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 classical swine fever virus E2 fusion protein produced in plants elicits a neutralizing humoral immune response in mice and pigs.

Classical swine fever (CSF) is one of the most important viral diseases of swine worldwide. Although live or attenuated virus vaccines have been used to control CSFV, it is difficult to distinguish vaccinated pigs from infected pigs; this leads to restrictions on import and export. Subunit vaccines based on the CSFV E2 glycoprotein have been developed using baculovirus or insect cell systems, but some weaknesses remain. Here, we describe production of an E2 recombinant protein using a Nicotiana benthamiana plant expression system. To do this, we took advantage of the ability of the swine Fc domain to increase solubility and stability of the fusion protein and to strengthen immune responses in target animals. N. benthamiana expressed high amounts of pFc2-fused E2 proteins, which were isolated and purified by affinity chromatography to yield a high pure recombinant protein in a cost-effective manner. Native-polyacrylamide gel electrophoresis and size exclusion chromatography confirmed that the pmE2:pFc2 fusion exists as a multimer rather than as a dimer. Injection of recombinant pmE2 protein into mice or piglets generated anti-pmE2 antibodies with efficient neutralizing activity against CSFV. These results suggest that a purified recombinant E2 protein produced in N. benthamiana generates high titers of neutralizing antibodies in vivo; as such, the protein could be developed as a subunit vaccine against CSFV.

Development of Recombinant Protein-Based Vaccine Against Classical Swine Fever Virus in Pigs Using Transgenic Nicotiana benthamiana.

Classical swine fever virus (CSFV) is highly contagious, and fatal to infected pigs. Vaccines against CSFV have been developed from attenuated or modified live viruses. These vaccines are effective for immunization of animals, but they are associated with problems such as the accidental spreading of viruses to animals in the field, and with barriers to trade following vaccination. Here, we report the generation of transgenic Nicotiana benthamiana plants for large-scale, cost-effective production of E2 fusion protein for use as a recombinant vaccine against CSFV in pigs. Transgenic N. benthamiana plants harboring an intergenic, single-copy insertion of a chimeric gene encoding E2 fusion protein had high levels of transgene expression. For large-scale production of E2 fusion protein from leaf tissues, we developed a protein-purification protocol consisting of cellulose-binding domain (CBD)-cellulose-based affinity purification and size-exclusion gel-filtration chromatography. E2 fusion proteins showed high immunogenicity in piglets and provided protection against CSFV challenge. The CBD in the E2 fusion protein was also highly immunogenic. These results suggest that plant-produced recombinant E2 fusion proteins can be developed into cost-effective vaccines against CSFV, with the CBD as a marker antigen to differentiate between vaccination and natural infection.

The Scaffolding Protein, Grb2-associated Binder-1, in Skeletal Muscles and Terminal Schwann Cells Regulates Postnatal Neuromuscular Synapse Maturation.

The vertebrate neuromuscular junction (NMJ) is considered as a "tripartite synapse" consisting of a motor axon terminal, a muscle endplate, and terminal Schwann cells that envelope the motor axon terminal. The neuregulin 1 (NRG1)-ErbB2 signaling pathway plays an important role in the development of the NMJ. We previously showed that Grb2-associated binder 1 (Gab1), a scaffolding mediator of receptor tyrosine kinase signaling, is required for NRG1-induced peripheral nerve myelination. Here, we determined the role of Gab1 in the development of the NMJ using muscle-specific conditional Gab1 knockout mice. The mutant mice showed delayed postnatal maturation of the NMJ. Furthermore, the selective loss of the gab1 gene in terminal Schwann cells produced delayed synaptic elimination with abnormal morphology of the motor endplate, suggesting that Gab1 in both muscles and terminal Schwann cells is required for proper NMJ development. Gab1 in terminal Schwann cells appeared to regulate the number and process elongation of terminal Schwann cells during synaptic elimination. However, Gab2 knockout mice did not show any defects in the development of the NMJ. Considering the role of Gab1 in postnatal peripheral nerve myelination, our findings suggest that Gab1 is a pleiotropic and important component of NRG1 signals during postnatal development of the peripheral neuromuscular system.

Achievable Rate Estimation of IEEE 802.11ad Visual Big-Data Uplink Access in Cloud-Enabled Surveillance Applications.

This paper addresses the computation procedures for estimating the impact of interference in 60 GHz IEEE 802.11ad uplink access in order to construct visual big-data database from randomly deployed surveillance camera sensing devices. The acquired large-scale massive visual information from surveillance camera devices will be used for organizing big-data database, i.e., this estimation is essential for constructing centralized cloud-enabled surveillance database. This performance estimation study captures interference impacts on the target cloud access points from multiple interference components generated by the 60 GHz wireless transmissions from nearby surveillance camera devices to their associated cloud access points. With this uplink interference scenario, the interference impacts on the main wireless transmission from a target surveillance camera device to its associated target cloud access point with a number of settings are measured and estimated under the consideration of 60 GHz radiation characteristics and antenna radiation pattern models.

GD3 accumulation in cell surface lipid rafts prior to mitochondrial targeting contributes to amyloid-ß-induced apoptosis.

Neuronal apoptosis induced by amyloid ß-peptide (Aß) plays an important role in the pathophysiology of Alzheimer's disease (AD). However, the molecular mechanism underlying Aß-induced apoptosis remains undetermined. The disialoganglioside GD3 involves ceramide-, Fas- and TNF-α-mediated apoptosis in lymphoid cells and hepatocytes. Although the implication of GD3 has been suggested, the precise role of GD3 in Aß-induced apoptosis is still unclear. Here, we investigated the changes of GD3 metabolism and characterized the distribution and trafficking of GD3 during Aß-induced apoptosis using human brain-derived TE671 cells. Extracellular Aß-induced apoptosis in a mitochondrial-dependent manner. GD3 level was negligible in the basal condition. However, in response to extracellular Aß, both the expression of GD3 synthase mRNA and the intracellular GD3 level were dramatically increased. Neosynthesized GD3 rapidly accumulated in cell surface lipid microdomains, and was then translocated to mitochondria to execute the apoptosis. Disruption of membrane lipid microdomains with methyl-ß-cyclodextrin significantly prevented both GD3 accumulation in cell surface and Aß-induced apoptosis. Our data suggest that rapidly accumulated GD3 in plasma membrane lipid microdomains prior to mitochondrial translocation is one of the key events in Aß-induced apoptosis.

Clinical implications of the topography and distribution of the posterior superior alveolar artery.

The aim of this study was to clarify the arrangement of the anatomic courses and distribution of the intraosseous branch (IObr) of posterior superior alveolar artery. The anatomic variations in the topographic relationships were described to provide beneficial data to minimize injury to the IObr during surgical procedure of the buccal wall of the maxillary sinus. The IObrs in 42 hemifaces of embalmed Korean cadavers were examined. The courses of the IObr of the posterior superior alveolar artery were classified into 2 categories: the straight (type 1) and the U-shaped (type 2). The type 1 was the most common (78.1%), and the type 2 was observed in 21.9% of the specimens. The minimum mean height from the cervix to the IObr was 21.1 mm in the first molar region. The IObr ran at the lowest level from the maxillary sinus floor at the first premolar region. These anatomic findings in the current study could represent useful information for the various surgical procedures of the maxilla.

Core-suction technique for the fabrication of optical fiber preforms.

A novel technique, named "core suction," for fabricating optical fiber preforms has been devised. The technique involves drawing the molten nonconventional core glass material into the cladding tube to form the preform. The developed technique is simple, inexpensive, and shows great potential for fabricating preforms of highly nonlinear nonconventional glasses as the core material. Preforms were made with Schott SF6 and a lead-tellurium-germanate glass in silica cladding tubes, and these preforms were then pulled into fibers.