• The Journal of the Korean Society for Microbiology
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• v.20 no.1
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• pp.79-89
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• 1985

It is important to discriminate between tuberculosis and tuberculosis-like disease by Mycobacteria other than tuberculosis in the serodiagnosis of tuberculosis. But because common antigens share among Mycobacteria, their antigenicities to human are similar. Therefore degree of cross-reactivity of antibody in the sera of patients with tuberculosis between M. tuberculosis and Mycobacteria other than tuberculosis should be checked to increase the specificity in the serodiagnosis of tuberculosis. The activity levels of IgG antibody in the sera of 106 patients confirmed as active pulmonary tuberculosis and 30 normal healthy control person to the pressate extract antigen (TE, BE, AE, and FE antigen) from M. tuberculosis, M. bovis, M. avium, and M. fortuitum were measured by enzyme-linked immunosorbent assay and the crossreactivity of IgG antibody with mycobacterial species was analysed. The results were as follows; 1. The activity level(O.D. at 492nm) of IgG to TE antigen in sera of patients with pulmonary tuberculosis was $0.228{\pm}0.167$ in minimal tuberculosis; moderately advanced, $0.556{\pm}0.616$; far advanced, $1.116{\pm}0.651$ and $0.315{\pm}0.245$ in miliary tuberculosis. 2. The activity level (O.D. at 492nm) of IgG to BE antigen in sera of patients with pulmonary tuberculosis was $0.190{\pm}0.162$ in minimal tuberculosis; moderately advanced, $0.337{\pm}0.361$; far advanced, $0.713[\pm}0.460$ and $0.204{\pm}0.162$ in miliary tuberculosis. 3. The activity level (O.D. at 492nm) of IgG to AE antigen in sera of patients with pulmonary tuberculosis was $0.165{\pm}0.114$ in minimal tuberculosis; moderately advanced, $0.392{\pm}0.494$; far advenced, $0.751{\pm}0.512$ and $0.233{\pm}0.191$ in miliary tuberculosis. 4. The activity level (O.D. at 492nm) of IgG to FE antigen in sera of patients with pulmonary tuberculosis was $0.280{\pm}0.227$ in minimal tuberculosis; moderately advanced, $0.460{\pm}0.564$ ; far advanced, $0.845{\pm}0.573$ and $0.257{\pm}0.103$ in miliary tuberculosis. 5. The activity level (O.D. at 492nm) of IgG in sera of healthy control person was $0.126{\pm}0.084$ to TE antigen. $0.105{\pm}0.041$ to BE antigen, $0.103{\pm}0.052$ to AE antigen, and $0.095{\pm}0.061$ to FE antigen. 6. Degree of correlation(r) in activity level of IgG between TE antigen and BE antigen was 0.905 ; between TE antigen and AE antigen, 0.760; between TE antigen and FE antigen, 0.790, and between AE antigen and FE antigen, 0.945. 7. As O.D. above 0.200 was determined positive for the serodiagnosis of pulmonary tuberculosis, the sensitivity and specificity in ELISA using TE antigen were 80% and 87% respectively, whereas in the case of using BE antigen, 66% and 100%; in the case of using AE antigen, 62% and 100%, and in the case of using FE antigen, 72% and 93%, respecitively.

• Journal of Life Science
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• v.19 no.4
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• pp.479-485
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• 2009

Bulnesia sarmienti (BS), a traditional South American herbal medicine native to Gran Chaco, has been used to treat various human ailments. We investigated the cytotoxic activities and the inhibitory effects of BS bark extract(0, 50, 100 and $200\;{\mu}g/\;mL$) on the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), cyclooxygenase (COX) and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) in the lipopolysaccharide (LPS) (100 ng/ml)-stimulated murine macrophage cell line RAW264.7. The levels of NO, COX, PGE2 production and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) were measured by ELISA kit. Cell viability, as measured by the MTT assay, showed that BS extract had no significant cytotoxicity in RAW264.7 cells. BS extract significantly inhibited the LPS-induced NO, $PGE_2$ and COX production accompanied by an attenuation of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ formation in macrophages. These results suggest that BS extract has potential as an herbal medicine for the treatment of inflammatory diseases.

• Tuberculosis and Respiratory Diseases
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• v.57 no.4
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• pp.336-344
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• 2004

Background : Immunotherapy for cancer has not been successful because of several obstacles in tumor and its environment. Inappropriate secretions of cytokines and growth factors by tumors cause substantial changes in the immune responses against tumors, affording the tumors some degree of protection from immune attack. Uteroglobin (UG, Clara cell secretory protein) has been known to have anti-inflammatory, immunomodulatory and anti-cancer activities. However, in lung cancer cells, UG expression is decreased. This study investigated the role of UG in the immunomodulation of lung cancer. Methods : The UG protein was overexpressed by Adenovirus(Ad)-UG transduction in non-small cell lung cancer cell lines. The concentration of Prostaglandin $E_2$ ($PGE_2$) was measured by Enzyme Immunoassay (EIA). Peripheral blood mononuclear cells (PBMC) from whole blood were prepared with Ficoll. PBMC were cultured in RPMI 1640, supernatant of A549, or A549 with UG or NS-398. Concentration of Th 1 type and Th 2 type cytokines from PBMC were measured by ELISA. Results : UG suppressed $PGE_2$, Cyclooxygenase-2 (COX-2) product. Both Th1 type such as Interleukin-2 (IL-2), Interferon-${\gamma}$ (IFN-${\gamma}$) and Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and Th2 type cytokines such as IL-10 and Tumor growth factor-${\beta}$ (TGF-${\beta}$) were increased when PBMC were cultured with supernatant of non small lung cancer cells. UG and COX-2 inhibitor, NS-398 induced normal immune response of PBMC. Although Th 1 type cytokines were increased, Th 2 type cytokines were reduced by UG. Conclusion : UG suppressed PGE2, COX-2 product. Supernatant of NSCLC induced imbalance of immune response of PBMC. However, UG reversed this imbalance. These results suggest that UG may be used in the development of immunotherapy for lung cancer.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.117-123
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• 2014

Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea. But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood. To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1). Methods : In the present research, cell viability was measured by MTS assay. pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and $NF-{\kappa}B$ in PMA+A23187-activated HMC-1 cells. Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and $250{\mu}g/ml$). SHWDS ($250{\mu}g/ml$) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells. Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression. In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2). Then, SHWDS suppressed activation of nuclear factor $NF-{\kappa}B$ in nuclear, degradation of IkB ${\alpha}$ in cytoplasm. Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and $NF-{\kappa}B$ activation, thereby decreasing the expression of pro-inflammatory genes.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.125-132
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• 2014

Objectives : Suryeon-hwan (SRH) exhibits potent anti-inflammatory activity with an unknown mechanism. However, there has been a lack of studies regarding the effects of SRH on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So, the investigation focused on whether SRH inhibited nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with 200 ng/mL of LPS 30 min prior to the addition of SRH. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The content of level of cytokines (PGE, IL-6) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. Results : We found that SRH inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, SRH suppressed the LPS-induced phosphorylation of MAPK and extracellular signal-regulated kinase 1/2 (ERK 1/2) activation. Conclusions : These results suggest that SRH has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the phosphorylation of MAPKs following activation.

• Journal of dental hygiene science
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• v.11 no.2
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• pp.91-97
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• 2011

The aim of this study was to investigate the prevention effect of oral disease on the school-based toothbrushing program(SBTBP) in Daejeon in 30 months. The experimental subjects were the 5th grade's 70 students who have gone to school with the school-based toothbrushing program since the 3th-grade. The control school was located in close geographical area with similar economical status. The questionnaire about oral health knowledge and behavior was done by self-recording. After one dentist examined the dental caries and periodontal status, the gingival crevicular fluid(GCF) was collected with #25 paperpoint for 1 miniutes. Matrix Metalloproteinase(MMP)-9 in GCF was analyzed by ELISA. The SBTBP group had the upper oral health knowledge than the control significantly(p<0.001), but, the SBTBP group had no difference of the oral health behavior from the control. Although the SBTBP group had the lower plaque index than the control significantly(p=0.02), the SBTBP group had no difference of the gingival index, calculus index and the concentration of MMP-9 in GCF from the control. In conclusion, The SBTBP had the effect to reduce the dental plaque and to improve the oral health knowledge. On the other hand, the effect to prevent dental caries and periodontal disease of SBTBP was not clear.

• IMMUNE NETWORK
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• v.8 no.1
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• pp.21-28
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• 2008

Background: A co-inhibitory molecule, B7-H4, is believed to negatively regulate T cell immunity by suppressing T cell proliferation and inhibiting cytokine production. However, the mechanism behind B7-H4-mediated tolerance remains unclear. Methods: Balb/c $(H-2^d)$ mice were fed with dendritic cell line, DC2.4 $(H-2^d)$ every day for 10 days. Meantime, mice were hydrodynamically injected with recombinant plasmid expressing B7-H4 fusion protein (B7-H4.hFc) or hFc via tail vein. One day after last feeding, mice were immunized with allogeneic B6 spleen cells. 14 days following immunization, mice were challenged with B6 spleen cells to ear back and the ear swelling was determined the next day. Subsequently, a mixed lymphocyte reaction (MLR) was also performed and cytokines profiles from the reaction were examined by sandwich ELISA. Frequency of immunosuppressive cell population was assayed with flow cytometry and mRNA for FoxP3 was determined by RT-PCR. Results: Tolerant mice given plasmid expressing B7-H4.hFc showed a significant reduction in ear swelling compared to control mice. In addition, T cells from mice given B7-H4.hFc plasmid revealed a significant hyporesponsiveness of T cells against allogeneic spleen cells and showed a significant decrease in Th1 and Th2 cytokines such as IFN-${\gamma}$, IL-5, and TNF-${\alpha}$. Interestingly, flow cytometric analysis showed that the frequency of CD4+CD25+FoxP3+ Tregs in spleen was increased in tolerant mice given recombinant B7-H4.hFc plasmid compared to control group. Conclusion: Our results demonstrate that B7-H4 synergistically potentiates oral tolerance induced by allogeneic cells by increasing the frequency of FoxP3+ CD4+CD25+ Treg and reducing Th1 and Th2 cytokine production.

• Journal of Life Science
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• v.30 no.3
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• pp.313-319
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• 2020

Recently, cattle epidemic diseases are caused by a pathogen such as a virus or bacterium. Such diseases can spread through various pathways, such as feed intake, respiration, and contact between livestock. Diagnosis based on the ELISA (Enzyme-linked immunosorbent assay) and PCR (Polymerase chain reaction) methods has limitations because these traditional diagnostic methods are time consuming assays that require multiple steps and dedicated equipment. In this review, we propose the use of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) Cas system based on DNA and RNA levels for early point-of-care diagnosis in cattle. In the CRISPR/Cas system, Cas effectors are classified into two classes and six subtypes. The Cas effectors included in class 2 are typically Cas9 in type II, Cas12 in type V (Cas12a and Cas12b) and Cas13 in type VI (Cas13a and Cas13b). The CRISPR/Cas system uses reporter molecules that are attached to the ssDNA strands. When the Cas enzyme cuts the ssDNA, these reporters either fluoresce or change color, indicating the presence of a specific disease marker. There are several steps in the development of a CRISPR/Cas system. The first is to select the Cas enzyme depending on DNA or RNA from pathogens (viruses or bacteria). Based on that, the next step is to integrate the optimal amplification, transducing method, and signal reporter. The CRISPR/Cas system is a powerful diagnostic tool using a gene-editing method, which is faster, better, and cheaper than traditional methods. This system could be used for early diagnosis of epidemic cattle diseases and help to control their spread.

• Journal of Nutrition and Health
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• v.48 no.4
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• pp.310-318
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• 2015

Purpose: The aim of this study is to investigate anti-arthritis activity using natural eggshell membrane (NEM). Methods: NEM was administered at 52 mg/kg, 200 mg/kg, and 400 mg/kg to SD-Rat, where arthritis was induced by monosodium iodoacetate (MIA) at 3 mg. NO production in serum was measured using Griess reagent. Cytokines including IL-$1{\beta}$, and IL-6 were measured by Luminex and $PGE_2$, MMP-2, MMP-9, TIMP-1, $LTB_4$, and hs-CRP were measured by ELISA. The cartilage of patella volume was examined and 3-D high-resolution reconstructions of the cartilage of patella were obtained using a Micro-CT system. Results: Production of NO, IL-$1{\beta}$, IL-6, $PGE_2$, MMP-2, MMP-9, TIMP-1, $LTB_4$, and hs-CRP in serum was decreased, respectively, in comparison with control. The cartilage of patella volume increased significantly. In addition, the NEM group showed a decrease in the cartilage of patella, synovial membrane, and transformation of fibrous tissue. Conclusion: The results for NEM showed significant anti-arthritis activity. These results may be developed as a raw material for new health food to ease the symptoms mentioned above.

• The Journal of Korean Academy of Prosthodontics
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• v.53 no.1
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• pp.9-18
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• 2015

Purpose: The purpose of this study is to examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell). Materials and methods: Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05. Results: In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained $0.08{\pm}0.04{\mu}m$ in the control group, $0.52{\pm}0.14{\mu}m$ in ASD group and $1.45{\pm}0.25{\mu}m$ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase. Conclusion: In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.