• Parasites, Hosts and Diseases
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• v.37 no.4
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• pp.277-283
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• 1999

To investigate the development of a subunit vaccine against theileriosis in cattle, the DNA fragments encoding piroplasm surface protein (p33) of Theileria sergenti of a Korean isolate were expressed in baculoviruses. The expressed p33 was characterized by indirect fluorescent antibody (IFA) and western blotting analysis. The expression of p33 was mainly detected on the surface of infected Sf21 cells by IFA. The immunoblotting analysis revealed the presence of a same molecular weight protein band of p33. The antigenicity of expressed polypeptide was further examined through the inoculation of a guinea pig. The sera of guinea pigs immunized with p33 expressed cell Iysate showed similar fluorescent antibody patterns and reacted with the same molecular weight protein of T. sergenti in immunoblotting analysis, thus indicating that this protein can be a promising candidate for a subunit vaccine in the future.

• Natural Product Sciences
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• v.6 no.3
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• pp.147-150
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• 2000

The extracts of Carica papaya (flower), Barringtonia macrostachya (leaves), Coleus tuberosus (tuber), Mangifera indica (fruit skin) and Eugenia polyantha (leaves) showed strong in vitro anti-tumor promoting activity when assayed using Raji cells (Mooi et al., 1999). The antitumor promoting activity of the crude extracts was further analyzed by immunoblotting analysis of Raji cells carving Epstein-Barr virus genome. The expression of early antigens diffuse (EA-D) and early antigens restricted (EA-R) was determined by performing western blotting of treated Raji cells with human sera of nasopharyngeal carcinoma patients. All the plant extracts were shown to be able to suppress both EA-D and EA-R.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.317-322
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• 2009

The proteins from 15 types of cultivar of hot peppers cultivated in Korea were extracted and its allergenicity was investigated by immunoblotting and enzyme-linked immunosorbent assay (ELISA). The immunoblotting of hot pepper proteins extracts (HPEs) against serum of hot pepper sensitized patients revealed dominant IgE binding to 14, 37, and 40 kDa molecules. The specific levels of IgE to HPEs sample No. 1, 3, and 7 were much higher than the other samples in patients. Also, IgE binding capacity of HPEs were not reduced by thermal processing and digestion in ELISA using human IgE antibody acquired from hot pepper sensitized patients. By means of Western blotting using anti-thaumatin IgY, thaumatin-like protein (TLP) acting as allergen in several plants and fruits was detected in tested hot peppers. This study demonstrates that the antigenic protein in hot peppers are present but are differently contained according to cultivars.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7537-7542
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• 2013

Background: Terpinen-4-ol, a monoterpene, is found as the main component of essential oil extracts from many plants. In this study apoptotic and autophagic types of cell death induced by terpinen-4-ol and associated mechanisms were investigated in human leukemic HL-60 cells. Materials and Methods: The cytotoxicity of human leukemic U937 and HL-60 cells was determined by MTT assay. Cytochrome c release, expression of Bax, Bcl-2, Bcl-xl and cleaved Bid were determined by Western blotting. Cell morphology was examined under a transmission electron microscope. LC3-I/II, ATG5 and Beclin-1 levels were detected by immunoblotting. Results: Terpinen-4-ol exhibited cytotoxicity to human leukemic HL-60 but not U937 cells. The apoptotic response to terpinen-4-ol in HL-60 cells was due to induction of cytochrome c release from mitochondria and cleavage of Bid protein after the stimulation of caspase-8. There was a slightly decrease of Bcl-xl protein level. The characteristic cell morphology of autophagic cell death was demonstrated with multiple autophagosomes in the cytoplasm. At the molecular level, the results from Western blot analysis showed that terpinen-4-ol significantly induced accumulation of LC3-I/II, ATG5 and Beclin-1, regulatory proteins required for autophagy in mammalian cells. Conclusions: Terpinen-4-ol induced-human leukemic HL-60 cell death was via both autophagy and apoptosis.

• Korean Journal of Veterinary Research
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• v.32 no.1
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• pp.83-90
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• 1992

Monoclonal antibodies against structural proteins of bovine viral diarrhea virus(BVDV) were derived by classical hybridoma techniques. These antibodies were characterized by serum neutralization, immunoblotting and immunoprecipitation. The neutralizing monoclonal antibody reacted with the 56kd to 54kd(M.W.) viral protein in western blotting and immunoprecipitation analysis. Although there was no neutralizing activity, another monoclanal antibody reacted with the 45kd protein by immunoprecipitation and with both the 45kd and 36kd proteins in immunoblotting analysis. respectively. Densitometer scanning of purified BVDV and the immunopreipitation of whole virus particles with neutralizing monoclonal antibody revealed the presence of more than twelve viral polypeptides. Although no possible precursor form of protein was identified with the neutralizing monoclonal antibody. the presence of intact virion was detected in the infected cell supernatant immediatelty after pulse labeling, indicating rapid translational processing as well as packaging of the virus. The partial peptide mapping of 45kd and 36kd proteins with Staphylococcus aureus V 8 protease showed that these two proteins are related.

• Biomedical Science Letters
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• v.3 no.2
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• pp.95-105
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• 1997

Currently, the laboratory diagnosis for lyme disease have been performed with the detection of antibodies against Borrelia burgdorferi. However there might be some difficulties in the interpretation of obtained results due to the usage of foreign isolates as an antigen in the test. Therefore the optimization of serological tests with Korean isolates of B. burgdorferi as an antigen would be needed to establish the standardized diagnostic method for the detection of antibodies against B. burgdorferi infection. In this study, the optimization of ELISA was investigated with experimentally challenged rabbit sera and sonicated B. burgdorferi antigens of Korean isolates. Of 217 human patient's sera with unknown fever, the mean seropositivities in ELISA, done under the optimized conditions obtained in this study, were found to be about 8％ against B. burgdorferi sensu late, showing the highest seropositivity of 14.3％ against B. afzelii. In immunoblotting assay with ELISA-positive human sera, the major reactive bands were 41kDa (flagellin) which might be the indication of early infection, and 27kDa, 31kDa (OspA), 34kDa (OspB) which are the characteristics of late infection. Obtained results in this study might strongly indicate the possibility of B. burgdorferi infections in Korea.

• Development and Reproduction
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• v.12 no.2
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• pp.183-194
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• 2008

A variety of stem cells has been emerging as therapeutic cells that can replace organ transplantation in human liver diseases. The present study focused on whether human eyelid adipose-derived stem cells (HAD) might differentiate into functional hepatocyte-like cells in vitro. HAD were isolated from human eyelid adipose tissue. Effect of dimethyl sulfoxide (DMSO), fibroblast growth factor (FGF)-2 and FGF-4 on the hepatic differentiation of HAD have been examined in vitro. Immunocytochemical analysis and PAS staining showed that HAD cultured in both DMSO and FGF-4 exhibited the most intense staining than HAD of the other experimental groups. These HAD expressed numerous hepatocyte-related genes. Immunoblotting analyses showed that HAD cultured in the presence of DMSO and FGF-4 secreted higher amount of human albumin than HAD cultured in other conditions. Urea analysis also demonstrated that these HAD produced higher amount of urea than any other groups of HAD. In conclusion, combined treatment of DMSO and FGF-4 could effectively induce the functional differentiation of HAD into hepatocyte-like cells.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.1971-1982
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• 2017

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

• The Journal of Internal Korean Medicine
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• v.41 no.1
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• pp.44-58
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• 2020

Objectives: This study was conducted to identify the protective effects of Chongmyunggongjin-dan (CMGJD) on Hydrogen peroxide (H₂O₂)-induced apoptosis mechanisms in C6 glial cells. Method: We used CMGJD after distilled water extraction, filtration, and lyophilization. The ROS scavenging effect was examined by fluorescence microscopy. Expression levels of proteins related to ROS generation were investigated by western blotting. Functional changes in organelles related to Reactive oxygen species (ROS) generation were investigated by immunoblotting and by verifying expression level of relevant enzymes. Results: The CMGJD extract protected the cells against H₂O₂-induced morphological changes and DNA fragmentation, inhibited the increase of Heme_oxygenase-1(HO^-1) and the decrease in catalase, protected against the loss of mitochondrial membrane potential, inhibited disturbances of lysosomal function, and induced an increase in peroxisomes. Conclusion: CMGJD was confirmed to have a protective effect on H₂O₂-induced C6 glial cell death possibly by blocking the pathways causing damage to subcellular organelles, such as mitochondria, lysosomes, and peroxisomes. We assume that CMGJD will be effective for the prevention and treatment of ischemic stroke in a clinical environment.

• Korean Journal of Veterinary Research
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• v.43 no.3
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• pp.383-392
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• 2003

Insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) are important regulators on the development of maternal tissues during pregnancy. This study was performed to examine the relationship between maternal IGFs/IGFBPs system (i.e: IGF-I, II, their receptors, and IGFBPs) in pre- and post-partum rats. The liver and kidney are important organs for the synthesis of IGFs and IGFBPs in adults. The levels of materanal IGFs and IGFBPs in serum, liver, and kidney were examined at 14 and 21 days of gestation and at 3, 7, 11, and 14 days after birth. The expression of IGFs and their receptors mRNA was also examined in fetal and maternal rat liver, kidney. IGF-I concentrations in maternal serum and liver were decreased during pregnancy. However, IGF-I concentration in maternal kidney was increased, having maximal effect at 14 days of gestation. IGF-I concentrations were decreased in serum, liver, and kidney of postpartum rat, compared to control (p < 0.05). On the other hand, IGF-II concentrations in serum, liver, and kidney were increased during pregnancy (p<0.05) and gradually decreased to control level in postpartum period. The levels of IGFBP-3 and IGFBP-2 are expressed in serum, liver, and kidney. However, IGFBP-3 is mainly expressed in serum and liver, and IGFBP-2 in kidney. The levels of IGFBP-3 and IGFBP-2 in maternal serum were markedly decreased during pregnancy and gradually recovered to control level during postpartum period by western ligand blotting. However, there was no change of IGFBP-3 and IGFBP-2 levels by western immunoblotting. The levels of IGFBP-3 and IGFBP-2 in maternal liver and kidney also showed the same pattern of serum, although the main IGFBP is different. In normal rat serum, IGF-I 150 kDa and 50 kDa carrier proteins were detected. The level of IGF-I 150 kDa carrier proteins in pregnant rat was decreased compared to normal rat, but that of 50 kDa carrier proteins was increased. IGFBP-3 protease activity was identified in pregnant rat serum and maternal placenta, and it was inhibited by EDTA ($Ca^{2+}$ chelating agent) and aprotinin (serine proteinase inhibitor). Taken together, these results suggest that the changes of IGFs and IGFBPs in maternal rats are regulated by liver and kidney IGFs and their receptors mRNA during the pregnancy.