• Animal cells and systems
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• v.13 no.4
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• pp.391-397
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• 2009

The house dust mite (Dermatophagoides pteronissinus) is an important factor in triggering allergic diseases. The function of eosinophils, particularly in the production of cytokine or chemokine, is critical in understanding the pathogenesis of inflammatory diseases. In this study, we examined whether D. pteronissinus extract (DpE) induces the expression of monocyte chemotactic protein 1 (MCP-1)/CCL2, IL-8/CXCL8, and IL-6 that mediate in the infiltration and activation of immune cells and in its signaling mechanism in the human eosinophilic cell line, EoL-1. DpE increased the mRNA and protein expression of MCP-1, IL-8, and IL-6 in a time- and dose-dependent course in EoL-1 cells. In our experiments using signal-specific inhibitors, we found that the increased expression of MCP-1, IL-8, and IL-6 due to DpE is associated with Src family tyrosine kinase and protein kinase C $\delta$ (PKC $\delta$). In addition, the activation of extracellular signal-regulated kinase (ERK) is required for MCP-1 and IL-8 expression while p38 mitogen-activated protein kinase (MAPK) is involved in IL-6 expression. DpE induced the phosphorylation of ERK and p38 MAPK. PP2, an inhibitor of Src family tyrosine kinase, and rottlerin, an inhibitor of PKC $\delta$, blocked the activation of ERK and p38 MAPK. DpE induces the activation of ERK and p38 MAPK via Src family tyrosine kinase and PKC $\delta$ for MCP-1, IL-8, or IL-6 production. Increased cytokine release due to the house dust mite and the characterization of its signal transduction may be valuable in understanding the eosinophil-related pathogenic mechanism of inflammatory diseases.

• Journal of Animal Science and Technology
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• v.62 no.6
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• pp.840-853
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• 2020

The present study was conducted to investigate the effect of a multi-protease on production indicators of broiler chickens fed a crude protein and amino acid deficient-diets for 35 days immediately after hatch. A total of 448 one-day-old Ross 308 male broiler chicks were allocated in a completely randomized design into one of eight dietary treatments (positive control [PC], negative control [NC: minus 0.5% from PC, and minus 2% of lysine, methionine, threonine and methionine plus cysteine], extreme negative control [ENC: minus 1% from PC, minus 4% of lysine, methionine, threonine and methionine plus cysteine], and plus multi-protease 150 or 300 g per ton [e. g., PC-150]; PC, PC-150, NC, NC-150, NC-300, ENC, ENC-150, ENC-300) to give eight replicates with seven birds in a battery cage. Body weight, average daily gain, average daily feed intake, feed conversion ratio, and mortality were measured every week. Carcass traits, proximate analysis of breast meat, and ileum digestibility were analyzed on day 21 and 35. Feeding a multi-protease (i.e., more than 150 g/ton) for 35 days immediately after hatching improved feed efficiency and ileum digestibility (i.e., dry matter, crude protein, and energy) compared to their counterparts (i.e., diets without multi-protease: PC, NC, and ENC). In conclusion, our results indicated that broiler chickens fed nutrients deficient-diet (i.e., crude protein and amino acids) supplemented a multi-protease had an ability to compensate and (or) improve their growth performance commensurate with increased ileal digestibility for 35 days immediately after hatch.

• Biomedical Science Letters
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• v.10 no.4
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• pp.375-383
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• 2004

It has been reported that osteoporosis induced by the deficiency of estrogens in menopause is associated with the unbalance of Ca/sup 2+/ and Pi levels. Proximal tubule is very important organ to regualte Ca/sup 2+/ and Pi level in the body. However, the effect of estrogens on Ca/sup 2+/ and Pi regulation was not elucidated. Thus, we examined the effect of 17-β estradiol (E₂) on Ca/sup 2+/ and Pi uptake in the primary cultured rabbit renal proxiaml tubule cells. In the present study, E₂(> 10/sup -9/M) decreases Ca/sup 2+/uptake and stimulates Pi uptake over 3 days. E₂-induced decrease of Ca/sup 2+/ uptake and stimulation of Pi uptake were blocked by actinomycin D (a gene transcription inhibitor), cycloheximide (a protein synthesis inhibitor). tamoxifen, and progesterone (estrogen receptor antagonists). E₂-induced decrease of Ca/sup 2+/ uptake and stimulation of Pi uptake were blocked by SQ22536 (an adenylate cyclase inhibitor), Rp-cAMP (a cAMP antagonist), and PKI (a protein kinase A inhibitor). Indeed, E₂ increased cAMP formation. In addition, E₂-induced decrease of Ca/sup 2+/ uptake and stimulation of Pi uptake were blocked by staurosporine, H-7, and bisindolylmaleimide I (protein kinase C inhibitors) and E₂ translocated PKC from cytoslic fraction to membrane fraction. In conclusion, E₂ decreased Ca/sup 2+/ uptake and stimulated Pi uptake via cAMP and PKC pathway in the PTCs.

• Journal of Ecology and Environment
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• v.41 no.9
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• pp.265-270
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• 2017

Background: Over the years, pine pollens have been excluded as an allergen due to its relatively large size, low protein content, and waxy hydrophobic layer, despite their abundance. However, recent studies suggest the possibilities of pine pollens being allergens, and it has been reported that allergy symptoms were highly prevalent in areas with considerably large pine forests and high possibility of exposure to the pollen. Therefore, we conducted a comparative analysis of the allergenicities of the pollens from the dominant species of Korean pines, red pine (Pinus densiflora), black pine (Pinus thunbergii), and pitch pine (Pinus rigida), in mice. Methods: The protein composition of the pollens from the three pine species was compared via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The pine pollens and proteins extracted from the pollens were introduced to BALB/c mice by nasal inhalation and application to exposed skin and the IgE produced by the mice were extracted from blood and analyzed via ELISA. Results: SDS-PAGE showed differing protein compositions of the pollens of the three pine species. Analysis of blood IgE compositions showed a similar amount of IgE produced when pollens were applied to skin. In contrast, when mice inhaled the pollens, P. densiflora was shown to induce significantly more IgE production than those of the other two species. Conclusions: The experimental results demonstrate that the pollens of all three South Korean pine species induce IgE production, and this production was more pronounced when the pollens were inhaled than when they were applied to the skin. Of the three species, the pollen of P. densiflora was found to induce the highest level of IgE production.

• Journal of Periodontal and Implant Science
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• v.38 no.1
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• pp.41-50
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• 2008

Purpose: Bone morphogenetic protein-2(BMP-2) has been shown to possess significant osteoinducitve potential. There have been attempts to overcome a limitation of mass production, and economical efficiency of BMP. The aim of this study was to produce recombinant human BMP-2(rhBMP-2) from E. coli in a large scale and evaluate its biological activity. Materials and Methods: The E.coli strain BL21(DE3) was used as a host for rhBMP-2 production. Dimerized rhBMP-2 was purified by affinity chromatography using Heparin column. To determine the physicochemical properties of the rhBMP-2 expressed in E. coli, we examined the HPLC profile and performed Western blot analysis. The effect of the purified rhBMP-2 dimer on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and representing morphological change using C2C12 cell. Results: E. coli was genetically engineered to produce rhBMP-2 in a non-active aggregated form. We have established a method which involves refolding and purifying a folded rhBMP-2 dimer from non-active aggregates. The purified rhBMP-2 homodimer was characterized by SDS-PAGE as molecular weight of about 28kDa and eluted at 34% acetonitrile, 13.27 min(retention time) in the HPLC profile and detected at Western blot. The purified rhBMP-2 dimer stimulated ALP activity and induced the transformation from myogenic differentiation to osteogenic differentiation. Conclusion: rhBMP-2 was produced in E. coli using genetic engineering. The purified rhBMP-2 dimer stimulated ALP activity and induced the osteogenic differentiation of C2C12 cells.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.15-21
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• 2013

We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

• BMB Reports
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• v.42 no.10
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• pp.691-696
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• 2009

The E2F gene family appears to regulate the proliferation and differentiation of events that are required for adipogenesis. Pref-1 is a transmembrane protein that inhibits adipocyte differentiation in 3T3-L1 cells. In this study, we found that the expression of pref-1 is regulated by the transcription factor E2F1. The expression of pref-1 and E2F1 was strongly induced in preadipocytes and at the late differentiation stage. Using luciferase reporter assay, ChIP assay and EMSA, we found that the -211/-194 region of the pref-1 promoter is essential for the binding of E2F1 as well as E2F1-dependent transcriptional activation. Knockdown of E2F1 reduced both pref-1 promoter activity and the level of pref-1 mRNA. Taken together, our data suggest that transcriptional activation of pref-1 is stimulated by E2F1 protein in adipocytes.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.807-811
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• 2007

Diabetes is marked by high glucose levels and is associated with decreased bone mass and increased fracture rates. To determine if [6]-gingerol could influence osteoblast dysfunction induced by 2-deoxy-D-ribose (dRib), osteoblastic MC3T3-E1 cells was treated with dRib and [6]-gingerol and markers of osteoblast function and oxidized protein were examined. [6]-Gingerol ($10^{-7}\;M$) significantly increased the growth of MC3T3-E1 cells in the presence of 30 mM dRib (p<0.05). [6]-Gingerol ($10^{-7}\;M$) caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and osteocalcin secretion in the cells. We then examined the effect of [6]-gingerol on the production of osteoprotegerin and protein carbonyl in osteoblasts. Treatment with [6]-gingerol ($10^{-9}$ and $10^{-7}\;M$) increased osteoprotegerin secretion in osteoblastic cells. Moreover, [6]-gingerol ($10^{-9}$ and $10^{-7}\;M$) decreased protein carbonyl contents of osteoblastic MC3T3-E1 cells in the presence of 30 mM dRib. Taken together, these results demonstrate that [6]-gingerol inhibits dRib-induced damage and may be useful in the treatment of diabetes related bone diseases.

• YAKHAK HOEJI
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• v.52 no.4
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• pp.293-298
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• 2008

Non-steroidal anti-inflammatory drug (NSAID)-induced gut damage and bacterial translocation (BT) have not been studies well, especially from the perspective of time after administration of NSAIDs. We therefore examined these changes in animals. The study was performed on 5 groups of rat; a control group (group A) and diclofenac groups (groups B, C, E, and F). Rats in the diclofenac groups were orally administered diclofenac sodium before intestinal permeability (IP) measurement (group B, 1 h before measurement; group C, 10 h before; group D, 22 h before; and group E, 52 h before). The IP, stool pellet number, serum biochemical profile, enteric bacterial number, and BT in the mesenteric lymph nodes (MLNs), liver, spleen, kidney and heart were measured. The administration of diclofenac resulted in significantly increased IP, caused intestinal protein loss, decreased stool pellet number, caused enteric bacterial overgrowth and increased BT in multiple organs in groups A, B, C, and D. IF, intestinal protein loss, and the BT in the liver and the spleen in group E were decreased than those in group D. There were no differences in the other parameters between group D and E. In the recovery phase of the diclofenac-induced gut damage, enteric bacterial overgrowth and BT in the kidneys and the heart did not change while the BT in the reticuloendothelial systems such as in the MLNs and liver was decreased.

• BMB Reports
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• v.36 no.2
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• pp.190-195
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• 2003

The Vitamin E (VE) effect was examined on oxidative damage to DNA, lipids, and protein in mice that were fed various levels of lipid diets after total body irradiation (TBI) with X-rays at 2 Gy. No increase of 8-hydroxydeoxyguanosine (8OHdG) by TBI was observed in the +VE group; however, in the case of the -VE group, a significantly higher 8OHdG level was observed in the high-lipid group than in the low- or basal-lipid group. In the groups with TBI, the concentration of thiobarbituric reactive substances (TBARS) only significantly increased in the high-lipid (-VE) group. These changes in TBARS, due to TBI, were not detected in other groups. The contents of protein carbonyls only increased in the (-VE) group. The contents of protein carbonyls was significantly different between the (+VE) and the (-VE) groups, regardless of the lipid levels. The concentrations of GSH, vitamins C and E in the liver were lower, and the concentration of non-heme iron in the liver was higher in the high-lipid group than in the low- and basal-lipid groups. These concentrations in the high-lipid group were significantly different between the (+VE) and the (-VE) groups. These results strongly suggest that mice that are fed a high-lipid diet are susceptible to TBI-induced oxidative damage. Also, decreases in the GSH levels and an increase in the iron level are involved in the mechanism of this susceptibility.