Field Application of a New CSF Vaccine Based on Plant-Produced Recombinant E2 Marker Proteins on Pigs in Areas with Two Different Control Strategies.

A classical swine fever virus (CSFV)-modified live LOM (low-virulence strain of Miyagi) vaccine (MLV-LOM) to combat CSF has been used in places where the disease is prevalent around the world, including in Korea, except in Jeju Island. In general, modified live virus-based vaccines (MLV) are known to be highly effective in inducing immune responses. At the same time, MLVs also have potential dangers such as a circulation in the field. There is still a need for safer and more effective vaccines to control CSF in the field. In this study, we applied a new CSF vaccine based on plant-produced recombinant E2 marker proteins at two different locations, Jeju Island and a suburb of Pohang, using different CSF control strategies. The result suggested that vaccinated sows in Jeju Island highly developed immunogenicity and maintained stably until 102 days post-vaccination (dpv). Its piglets that received maternal antibodies were shown to carry high serological values and maintained them until 40 days of age, which was the end of the follow-up. Naïve piglets vaccinated at 40 days of age showed high serological values and these were maintained until 100 days of age (60 dpv), which was the end of the follow-up. The vaccine was also effective in inducing immune responses in newborn piglets that carried maternal antibodies received from MLV-LOM vaccine-immunized mother sows.

A Novel E2 Glycoprotein Subunit Marker Vaccine Produced in Plant Is Able to Prevent Classical Swine Fever Virus Vertical Transmission after Double Vaccination.

The efficacy of a novel subunit vaccine candidate, based in the CSFV E2 glycoprotein produced in plants to prevent classical swine fever virus (CSFV) vertical transmission, was evaluated. A Nicotiana benthamiana tissue culture system was used to obtain a stable production of the E2-glycoprotein fused to the porcine Fc region of IgG. Ten pregnant sows were divided into three groups: Groups 1 and 2 (four sows each) were vaccinated with either 100 µg/dose or 300 µg/dose of the subunit vaccine at 64 days of pregnancy. Group 3 (two sows) was injected with PBS. Groups 1 and 2 were boosted with the same vaccine dose. At 10 days post second vaccination, the sows in Groups 2 and 3 were challenged with a highly virulent CSFV strain. The vaccinated sows remained clinically healthy and seroconverted rapidly, showing efficient neutralizing antibodies. The fetuses from vaccinated sows did not show gross lesions, and all analyzed tissue samples tested negative for CSFV replication. However, fetuses of non-vaccinated sows had high CSFV replication in tested tissue samples. The results suggested that in vaccinated sows, the plant produced E2 marker vaccine induced the protective immunogenicity at challenge, leading to protection from vertical transmission to fetuses.

Correlation between surface hydrophilicity and osteoblastic differentiation on microgrooved titanium substrata.

Surface microgrooves and acid etching on titanium (Ti) have been proposed to enhance various cell behaviors. In this study, surface hydrophilicity, protein adsorption, and alkaline phosphatase activity of osteoblasts were analyzed and compared between microgrooved Ti, Ti with microgrooves and further acid etching, smooth Ti, and acid-etched smooth Ti. Correlations between the results of each experiment were analyzed using Pearson's correlation analysis, and the influential factor on alkaline phosphatase activity was determined using multiple stepwise regression analysis. Among groups, the Ti substrata with microgrooves and subsequent acid etching showed significantly greater surface hydrophilicity and alkaline phosphatase activity compared with smooth Ti, whereas the Ti substrata with only microgrooves showed the greatest protein adsorption. Multiple stepwise regression analysis determined the surface hydrophilicity of Ti as the influential factor on alkaline phosphatase activity. This study indicates that surface microgrooves and acid etching on Ti substrata enhance surface hydrophilicity, leading to increased alkaline phosphatase activity.

Effect of etched microgrooves on hydrophilicity of titanium and osteoblast responses: A pilot study.

PURPOSE: The aim of this pilot study was to investigate the effect of etched microgrooves on the hydrophilicity of Ti and osteoblast responses. MATERIAL AND METHODS: Microgrooves were applied on Ti to have 15 and 60 µm width, and 3.5 and 10 µm depth by photolithography, respectively. Further acid etching was applied to create Ti surfaces with etched microgrooves. Both smooth- and acid-etched Ti were used as the controls. The hydrophilicity of Ti was analyzed by determining contact angles. Cell proliferation and osteogenic activity of MC3T3 mouse preosteoblasts were analyzed by bromodeoxyuridine assay and alkaline phosphatase (ALP) activity test, respectively. One-way ANOVA, Pearson's correlation analysis and multiple regression analysis were used for statistics. RESULTS: Etched microgrooves significantly increased the hydrophilicity of Ti compared to the smooth Ti. 60 µm-wide etched microgrooves significantly enhanced cell proliferation, whereas the osteogenic activity showed statistically non-significant differences between groups. Result of the osteogenic activity significantly correlated with those of hydrophilicity and cell proliferation. Hydrophilicity was determined to be an influential factor on osteogenic activity. CONCLUSION: This study indicates that increase in hydrophilicity of Ti caused by etched microgrooves acts as an influential factor on osteogenic activity. However, statistically non-significant increase in the ALP activity suggests further investigation.

Anti-inflammatory effects of BT-201, an n-butanol extract of Panax notoginseng, observed in vitro and in a collagen-induced arthritis model.

BACKGROUND & AIMS: Although there has been some success with protein-based anti-tumor necrosis factor alpha (TNF-alpha) therapeutics, the problems associated with protein-based drugs demand alternative approaches. We screened various herbal extracts for their ability to inhibit TNF-alpha secretions and found that BT-201, an n-butanol extract of Panax notoginseng (Burk.) F. H. Chen (P. notoginseng) has such an ability. METHODS: The purpose of this study has been to evaluate the anti-inflammatory and anti-rheumatic effects of BT-201. The anti-inflammatory effects were evaluated by measuring the effects of BT-201 on the production of TNF-alpha, interleukin (IL)-1beta, inducible nitric oxide (iNO), and matrix metalloproteinase-13 (MMP-13), in vitro. The anti-rheumatic effects were evaluated by treating mice with collagen-induced arthritis (CIA) using a daily oral administration of BT-201 at 15 mg/kg/day. In addition, the effects on NF-kappaB and mitogen-activated protein kinase (MAPK) pathways were evaluated by Western blotting using phospho-specific antibodies. RESULTS: BT-201 significantly inhibited all the inflammatory parameters evaluated in vitro and delayed the onset and progression of CIA. BT-201 inhibited the activation of NF-kappaB, ERK, p38, and JNK pathways. CONCLUSIONS: Our results demonstrated that BT-201 can modulate various aspects of inflammation in vitro and that it has disease-modifying, anti-rheumatic effects in vivo, suggesting that it can be a potential alternative to the current anti-TNF-alpha therapeutics for rheumatoid arthritis and other inflammatory disease.

OmpA genotyping of Chlamydia trachomatis from Korean female sex workers.

OBJECTIVES: To determine ompA genotypes of Chlamydia trachomatis in female commercial sex workers (FCSW) in Korea. MATERIALS AND METHODS: To accomplish this, 40 endo-cervical chlamydial PCR positive samples from FCSWs in one city of Korea were collected from April 2004 to August 2004. OmpA gene of C. trachomatis from the extracted genomic DNA was amplified and read whole variable domains and some constant domains sequences. RESULTS: The deduced serovars found, in descending order of prevalence, were E (45%), F (20%), G (15%), D (5%), H (5%), J (2.5%), and mixed type (7.5%). While we found various serovars in limited FCSWs, only one genotype was found in each E, F, G, D, H, and J type. CONCLUSIONS: The serotype E was the most prevalent type in commercial sex workers and ompA DNA polymorphism in each serovar was not a typical finding in FCSWs. Interestingly, the prevalence of serovar G, KS_G1 in our study, was higher in Pacific cities of U.S.A. than that in European studies or other cities in U.S.A. It may suggest that the high prevalence of KS_G1 in Korea has a relation with the high incidence of serovar G in Pacific cities in U.S.A.