• Journal of Veterinary Science
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• v.22 no.2
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• pp.15.1-15.13
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• 2021

Background: Attenuated Salmonella strain can be used as a vector to transport immunogens to the host antigen-binding sites. Objectives: The study aimed to determine the protective efficacy of attenuated Salmonella strain expressing highly conserved Brucella immunogens in goats. Methods: Goats were vaccinated with Salmonella vector expressing individually lipoprotein outer-membrane protein 19 (Omp19), Brucella lumazine synthase (BLS), proline racemase subunit A (PrpA), Cu/Zn superoxide dismutase (SOD) at 5 × 10⁹ CFU/mL and challenge of all groups was done at 6 weeks after vaccination. Results: Among these vaccines inoculated at 5 × 10⁹ CFU/mL in 1 mL, Omp19 or SOD showed significantly higher serum immunoglobulin G titers at (2, 4, and 6) weeks post-vaccination, compared to the vector control. Interferon-γ production in response to individual antigens was significantly higher in SOD, Omp19, PrpA, and BLS individual groups, compared to that in the vector control (all p < 0.05). Brucella colonization rate at 8 weeks post-challenge showed that most vaccine-treated groups exhibited significantly increased protection by demonstrating reduced numbers of Brucella in tissues collected from vaccinated groups. Real-time polymerase chain reaction revealed that Brucella antigen expression levels were reduced in the spleen, kidney, and parotid lymph node of vaccinated goats, compared to the non-vaccinated goats. Besides, treatment with vaccine expressing individual antigens ameliorated brucellosis-related histopathological lesions. Conclusions: These results delineated that BLS, Omp19, PrpA, and SOD proteins achieved a definite level of protection, indicating that Salmonella Typhimurium successfully delivered Brucella antigens, and that individual vaccines could differentially elicit an antigen-specific immune response.

• Fisheries and Aquatic Sciences
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• v.24 no.4
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• pp.153-162
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• 2021

In this study, a fish metabolic accelerator (a combination of butaphosphan and cyanocobalamin [BPC]) was injected into the muscle of the olive flounder, Paralichthys olivaceus, to investigate its effect on immunity and stress in fish maintained at low temperatures. A single dose of BPC was injected (100 mg/kg body weight) into the olive flounder, and its immunity and stress were observed after one and two weeks. Immunity tests revealed the presence of lysozyme (LZM), nitroblue tetrazolium (NBT), myeloperoxidase (MPO), anti-protease (AP), glutathione peroxidase (GPx), and total immunoglobulin (TIg). BPC injection was found to increase immunity activity compared to the control group. In particular, there was significantly high GPx activity. There was similarly high activity for MPO and GPx in the first week following the injection, followed by significant differences between the BPC-injected and control groups in the second week. There was a reduced low water-temperature stress response in the BPC-injected fish, as evidenced by the cortisol and glucose levels of the control and BPC groups. Lower levels were also observed in the BPC group than the control group during the second week. Cortisol levels were significantly lower in the BPC group than the control group. Histological examinations were conducted in the first and second weeks after the intramuscular injection of the recommended BPC dose to confirm the safety of BPC in aquaculture. There were no abnormalities observed in any tissue samples. This study confirms that the injection of BPC is safe even when used in a culture situation. BPC helps relieve stress and improves non-specific immune responses (innate immunity) in the olive flounder.

• Journal of Veterinary Clinics
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• v.38 no.3
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• pp.120-126
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• 2021

Intradermal skin test (IDST) is generally considered a useful tool in identifying causal allergens in canine atopic dermatitis. Currently, multiple allergen simultaneous test (MAST), an in vitro testing method for allergen-specific immunoglobulin E, is being used as an alternative method. However, there are no reports comparing the IDST and MAST results in the same dogs. This study compared the results of both tests to evaluate the agreement and correlation between them. The sensitivity, specificity, and accuracy of the MAST were 76.2%, 64%, and 66.7%, respectively. Moderate positive predicted value (PPV, 50-75%) or high sensitivity (80-100%) were identified for indoor allergens, such as cat epithelia, house dust, and house dust mites. In contrast, high negative predicted value (NPV, 93.3-100%) and specificity (60-100%) were observed for environmental allergens and fungi. Although the agreement between IDST and MAST for all allergens was fair (κ = 0.301), that for each allergen was poor (κ < 0.01), except for birch (κ = 0.158). Spearman's rank correlation analysis revealed a low correlation between the MAST and IDST results (ρ = 0.308, p = 0.001). As compared to the IDST results, the MAST results did not identify the causative allergens sufficiently. IDST may not be performed for environmental allergens and fungi with high NPV and specificity if the MAST result is negative, but it may have to be performed for indoor allergens with moderate PPV and high sensitivity when the MAST result is positive.

• IMMUNE NETWORK
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• v.22 no.6
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• pp.48.1-48.13
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• 2022

With the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, which are randomly mutated, the dominant strains in regions are changing globally. The development of preclinical animal models is imperative to validate vaccines and therapeutics against SARS-CoV-2 variants. The objective of this study was to develop a non-human primate (NHP) model for SARS-CoV-2 Delta variant infection. Cynomolgus macaques infected with Delta variants showed infectious viruses and viral RNA in the upper (nasal and throat) and lower respiratory (lung) tracts during the acute phase of infection. After 3 days of infection, lesions consistent with diffuse alveolar damage were observed in the lungs. For cellular immune responses, all macaques displayed transient lymphopenia and neutrophilia in the early stages of infection. SARS-CoV-2 Delta variant spike protein-specific IgM, IgG, and IgA levels were significantly increased in the plasma of these animals 14 days after infection. This new NHP Delta variant infection model can be used for comparative analysis of the difference in severity between SARS-CoV-2 variants of concern and may be useful in the efficacy evaluation of vaccines and universal therapeutic drugs for mutations.

• The Journal of Internal Korean Medicine
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• v.38 no.6
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• pp.955-970
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• 2017

Objectives: The aim of this observational study was to identify and analyze the patterns to compare the characteristics of asthma patients. Methods: The subjects were 40 asthma patients who had satisfied the inclusion and exclusion criteria. They were divided into deficiency syndrome and excess syndrome groups, and cold syndrome and heat syndrome groups. Their quality of life was measured by the quality of life questionnaire for adult Korean asthmatics (QLQAKA) and VAS. Heart rate variability (HRV) was measured, and the degree of obesity was evaluated by body mass index (BMI). Hematological, biochemical, and immunoglobulin (Ig) E laboratory tests were included. Results: Based on pattern identification, the 40 asthma patients could be divided into two categories of groups: 1) the deficiency syndrome (N=18) and the excess syndrome (N=22) groups: 2) the cold syndrome (N=35) and the heat syndrome (N=5) groups. The mean value of HF differed significantly between the deficiency and excess syndrome groups. The mean value of IgE in blood tests of asthmatics was greater than four times the reference value. For BMI, the subjects were classified into three groups: normal weight (N=12), overweight (N=12), and obese (N=16). Conclusions: Development of a more accurate asthma-specific pattern identification tool could play a crucial role in asthma control. In addition, good control of asthma can improve the quality of life. Obesity is one of the factors associated with asthma exacerbation.

• Development and Reproduction
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• v.21 no.4
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• pp.407-415
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• 2017

In the present study, we investigated the role of binding immunoglobulin protein/glucose-regulated protein, 78-kDa (BIP/GRP78)-regulated endoplasmic reticulum (ER)-stress on meiotic maturation and cumulus cells expansion in porcine cumulus-oocyte complexes (COCs). Previously, it has been demonstrated that unfolded protein response (UPR)-related genes, such as molecules involved in ER-stress defense mechanisms, were expressed in matured oocytes and cumulus cells during in vitro maturation (IVM) of porcine oocytes. However, BIP/GRP78-mediated regulation of ER stress in porcine oocytes has not been reported. Firstly, we observed the effects of knockdown of BIP/GRP78 (an UPR initiation marker) using porcine-specific siRNAs (#909, #693, and #1570) on oocyte maturation. Among all siRNAs, siRNA #693 significantly reduced the protein levels of UPR marker proteins (BIP/GRP78, ATF4, and P90ATF6) in porcine COCs observed by Western blotting and immunofluorescence analysis. We also observed that the reduction of BIP/GRP78 levels by siRNA#693 significantly inhibited the meiotic maturation of oocytes (siRNA #693: $32.5{\pm}10.1%$ vs control: $77.8{\pm}5.3%$). In addition, we also checked the effect of ER-stress inhibitors, tauroursodeoxycholic acid (TUDCA, $200{\mu}M$) and melatonin ($0.1{\mu}M$), in BIP/GRP78-knockdown oocytes. TUDCA and melatonin treatment could restore the expression levels of ER-stress marker proteins (BIP/GRP78, $p-eIF2{\alpha}$, $eIF2{\alpha}$, ATF4, and P90ATF6) in siRNA #693-transfected matured COCs. In conclusion, these results demonstrated that BIP/GRP78-mediated regulation of UPR signaling and ER stress plays an important role in in vitro maturation of porcine oocytes.

• Fisheries and Aquatic Sciences
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• v.26 no.12
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• pp.726-737
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• 2023

Piperine, the main bioactive component of black pepper (Piper nigrum Linn.), has anti-inflammatory, antifungal, and antibacterial properties. This study evaluated the supplemental effects of piperine or black pepper on innate immunity, growth, feed utilization efficiency, and intestinal morphology in red seabream (Pagrus major). Six experimental diets were formulated, supplementing piperine at 0.0, 0.25, 0.5, 1.0, and 2.0 g/kg levels (Con, P25, P50, P100, and P200) or 1.0 g/kg black pepper (BP100). Juvenile fish (7.6 ± 0.1 g) were randomly stocked into 18 circular tanks (220 L), including 30 fish per tank. Each diet was randomly assigned to triplicate groups, and the feeding trial was conducted for 8 weeks. The results showed that final body weight, specific growth rate, weight gain, and feed utilization efficiency were significantly improved (p < 0.05) when piperine was supplemented into diets at 0.25-2.0 g/kg levels compared to the Con group. Compared to the Con diet, condition factor was significantly increased (p < 0.05) in fish fed with dietary piperine at 0.25-2.0 g/kg or BP100 diet. Serum myeloperoxidase activity was increased (p < 0.05) in P25 and P100 groups and antiprotease activity was increased (p < 0.05) in P100 group compared to the Con group. Significantly higher (p < 0.05) lysozyme activity was observed in P50, P100, P200 and BP100 groups, while total immunoglobulin level was increased in P50, P100 and BP100 groups than Con group. Superoxide dismutase activity was increased (p < 0.05) by dietary piperine at 0.25-2.0 g/kg levels and BP100 diet compared to Con diet. Plasma cholesterol was significantly lower (p < 0.05) in fish fed with piperine (0.5-2.0 g/kg) or BP100 compared to the Con diet. Compared to the Con diet significantly longer (p < 0.05) intestinal villi were observed in fish fed with piperine at 0.25-1.0 g/kg levels, and higher goblet cell count was observed in P25 and BP100 groups. Dietary inclusion of piperine would be a potent immunostimulant in fish diet and the optimum supplementation level would be 0.25-1.0 g/kg.

• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Allergy, Asthma & Immunology Research
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• v.10 no.6
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• pp.648-661
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• 2018

Purpose: Pollen-food allergy syndrome (PFAS) is an immunoglobulin E (IgE)-mediated allergy in pollinosis patients caused by raw fruits and vegetables and is the most common food allergy in adults. However, there has been no nationwide study on PFAS in Korea. In this study, we investigated the prevalence and clinical characteristics of PFAS in Korea. Methods: Twenty-two investigators participated in this study, in which patients with allergic rhinoconjunctivitis and/or bronchial asthma with pollen allergy were enrolled. The questionnaires included demographic characteristics, a list of fruits and vegetables, and clinical manifestations of food allergy. Pollen allergy was diagnosed by skin prick test and/or measurement of the serum level of specific IgE. Results: A total of 648 pollinosis patients were enrolled. The prevalence of PFAS was 41.7% (n = 270). PFAS patients exhibited cutaneous (43.0%), respiratory (20.0%), cardiovascular (3.7%) or neurologic symptoms (4.8%) in addition to oropharyngeal symptoms. Anaphylaxis was noted in 8.9% of the PFAS patients. Seventy types of foods were linked to PFAS; e.g., peach (48.5%), apple (46.7%), kiwi (30.4%), peanut (17.4%), plum (16.3%), chestnut (14.8%), pineapple (13.7%), walnut (14.1%), Korean melon (12.6%), tomato (11.9%), melon (11.5%) and apricot (10.7%). Korean foods such as taro/taro stem (8.9%), ginseong (8.2%), perilla leaf (4.4%), bellflower root (4.4%), crown daisy (3.0%), deodeok (3.3%), kudzu root (3.0%) and lotus root (2.6%) were also linked to PFAS. Conclusions: This was the first nationwide study of PFAS in Korea. The prevalence of PFAS was 41.7%, and 8.9% of the PFAS patients had anaphylaxis. These results will provide clinically useful information to physicians.

• Clinical and Experimental Pediatrics
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• v.52 no.7
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• pp.818-823
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• 2009

Purpose : We aimed to evaluate the frequency and characteristics of minor clinical manifestations of atopic dermatitis (AD) in Korean children to aid the diagnosis and treatment of AD. Methods : From April 2007 to December 2007, we enrolled 106 children (aged 1 month [infants] to 15 years) diagnosed with AD at the Pediatric Allergy Respiratory Center in Soonchunhyang University Hospital. Clinical manifestations were examined and laboratory findings (total and specific immunoglobulin E [IgE] levels and peripheral blood eosinophil count) were analyzed and compared. Results : Minor symptoms, in order of frequency, included xerosis (78.3%), aggravation due to environmental or emotional stress (43.4%), lichenification (35.8%), orbital darkening (34.0%), periauricular eczema (33.0%), and cutaneous infection (31.1%). Older children (${\geq}2$ years) showed more orbital darkening (P=0.01), horizontal crease (P=0.01), and lichenification (P=0.001) than infants. Patients with severe AD (scoring atopic dermatitis [SCORAD] score, ${\geq}40$) showed higher frequencies of xerosis (P= 0.04), cutaneous infection (P=0.03), ichthyosis (P=0.18), keratosis pilaris (P=0.02), pityriasis alba (P=0.07), recurrent conjunctivitis (P=0.02), orbital darkening (P=0.001), aggravation due to environmental or emotional stress (P=0.05), facial eczema (P=0.001), lichenification (P=0.001), and hand/foot eczema (P=0.04) than those with mild-to-moderate AD. Children with atopic eczema showed more facial eczema (P=0.01) and lichenification (P=0.04) than those with non-atopic eczema. Conclusion : The clinical manifestations of AD were similar to those established by Hanifin and Rajka. However, we need to develop our own diagnostic criteria for AD, because the frequencies shown by our subjects differed from those observed in other countries.