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.
Kim, In-Seop;Eo, Ho-Gueon;Park, Chan-Woo;Chong E. Chang;Lee, Soungmin
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.
Park, Tae-Joon;Kang, Jung-Mi;Na, Byoung-Kuk;Sohn, Woon-Mok
Parasites, Hosts and Diseases
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v.47
no.4
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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
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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.
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.
Shin, Ha-Hyeon;Moon, Sung-Jin;Lim, Byung-Kwan;Kim, Jin Hee
Korean Journal of Pharmacognosy
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v.49
no.4
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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.
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.
Aleyas, Abi G.;George, Junu A.;Han, Young-Woo;Kim, Hye-Kyung;Kim, Seon-Ju;Yoon, Hyun-A;Eo, Seong-Kug
IMMUNE NETWORK
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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.
From January 2020 to October 2021, more than 500,000 academic studies related to COVID-19 (Coronavirus-2, a fatal respiratory syndrome) have been published. The rapid increase in the number of papers related to COVID-19 is putting time and technical constraints on healthcare professionals and policy makers to quickly find important research. Therefore, in this study, we propose a method of extracting useful information from text data of extensive literature using LDA and Word2vec algorithm. Papers related to keywords to be searched were extracted from papers related to COVID-19, and detailed topics were identified. The data used the CORD-19 data set on Kaggle, a free academic resource prepared by major research groups and the White House to respond to the COVID-19 pandemic, updated weekly. The research methods are divided into two main categories. First, 41,062 articles were collected through data filtering and pre-processing of the abstracts of 47,110 academic papers including full text. For this purpose, the number of publications related to COVID-19 by year was analyzed through exploratory data analysis using a Python program, and the top 10 journals under active research were identified. LDA and Word2vec algorithm were used to derive research topics related to COVID-19, and after analyzing related words, similarity was measured. Second, papers containing 'vaccine' and 'treatment' were extracted from among the topics derived from all papers, and a total of 4,555 papers related to 'vaccine' and 5,971 papers related to 'treatment' were extracted. did For each collected paper, detailed topics were analyzed using LDA and Word2vec algorithms, and a clustering method through PCA dimension reduction was applied to visualize groups of papers with similar themes using the t-SNE algorithm. A noteworthy point from the results of this study is that the topics that were not derived from the topics derived for all papers being researched in relation to COVID-19 (
) were the topic modeling results for each research topic (
) was found to be derived from For example, as a result of topic modeling for papers related to 'vaccine', a new topic titled Topic 05 'neutralizing antibodies' was extracted. A neutralizing antibody is an antibody that protects cells from infection when a virus enters the body, and is said to play an important role in the production of therapeutic agents and vaccine development. In addition, as a result of extracting topics from papers related to 'treatment', a new topic called Topic 05 'cytokine' was discovered. A cytokine storm is when the immune cells of our body do not defend against attacks, but attack normal cells. Hidden topics that could not be found for the entire thesis were classified according to keywords, and topic modeling was performed to find detailed topics. In this study, we proposed a method of extracting topics from a large amount of literature using the LDA algorithm and extracting similar words using the Skip-gram method that predicts the similar words as the central word among the Word2vec models. The combination of the LDA model and the Word2vec model tried to show better performance by identifying the relationship between the document and the LDA subject and the relationship between the Word2vec document. In addition, as a clustering method through PCA dimension reduction, a method for intuitively classifying documents by using the t-SNE technique to classify documents with similar themes and forming groups into a structured organization of documents was presented. In a situation where the efforts of many researchers to overcome COVID-19 cannot keep up with the rapid publication of academic papers related to COVID-19, it will reduce the precious time and effort of healthcare professionals and policy makers, and rapidly gain new insights. We hope to help you get It is also expected to be used as basic data for researchers to explore new research directions.
Theileria sergenti were isolated from infected erythrocytes by hypotonic Iysis, and soluble meroBoite antigens were purified by sonication and differential centrifugation. The preparation contained 29, 34, 35 and 105 kD immuno-dominant polypeptides. The soluble antigens (0.5 mg/ml) were prepared and fortified with Freund's adjuvant. Five month old naive Korean calves were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later Nine weeks after the booster dose, vaccinates and controls were challenged with a homologous stabilate (5.6×106 RBC/dose, 40% Parasitemia). All animals were monitored for hematocrit, total erythrocyte count, parasitemia and for the specific antibody by Western immuno- blot (WB) and indirect immuno-auorescent antibody(IFA) test. By 18 weeks after vaccination (6 weeks after the challenge), vaccinated cattle had an average IFA titer of 1 : 10,240 compared with 1 : 1,280 of the controls. The vaccinates showed ne91igib1e change in hematocrit and total RBC count whereas control animals showed significant (P<0.05) hematological chanties and associated anemia. After vaccination and challenge, the antibody responses demonstrated that vaccination had induced significant production of antibody to the 29 and 35 kD polypeptides. The latter polypeptide was much more strongly recognized by the vaccinated animals, and thus it may be a potential candidate for the vaccine.
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