• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.12
• /
• pp.1741-1745
• /
• 2005

Zinc is a trace element that is associated with a stimulation of immune function and regulation of ion balance for livestock production. In this study, the effect of zinc as inhibitor to apoptosis-induced cells was examined in vitro using mammary epithelial cell line, HC11 and macrophage cell line, NCTC3749. Cell viability, measured by MTT assay, indicated that 10 g/ml of zinc had a negative impact on cellular activity and 50 ng/ml was chosen for further testing. Apoptosis was induced in cells treated with C2-ceramide in serum-free media. DNA fragmentation and gene expression of acidic sphingomyelinase (a gene responsible for the progress of apoptosis) were distinctively low in zinc treated cells compared with those in non-treated controls. In conclusion, zinc is involved in the regulation of cell proliferation and apoptosis in mammary epithelial cells and macrophages.

• Microbiology and Biotechnology Letters
• /
• v.14 no.6
• /
• pp.455-460
• /
• 1986

Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted front Dane particle core after a DNA polymerase reaction with $\alpha$-($^{32}$P) dNTP, was identified as HBV DNA by liquid scintillation counter and agarose gel electrophoresis-G.M. counting. To produce Hepatitis B surface antigen for use as a vaccine against Hepatitis B virus infection, yeast strains harboring recombinant plasmid with Apase promoter was used. Recombinant plasmid was construced from pHBV 130 and pAN 82, transformed into E coli, and then transferred into yeast strains. HBsAg was produced by derepression in Burkholder minimal medium with controlled inorganic phosphate concentration. The kinetics of HBsAg production was also investigated. Total HBsAg activity increased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphate-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.

• Journal of Plant Biotechnology
• /
• v.36 no.3
• /
• pp.268-274
• /
• 2009

Porphyromonas gingivalis, the gram-negative anaerobic oral bacterium, initiates periodontal disease by binding to saliva-coated oral surface. The cholera toxin B subunit (CTB) genetically linked to FimA1 (1-200 aa) or FimA2 (201-337 aa) of the P. gingivalis fimbrial antigen were introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation method. The integration of CTB-FimA1 or CTB-FimA2 fusion genes were confirmed in the chromosome of transformed leaves by genomic DNA PCR amplification method. Synthesis and assembly of the CTB-FimA fusion proteins into oligomeric structures with pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding activities of CTB-FimA fusion proteins to intestinal epithelial cell membrane receptors were confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA showed that the expression levels of the CTB-FimA1 or CTB-FimA2 fusion proteins were 0.0019, 0.002% of the total soluble protein in transgenic tuber tissues, respectively The synthesis of CTB-FimA monomers and their assembly into biologically active oligomers in transformed potato tuber tissues demonstrates the feasibility of using edible plants for the production of enterocyte targeted fimbrial antigens that could elicit mucosal immune responses.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.1
• /
• pp.25-30
• /
• 2001

Viral safety is a prerequisite for manufacturing clinical albumin and immunoglobulins from human plasma pools. This study was designed to evaluate the efficacy of cold ethanol fractionation and pasteurization (60$\^{C}$ heat treatment for 10h) for the removal/inactivation of human immunodeficiency virus type 1 (HIV-1) during the manufacturing of albumin and immunoglobulins. Samples from the relevant stages of the production process were spiked with HIV-1, and the amount of virus in each fraction was quantified by the 50% tissue culture infectious dose(TCID(sub)50). Both fraction IV fractionation and pasteurization steps during albumin processing were robust and effective in inactivating HIV-1, titers of which were reduced from an initial 8.5 log(sub)10 TCID(sub)50 to undetectable levels. The log reduction factors achieved were $\geq$ 4.5 and $\geq$ 6.5, respectively. In addition, fraction III fractionation and pasteurization during immunoglobulins processing were robust and effective in eliminating HIV-1. HIV-1 titers were reduced from an initial 7.3 log(sub)10 TCID(sub)50 to undetectable levels. The log reduction factors achieved in this case were $\geq$ 4.9 and $\geq$ 5.3, respectively. These results indicate that the process investigated for the production of albumin and immunoglobulins have sufficient HIV-1 reducing capacity to achieve a high margin of safety.

• Parasites, Hosts and Diseases
• /
• v.47 no.4
• /
• pp.359-367
• /
• 2009

Paramyosin is a myofibrillar protein present in helminth parasites and plays multifunctional roles in host-parasite interactions. In this study, we identified the gene encoding paramyosin of Clonorchis sinensis (CsPmy) and characterized biochemical and immunological properties of its recombinant protein. CsPmy showed a high level of sequence identity with paramyosin from other helminth parasites. Recombinant CsPmy (rCsPmy) expressed in bacteria had an approximate molecular weight of 100 kDa and bound both human collagen and complement 9. The protein was constitutively expressed in various developmental stages of the parasite. Imunofluorescence analysis revealed that CsPmy was mainly localized in the tegument, subtegumental muscles, and the muscle layer surrounding the intestine of the parasite. The rCsPmy showed high levels of positive reactions (74.6%, 56/75) against sera from patients with clonorchiasis. Immunization of experimental rats with rCsPmy evoked high levels of IgG production. These results collectively suggest that CsPmy is a multifunctional protein that not only contributes to the muscle layer structure but also to non-muscular functions in host-parasite interactions. Successful induction of host IgG production also suggests that CsPmy can be applied as a diagnostic antigen and/or vaccine candidate for clonorchiasis.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.1
• /
• pp.128-133
• /
• 2000

This study was designed to verify the efficacy of garlic extracts for protecting the infecton of influenza and Japanese B encephalitis virus. Influenza virus (AO/PR8 strain) and Japanese B encephalitis virus (JaGAr O1 strain) were used to attack mouse through nasal route and each vaccines were injected subcutaneously. 0.002 and 0.2 mL/day of garlic extracts were orally administered to mice. The blood and serum samples were taken from the mice to measure LD50, Defense Index (DI), virus-neutralizing antibody for comparing virus influence inhibiting activities. Defense indices of the male and female mice were not significantly different at every experiment. Vaccination effectively inhibited the influence of influenza virus and 0.002 mL/day garlic extract (0.55$\pm$0.05) resulted in significantly higher DI than the control (0$\pm$0.05) (p<0.05). Although 0.002 mL/day garlic extract (0.55$\pm$0.05) resulted in significantly lower DI than the vaccination (1.10$\pm$0.05), 0.2 mL/day garlic extract (2.05$\pm$0.05) resulted in 10 times higher DI than the vaccination (1.10$\pm$0.05). Garlic extract did not affect DI in Japanese B encephalitis virus influence of the vaccinated mouse, but significantly reduced DI of the non-vaccinated mouse (p<0.05). Garlic extracts did not affect the production of the neutralizing antibody against influenza by vaccination. However, neutralizing antibody production of Japanese B encephalitis was accelerated by vaccination. Consequently, the current study proved the efficacy of garlic on inhibition of influenza virus. Finally, it is very hard to show the higher preventing effect on flu through ingestion of garlic as a food than vaccination.

• Korean Journal of Veterinary Service
• /
• v.32 no.1
• /
• pp.27-32
• /
• 2009

We studied the seroprevalence of four respiratory pathogens in Korean swine farms located in Chungnam, Chungbuk, Gyeongnam and Gyeongbuk provinces during the period of spring of 2007 to winter of 2008. Serological tests were performed using commercial ELISA kits. A total of 530 serum samples were tested for the antibodies against porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae (M. hyo) and Actinobacillus pleuropneumoniae (APP). Seroprevalence for four respiratory pathogens were estimated by ELISA-positive rates of the submitted samples. The overall seropositive rates of PRRSV, APP, M. hyo and PCV2 were 32.6%, 10.6%, 38.4% and 88.5%, respectively. By production stage, the seropositive rate for PRRSV was highest in nursery pig populations (46.2%). In contrast, the highest seropositive rates of APP and M. hyo were observed in sow and growing pigs. However, the seroprevalence of PCV2 was ranged from 85.7% to 89.6%, showing no significant difference among the production stages. In the seroprevalence by season, PRRSV, APP and M. hyo infections revealed typical seasonal patterns that the peaks of the seropositive rates were observed between early winter and late spring. In case of PCV2, no particular seasonal patterns were noticed. The pig herds in Gyeongbuk province where PMWS was endemic during the period of survey showed the highest seropositive rates for PRRSV (44.6%), M. hyo (47.5%), and PCV2 (92.7%). Seropositive rates for APP of four provinces were approximately 10%. These results might be valuable for control and prevention of the respiratory diseases and helpful to define strategies related to vaccine applications.

• Korean Journal of Pharmacognosy
• /
• v.49 no.4
• /
• pp.291-297
• /
• 2018

Coxsackievirus B3 (CVB3) is a very well-known causative agent for viral myocarditis and meningitis in human. However, the effective vaccine and therapeutic drug are not developed yet. CVB3 infection activates host cell AKT signaling. Inhibition of AKT signaling pathway may attenuate CVB3 replication and prevent CVB3-mediate viral myocarditis. In this study, we determined antiviral effect of the selected natural plant extract to develop a therapeutic drug for CVB3 treatment. We screened several chemically extracted natural compounds by using HeLa cell-based cell survival assay. Among them, Linum usitatissimum L. extract was selected for antiviral drug candidate. L. usitatissimum extract significantly decreased CVB3 replication and cell death in CVB3 infected HeLa cells with no cytotoxicity. CVB3 protease 2A induced eIF4G1 cleavage and viral capsid protein VP1 production were dramatically decreased by L. usitatissimum extract treatment. In addition, virus positive and negative strand genome amplification were significantly decreased by 1 mg/ml L. usitatissimum extract treatment. Especially, L. usitatissimum extract was associated with inhibition of AKT signal and maintain mTOR activity. In contrast, Atg12 and LC3 expression were not changed by L. usitatissimum extract treatment. In this study, the potential AKT signal inhibitor, L. usitatissimum extract, was significantly inhibited viral genome replication and protein production by inhibition of AKT signal. These results suggested that L. usitatissimum extract is a novel therapeutic agent for treatment of CVB3-mediated diseases.

• Korean Journal of Poultry Science
• /
• v.32 no.1
• /
• pp.43-48
• /
• 2005

An experiment was conducted to investigate the effects of mineral extract from granite on the performance, ammonia production from the litter, components of blood, Newcastle Disease (ND) titer and intestinal microflora in broiler chickens. Nine hundred sixty one-day-old broiler chickens (Ross) were assigned to five treatments: C; control, Zeolite; control + zeolite 1$\%$, AM10: control + active mineral water $10\%$ adsorbed zeolite $1\%$, AM20; control + active mineral water $20\%$ adsorbed zeolite $1\%$ and AM30; control + active mineral water $30\%$ adsorbed zeolite $1\%$. Each treatment consisted of four replicates with 48 broiler chicks for feeding trial. In order to test the effect of ND vaccine on the components of blood, ND titer and intestinal microflora, a separate group of 48 broiler chicks were assigned to the same 5 treatment as the feeding trial plus one negative control (No ND vaccine). Weight gain, feed intake, feed conversion and mortality were not significantly affected by dietary treatments but AM30 tended to be higher than other treatments in weight gain and feed intake, especially during later period (4 to 5 weeks of age). Ammonia production from the litter of AM30 treatment was significantly (P<0.01) lower than the control. Components of blood and ND titer in serum of broiler chickens were not significantly affected by treatments but MCHC (mean corpuscular hemoglobin concentration) of blood was significantly lower (P<0.05) in Zeolite treatment compared to others. The colony forming unit (CFU) of Clostridium perfringens in the small intestinal content of all zeolite and AM treated groups was significantly (P<0.01) lower than the control while the CFU of Escherichia coli was not significantly affected. The CFU of Lactobacilli in AM30 treatment was significantly (P<0.05) higher than the control. In conclusion, dietary supplement of active mineral water adsorbed to zeolite at $30\%$ level (AM30) tended to improve growth performance of broiler chickens and significantly reduced ammonia production from the litter. It also significantly increased CFU of intestinal Lactobacilli.

• IMMUNE NETWORK
• /
• v.7 no.2
• /
• pp.66-74
• /
• 2007

Background: The genus Flavivirus consists of many emerging arboviruses, including Dengue virus (DV), Japanese encephalitis virus (JEV) and West Nile virus (WNV). Effective preventive vaccines remain elusive for these diseases. Mice are being increasingly used as the animal model for vaccine studies. However, the pathogenic mechanisms of these viruses are not clearly understood. Here, we investigated the interaction of DV and JEV with murine bone marrow-derived dendritic cells (bmDC). Methods: ELISA and FACS analysis were employed to investigate cytokine production and phenotypic changes of DCs obtained from bone marrow following flavivirus infection. Results: We observed that these viruses altered the cytokine profile and phenotypic markers. Although both viruses belong to the same family, JEV-infected bmDC produced anti-inflammatory cytokine (IL-10) along with pro-inflammatory cytokines, whereas DV infection induced production of large amounts of pro-inflammatory cytokines (IL-6 and TNF-${\alpha}$) and no IL-10 from murine bmDCs. Both flaviviruses also up-regulated the expression of co-stimulatory molecules such as CD40, CD80 and CD86. JEV infection led to down-regulation of MHC II expression on infected bmDCs. We also found that cytokine production induced by JEV and DV is MyD88-dependent. This dependence was complete for DV, as cytokine production was completely abolished in the absence of MyD88. With regard to JEV, the absence of MyD88 led to a partial reduction in cytokine levels. Conclusion: Here, we demonstrate that MyD88 plays an important role in the pathogenesis of flaviviruses. Our study provides insight into the pathogenesis of JEV and DV in the murine model.