• Toxicological Research
• /
• v.21 no.3
• /
• pp.241-247
• /
• 2005

HM10760 is a recombinant human erythropoietin that has beer developed as a drug for anemia. In this study, antigenic potential of HM10760 was examined by active systemic anaphylaxis in guinea pigs and passive cutaneous anaphylaxis in a guinea pig-guinea pig system. HM10760 was subcutaneously administered at 0, 2, and $20{\mu}g/kg$ and also as a suspension with adjuvant ($20{\mu}g/kg$ + FCA). Ovalbumin as a suspension with adjuvant was administered to induce positive control responses. In the active systemic anaphylaxis test, no symptoms except rubbing or licking nose and urination that were considered as physiological phenomena were observed at $0{\mu}g/kg$. Four of 5 animals at $2{\mu}g/kg$ and all the 5 animals at $20{\mu}g/kg$ showed cyanosis and lying on side. All animals in the adjuvant mixture group showed relatively mild symptoms such as rubbing or licking nose, urination, and evacuation. In the passive cutaneous anaphylaxis test, 0/5, 3/5, and 5/5 serum samples from the animals immunized with 0, 2, and $20{\mu}g/kg$, respectively, showed positive reactions against HM10760. All 5 sera collected from the animals immunized with an adjuvant mixture contained HM10760-specific antibodies. These results suggest HM10760 have antigenicity in guinea pigs.

• Parasites, Hosts and Diseases
• /
• v.49 no.3
• /
• pp.207-212
• /
• 2011

Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.

• Molecular & Cellular Toxicology
• /
• v.2 no.1
• /
• pp.67-73
• /
• 2006

In previous studies, it has been discovered that cancer cells not only overexpress regulatory subunit I (Rl)/protein kinase type I (PKA-I) but also secrete outside the cell an extracellular form of PKA (ECPKA) and that the ECPKA secretion detected in patients' serum is obviously greater than that found in non-cancer patients or healthy subjects. We now found that ECPKA elicits the formation of serum autoantibodies that can serve as a cancer diagnostic and prognostic marker. To measure the presence of anti-ECPKA autoantibody in the human sera, basic methodology for ECPKA assay was established an enzyme-linked immunosorbent assay (ELISA). We obtained serum samples from 199 patients with different types of cancer, and also obtained 31 serum samples to compare with ECPKA concentrations from non-cancer patients and 119 normal volunteers. Compared with normal or non-cancer patient sera, we found that the frequency of anti-ECPKA autoantibody was significantly higher in cancer patients (88%) than in those without cancer (17%). Furthermore the presence of anti-ECPKA autoantibodies in the serum of cancer patients was highly correlated with the site of metastasis. The immunoassay developed for anti-ECPKA antibodies is highly sensitive and specific. Therefore, this discovery of an autoantibody-based cancer diagnostic may have serious clinical application and may become an important advance over current technology.

• Korean journal of food and cookery science
• /
• v.10 no.3
• /
• pp.254-259
• /
• 1994

Meat provides high quality proteins, lipids, minerals and vitamins. The meat protein is especially high in essential amino acids that are crucial for human health, growth & development and for the formation of enzymes, hormones and antibodies. Relatively cheap and nutritionally sound vegetable proteins that are similar to animal proteins are being developed to replace the animal proteins in texture, nutrition and food characteristics. In this study a nutritionally sound meat lipid replacing food Oatrim that has been produced by converting oat starch into maltodextrin by ${\alpha}$-amylase, have been partially substituted for beef and general component analysis, texture measurement and sensory tests have been conducted. The results are 1. Water content of the non-treated (0% treated) was 67.1% and the treated (10% treated) was 77%. The treated showed better water holding capacity. 2. Protein content of the non-treated was 21.2 g/100 g; the 4% treated, 18.4 g/100 g; the 6% treated, 18.2 g/100 g; the 8% treated, 17.2 g/100g; and the 10% treated, 16.0 g/100 g. The protein content tended significant. 3. Amino acid analysis results showed that glutamic acid content was the highest in Oatrim and as its amino acid make up is exellent, it is valuable as a fine low fat protein food. 4. Sensory tests show that the increased Oatrim content increased the appearance quality but food characteristics were high only in the 4% and 6% treated groups, indicating that the replacement ratio should not exceed 10%. 5. Texture measurement analysis results show that the higher the replacement content, lower the springness, cohesiveness, hardness, chewiness and gumminess, resulting in relatively soft overall texture. However, in order to better the food characteristics, more studies must be continuously done, and so by being able to increase vegetable substitution over meat, it may be able to contribute to the prevention of adult disease.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.6
• /
• pp.2909-2916
• /
• 2016

Background: Canine malignant lymphoma is classified into B- or T-cell origin, as in the human case. Due to differences in prognosis, a suitable method needs to be developed for lineage identification. Aims: To determine the accuracy of immunophenotypic and molecular information between three methods: immunocytochemistry (ICC), immunohistochemistry (IHC) and heteroduplex polymerase chain reaction for antigen receptor rearrangements (hPARR) in spontaneous canine lymphomas. Materials and Methods: Peripheral blood, fine needle aspiration and tissue biopsies from enlarged peripheral lymph nodes prior to treatment of 28 multicentric lymphoma patients were collected. Cytopathology and histopathology were examined and classified using the updated Kiel and WHO classifications, respectively. Anti-Pax5 and anti-CD3 antibodies as B- and T-cell markers were applied for immunophenotyping by ICC and IHC. Neoplastic lymphocytes from lymph node and white blood cell pellets from peripheral blood were evaluated by hPARR. Results: In this study, low grade B-cell lymphoma accounted for 25% (7/28), high grade B-cell lymphoma for 64.3% (18/28) and high grade T-cell lymphoma for 10.7% (3/28). According to the WHO classification, 50% of all cases were classified as diffuse large B-cell lymphoma. In addition, ICC showed concordant results with IHC; all B-cell lymphomas showed Pax5+/CD3, and all T-cell lymphomas exhibited Pax5-/CD3+. In contrast to hPARR, 12 B-cell lymphomas featured the IgH gene; seven presented the $TCR{\gamma}$ gene; five cases showed both IgH and $TCR{\gamma}$ genes, and one case were indeterminate. Three T-cell lymphomas showed the $TCR{\gamma}$ gene. The percentage agreement between hPARR and ICC/IHC was 60%. Conclusions: Immunophenotyping should not rely on a single method. ICC or IHC with hPARR should be used concurrently for immunophenotypic diagnosis in canine lymphomas.

• Parasites, Hosts and Diseases
• /
• v.57 no.3
• /
• pp.283-290
• /
• 2019

A rapid diagnostic test (RDT) kit was developed to detect non-structural protein 1 (NS1) of yellow fever virus (YFV) using monoclonal antibody. NS1 protein was purified from the cultured YFV and used to immunize mice. Monoclonal antibody to NS1 was selected and conjugated with colloidal gold to produce the YFV NS1 RDT kit. The YFV RDTs were evaluated for sensitivity and specificity using positive and negative samples of monkeys from Brazil and negative human blood samples from Korea. Among monoclonal antibodies, clones 3A11 and 3B7 proved most sensitive, and used for YFV RDT kit. Diagnostic accuracy of YFV RDT was fairly high; Sensitivity was 0.0% and specificity was 100% against Dengue viruses type 2 and 3, Zika, Chikungunya and Mayaro viruses. This YFV RDT kit could be employed as a test of choice for point-of-care diagnosis and large scale surveys of YFV infection under clinical or field conditions in endemic areas and on the globe.

• Journal of Life Science
• /
• v.29 no.10
• /
• pp.1159-1163
• /
• 2019

Although there is a growing interest in male menopause, a phenomenon associated with male hormone depletion, current kits using antibodies to quantify male hormones are expensive. In this study, we constructed an in vitro system for verifying the activity or concentration of male steroid hormones using a transcriptional activity test. A reporter plasmid, pGL2-Neo-ARE-AdE1BTATA, which reacts to testosterone, was constructed. In this plasmid, the ARE-AdE1bTATA sequences can be bounded by the testosterone - androgen receptor complex to express luciferase as a reporter. Then, a stable transfection was performed on the human prostate cancer cell line, LNcap-LN3. The constructed LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA testosterone compound detection (TCD) system showed quantitatively proportional luciferase activities to concentrations of $10^{-13}$ to $10^{-8}M$ of standard testosterone. The established in vitro TCD system will contribute to the development of materials for health/functional foods and drugs as it will be possible to search en masse for testosterone-like or testosterone-inhibiting substances derived from natural materials.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.5
• /
• pp.813-819
• /
• 2019

Middle East respiratory syndrome coronavirus (MERS-CoV) induces severe respiratory impairment with a reported mortality rate of ~36% in humans. The absence of clinically available MERS-CoV vaccines and treatments to date has resulted in uncontrolled incidence and propagation of the virus. In vaccine design, fusion with the IgG Fc domain is reported to increase the immunogenicity of various vaccine antigens. However, limited reports have documented the potential negative effects of Fc fusion on vaccine antigens. To determine whether Fc fusion affects the immunogenicity of MERS-CoV antigen, we constructed a Fcassociated MERS-CoV spike protein (eS770-Fc, 110 kDa), whereby human IgG4 Fc domain was fused to MERS-CoV spike protein (eS770) via a Gly/Pro linker using baculovirus as the expression system. For comparative analyses, two eS770 proteins lacking the IgG4 Fc domain were generated using the IdeS protease ($eS770-{\Delta}Fc$) or His tag attachment (eS770-His) and the immunogenicity of the above constructs were examined following intramuscular immunization in mice. Contrary to expectations, non-Fc spike proteins ($eS770-{\Delta}Fc$, eS770-His; 90 kDa) showed higher immunogenicity than the Fc fusion protein (eS770-Fc). Moreover, unlike non-Fc spike proteins, eS770-Fc immunization did not elicit neutralizing antibodies against MERS-CoV. The lower immunogenicity of Fc-fused eS770 was related to alterations in the structural conformation of the spike protein. Taken together, our results indicate that IgG Fc fusion reduces the immunogenicity of eS770 by interfering with the proper folding structure.

• Journal of the Korea Institute of Military Science and Technology
• /
• v.24 no.3
• /
• pp.348-355
• /
• 2021

Hantaan virus(HTNV) causes hemorrhagic fever with renal syndrome(HFRS) with a case fatality rate ranging from <1 to 15 % in human. Hantavax^Ⓡ is a vaccine against the Hantavirus, which has been conditionally approved by the Ministry of Food and Drug Safety(MFDS). However, only 50 % of volunteers had neutralizing antibodies 1 year following the boost. Effective antiviral treatments against HTNV infection are limited. Hantaviruses generally cause asymptomatic infection in adult mice. On the other hand, infection of suckling and newborn mice with hantaviruses causes lethal neurological diesease or persistant infection, which is different from the disease in humans. The development of vaccines and antiviral strategies for HTNV has been partly hampered by the lack of an efficient lethal mouse model to evaluate the efficacy of the candidate vaccines or antivirals. In this report, we established a lethal mouse model for HTNV, which may facilitate in vivo studies on the evaluation of candidate drugs against HTNV. The median lethal dose value of HTNV was calculated by probit analysis of deaths occurring within two weeks. Five groups of ten ICR mice were injected intracranially with serial 2-fold dilutions (from 50 to 3.125 PFU/head) of HTNV. Mice injected with HTNV began to die at 8 days post-infection. The lethal dose required to kill 50 % of the mice (LD₅₀) was calculated to be 2.365 PFU/head.

• Korean Journal of Veterinary Service
• /
• v.44 no.1
• /
• pp.27-33
• /
• 2021

Q fever is a worldwide zoonotic disease caused by Coxiella burnetii. Domestic ruminants are considered to be major source of human infection. The aim of this survey was to investigate seroprevalence of C. burnetii in ruminants in Gwangju area. A total of 1,000 samples (serum and lactoserum) were collected from 987 Korean native cattle, 5 Korean native goats, 2 beef cattle, 6 bulk-tank milk from each dairy farm in Gwangju area from January to October 2020 and analyzed by ELISA. The seroprevalence of C. burnetii in bulk-tank milk from each dairy farms was 50.0%. Korean black goat and beef cattle had negative antibody test results for C. burnetii. The seroprevalence of C. burnetii in Korean native cattle in Gwangju area was 7.1% and was higher in female (7.8%) than in male (3.4%) (P=0.024). The seroprevalence of C. burnetii in Korean native cattle appeared to increase with age (3.8% in 1 year-old, 7.1% in 3 year-old, and 10.7% in more than 5 year-old) (P<0.001). The seroprevalence of C. burnetii of Korean native cattle increased in spring and May was the highest in particular (P<0.001). As the distribution and density of tick-habitat are expected to increase due to climate crisis, this survey highlights the need for monitoring C. burnetii in domestic ruminants, including surveillance of C. burnetii infection in people working for livestock industry.