary intake, protein degradability and in vitro assays to simulate starch digestion. The potential to monitor the diets of grazing animals by using faecal NIR spectra is also now being realized. NIR measurements on animal carcasses and even live animals have also been attempted, with varying degrees of success, The use of discriminant analysis in these fields is proving a useful tool. The latest giant leap is likely to be the advent of relatively low-cost, portable and ultra-fast diode array NIR instruments, which can be used “on-site” and also be fitted to forage or grain harvesters. The fodder and livestock industries are no longer satisfied with what we once thought was revolutionary: a 2-3 day laboratory turnaround for fred quality testing. This means that the instrument needs to be taken to the samples rather than vice versa. Considerable research is underway in this area, but the challenge of calibration transfer and maintenance of instrument networks of this type remains. The animal world is currently facing its biggest challenges ever; animal welfare, alleged effects of animal products on human health, environmental and economic issues are difficult enough, but the current calamities of BSE and foot and mouth disease are “the last straw” NIR will not of course solve all these problems, but is already proving useful in some of these areas and will continue to do so.

• 한국근적외분광분석학회:학술대회논문집
• /
• 한국근적외분광분석학회 2001년도 NIR-2001
• /
• pp.1141-1141
• /
• 2001

The aim of the study was to evaluate the performance of 3 calibration methods, modified partial least squares (MPLS), local PLS (LOCAL) and artificial neural network (ANN) on the prediction of chemical composition of forages, using a large NIR database. The study used forage samples (n=25,977) from Australia, Europe (Belgium, Germany, Italy and Sweden) and North America (Canada and U.S.A) with information relative to moisture, crude protein and neutral detergent fibre content. The spectra of the samples were collected with 10 different Foss NIR Systems instruments, which were either standardized or not standardized to one master instrument. The spectra were trimmed to a wavelength range between 1100 and 2498 nm. Two data sets, one standardized (IVAL) and the other not standardized (SVAL) were used as independent validation sets, but 10% of both sets were omitted and kept for later expansion of the calibration database. The remaining samples were combined into one database (n=21,696), which was split into 75% calibration (CALBASE) and 25% validation (VALBASE). The chemical components in the 3 validation data sets were predicted with each model derived from CALBASE using the calibration database before and after it was expanded with 10% of the samples from IVAL and SVAL data sets. Calibration performance was evaluated using standard error of prediction corrected for bias (SEP(C)), bias, slope and R2. None of the models appeared to be consistently better across all validation sets. VALBASE was predicted well by all models, with smaller SEP(C) and bias values than for IVAL and SVAL. This was not surprising as VALBASE was selected from the calibration database and it had a sample population similar to CALBASE, whereas IVAL and SVAL were completely independent validation sets. In most cases, Local and ANN models, but not modified PLS, showed considerable improvement in the prediction of IVAL and SVAL after the calibration database had been expanded with the 10% samples of IVAL and SVAL reserved for calibration expansion. The effects of sample processing, instrument standardization and differences in reference procedure were partially confounded in the validation sets, so it was not possible to determine which factors were most important. Further work on the development of large databases must address the problems of standardization of instruments, harmonization and standardization of laboratory procedures and even more importantly, the definition of the database population.

• 대한한방부인과학회지
• /
• 제24권3호
• /
• pp.28-47
• /
• 2011

Objectives: This study was performed to investigate the effects of Angelicae gigantis Radix (AGR) which is one of the most useful herbal-drug to treat patients with Polycystic Ovary Syndrome (PCOS) in Oriental medicine on gene expression of ovary tissue. Methods: The effects of AGR on gene expression of ovary tissue resected from PCOS induced rats using single injection of $\ss$-Estradiol 17-valerate (EV) was measured using microarray technique, and the functional analysis on these genes was conducted. Results: Total 2,812 genes were up-regulated or down-regulated, 1,421 genes were up-regulated, 1,391 genes were down-regulated by induction of PCOS. Up-regulated genes were mainly involved in biological function such as cell signalling pathways and inflammatory response. Expression levels of 1,442 genes were restored to those of naive animals by administration of AGR. 558 genes were restored to those of naive animals, which were lowered by induction of PCOS. 884 genes were lowered to naive levels, which were elevated by induction of PCOS. The functions of restored genes were partially involved in the restoration of expression levels, which were changed by induction of PCOS. Especially, up-regulated gene by induction of PCOS were mainly involved in these changes. These results mean restorative effects of AGR on damaged functions by induction of PCOS. The network of total protein interactions was measured using cytoscape program, and some key molecules, such as IRS2, MCM10, ORC2L related in up-regulated genes, CTBP2, CD44, RHOA, related in down-regulated genes that can be used for elucidation of therapeutical mechanism of medicine in future were identified. Conclusion: Restored genes by AGR were thought to have common pathways related in regulation of gene expressions. Especially, genes in restored expression levels by AGR, which were up-regulated by induction of PCOS, were regulated by 9 of common transcription factors, genes in restored expression levels by AGR, which were down-regulated by induction of PCOS, were involved in 25 of common transcription factors.

• IMMUNE NETWORK
• /
• 제3권3호
• /
• pp.219-226
• /
• 2003

Background: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than $10^9$ members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to $10^{14}$ molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. Methods: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. Results: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. Conclusion: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

• IMMUNE NETWORK
• /
• 제12권2호
• /
• pp.58-65
• /
• 2012

Background: A cell line with transfected Wilms' tumor protein 1 (WT1) is has been used for the preclinical evaluation of novel treatment strategies of WT1 immunotherapy for leukemia due to the lack of appropriate murine leukemia cell line with endogenous WT1. However, silencing of the transgene occurs. Regarding the effects of hypomethylating agents (HMAs) on reactivation of silenced genes, HMAs are considered to be immune enhancers. Methods: We treated murine WT1- transfected C1498 (mWT1-C1498) with increasing doses of decitabine (DAC) and azacitidine (AZA) to analyze their effects on transgene reactivation. Results: DAC and AZA decreased the number of viable cells in a dose- or time-dependent manner. Quantification of WT1 mRNA level was analyzed by real-time polymerase chain reaction after mWT1-C1498 treated with increasing dose of HMA. DAC treatment for 48 h induced 1.4-, 14.6-, and 15.5-fold increment of WT1 mRNA level, compared to untreated sample, at 0.1, 1, and $10{\mu}M$, respectively. Further increment of WT1 expression in the presence of 1 and $10{\mu}M$ DAC was evident at 72 h. AZA treatment also induced up-regulation of mRNA, but not to the same degree as with DAC treatment. The correlation between the incremental increases in WT1 mRNA by DAC was confirmed by Western blot and concomitant down-regulation of WT1 promoter methylation was revealed. Conclusion: The in vitro data show that HMA can induce reactivation of WT1 transgene and that DAC is more effective, at least in mWT1-C1498 cells, which suggests that the combination of DAC and mWT1-C1498 can be used for the development of the experimental model of HMA-combined WT1 immunotherapy targeting leukemia.