To evaluate alterations in fecal bacterial compositions in weaning piglets after supplementation with Phellodendron Cortex extract (PCE), piglets were allowed freely to consume feed containing 10 ppm of PCE for 7 weeks. During the feeding period, fecal samples were collected from individual piglets. A total of 578 valid sequence reads were generated from 40 fecal samples of normal piglets before supplementation with PCE. Fifteen classes were identified. Approximately 90% of classifiable sequences belonged to Clostridia and Bacilli classes. The abundance of Clostridium spp. and Lactobacillus spp. was determined based on the results of the sequencing. The supplementation with 10 ppm PCE for 3 weeks increased the numbers of Clostridium spp. and Lactobacillus spp. whereas seven weeks of supplementation with PCE reduced their abundance. In vitro, PCE inhibited the growth of E. coli and Salmonella enteritidis in a dose-dependent manner.
We demonstrated an effective way of preparing melt spinnable mesophase pitches via catalytic hydrogenation of petroleum residue (fluidized catalytic cracking-decant oil) and their subsequent thermal soaking. The mesophase pitches thus obtained were analyzed in terms of their viscosity, elemental composition, solubility, molecular weight, softening point and optical texture. We found that zeolite-induced catalytic hydrogenation under high hydrogen pressure contributed to a large variation in the properties of the pitches. As the hydrogen pressure increased, the C/H ratio decreased, and the solubility in n-hexane increased. The mesophase pitch with entirely anisotropic domains of flow texture exhibited good meltspinnability. The mesophase carbon fibers obtained from the catalytically hydrogenated petroleum residue showed moderate mechanical properties.
In recent years, flame synthesis has absorbed a great deal of attention as a combustion method for the production of metal oxide nanoparticles, carbon nanotubes, and other related carbon nanostructures, over the existing conventional methods. Flame synthesis is an energyefficient, scalable, cost-effective, rapid and continuous process, where flame provides the necessary chemical species for the nucleation of carbon structures (feed stock or precursor) and the energy for the production of carbon nanostructures. The production yield can be optimized by altering various parameters such as fuel profile, equivalence ratio, catalyst chemistry and structure, burner configuration and residence time. In the present report, diffusion and premixed flame synthesis methods are reviewed to develop a better understanding of factors affecting the morphology, positioning, purity, uniformity and scalability for the development of carbon nanotubes along with their correlated carbonaceous derivative nanostructures..
The objectives of this study were to evaluate the antimicrobial effects of ε-poly-L-lysine mixture (EPM) on cheon-ggukjang and to determine the optimal mixing ratio in cheonggukjang. Increasing the EPM up to 2% decreased via-ble cell counts at both 15oC and 30oC storage. The pH of cheonggukjang without EPM increased slowly over thestorage periods, but cheonggukjang with EPM showed a slight increasing tendency, with the highest pH at 2% EPM.The sensory scores, such as texture and flavor, were highest in samples containing 2% EPM; however, overall pref-erence was not significantly different when compared to the control. For the optimum cheonggukjang processingconditions, 13 experimental points were selected. Soybean and EPM were chosen as independent variables. Viablecell count, pH, texture, and overall preference were measured. The optimum formulation of cheonggukjang usingthe numerical analysis was found to be 98.52% soybean and 1.48% EPM, resulting in a 0.722 desirability value.
Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of various polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the previously reported minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The marked increase of EGFP by these fusion expressions was confirmed through protein and fluorescence intensity analyses. Among the fusion-expressed protein in nucleus and cytoplasm, the most hyper-expression was observed in the fusion of amino acids 19 to 110 and 32 to 59 of polyhedrin. Also these fragments, some degradation of only the fused polyhedrin was observed in the fusion of amino acids 19 to 85 and 32 to 85. The production of E2 protein, which is a major antigen of classical swine fever virus, was dramatically increased by fusion expression with polyhedrin amino acids 19 to 110, and its preliminary immunogenicity was verified using experimental guinea pigs. The production of luciferase was approximately two folds increased by fusion expression with polyhedrin amino acids 32 to 59, and its activity was measured using Luminometer. This study suggests a new option for higher expression of useful foreign recombinant protein using the partial polyhedrin fusion expression in baculovirus.
It has been demonstrated in a previous study that carbon nanotube (CNT)/epoxy/basalt composites produce better flexural properties than epoxy/basalt composites. In this study, mode I fracture tests were conducted using CNT/epoxy/basalt composites with and without seawater absorption in order to investigate the effect of the seawater absorption on the mode I fracture toughness (GIC) of the CNT/epoxy/basalt composites. The results demonstrated that the compliance of the seawater-absorbed specimen was larger than that of the dry specimen at the same crack length, while the opposite result was obtained for the fracture load. The GIC value of the seawater-absorbed CNT/epoxy/basalt composites was approximately 20% lower than that of the dry CNT/epoxy/basalt composites.
To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of polyhedrin fragments were investigated by fusion expression them with the enhanced green fluorescence protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The increase of EGFP production by fusion expressions was confirmed through protein and fluorescence intensity analyses. The importance of nuclear localization for enhanced production of EGFP was shown by the mutation of the NLS within the fused polyhedrin fragment. Among the fusion expressed protein in cytoplasm, the most hyper-expression was observed in the fusion of amino acids 32 to 59 of polyhedrin. Polyhedrin fragment fusion expression with classical swine fever virus E2 protein also resulted hyper-enhanced expression of E2 protein. However, the fusion expression of porcine circovirus ORF2 with polyhedrin fragment did not show significant enhance of ORF2 production. These results suggested that the enhancement of foreign protein production when fused with polyhedrin is caused by the enhanced stability of expressed protein.
The green peach aphid (Myzus persicae) is a cosmopolitan pest of agricultural and horticultural crops and causes serious economic damages. M. persica has rapidly developed resistance to a wide variety of insecticides, including pyrethroids. Target site insensitivity mechanism mediated by two mutations (L1014F and M918T) on the para-type voltage-sensitive sodium channel (vssc) is mainly responsible for pyrethroid resistance. To predict the vssc resistance allele frequency, quantitative sequencing (QS) protocol was established. Frequency prediction equations generated from the plots of signal ratios and amplification critical time showed a high correlation coefficient (r2>0.993), indicating its high accuracy in prediction. QS results revealed that the kdr-type L1014F mutation is only present in Pyeongchang strain. No field strains of M. persicae possessed the super-kdr type M918T mutation. However, a novel M918L mutation was found by genotyping approach. The allele frequencies of M918L and L1014F were 0% to 53% in populations examined, and the level of M918L mutation frequency was closely related with pyrethroid resistance. Therefore, QS-based detection of M918L mutation frequency should faciltate the monitoring of pyrethroid resistance in the field.
The green peach aphid (Myzus persicae) is a serious pest of agricultural and horticultural crops all over the world. M. persica has rapidly developed resistance to a wide variety of insecticides, including carbamates. The E4/FE4 carboxylesterase is known to be involved in carbamate resistance. To compare the E4/FE4 carboxylesterase gene copy number, as a genetic resistance marker, between seven field strains, quantitative real-time PCR (qPCR) was performed. In addition, quantitative sequencing (QS) was employed to predict the frequencies of acetylcholinesterase (AChE) mutations (A301S and S431F) that are associated with target site insensitivity. All M. persica strains examined possessed the S431F mutation in the heterozygous state except for a susceptible strain, implying the possibility of AChE duplication. In contrast, no A301S mutation was found. Frequency prediction equation was generated from the plots of signal ratios and amplification critical time, which showed a high correlation (r2>0.996). QS analysis of M. persicae populations revealed that the allele frequency of S431F ranged 4% to 63%. Taken together, the AChE resistance allele frequencies determined by QS and the E4/FE4 gene copy number by qPCR should facilitate the detection and monitoring of carbamate resistance in M. persicae in the field.
Porcine Circovirus Type2 (PCV2), a single-stranded DNA virus associated with Postweaning multisystemic wasting syndrome(PMWS) of swine, has two major open reading frames, ORF1 and ORF2. The genomic size and molecular weight of ORF2 is respectively 699bp, 28kDa. ORF2 encodes the capsid protein (structural protein) that has type-specific epitopes and is very immunogenic and associated with the induction of neutralizing antibodies, suggesting its potential use in diagnostic assays as well as vaccine development. For efficient production of the capsid proteins, we expressed the PCV2 ORF2 gene with baculovirus in the insect cells. In this study, PCV2 ORF2 was appropriately ligated into the baculovirus transfer vector, pBacPAK9 and pB9-Acpol19-110-EK. Sf21 cells were transfected with a mixture of the purified recombinant transfer vector and bAcGOZA. We generated and purified recombinant viruses containing PCV2 ORF2, and named rAc-B9-PCV2ORF2 and rAc-B9-19-110-EK-PCV2ORF2, respectively. Expression levels of capsid fusion proteins with a partial polyhedrin region of AcNPV more increased than recombinant proteins from non-fusion expressed. Also, expression efficiency increased over time and differed at MOI. As a results, fusion expression of porcine circovirus type2 ORF2 using baculovirus could be utilized as an alternative expression method to produce recombinant antigen against PCV2 infection and is worthy of further investigation.
Aujeszky's disease (AD), also called pseudorabies, is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Aujeszky's disease virus (ADV) virions contain several envelope glycoproteins. Among them, glycoproteins gB, gC and gD are regarded as the major immunogenicity proteins and the antibodies induced by them can neutralize virus in vitro or in vivo. In this study, we investigated expression of these glycoproteins using the bacterial and baculovirus expressionn system. Successful expression of ADV glycoproteins in E. coli was confirmed by SDS-PAGE and Western blot analysis and their optimal expression condition was determined. However, the recombinant proteins generated in the bacterial expression system which lacks glycosylation process frequently lose their biological activity. We tried to express the ADV glycoproteins using the baculovirus expression vector system. The recombinant gB, gC and gD were detected at approximately 100, 60 and 50 kDa on SDS-PAGE and Western blotting, respectively. The optimal expression conditions were determined for MOI(multiplicity of infection) and post-infection days. One MOI and 4 or 5 days post-infection were the best conditions for the expression of the ADV glycoproteins in Sf21 cells. We are currently investigating the antigenicity of recombinant proteins using experimental animals.
Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). To enhance the expression level of baculovirus vector system, we constructed several fusion vectors using various fragments of the polyhedrin. The polyhedrin fragments were genetically fused to the enhanced green fluorescent protein (eGFP) under the control of polyhedrin promoter, and their expressions were analyzed in Sf21 insect cells. Expression of the fusion protein was identified by SDS-PAGE and Western blot analysis using anti-GFP and anti-Polyhedrin. The expression level of eGFP was markedly increased by the fusion of partial polyhedrin. Also, the fluorescence intensity of fusion proteins was higher than that of non-fusion protein. Confocal laser scanning microscopy demonstrated that fusion proteins were localized to the cytosol or nucleus of insect cells. In additional, the glycoprotein E2 (gE2) of classical swine fever virus (CSFV) expressed by the these vectors was dramatically increased and its immunogenicity was proofed using experimental animal guinea pigs that were immunized with the partial polyhedrin containing gE2. This study provides a new option for the higher expression of useful foreign recombinant protein by using the partial polyhedrin in BEVS.
Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.
Background : As the farmers who grow Chinese matrimony vine (Lycium chinense Miller) in rain-shelter greenhouses increase, it is necessary to develop a cultivar with self-compatibility and high quality. Methods and Results : The new boxthorn line, CBP06419-243 was selected from the cross between CBP03310-250(IT232705) and CBP05399-1 to breed the cultivar with self-compatibility and high quality. These lines used for crossing was tetradilpoid induced by the colchicine treatment on bud. Its preliminary yield test was performed from 2009 to 2011 and the selected line was named Cheongyang 18. Its regional yield trials were carried out in Cheongyang, Yesan and Jindo from 2012, 2014, 2015 and Cheongyang 18 was registered as the new variety, Whasu, in 2015. The specific characteristics were summarized as follows; Tree shape is semi-open type and the leaf is lanceolate. The flowers are distinctly larger. The fruit type is medium size, long oblong and yellowish red. The flowering was June 21 with medium flowering. Branching by pruning its sprout was generated less than the check variety, Cheongmeong. The infection rates on leaves to Eriophys macrodonis Keifer was as strong as 7.7 percent. Anthracnose on fruits in open field was slightly severe compared to the check variety, but the infection rates in rain shelter greenhouses was similar. The content of betaine and free sugar in dried-fruits were higher than that of the check. The dried-fruit yield was decreased about 13 percent in open field, but increased 18 percent in rain shelter greenhouses. Conclusion : This variety ‘Whasu’ was suitable for cultivation in plastic film greenhouses because of self-compatibility and high-yielding. It is not necessary the companion variety.
Recently there are increasing numbers of people participating in sports activities. Accordingly, the rate of sports injuries is increasing as well. In this light, this study was conducted to investigate the extent of acute damage and its forms by examining the current state of patients with sports injuries. This research study was conducted between January of 2008 and December
of 2010 in Chosun University Hospital through retrospective review of the medical records of patients with sportsrelated injuries. The total number of patients with sports injuries who visited the emergency center was 887. The number of male patients was 726 (81.85%), far more than the number of female patients (18.15%). More than half of the patients, accounting for 419 (47.23%), had injuries on their face; second and common parts, in which injuries occurred, were upper and
lower body, accounting for 146 (16.45%) and 193 (21.75%), respectively. The majority of patients did not undergo surgical treatment. Only 113 (12.73%) patients underwent surgical treatment. In addition, more than 80% of them were not hospitalized but 170 of them needed to be hospitalized. Most sports injuries occurred in young male patients. This can interrupt participation in sports activities for a long time and result in lower quality of life. Hence proper preventative measures and scientific treatment methods are needed.