• Biomolecules & Therapeutics
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• v.13 no.2
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• pp.101-106
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• 2005

The generation of secretory IgA antibodies (Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the heat-labile Escherichia coli enterotoxin A2B (ltxa2b) gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ltxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and westem blotting using antibodies to the maltose binding protein (MBP) or the heat labile E. coli subunit B (LTXB), plus the N-terminal amino acid sequence was analyzedd. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/LTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of LTXB. thisstudy also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA (sIgA) and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked LTXA2B acts as a useful mucosal adjuvant, and that adhesin/LTXA2A chimeric protein might be a potential antigen for oral immunization against UPEC.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.2 no.2
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• pp.147-152
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• 1999

Purpose: The aim of this study was to evaluate the prevalence of H. pylori infection in normal preschool children. Methods: Study population consisted of 220 preschool children aged 2 to 5 years in Seoul, Korea. We measured H. pylori specific IgG antibody using the GAP test IgG kit, and samples with concentrations greater than 20 U/ml were considered positive. Results: Of the 220 children, 5 (2.3%) were positive, and thirteen (5.9%) of 220 were equivocally probable positive. Conclusion: The positive rate of H. pylori infection detected by specific IgG antibody in normal preschool children aged 2 to 5 years was approximately 2.3% to 8.2%. This prevalence rate is still high, compared to that of the West. A better knowledge of the transmission of H. pylori is crucial to formulate recommendations for lowering the rate.

• Parasites, Hosts and Diseases
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• v.53 no.5
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• pp.605-610
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• 2015

Toxoplasmosis is considered as an important risk factor for bad obstetric history (BOH) and one of the major causes of congenitally acquired infections. The present study aimed to estimate the seropositivity of T. gondii infection and associated risk factors among the attendees of high risk pregnancy and low risk antenatal care clinic of Minia Maternity and Pediatric University Hospital, Minia, Egypt. The study was carried out from April 2013 to April 2014 through 2 phases, the first phase was case-control study, and the second phase was follow-up with intervention. A total of 120 high risk pregnant and 120 normal pregnant females were submitted to clinical examinations, serological screening for anti-Toxoplasma IgM and IgG antibodies by ELISA, and an interview questionnaire. Seropositive cases were subjected to spiramycin course treatment. The results showed that the seroprevalence of toxoplasmosis in high-risk pregnancy group was 50.8%, which was significantly different from that of normal pregnancy group (P<0.05). Analysis of seropositive women in relation to BOH showed that abortion was the commonest form of the pregnancy wastage (56.5%). The high prevalence of T. gondii seropositive cases was observed in the age group of 21-30 years. Post-delivery adverse outcome was observed in 80.3% of high-risk pregnancy group compared to 20% of normal pregnancy group. There was a statistically significant relationship between seropositivity and living in rural area, low socioeconomic level, and undercooked meat consumption (P<0.05). Serological screening for anti-Toxoplasma antibodies should be routine tests especially among high-risk pregnant women.

• Journal of Nutrition and Health
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• v.43 no.4
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• pp.367-373
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• 2010

The present study was to investigate anti-oxidative and anti-inflammatory activity of Perillae semen in RBL-2H3 basophilic leukemia cells. Inhibitory effect of Perillae semen onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activities in vitro. Anti-inflammatory actions of Perillae semen extracts (100, 250, $500\;{\mu}g/mL$) were assessed by testing their effects on the degranulation of mast cells. For this, $\beta$-hexosaminidase released from RBL-2H3 cells was used and proinflammatory cytokines were measured by an ELISA kit. Our results indicated that Perillae semen water extracts effectively inhibited free radical generation. At the concentration of $500\;{\mu}g/mL$ of water extract, the degranulation of RBL-2H3 cells were inhibited by 42.1%. The IgE-antigen complex increased the accumulation of IL-4 and TNF-$\alpha$ secretion in RBL-2H3 cells and treatments with 250 and $500\;{\mu}g/mL$ of Perillae semen extracts suppressed the IgE induced secretion of IL-4 and TNF-$\alpha$ protein by 20.5, 26.9% and 14.5, 16.5% respectively. We observed that Perillae semen water extract reduced $\beta$-hexosaminidase, IL-4, and TNF-$\alpha$ secretion in RBL-2H3 cells. These results provide that Perillae semen may be beneficial in the treatment of allergic inflammatory disease.

• Journal of Ginseng Research
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• v.46 no.4
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• pp.550-560
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• 2022

Background: The effect of ginsenoside Rh2 (G-Rh2) on mast cell-mediated anaphylaxis remains unclear. Herein, we investigated the effects of G-Rh2 on OVA-induced asthmatic mice and on mast cell-mediated anaphylaxis. Methods: Asthma model was established for evaluating airway changes and ear allergy. RPMCs and RBL-2H3 were used for in vitro experiments. Calcium uptake, histamine release and degranulation were detected. ELISA and Western blot measured cytokine and protein levels, respectively. Results: G-Rh2 inhibited OVA-induced airway remodeling, the production of TNF-α, IL-4, IL-8, IL-1β and the degranulation of mast cells of asthmatic mice. G-Rh2 inhibited the activation of Syk and Lyn in lung tissue of OVA-induced asthmatic mice. G-Rh2 inhibited serum IgE production in OVA induced asthmatic mice. Furthermore, G-Rh2 reduced the ear allergy in IgE-sensitized mice. G-Rh2 decreased the ear thickness. In vitro experiments G-Rh2 significantly reduced calcium uptake and inhibited histamine release and degranulation in RPMCs. In addition, G-Rh2 reduced the production of IL-1β, TNF-α, IL-8, and IL-4 in IgE-sensitized RBL-2H3 cells. Interestingly, G-Rh2 was involved in the FcεRI pathway activation of mast cells and the transduction of the Lyn/Syk signaling pathway. G-Rh2 inhibited PI3K activity in a dose-dependent manner. By blocking the antigen-induced phosphorylation of Lyn, Syk, LAT, PLCγ2, PI3K ERK1/2 and Raf-1 expression, G-Rh2 inhibited the NF-κB, AKT-Nrf2, and p38MAPK-Nrf2 pathways. However, G-Rh2 up-regulated Keap-1 expression. Meanwhile, G-Rh2 reduced the levels of p-AKT, p38MAPK and Nrf2 in RBL-2H3 sensitized IgE cells and inhibited NF-κB signaling pathway activation by activating the AKT-Nrf2 and p38MAPK-Nrf2 pathways. Conclusion: G-Rh2 inhibits mast cell-induced allergic inflammation, which might be mediated by the AKT-Nrf2/NF-kB and p38MAPK-Nrf2/NF-κB signaling pathways.

• Parasites, Hosts and Diseases
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• v.30 no.1
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• pp.33-42
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• 1992

Naegleria fowleri, a free-living amoeba commonly found in moist soil and fresh water, enters the body via the nasal mucosa and migrates along the olfactory nerve to t he brain, where it causes acute amoebic meningoencephalitis. In the present study 7 clones secreting monoclonal antibodies (McAbs) against N. fowleri were produced and the effector function of them was investigated. Their isotopes were IgGl (Nf 1, Nf 154), 19G3 (Nf 137) and 19A (Nf 1, Nf 2, Nf 256, Nf 279). Five McAbs (McAb Nf 2, Nf 279, Nf 27, Nf 154, Nf 137) were specific for N. fowleri by ELISA and recognized the antigenic determinants located on the trophoBoite surface by IFAT and immunoperoxidase stain. These aye McAbs had capacity to agglutinate N. fowleri trophozoites and inhibited the growth of the amoeba in culture medium. McAb Nf 2 inhibited proliferation of trophozoites in vitro significantly. Also the cytotoxicity of JV. fowleri against CHO cell was reduced in the presence of McAb Nf 2 and McAb Nf 154. From these results McAb Nf 2 was confirmed to weaken the virulence of the amoeba among 7 screened McAbs.

• Parasites, Hosts and Diseases
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• v.30 no.2
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• pp.83-90
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• 1992

Sensitivity of anti-Texoplasma antibody (IgG) test by enzyme·linked immunosorbent assay (ELISA) was evaluated in comparison with indirect laten agglutination (ILA) using 2,016 paired human samples of serum and cerebrospinal quid (CSF) . The samples were collected from neurologic patients in Korea with mass lesions in central nervous system(CNS) as revealed by imaging diagnosis(CTIMRI). When the sera were screened for anti-Toxoplasma antibody by ILA, 76 cases (3.8%) were positive (1:32 or higher titers). In the pairs samples of CSF, no positive reactions were observed. When ELISA was performed using PBS extract of Percoll purified tachysoites as antigen, cut-off absorbance was determined as 0.40 for serum and 0.27 for CSF tests. The antibody positive rates by ELISA were 7.0% in serum and 5.6% in CSF Of them, 40 cases(2.0%) showed positive reactions in both serum and CSF, The antibody positive rates were higher in groups older than 40 years, The rates were higher in male(4.7% by ILA, 8.3% by ELISA) than in female(2.2% by ILA, 5.0% by ELISA). The rates in CSF showed no such sex difference. ELISA showed twice higher positive rates when serum was tested, and was sensitive enough to detect specific antibodies in CSF. Etiologic relations between positive antibody tests and CNS lesions remained unknown.

• Pediatric Infection and Vaccine
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• v.3 no.2
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• pp.175-184
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• 1996

Purpose : This study was intended to measure seropositivities and the levels of mumps- and rubella-specific IgG of MMR vaccinees over 17 months of age in Korea. Materials and Methods : From June 1994 to April 1995 we obtained sera from visitors of well baby clinic and patients in Korea University Hospital, who were MMR vaccinees over 17 months of age and had no evidence of immunodeficiency. These 275 study population include 145 males and 130 females. Mumps- and rubella-specific IgG antibody levels were measured by ELISA. Cut-off values for seropositivity were 20 GU(Gamma Unit) in mumps and 0.17 in rubella. Results : 1) As age increased, seropositivities of mumps-specific IgG increased significantly, being 69.0% in 1.5~2 year, 75.0% in 3~4 year, 76.0% in 5~6 year, 90.0% in 7 year, 100% in 8 year, 96.9% in 9 year, 97.4% in 10 year, 97.4% in 11 year, and 96.6% in 12 year of age(p<0.001). 2) As age increased, the levels of mumps-specific IgG antibody(mean${\pm}$standard deviation, GU) increased significantly, being $64.9{\pm}66.5$ in 1.5-2 year, $117.7{\pm}126.4$ in 3~4 year, $152.3{\pm}147.1$ in 5~6 year, $194.3{\pm}168.2$ in 7 year, $258.1{\pm}190.6$ in 8 year, $193.1{\pm}130.1$ in 9 year, $225.7{\pm}119.6$ in 10 year, $220.7{\pm}114.3$ in 11 year, and $222.3{\pm}127.1$ in 12 year of age(p<0.001). There was positive correlation between age and mumps-specific antibody level (r=0.3282, p<0.001). 3) As age increased, seropositivities of rubella-specific IgG decreased significantly, being 72.4% in 1.5~2 year, 75% in 3~4 year, 72% in 5~6 year, 60% in 7 year, 44.4% in 8 year, 40.6% in 9 year, 28.2% in 10 year, 23.1% in 11 year, and 17.2% in 12 year of age(p<0.001). 4) As age increased, rubella-specific IgG decreased significantly, being $0.462{\pm}0.356$ in 1.5~2year, $0.438{\pm}0.306$ in 3~4 year, $0.287{\pm}0.179$ in 5~6 year, $0.204{\pm}0.139$ in 7 year, $0.189{\pm}0.153$ in 8 year, $0.124{\pm}0.121$ in 9 year, $0.093{\pm}0.114$ in 10 year, $0.104{\pm}0.135$ in 11 year, and $0.080{\pm}0.001$ in 12 year of age(p<0.001). There was negative correlation between age and rubella-specific IgG titer (r=-0.551, p<0.001). Conclusions : Eventhough seropositivities and the level of mumps-specific IgG increased as age increased, they are not enough to prevent mumps infection in 1.5 to 6 years of age. Seropositivities and the level of rubella-specific IgG decreased as age increased. Appropriate change in vaccine schedule may be needed to decrease the risks of mumps and rubella infection.

• Food Science of Animal Resources
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• v.23 no.2
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• pp.161-167
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• 2003

Identification of salmonellosis-infected commercial poultry flocks has become a pivotal component of efforts to reduce incidence of egg-associated transmission of S. enteritidis and S. typhimurium to humans. As a basic study for sanitary control of S. enteritidis and S. typhimurium, main food-borne pathogenic bacteria in eggs produced by domestic hens, commercial egg samples were tested for specific antibodies to whole cells of S. enteritidis and S. typhimurium and outer membrane protein(OMP) of S. typhimurium by ELISA to detect infection of S. enteritidis and S. typhimurium in various groups of hens. When the antibody titers of yolks from three commercial brand eggs were tested after diluting in the ratio from 1:100 to 1:1,600 with double dilution method, ELISA values of the specific antibodies could be shown as differences in dilution patterns by comparing with negative control egg. When the antibody titers of the yolks from two commercial brand eggs were tested after diluting in the ratio of 1:200 and 1:1,000, ELISA values of specific antibodies were different among same brand eggs. When the antibody titers of yolks from five eggs sampled randomly from twenty one commercial brand eggs were tested after diluting in the ratio of 1:1,000, ELISA value of the specific antibodies were shown generally high. ELISA values of 28.5, 30, and 28.5% of yolks from 21 brand eggs were shown low and similar to negative control egg in antibody titers to whole cells of S. enteritidis and S. typhimurium and OMP of S. typhimurium, respectively. The results demonstrated that ELISA test of egg yolk antibody could provide a highly sensitive indicator to detect contamination of S. typhimurium and S. enteritidis in poultry, and could be used effectively to reduce incidence of S. typhimurium and S. enteritidis infection in poultry.

• Parasites, Hosts and Diseases
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• v.47 no.4
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• pp.387-391
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• 2009

Specific IgE against Acarus siro, Glycphagus domesticus, Tyrophagus putrescentiae, and Lepidoglyphus destructor have been investigated by ELISA in sera of 92 children. Of them, 41 were found to be specific IgE positive ($\geq$ 0.35 IU/ml) against at least one of house dust mite species, Dermatophagoides pteronyssinus and Dermatophagoides farinae, by an immunoblot. In 65.9% of the dust mite-sensitized children, specific IgE against at least one of these mite species was found. Sensitization levels, including co-sensitization cases were found to be 35.7% against A. siro, 24.4% against T. putrescentiae, 31.7% against L. destructor, and 26.8% against G. domesticus. In non-sensitized children, dust mite sensitization level was found to be 25.5%. Breakdown of sensitization by individual species in this group was; against A. siro and T. putrescentiae at 7.8%, against L. destructor at 13.7%, and against G. domesticus at 9.8%. When all children were reckoned, 43.5% was found to be sensitized against at least one storage mite species, with sensitizations against A. siro at 18.5%, T. putrescentiae at 26.1%, L. destructor at 21.7%, and G. domesticus at 17.4%. In dust samples collected from the dwellings of children, distribution of species was found to be A. siro (17%), G. domesticus (23%), T. putrescentiae (29%), L. destructor (25%), and unidentified (6%). In Fisher's chi-square test on SPSS program, there was a relationship between dust mite sensitization and storage mite sensitization (P < 0.05), but no meaningful relationship was found on the basis of individual mite species.