• Toxicological Research
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• v.22 no.4
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• pp.357-364
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• 2006

This study was undertaken to develop a reliable mice model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. BALB/C mice and NC/Nga mice were sensitized twice with $100{\mu}l$ of 1% 2,4-dinitrochlorobenzene (DNCB) or vehicle (acetone : olive oil=4:1 mixture) in a week and challenged twice with $100{\mu}l$ of 0.2% DNCB or the vehicle at the following week. Mice were sacrificed at 19 days following the second DNCB or vehicle challenge for NC/Nga mice and at 28 days following the second DNCB or vehicle challenge for BALB/c mice. Upregulation of plasma 1gE, a hallmark of atopic dermatitis occurrence, was evident in the plasma obtained 4 day after the second DNCB challenge from BALB/c mice (approximately 4-fold) and NC/Nga mice (approximately 6-fold) treated with DNCB in comparison with that of the vehicle treated-control mice, and remain higher $3{\sim}4$ week after the second challenge. Ratio of plasma IgG1 versus IgG2a concentration was significantly higher in the mice treated with DNCB than the control mice, which also implies the skewed type-2 reactivity in vivo. Ratio of interleukin-4 versus interferon gamma produced in the splenic T cell culture supernatants was approximately 3-fold higher in the both strains of mice treated with DNCB than their control mice, respectively. The DNCB-treated mice demonstrated atopic dermatitis-like skin legions characterized with erythma, scaling, and hemorrhage, which was not observed with the control mice. Scratching on face or dorsal area was significantly more frequent (approximately 25-fold) in the DNCB-treated mice than the control at next day of the second DNCB challenge, and scratching frequency remains higher (approximately 4-fold) in the mice treated with DNCB than the control at 14 day following the second DNCB challenge. Overall, the mice model developed through sensitization and challenge with DNCB may be useful for research on atopic dermatitis and development of treatment materials for atopic dermatitis.

• Korean Journal of Veterinary Research
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• v.47 no.2
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• pp.175-185
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• 2007

This study was carried out to investigate the pathogenesis and pathogenicity of the porcine circovirus type 2 (PCV2) Korean isolate from weaned pigs. Twenty four weaned pigs, PCV2, porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) antibodies free, were allocated to 4 groups (n = 6). Six pigs were inoculated intranasally with PCV2 alone, 6 with PCV2 and PRRSV, 6 with the combined PCV2/PRRSV/PPV inoculum, and 6 were remained as a uninoculated negative control. Pigs were killed 3 and 6 weeks after inoculation and tissue samples examined for gross and microscopic lesions and for the presence of PCV2 antigens and nucleic acids. Experimentally inoculated pigs were evaluated for 3 considerations: 1. development of postweaning multisystemic wasting syndrome (PMWS), 2. distribution of viral antigens by immunohistochemistry and polymerase chain reaction (PCR), and 3. cytokine mRNA levels in lymph nodes. Pigs inoculated with PCV2/PRRSV/PPV showed typical clinical signs, gross findings, and histopathologic characteristics of PMWS. In the PCV2/PRRSV/PPV inoculated group, the PCV2 antigen was widely distributed in various parenchymal organs such as brain, spinal cord, tonsil, lymph nodes, lung, heart, liver, kidney, spleen, and peyer's patch. Lymph node mRNA expression of IL-$1{\alpha}$, IL-2R and IL-8 was determined by real-time PCR. The pigs of PCV2/PRRSV and PCV2/PRRSV/PPV inoculation group, the mRNA expression was characterized by a decrease of IL-$1{\alpha}$, IL-2R and IL-8. The decrease of cytokine mRNA represent the state of T cell immuno-suppression in pig, and nicely support the evidence for the impairment of immune system in pigs with PMWS. In conclusion, PCV2 infection and some additional infectious causes such as PRRSV and/or PPV are warranted for the presence of PMWS in weaned pigs in Korea.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.249-253
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• 2016

The inky cap, Coprinellus congregatus, produces several laccase isozymes during its life cycle: both hyphal tip laccase and sclerotial laccase are involved in the fungal development. When this fungus was transferred to an acid liquid medium (pH 4.0-4.5), a new laccase was synthesized and secreted into the culture supernatant. In order to examine its regulation by external pH, green fluorescent protein gene was ligated at the downstream of the promoters having different lengths. These expression vectors having different promoter lengths were inserted into the fungal transformation vector, pBARGEM7-1. These expression vectors were introduced to the mating type a1 and a2 monokaryons, and the transformants were selected by the phosphinothricin resistance. Transformant a1 (a1TF) and transformant a2 (a2TF) were mated with each other to generate homozygotic dikaryon transformants. All these transformants were grown in neutral liquid medium for 5 days, and then the whole cell homogenates were transferred to the acidic liquid medium (pH 4.1). After 36 h incubation at $25^{\circ}C$, cells were harvested for the analysis of GFP expression. GFP expression was detected in the transformant having full-length promoter (2.0 kb), but other transformants having shorter length promoter (shorter than 1.29 kb) failed to show the fluorescence. Therefore, the acid-responsive element in the laccase promoter should be localized between -2.0 kb ~ -1.29 kb region.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.211-218
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• 2001

The $M_r$ 63,000 glycoprotein (GP63) and lipophosphoglycan (LPG) of Leishmania donovani were evaluated as vaccine candidates against visceral leishmaniasis. Mice were immunized with liposomeencapsulated GP63 and/or LPG that were purified from the soluble extract of L. donovani promastigotes, and were challenged with virulent amastigotes. Mice immunized with GP63/LPG in liposomes plus BCG resulted in a 27.4% reduction of amastigotes in the liver compared to the control group (liposomes plus BCG), and mice immunized with liposome-GP63 plus BCG failed to induce a protective immune response against the challenge infection. Immunization of mice with GP63 fused to the Schistosoma japonicum glutathione S-transferase (GP63-GST) plus BCG also failed to elicit protective immunity. To analyze the cause of failure to induce protection, cytokine release from the spleen cells of immunized mice and Leishmania-specific serum antibodies were analyzed. Spleen cells from mice immunized with GP63-GST plus BCG that were exposed to soluble extract of L. donovani in vitro produced 10-fold greater quantities of IFN-gamma and 3-fold greater quantities of IL-5 than cells from mice receiving BCG only or saline. Western blot analysis revealed that sera from these mice had Leishmania-specific antibodies recognizing 1 to 3 antigens of L. donovani with M. W. of 60-65 kDa. Although immunization of mice with GP63-GST induced a strong Th1 response, this study indicated that GP63-GST simultaneously elicited the Th2 response of the CD4+ T-cell, which was known to abrogate the protective immune response conferred by the Th1 effector function.

• The Korean Journal of Food And Nutrition
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• v.29 no.2
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• pp.178-185
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• 2016

This study was designed to develop and to qualify a coffee alternative beverage using a mixture of coffee beans and roasted black beans (Rhynchosia nulubilis). Therefore, the total isoflavone content (TIC), total phenol content (TPC), antioxidant activity, anti-inflammatory activity, NFATc1 (Nuclear factor of activated T-cells c1) expression in RANKL (receptor activator of nuclear factor kappa-B ligand)-stimulated RAW264.7 cells and sensory evaluation were measured for 5 different Cb (coffee bean)-RoS (roasted seomoktae) mixture extracts (Cb100RoS0, Cb75RoS25, Cb50RoS50, Cb25RoS75, and Cb0RoS100). Cb0RoS100 had the highest TIC ($516.83{\pm}36.61mg/100g$) and TPC ($18.11{\pm}1.77mg$ TAE/100 g) along with the highest antioxidant activity as measured by DPPH radical scavenging activity ($73.55{\pm}8.11%$) and ABTS radical scavenging activity ($63.27{\pm}7.27%$). Also, Cb0RoS100 showed the highest anti-inflammatory activity as measured by NO production ($13.57{\pm}2.21{\mu}M$) and PGE2 production ($3.25{\pm}0.21ng/mL$). The more the RoS ratio was increased in the mixtures of Cb-RoS, the more the NFATc1 protein expression was decreased in RANKL-stimulated RAW264.7 cells. In case of sensory evaluation, Cb50RoS50 had the highest scores for flavor, delicate flavor and overall quality, which were similar to those in Cb alone (Cb100RoS0). We suggest that the use of RoS replacement instead of Cb in/as a coffee alternative beverage may help to reduce the risk of caffeine-related bone loss and/or bone disease by effectively blocking NFATc1 expression in RANKL-stimulated RAW264.7 cells compared with Cb alone.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.91-97
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• 2000

Objective: The presence of the various cytokines in human peritoneal fluid has been evaluated incompletely. Changes in cytokine levels may be related to activation of peritoneal macrophage and T-lymphocyte, development of endometriosis, and infertility. This study assesses peritoneal fluid levels of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in infertile women with endometriosis and normal women without endometriosis. Design: Prospective and case-control study in university hospital. Materials and Methods: Cytokine levels in peritoneal fluid obtained during laparotomy or laparoscopy from 21 patients in infertile patients with endometriosis and 24 controls undergoing laparotomy or laparoscopy with no evidence of pelvic endometriosis were determined by enzyme-linked immunosorbent assay. Results: The mean levels of interleukin-6 in infertile patients with endometriosis and controls were $72.7{\pm}23.7$ pg/ml and $18.5{\pm}9.7$ pg/ml respectively (p=0.02). Similarly, the mean levels of interleukin-8 in infertile patients with endometriosis was significantly higher than that of controls ($445.0{\pm}89.6$, vs $45.1{\pm}48.4$, p=0.04). The mean concentration of interleukin-10 in infertile patients with endometriosis was significantly lower than that of controls ($1.09{\pm}0.04$ vs $2.19{\pm}0.03$, p=0.03). The level of tumor necrosis factor-${\alpha}$ was not significantly different between the two study groups. Conclusions: Increased IL-6 and IL-8 and decreased IL-10 levels in the peritoneal fluid may be related to pathogenesis in the endometriosis and infertility, suggesting that partially contribute to the disturbed immune regulation observed in infertili women with endometriosis.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.133-142
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• 2002

The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.

• Journal of Bio-Environment Control
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• v.12 no.1
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• pp.7-11
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• 2003

Cellular glass foam (CGF), the reprocessed glass, has a possibility to be used as a medium component in plug culture of horticultural crops due to the its excellent air and water permeability as comparable to perlite. An experiment was conducted to investigate growth of plug seedlings of Lycopersicum esculentum 'Segye' as influenced by irrigation frequency in various medium combinations of CGF (2.0-4.0 mm particle size) and peatmoss. Seeds were sown in 200-cell plug trays, filled with mixtures of CGF and peatmoss either at 33：67 or 25：75 (％. v/v) and were germinated on a fogged propagation bed. The irrigation frequencies used were one, two or three times per every two days. A commercial plug medium (Tosilee medium) was used as the control, and the irrigation frequency in the control was one time per day. Growth of seedlings, and medium pH and EC were measured at 33 days after sowing. The medium composition had little influence on overall growth of seedlings. Irrigation frequency very significant affected number of leaves, leaf area, chlorophyll concentration, fresh and dry weights of shoots and roots, and dry matter. Growth of seedlings was the greatest with the highest irrigation frequency in the 25% CGF＋75% peatmoss mixture.

• BMB Reports
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• v.52 no.4
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• pp.289-294
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• 2019

The present study evaluated the role of AHNAK in Bartonella henselae infection. Mice were intraperitoneally inoculated with $2{\times}10^8$ colony-forming units of B. henselae Houston-1 on day 0 and subsequently on day 10. Blood and tissue samples of the mice were collected 8 days after the final B. henselae injection. B. henselae infection in the liver of Ahnak-knockout and wild-type mice was confirmed by performing polymerase chain reaction, with Bartonella adhesion A as a marker. The proportion of B. henselae-infected cells increased in the liver of the Ahnak-knockout mice. Granulomatous lesions, inflammatory cytokine levels, and liver enzyme levels were also higher in the liver of the Ahnak-knockout mice than in the liver of the wild-type mice, indicating that Ahnak deletion accelerated B. henselae infection. The proportion of CD4+interferon-${\gamma}$ ($IFN-{\gamma}^+$) and $CD4^+$ interleukin $(IL)-4^+$ cells was significantly lower in the B. henselae-infected Ahnak-knockout mice than in the B. henselae-infected wild-type mice. In vitro stimulation with B. henselae significantly increased $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected wild-type mice, but did not increase $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected Ahnak-KO mice. In contrast, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, IL-10, RANTES, and tumor necrosis $factor-{\alpha}$ secretion was significantly elevated in the splenocytes obtained from both B. henselae-infected wild-type and Ahnak-knockout mice. These results indicate that Ahnak deletion promotes B. henselae infection. Impaired $IFN-{\gamma}$ and IL-4 secretion in the Ahnak-knockout mice suggests the impairment of Th1 and Th2 immunity in these mice.

• Journal of Plant Biotechnology
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• v.48 no.2
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• pp.106-114
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• 2021

We evaluated the anti-cancer activity against human prostate cancer cells and the associated molecular mechanism of extracts from the branches of Rhamnus yoshinoi (RYB). Treatment with RYB suppressed viability of human prostate cancer cells (PC-3) and decreased protein levels of both β-catenin and T-cell factor 4 (TCF4). This was reflected in reduced TCF4 mRNA, but not decreased β-catenin mRNA. PC-3 cells were pretreated with the proteosome inhibitor MG132 before treatment with RYB, which blocked RYB-mediated down regulation of β-catenin in PC-3 cells, thus confirming that RYB promotes the proteasomal degradation of β-catenin. RYB induced β-catenin phosphorylation, and GSK-3β inhibition by LiCl blocked the phosphorylation and proteasomal degradation of β-catenin by RYB. These results suggest that GSK-3β may be an important upstream kinase for RYB-mediated regulation of β-catenin. Finally, GC/MS analysis of RYB identified 18 compounds. Based on these findings, RYB shows potential for development as a therapeutic agent for prostate cancer.