• Journal of Life Science
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• v.32 no.7
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• pp.491-500
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• 2022

HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient individually approved by the Korea Ministry of Food and Drug Safety for liver health. The anti-inflammatory effect of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in RAW 264.7 macrophage cells treated with lipopolysaccharide (LPS). An active hexose correlated compound (AHCC) was used as a positive control. LPS-activated RAW 264.7 cells were treated with HKSMM50 and AHCC (0, 20, 100, 500 ㎍/ml) and cultured for 24 hr. Inflammation-related elements in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in the cells was analyzed by Western blotting. The HKSMM50 lowered iNOS and COX-2 protein expressions, and nuclear factor-kappa B (NF-κB), nitric oxide (NO) and prostaglandin E₂ (PGE₂) contents in a concentration-dependent manner as compared to LPS treatment. Similarly, the HKSMM50 lowered the content of pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-6 (IL-6) contents and increased the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The efficacy of the AHCC treatment was similar to that of the HKSSM50 treatments. These results indicate that HKSMM50 showed an anti-inflammatory effect in LPS-treated RAW 264.7 cells by down-regulation of NF-κB signaling and suggest that HKSMM could be used as a health functional food ingredient to help improve immune function.

• Journal of Veterinary Science
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• v.21 no.3
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• pp.36.1-36.9
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• 2020

Background: Pseudorabies, also known as Aujeszky's disease, is caused by the pseudorabies virus (PRV) and has been recognized as a critical disease affecting the pig industry and a wide range of animals around the world, resulting in great economic losses each year. Shandong province, one of the most vital food animal-breeding regions in China, has a very dense pig population, within which pseudorabies infections were detected in recent years. The data, however, on PRV epidemiology and coinfection rates of PRV with other major swine diseases is sparse. Objectives: This study aimed to investigate the PRV epidemiology in Shandong and analyze the current control measures. Methods: In this study, a total number of 16,457 serum samples and 1,638 tissue samples, which were collected from 362 intensive pig farms (≥ 300 sows/farm) covered all cities in Shandong, were tested by performing enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results: Overall, 52.7% and 91.5% of the serum samples were positive for PRV-gE and -gB, respectively, based on ELISA results. In addition, 15.7% of the tissue samples were PCR positive for PRV. The coinfection rates of PRV with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus, and classical swine fever virus were measured; coinfection with PCV2 was 35.0%, higher than those of the other two viruses. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Conclusions: The results demonstrate the PRV prevalence and its coinfection rates in Shandong province and indicate that pseudorabies is endemic in pig farms in this region. This study provides epidemiological data that can be useful in the prevention and control of pseudorabies in Shandong, China.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.27 no.1
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• pp.26-36
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• 2024

Purpose: We investigated the role of CD8⁺T cells as host immune factors in pediatric patients with Helicobacter pylori gastritis. Methods: Gastric mucosal tissue and blood samples were collected from 39 children, including 11 children with H. pylori infection and 28 children as controls. Anti-CD8 and anti-T-bet antibodies were used for immunohistochemistry of the gastric mucosa. For the cell surface and intracellular staining, peripheral blood mononuclear cells were stained with anti-IL7Rα, anti-CX3CR1, anti-CD8, anti-T-bet, and anti-IFN-γ antibodies. Cytokines of sera such as tumor necrosis factor alpha (TNF-α) and CX3CL1 were analyzed using enzyme- linked immunosorbent assay (ELISA). Results: In the immunohistochemistry of gastric mucosa, the frequency of CD8⁺ and T-bet⁺ T cells cells was higher in the H. pylori-positive group than in the control group (26.9± 7.8% vs. 16.9±3.3%, p<0.001; 5.0±2.5% vs. 2.2±0.7%, p=0.001). Between the control and H. pylori-positive groups, the frequency of IL-7Rα^lowCX3CR1⁺ CD8⁺ and T-bet⁺ INF-γ⁺ CD8⁺ T cells were not significantly different between surface and intracellular staining, respectively (40.4±24.0% vs. 38.2±17.8%, p=0.914; 40.4±24.0% vs. 38.2±17.8%, p=0.914). In the ELISA, no significant differences in TNF-α and CX3CL1 concentrations were observed between the control and H. pylori-positive groups (34.3±12.1 pg/mL vs. 47.0±22.6 pg/mL, p=0.114/0.5± 0.1 pg/mL vs. 0.5±0.1 pg/mL, p=0.188). Conclusion: CD8⁺ T and Th1 cells, which secrete IFN-γ, might play important roles in the mucosal immunity of the stomach in children with H. pylori infection.

• IMMUNE NETWORK
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• v.16 no.5
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• pp.296-304
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• 2016

It has been reported that fatty acid binding proteins (FABPs) do not act only as intracellular mediators of lipid responses but also have extracellular functions. This study aimed to investigate whether extracellular liver type (L)-FABP has a biological activity and to determined serum L-FABP levels in patients with end-stage renal disease (ESRD). We isolated L-FABP complementary deoxyribonucleic acid (cDNA) from the Huh7 human hepatocarcinoma cell line and expressed the recombinant L-FABP protein in Escherichia coli. A549 lung carcinoma and THP-1 monocytic cells were stimulated with the human recombinant L-FABP. Human whole blood cells were also treated with the human recombinant L-FABP or interleukin (IL)-1α. IL-6 levels were measured in cell culture supernatants using IL-6 enzyme-linked immunosorbent assay (ELISA). Human recombinant L-FABP induced IL-6 in a dose-dependent manner in A549, THP-1 cells, and whole blood cells. The blood samples of healthy volunteers and patients with ESRD were taken after an overnight fast. The serum levels of L-FABP in healthy volunteers and ESRD patients were quantified with L-FABP ELISA. The values of L-FABP in patients with ESRD were significantly lower than those in the control group. Our results demonstrated the biological activity of L-FABP in human cells suggesting L-FABP can be a mediator of inflammation.

• Korean Journal of Poultry Science
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• v.18 no.3
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• pp.183-196
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• 1991

In order to establish ELISA method to detect antibody against IBV various factors involved were examined. Antigen was prepared from Massachusetts type IBV which is known to be one of serotypes distributed most widely. The virus was grown in embryonated SPF chicken eggs. Allantoic fluid harvested was processed to ultracentrifugation and sucrose density gradient centrifugation to produce a purified antigen The antisera selected from the field samples based on hemagglutination inhibition test were used as the standard positive and negative sera for this study and the results obtained were summarized as follows. 1 , It was found that ELISA test was satisfactory when the purified antigen was coated on the plate in the amount of about 40ng protein per well. In case of the phospholipase treated hemagglutinating antigen it gave satisfactory results when the each well wns coated with 1.2 to 2.5 hemagglutinating unit which was equivalent to 40 to 90ng of protein. 2. There was no significant difference in the ratio of optical density of positive to that of negative serum whether the coated antigen was held for 1 hour at 37$^{\circ}C$ or it was held overnight at 4$^{\circ}C$. The coated antigen could be kept in dried state without change of antigenecity for at least one month of experimental period at 4$^{\circ}C$. 3. There was a big variation in the optical density and P/N values depending on the maker of the plates and on the plate of the same maker. 4. It was found that background optical density was negligible when serum was diluted more than 1：50 and serum dilution of 1：100 appeared to be appropriate as a routine test dilution to screen the antibody. 5. Optical density was fairly constant 15 minutes afterward from the time substrate was treated and during the 4 hours after stopper was treated. 6. There was a low correlation(r＝0.42) between ELISA and HI test. However, when 74serum samples were tested for the IBV antibody, 98.7% were found to be positive by both tests in which titers of 2$^{6}$ or more by HI test and P/N values of 1.4 or more by ELISA were considered to be positive, 7 Day-old IBV vaccinated chickens shows a similar antibody decay and rising pattern until 8 weeks of age by the two tests, ELISA and HI.

• Tuberculosis and Respiratory Diseases
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• v.45 no.2
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• pp.271-279
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• 1998

Background: Diagnosis by direct microscopy and/or by culture of the Mycobacterium tuberculosis from body fluids or biopsy specimens is "Gold standard". However, the sensitivity of direct microscopy after Ziehl-Neelsen staining is relatively low and culture of mycobacteria is time consuming. Detection of mycobacterial DNA in clinical samples by the polymerase chain reaction is highly sensitive but laborious and expensive. Therefore, rapid, sensitive and readily applicable new tests need to be developed. So we had evaluated the clinical significance of serologic detection of antibody to 38 kDa antigen, which is known as the most specific to the M. tuberculosis complex, and culture filtrate antigen by ELISA in sputum AFB smear negative patients. Method: In this study, culture tests for acid fast bacilli with sputa or bronchial washing fluids of 183 consecutive patients who were negative of sputum AFB smear were performed. Simultaneously serum antibodies to 38 kDa antigen and unheated culture filtrate of M. tuberculosis were detected by an ELISA method. Results: The optical densities of ELISA test with 38 kDa and culture filtrate antigen were significantly higher in active pulmonary tuberculosis cases than in non tuberculous pulmonary diseases (p<0.05), but in patients with active pulmonary tuberculosis, those of the sputum culture positive patients for M. tuberculosis were not significantly different from those of the sputum culture negative cases(p>0.05). In the smear-negative active pulmonary tuberculosis patients, the sensitivity of the ELISA using 38 kDa antigen and culture filtrate was 20.0% and 31.4%. respectively. The specificity was 95.3% and 93.9%. respectively. Conclusion : In active pulmonary tuberculosis but smear negative, the serologic detection of antibody to 38 kDa antigen and culture filtrate by ELISA cannot substitute traditional diagnostic tests and does not have clinically significant role to differenciate the patient with active pulmonary tuberculosis from other with non-tuberculous pulmonary diseases.

• Journal of Preventive Medicine and Public Health
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• v.29 no.3 s.54
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• pp.669-678
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• 1996

Helicobacter pylori is now recognized as causative agent of chronic gastritis and peptic ulcer, and strongly associated with development of gastric carcinoma. With development of sensitive and specific serologic tests to identify individuals infected with Helicobacter pylori, the epidemiologic study of this diseases has been investigated. But it's transmission route is not established, yet. The purpose of this study is to measure the prevalence of Helicobacter pylori infection in healthy children and young adults and to evaluate related factors for Helicobacter pylori infection in Korea. The seroprevalence of Ig G antibodies to Helicobacter pylori was determined using a Enzyme Linked Immunosorbent Assay and we obtained the information, such as demographic characteristics, monthly household income, numbers of family members in the house, numbers of persons using same room, type of house, and type of drinking water through the questionnaire survey. The observed overall seropositivity rate was 25.7%. The rate is increased progressively from 5.8% in the age group $1\sim3$ years to 44.4% in the age group $20\sim29$years($\chi^2$ for trend, p<0.001). Especially, the rate increased steeply from 6.5% in the age group $4\sim6$ years to 20.8% in the age group $7\sim9$ years, and this suggested that elementary school age was the major acquisition time of Helicobacter pylori infection. In multivariate logistic regression model, age, numbers of family members in the house, and type of house was statistically significant variables for Helicobacter pylori infection. Each odds ratio(93% CI) were as follows; base to age group $1\sim9$ years, age group $10\sim19$ years $3.6(2.0\sim6.4)$, age group $20\sim29$ years $7.3(4.1\sim13.1)$ and base to group of $1\sim3$ family members, group of $4\sim5$ family members $2.1(1.1\sim4.0)$, group of 6 or more family members $2.7(1.3\sim5.4)$ and base to apartment, single and multihouse $1.9(1.1\sim3.5)$. Sex, monthly household income, numbers of persons using same room, and type of drinking water was not statistically significant for Helicobacter pylori infection.

• Journal of fish pathology
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• v.24 no.2
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• pp.153-160
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• 2011

The specific antibody response of olive flounder, Paralichthys olivaceus to change in rearing-environmental conditions post immunization with antigen (bovine serum albumin, BSA) and different routes of antigen administration were investigated by enzyme-linked immunosorbent assay (ELISA). To test the effect of routes of antigen administration, flounder were injected intraperioneally or intramuscularlly with 1 mg of BSA. In addition, to test the effect of change in environmental condition post immunization, flounder were injected intraperioneally with 1 mg of antigen, and then were exposed to acute thermal change (the water temperature (WT) was decreased from $21^{\circ}C$ to $15^{\circ}C$ within 30 min and maintained at $15^{\circ}C$ for 3 h), handling (fish were caught and subsequently held out of water for 1 min) or heavy oil (76 g/200 L for 2 days). Consequently, there was no significant difference between intraperioneal (IP) and intramuscular (IM) injections except at 10 days post-immunization. With these results, it suggests that both 1M and IP injections may be used as route of vaccination. Furthermore, no significant difference was observed in the antibody response among the groups exposed to heavy oil, handling, sudden drop of WT and positive control except at 10 days post-immunization. From these results, it was confirmed that specific antibody response was not affected by the above mentioned rearing-environmental conditions, suggesting that vaccination can be employed at changing rearing-environmental conditions.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.447-453
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• 1986

To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.3
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• pp.220-226
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• 2017

Dopamine (DA) and serotonin (5-Hydroxytryptamine, 5-HT) are neurotransmitters and hormones that exist in small amounts but have important role in the body. Serum and 24-hour urine are used as specimens, and are usually examined by HPLC-MS. In this study, we tried to detect DA and 5-HT by competitive ELISA using antigen-antibody (Ab) reaction. After immobilizing $5{\mu}g/mL$ BSA conjugate on a 96-well surface, hormone and primary Ab, which are respectively diluted to different concentrations, were treated. Then, HRP-conjugated secondary Ab and TMB were added to measure absorbance. The regression equation and $R^2$ value were calculated based on absorbance, and sensitivity of Ab to hormone as well as the correlation between hormone concentration and absorbance were determined. In DA ELISA, $R^2$, the correlation between the concentration of hormone and absorbance, was the highest by 0.91 when anti-dopamine Ab was diluted 6,000 times and 7,000 times. In 5-HT ELISA, $R^2$ was bigger than 0.90 in every concentration except 3,000 times and 6,000 times. Both DA and 5-HT were not effectively detected at low concentrations (less than $1.0{\times}10^{-7}M$); and because reference value of serum DA is lower than this, HPLC-MS was required to detect serum DA. However, competitive ELISA may be effective in detecting 24-hour urine DA, serum, and 24-hour 5-HT. Further studies are needed to detect hormones more accurately at lower concentrations.