• Journal of Environmental Health Sciences
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• v.38 no.6
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• pp.541-549
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• 2012

Objectives: This study was undertaken in order to evaluate a potential mechanism involved in gastro-intestinal problems observed in autistic subjects and uses an animal model of autism investigation. Methods: BTBR T+tf/J, a mouse strain with typical socio-behavioral characteristics of autistic subjects and FVB mice with highly social behaviors as the control strain were used. Both genders of mice aged three weeks and six months were used from four separate litters for each strain. Serum was prepared following cardiac puncture, and mesenteric lymph nodes were collected for in vitro stimulation and enumeration of major immune cell proportion. Results: The level of serum IgA was significantly enhanced in six-month-old BTBR mice compared with three-week-old BTBR, which was not observed with the FVB control mice. The serum IgE level was also higher among BTBR mice than among age-sex matched FVB mice, respectively. Considering the ratio of interleukin-4 vs interferon-gamma production from mesenteric lymph node T cells, skewedness toward type-2 reactivities was observed. In addition, the proportion of B cells in mesenteric lymph nodes was significantly higher in BTBR mice than in FVB mice. Conclusion: Upregulation of mucosal immunity related with enhanced type-2 immune reactivity observed in BTBR mice could be involved with the etiology of gastro-intestinal abnormalities in autism.

• Journal of Microbiology and Biotechnology
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• v.31 no.11
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• pp.1583-1590
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• 2021

Studies have demonstrated that PE_PGRS45 is constitutively expressed under various environmental conditions (such as nutrient depletion, hypoxia, and low pH) of the in vitro growth conditions examined, indicating that PE_PGRS45 protein is critical to the basic functions of Mycobacterium tuberculosis. However, there are few reports about the biochemical function and pathogenic mechanism of PE_PGRS45 protein. The fact that this M. tuberculosis gene is not easily expressed in E. coli may be mainly due to the high content of G+C and the use of unique codons. Fusion tags are indispensable tools used to improve the soluble expression of recombinant proteins and accelerate the characterization of protein structure and function. In the present study, His6, Trx, and His6-MBP were used as fusion tags, but only MBP-PE_PGRS45 was expressed solubly. The purification using His6-MBP tag-specific binding to the Ni column was easy to separate after the tag cleavage. We used the purified PE_PGRS45 to immunize New Zealand rabbits and obtained anti-PE_PGRS45 serum. We found that the titer of polyclonal antibodies against PE_PGR45 was higher than 1:256000. The result shows that purified PE_PGRS45 can induce New Zealand rabbits to produce high-titer antibodies. In conclusion, the recombinant protein PE_PGRS45 was successfully expressed in E. coli and specific antiserum was prepared, which will be followed by further evaluation of these specific antigens to develop highly sensitive and specific diagnostic tests for tuberculosis.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.38-51
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• 2022

Whole-plant corn (Zea may L.) and sorghum-sudangrass hybrid [Sorghum bicolor (L.) Moench] are major summer crops that can be fed as direct-cut or silage. Proso millet is a short-season growing crop with distinct agronomic characteristics that can be productive in marginal lands. However, information is limited about the potential production, feed value, and ensilability of proso millet forage. We evaluated proso millet as a silage crop in comparison with conventional silage crops. Proso millet was sown on June 8 and harvested on September 5 at soft-dough stage. Corn and sorghum-sudangrass hybrid were planted on May 10 and harvested on September 10 at the half milk-line and soft-dough stages, respectively. The fermentation was evaluated at 1, 2, 3, 5, 10, 15, 20, 30, and 45 days after ensiling. Although forage yield of proso millet was lower than corn and sorghum-sudangrass hybrid, its relative feed value was greater than sorghum-sudangrass hybrid. Concentrations of dry matter (DM), crude protein, and water-soluble carbohydrate decreased commonly in the ensiling forage crops. The DM loss was greater in proso millet than those in corn and sorghum-sudangrass hybrid. The in vitro dry matter digestibility declined in the forage crops as fermentation progressed. In the early stages of fermentation, pH dropped rapidly, which was stabilized in the later stages. Compared to corn and sorghum-sudangrass hybrid, the concentration of ammonia-nitrogen was greater in proso millet. The count of lactic acid bacteria reached the maximum level on day 10, with the values of 6.96, 7.77, and 6.95 Log₁₀ CFU/g fresh weight for proso millet, corn, and sorghum-sudangrass hybrid, respectively. As ensiling progressed, the concentrations of lactic acid and acetic acid of the three crops increased and lactic acid proportion became higher in the order of sorghum-sudangrass hybrid, corn, and proso millet. Overall, the shorter, fast-growing proso millet comparing with corn and sorghum-sudangrass hybrid makes this forage crop an alternative option, particularly in areas where agricultural inputs are limited. However, additional research is needed to evaluate the efficacy of viable strategies such as chemical additives or microbial inoculants to minimize ammonia-nitrogen formation and DM loss during ensiling.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.1
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• pp.39-44
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• 2013

In Korea, wide spread use of whole cottonseed, which is primarily a GMO plant imported from foreign countries and being fed to animals as raw state, has aroused concern that it may disturb the existing ecology of the country unless dispersion of the seed is under proper control. The objective of this study was to elucidate the changes in various nutritive parameters due to heat treatment and to determine the effective condition for removing germination ability of whole cottonseed (WCS). Of the various temperatures applied (76, 78, 80, 85, $100^{\circ}C$/30 min) $85^{\circ}C$ for 30 min was confirmed to be the lowest temperature treatment which resulted in a complete removal of the germination ability of WCS. Therefore, based on the determined temperature condition ($85^{\circ}C$ 30 min) we tried to examine the changes of various nutritional parameters, including nutrient composition, in vitro digestibilities and ruminal protein degradabilities, comparing raw whole cotton seed (RWCS) and heated whole cotton seed (HWCS). Some changes in amino acid composition were observed with heat treatment of WCS, but these were regarded to originate from the variation in plant quality and seed morphology, which are usually affected by different environmental factors during the vegetation period. As for fatty acid composition, no significant differences were observed to occur during heat treatment. However, WCS heated at $85^{\circ}C$ for 30 min in a circulating oven showed a significant decrease (p<0.05) of in situ rumen degradability in both dry matter (DM) and crude protein (CP), as compared to raw WCS. Overall results obtained in the study indicate that the heating condition used in this study, which was proven to be the most appropriate and economic to remove germination ability of WCS, may also improve the nutritional value of the ruminant with regard to reducing its protein degradability within the rumen.

• Journal of Applied Biological Chemistry
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• v.57 no.4
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• pp.365-371
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• 2014

In the present study, we investigated the biological activities of Xylosma congesta leaf ethanol extract (XCO) using a variety of in vitro and cell culture model systems for anti-melanogenic, anti-wrinkle, anti-inflammatory and anti-oxidant activities. First, XCO markedly inhibited ${\alpha}$-melanocyte stimulating hormone-stimulated melanin synthesis in B16F10 cells. Secondly, XCO marginally induced procollagen synthesis in CCD-986SK cells. Thirdly, XCO dose-dependently suppressed lipopolysaccharide-induced nitric oxide (NO) production in RAW 264.7 cells. XCO did not affect cell viability at different concentrations used in this study, indicating that XCO-mediated inhibition of melanin, procollagen and NO synthesis is not mediated by cytotoxicity. Finally, XCO was found to exert anti-oxidant effect. Taken together, these findings demonstrate for the first time that XCO possesses anti-melanogenic, anti-wrinkle, anti-inflammatory and anti-oxidant activities, and suggest further evaluation and development of XCO as a functional supplement or cosmetic that may be useful for whitening skin, reducing wrinkles and treating inflammatory responses.

• The korean journal of orthodontics
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• v.27 no.6 s.65
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• pp.979-995
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• 1997

The purpose of this study was to identify the preventive and the progressive inhibitory effects of enamel demineralization with fluoride releasing light-and self-cured orthodontic sealants(FluoroBond), in vitro, under the polarizing light microscope and the scanning electon microscope. The polarizing light microscopic group was subdivided into seven groups(Group A-Group G). The scanning electron microscopic group was also subdivided into seven groups(Group A'-Goup G'). For polarizing light microscopic evaluation, longitudinal sections were made longitudinally by Maruto cutter(Maruto Co., Japan) and Maruto grinding machine(Maruto Co., Japan). Sections were examined and photographed by the polarizing light microscope(Olympus Optical Co., Japan) using crossed polars and with the enamel rod longitudinal axis oriented at $45^{\circ}$ to the extinction position. For scanning electron microscopic evaluation, the specimens were coated with a highly conducting layer of gold palladium in a model Hus-4 high-vacuum evaporator and examined in an ISI-100B scanning electron microcope operated at 20kV. The results of this study were as follows : 1. The mean depths of artificial carious lesions under a polarized light microscope were $Group\;A(5.08{\mu}m),\;Group\;B(47.82{\mu}m,\;Group\;C(8.42{\mu}m),\;Group\;D(7.20{\mu}m),\;Group\;E(85.41{\mu}m),\;Group\;F(60.38{\mu}m),\;Group\;G(60.13{\mu}m)$. 2. There were statistically significant differences in Group B compared with Group A, C, and D(p<0.05), and also, in Group I compared with Group F and Group G(p<0.05). 3. Light-and self-cured orthodontic sealants had the preventive effects of enamel demineralization. 4. Light-and self-cured orthodontic sealants had the progressive inhibitory effects of enamel demineralization. 5. The time progress of demineralizing agent had no influence on the samples of light-and self-cured orthodontic sealants under the scanning electron microscope. 6. There was no difference between the specimens of light-and self-cured orthodontic sealants both in the polarized light microscopic group and in the scanning electron microscopic group.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.3
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• pp.217-224
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• 2008

Angiogenesis plays an important role in bone development and postnatal bone fracture repair. Vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptors (VEGFRs) have been thought to be primarily involved in promoting angiogenesis. It is well known that VEGF and its receptors have been reported to play an important role in the regulation of the interaction between angiogenesis and osteogenesis during bone repair processes. Dexamethasone, a potent synthetic glucocorticoid, promotes phenotype markers of osteoblast differentiation, such as ALP and osteocalcin. It stimulates in vitro osteogenesis of human bone marrow osteogenic stromal cells. Dexamethasone has been reported to suppress VEGF gene expression in some cells. However, our previous study demonstrated VEGF quantification increased in a time-dependent manner in periosteal-derived osteogenesis under dexamethasone. So, the purpose of this study was to examine the angiogenic phenotypes in cultured human periosteal-derived cells under high-dose dexamethasone. Periosteal-derived cells were cultured using a technique previously described. After passage 3, the periosteal-derived cells were further cultured for 28 days in an osteogenic inductive culture medium containing ascorbic acid, ${\beta}$-glycerophosphate and high-dose dexamethasone, We evaluated the expression of VEGF isoforms, VEGFR-1, VEGFR-2, and neuropilin-1, ALL VEGF isoforms ($VEGF_{121},\;VEGF_{165},\;VEGF_{189}$, and $VEGF_{206}$) expression was observed by RT-PCR analysis. VEGFR-1, VEGFR-2 and neuropilin-1 expression increased up to day 14, particularly during the early stage of mineralization. Our results suggest the involvement of direct VEGFs/VEGFRs system on periosteal-derived cells during early mineralization phase under high-dose of dexamethasone. These also suggest that VEGF might act as an autocrine growth molecule during osteoblastic differentiation of cultured human periosteal-derived cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.1
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• pp.1-16
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• 2004

The use of artificial nerve conduit containing viable Schwann cells is one of the most promising strategies to repair the peripheral nerve injury. To fabricate an effective nerve conduit whose microstructure and internal environment are more favorable in the nerve regeneration than existing ones, a new three-dimensional Schwann cell culture technique using $Matrigel^{(R)}$. and dorsal root ganglion (DRG) was developed. Nerve conduit of three-dimensionally arranged Schwann cells was fabricated using direct seeding of freshly harvested DRG into a $Matrigel^{(R)}$ filled silicone tube (I.D. 1.98 mm, 14 mm length) and in vitro rafting culture for 2 weeks. The nerve regeneration efficacy of three-dimensionally cultured Schwann cell conduit (3D conduit group, n=6) was assessed using SD rat sciatic nerve defect of 10 mm, and compared with that of silicone conduit filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method (2D conduit group, n=6). After 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were examined using image analyzer and electromicroscopic methods. The SFI and ankle stance angle (ASA) in the functional evaluation were $-60.1{\pm}13.9$, $37.9^{\circ}{\pm}5.4^{\circ}$ in 3D conduit group (n=5) and $-87.0{\pm}12.9$, $32.2^{\circ}{\pm}4.8^{\circ}$ in 2D conduit group (n=4), respectively. And the myelinated axon was $44.91%{\pm}0.13%$ in 3D conduit group and $13.05%{\pm}1.95%$ in 2D conduit group to the sham group. In the TEM study, 3D conduit group showed more abundant myelinated nerve fibers with well organized and thickened extracellular collagen than 2D conduit group, and gastrocnemius muscle and biceps femoris tendon in 3D conduit group were less atrophied and showed decreased fibrosis with less fatty infiltration than 2D conduit group. In conclusion, new three-dimensional Schwann cell culture technique was established, and nerve conduit fabricated using this technique showed much improved nerve regeneration capacity than the silicone tube filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method.

• The Korea Journal of Herbology
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• v.30 no.4
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• pp.37-44
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• 2015

Objectives : The present study was designed to evaluate the anti-inflammatory and anti-oxidative stress activities through regulation of Nrf2-mediated genes by Rhei rhizoma and Glycyrrhiza rhizoma combined extract (RGE) in reflux esophagitis.Methods : The antioxidant activity of RGE in vitro was measured in terms of radical scavenging capacity such as DPPH and ABTS. RGE was administered at 350 mg/kg body weight prior to induction of reflux esophagitis. Reflux esophagitis was induced that tied the pylorus and the transitional junction between the forestomach and the corpus in Sprague-Dawley rats.Results : RGE scavenged DPPH and ABTS effectively and IC₅₀of RGE each were 4.9 μg/ml and 45.6 μg/ml. Our results show that RGE administration markedly ameliorated mucosal damage upon histological evaluation. In serum and esophagus tissue, RGE significantly suppressed the oxidative stress biomarkers. Reflux esophagitis induced rats exhibited down-regulation of antioxidant-related proteins in the esophagus; however, the levels with treatment of RGE were significantly higher than those of vehicle reflux esophagitis rats. RGE treatment caused significant reductions in activation of NF-κB transcription factor. Thus, RGE significantly exhibited potent anti-inflammatory activities by suppressing the protein expression levels of pro-inflammatory proteins such as COX-2 and iNOS and inflammatory cytokines such as TNF-αin the esophagus tissue.Conclusions : Reflux esophagitis caused considerable levels of oxidative stress in the esophageal mucosa and the administration of RGE reduced the esophageal mucosa damage through the regulation of Nrf2 and NF-κB pathways. Our findings can considered as supplementary therapy in the prevention or treatment of reflux esophagitis.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.4037-4043
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• 2012

Despite progress in elucidating mechanisms associated with colorectal cancer and improvement of treatment methods, it remains a frequent cause of death worldwide. New and more effective therapies are therefore urgently needed. Recent studies have shown that immunogenicity of whole ovarian tumor cells and subsequent T cell response were potentiated by oxidation modification with hypochlorous acid (HOCl) in vitro and ex vivo. These results prompted us to investigate the protective antitumor response with an HOCl treated CT26 colorectal cancer cell vaccine in an in vivo mouse model. Administration of HOCl modified vaccine triggered robust antitumor immunity to autologous tumor cells in mice and prolonged survival period significantly. In addition, increased necrosis and apoptosis were found in tumor tissue from the oxidation group. Interestingly, ELISPOT assays showed that specific T cell responses were not elicited in response to the immunizing cellular antigen, in contrast to raising sera antibody titer and antibody binding activity shown by ELISA assay and flow cytometry. Further evaluation of the mechanisms underlying HOCl modified vaccine mediated humoral immunity highlighted the role of antibody-dependent cell-mediated cytotoxicity. These results combined with previous studies suggest that HOCl oxidation modified whole cell vaccine has wide applicability as a cancer vaccine because it can target both T cell- and B cell-specific responses. It may thus represent a promising approach for the immunotherapy of colorectal cancer.