• Food Science and Biotechnology
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• v.17 no.5
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• pp.919-924
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• 2008

The aim of this study was to observe the changes in allergenicity of saeujeot (salted and fermented shrimp) using a competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). The fermentation conditions tested for saeujeot consisted of various temperatures (25, 15, and $5^{\circ}C$) and salt concentrations (25, 15, and 10%). When saeujeot was fermented at a low salt concentration and high temperature, the binding ability of mAb and shrimp-allergic patient serum to allergen was significantly decreased. In particular, the binding ability of mAb to allergen in saeujeot fermented with 10% salt at $25^{\circ}C$ for 5 days decreased to 5%. Also, the binding ability of shrimp-allergic patient serum to allergen in saeujeot fermented for 5 days with 10% salt at $25^{\circ}C$ was 8%. In conclusion, the binding of mAb and shrimp-allergic patient serum to tropomyosin in saeujeot decreased with longer fermentation periods, lower salt concentrations (10%), and higher temperatures ($25^{\circ}C$).

• The Journal of Internal Korean Medicine
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• v.39 no.3
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• pp.302-312
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• 2018

Objectives: This study was performed to investigate the effects of two herbal medicines, Agastachis Herba and Lysimachiae Herba, on a cellular model of non-alcoholic fatty liver diseases (NFLDs). Methods: HepG2 cells were treated with palmitic acid and with various concentrations of Agastachis Herba (AH) or Lysimachiae Herba (LH) extract in water. The lipotoxicity was assessed using EZ-cytox, and the lipoapoptosis was assessed using cell death detection ELISA. Intracellular lipids were measured by oil red O staining. The efficacy of AH and LH on sterol regulatory element-binding transcription factor-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC) in HepG2 cells was measured by reverse transcription polymerase chain reaction (RT-PCR). Results: Both AH and LH extracts increased lipoapoptosis and decreased lipotoxicity and levels of SREBP-1c, ACC, and FAS (SREBP-1c, ACC, and FAS are factors in lipid synthesis). In the oil red O staining experiment, both extracts also reduced intracellular lipid accumulation; in this instance, LH's efficacy was superior to that of AH. Conclusions: According to the results, both AH and LH are likely to contribute to non-alcoholic fatty liver disease, as both interfere with lipid synthesis.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1076-1080
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• 2008

Glucagon, a peptide hormone produced by alpha-cells of Langerhans islets, is a physiological antagonist of insulin and stimulator of its secretion. In order to improve its bioactivity, we modified its structure at the C-terminus by amidation catalyzed by a recombinant amidase in bacterial cells. The human gene coding for glucagon-gly was PCR amplified using three overlapping primers and cloned together with a rat ${\alpha}$-amidase gene in plasmid pMGA. Both genes were expressed under control of the strong constitutive promoter of aph and secretion signal melC1 in Streptomyces lividans. With Phenyl-Sepharose 6 FF, Q-Sepharose FF, SP-Sepharose FF chromatographies and HPLC, the peptide was purified to about 93.4% purity. The molecular mass of the peptide is 3.494 kDa as analyzed by MALDI TOF, which agrees with the theoretical mass value of the C-terminal amidated glucagon. The N-terminal sequence of the peptide was also determined, confirming its identity with human glucagon at the N-terminal part. ELISA showed that the purified peptide amide is bioactive in reacting with glucagon antibodies.

• Journal of Sasang Constitutional Medicine
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• v.23 no.3
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• pp.374-390
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• 2011

1. Objectives: The purpose of this study is to investigate the effect of Suhwagije-tang(SGT) distillate on the immune activity of spleen cells of aged SD rats. 2. Methods: We used 10, 50, 72 weeks old SD rats in this study. Spleen cells from SD rats were stimulated with ConA and treated with 1% Vitamin C(Vit.C) or Suhwagijetang distillate(SGT). After 24 hours, the concentrations of IL-2, IL-4, IL-10, IFN-${\gamma}$ in the cell culture supernatant were measured by ELISA. 3. Results and Conclusions 1) At all concentration of SGT distillate, survival rates of liver cells were higher than the control group. In addition, 50% SGT distillate group's cell survival rates were significantly higher than other groups. 2) In 10 weeks SD rats(SGT group), the concentration of IL-2 significantly decreased in comparison with ConA group, Vit.C group. In 52 weeks SD rats(SGT group), the concentration of IL-2 significantly decreased in comparison with ConA group. 3) In 10, 52 weeks SD rats(SGT group), the concentration of IL-4 significantly decreased in comparison with ConA group. 4) In 10 weeks SD rats(SGT group), the concentration of IL-10 significantly decreased in comparison with ConA group. And in 72 weeks SD rats(SGT group), the concentration of IL-10 significantly increased in comparison with Vit.C group. 5) In 52, 72 weeks SD rats(SGT group), the concentration of IFN-${\gamma}$ significantly decreased in comparison with 10 weeks SD rats(SGT group). These results suggest that Suhwagije-tang(SGT) distillate has the effect of increasing the immune activity of spleen cells of aged SD rats.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.281-286
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• 2003

Background: Hepatitis B virus (HBV) infection is one of the worldwide public health problem affecting about 300 million people. The envelope protein of HBV consists of three components known as preS1, preS2, and S antigen. According to the recent study, anti-HBs Ab showed effective neutralization ability against HBV from chronic hepatitis B and liver transplant patients, suggesting the possible development of therapeutic antibody. Methods: Spleen cells immunized with S antigen of HBV were fused with myeloma cell line to obtain HBsAg specific monoclonal antibodies. High affinity antibodies against HBsAg (adr, ad and ay type) were selected by competitive ELISA method. Nucleotide sequence of the variable regions of monoclonal antibodies was analyzed by RT-PCR followed by conventional sequencing method. Results: We produced 14 murine monoclonal antibodies which recognize S antigen of HBV. Two of them, A9-11 and C6-9 showed the highest affinity. The sequence analysis of A9-11 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain I (B) and light chain lambda 1, respectively. Likewise, the sequence analysis of C6-9 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain II (B) and light chain kappa 1, respectively. Neutralization assay showed that A9-11 and C6-9 effectively neutralize the HBV infection. Conclusion: These results suggest that A9-11 and C6-9 mouse monoclonal antibodies can be used for the development of therapeutic antibody for HBV infection.

• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.541-560
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• 1994

The present study was performed to evaluate the relationship between the serotype or the genotype of Actnobacillus actinomycetemcomitans (A. a.) and the severity of periodontal disease. Total 64 A. a. clinical isolates were sampled from 46 sites of 20 subjects classified into the group I (1 periodontally healthy subject, 2 gingivitis patients, 5 ealry adult periodontitis patients), group II (3 moderatelly adult periodontitis patients) and group III (1 advanced adult periodontitis patient, 8 RPP patients). Southern bolt hybridization (fingerprinting) patterns of the five reference strains, A. a. strain ATCC 29523 (serotype a), ATCC 29522 (Serotype b), ATCC 43719 (serotype c), IDH 781 (serotype d) and IDH 1705 (serotype e), were used as the five basic genotypic patterns (A, B, C, D, E). NT type was designated as one which did dnot represent any of those five basic types. The serotypes were determined by ELISA technique with the serum samples from pre-immunized rabbit. Based on subject-based analysis, it was noted that genotypes A and C, NT, and B, D, E were significantly related to the disease groups I, II, and III, respectively. It was also noted that both the serotypes a and c were significantly related to the disease group I and II, while serotypes were significantly related bm), and serotypes b and nd were frequently found in sites with severe attachment loss (LA>6mm). The results indicated that the significant relationship can be delineated beteen the genotypes and the serotypes of Acinobacillus actinomycetemcomitans and the periodontal disease severity. The results also indicated that genotyping can provide more detailed information on its relationship with the disease severity based on both the patient-based and the site-based analyses.

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.65-69
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• 1999

Effect of chemo- and thermotherapy on LSV elimination was investigated in Lilium Oriental Hybrid cv. Casa Blanca which was infected by LSV. Either 20 mg/L ribavirin or the treatment of 35$^{\circ}C$/$25^{\circ}C$ (day/night) thermotherapy resulted in the desired growth rate and high percentage (86%, 80% respectively) of LSV elimination. When we combinated those application, the rate of LSV elimination was increased with the time of heat treatment, and was 100% by all heat treatment of 16 weeks. After all of the LSV-free plants transferred into the soil, the number of LSV-free plants was 8 plants out of 12 plants and the efficiency of LSV elimination was best in the combination of 20 mg/L ribavirin and 8 weeks of heat treatment. About 21 % of plants kept LSV-free after transferred all of the LSV-free plants into the soil, the others became LSV-infected again.

• Molecules and Cells
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• v.41 no.3
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• pp.234-243
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• 2018

In recent years, the interest towards the relationship between incretins and bone has been increasing. Previous studies have suggested that glucagon-like peptide-1 (GLP-1) and its receptor agonists exert beneficial anabolic influence on skeletal metabolism, such as promoting proliferation and differentiation of osteoblasts via entero-osseous-axis. However, little is known regarding the effects of GLP-1 on osteoblast apoptosis and the underlying mechanisms involved. Thus, in the present study, we investigated the effects of liraglutide, a glucagon-like peptide-1 receptor agonist, on apoptosis of murine MC3T3-E1 osteoblastic cells. We confirmed the presence of GLP-1 receptor (GLP-1R) in MC3T3-E1 cells. Our data demonstrated that liraglutide inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as detected by Annexin V/PI and Hoechst 33258 staining and ELISA assays. Moreover, liraglutide upregulated Bcl-2 expression and downregulated Bax expression and caspase-3 activity at intermediate concentration (100 nM) for maximum effect. Further study suggested that liraglutide stimulated the phosphorylation of AKT and enhanced cAMP level, along with decreased phosphorylation of $GSK3{\beta}$, increased ${\beta}-catenin$ phosphorylation at Ser675 site and upregulated nuclear ${\beta}-catenin$ content and transcriptional activity. Pretreatment of cells with the PI3K inhibitor LY294002, PKA inhibitor H89, and siRNAs GLP-1R, ${\beta}-catenin$ abrogated the liraglutide-induced activation of cAMP, AKT, ${\beta}-catenin$, respectively. In conclusion, these findings illustrate that activation of GLP-1 receptor by liraglutide inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation through $cAMP/PKA/{\beta}-catenin$ and $PI3K/Akt/GSK3{\beta}$ signaling pathways.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.87-94
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• 1997

Clonorchis sinearis is a liver fluke which is the most prevalent helminth of humans in Korea. The better diagnostic measure of clonorchiasis is required for its nationwide control program. The present study observed antigenic bands of C. sinensis and reacting immunoglobulins in serum of infected residents. Adult C. sinensis were recovered from experimentally infected rabbits and soluble crude extract of the worms was used as the antigen after supplementation of E-64, a cysteine proteinase inhibitor. SDS- PAGE of the crude antigen resolved more than 20 protein bands between 200 and 14 kDa. The sera of infected humans collected at an endemic village showed specific IgG and IgE antibodies but little IgM and IgA antibodies. The protein bands of 94, 80, 72, 68, 52, 47, 43, 37, 34, and 28-25 kDa strongly reacted with serum Ig(GMA) or IgG antibody and 64, 62, 52, 47,44, 34,28, and 26 kDa bands reacted with serum IgE antibody. However, the 94, 80, 72, 68, 64, 62, 52, 47, and 40 kDa bands of C. sinensis antigen were found non specific. The protein bands of 43, 34, and 28-25 kDa of C. sinensis are primary target molecules of further analysis.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.1
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• pp.431-437
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• 2016

In this investigation, global DNA methylation patterns and the specific methylation status of 5 genes were studied in DNA from peripheral blood (PB) and impact on progression free survival (PFS) and overall-survival (OS) in patients with de novo or relapsed acute myeloid leukemia (AML) treated with decitabine-based regimens waas assessed. DNA was isolated from PB samples at the time of -1, 1, and 7 days of chemotherapy. Global methylation was determined by ELISA, and the CpG island DNA methylation profile of 5 genes using a DNA methylation PCR system. Our data demonstrated that patients with a high level of 5-mC had a poor prognosis after demethylation therapy and those who have low levels of 5-mC in PB achieved higher CR and better SO, but there was no significant correlation found between the 5-mC levels and other clinical features before treatment except the disease status. Higher methylation status of Sox2 and Oct4 genes was associated with differential response to demethylation therapy. A relatively low methylation percentage in one or both of these two genes was also associated with longer OS after decitabine based chemotherapy. We also suggest that global DNA and Oct-4/Sox2 methylation might impact on the pathogenesis of leukemia and play an important role in the initiation and progression. Moreover, dynamic analysis of 5-mC and Oct-4/Sox2 in peripheral blood nucleated cells of leukemia patients may provide clues to important molecular diagnostic and prognostic targets.