Three-dimensional porous collagen/chitosan complex sponge for tissue engineering

A three-dimensional, porous collagen/chitosan complex sponge was prepared to closely simulate basic extracellular matrix (ECM) constitutes, collagen and glycosaminoglycan. The complex sponge was prepared by a lyophilization method and had the regular network with highly porous structure, suitable for cell adhesion and growth. The pores were well interconnected, and their distribution was fairly homogeneous. The complex sponge was crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to increase its biological stability and enhance its mechanical properties. The crosslinking medium had a great effect on the inner structure of the sponge. The homogeneous, porous structure of the sponge was remarkably collapsed in an aqueous crosslinking medium. However, the morphology of the sponge remained almost intact in a water/ethanol mixture crosslinking milieu. Mechanical properties of the collagen/chitosan sponge were significantly enhanced by EDC-mediated crosslinking. The potential of the sponge as a scaffold for tissue engineering was investigated using a Chinese hamster ovary cell (CHO-K1) line.     

Expression of 4 truncated fragments of Pasteurella multocida toxin and their immunogenicity

Pasteurella multocida toxin (PMT) is a poor antigen that becomes more immunogenic after its native structure has been destroyed. In contrast, partially truncated PMT proteins, which are predicted to be good antigens when used as a vaccine, might be used to improve the control of atrophic rhinitis in pigs. In this study, 4 truncated PMT fragments were expressed in Escherichia coli, and those 4 fragments were inoculated into mice to produce the polyclonal antibodies. The results of an enzyme-linked immunosorbent assay (ELISA) revealed that #1 and #4 fragments were the most immunogenic. Immunized mice were subsequently challenged intraperitoneally with P. multocida type D. Five of the eight #1 fragment-immunized mice showed some protection against death and bacterial clearance. Pigs immunized with #1 fragment produced no or mild atrophic rhinitis (turbinate conchal score) after challenge, suggesting that this #1 fragment could be a good candidate for a subunit recombinant-type vaccine.     

Reporting 678 putative cSNPs from full-length enriched cDNA sequences of the Korean native pig

Sequences from the clones of full-length enriched cDNA libraries serve as valuable resources for functional genomic studies. We have analysed 1970 high-quality chromatograms (Phred value ≥ 30) that were obtained from sequencing the 5' ends of brainstem, liver, neocortex and spleen clones derived from full-length enriched cDNA libraries from Korean native pigs. In addition, 50 000 pig expressed sequence tag (EST) sequence trace files were obtained from Genbank and combined with our sequencing information to facilitate SNP identification in silico . The process generated 8118 contigs, of which 239 included minimum one sequence from Korean native pig and contained 678 putative coding single nucleotide polymorphisms (cSNPs). Of these, 33 putative cSNPs were randomly selected for confirmatory analysis and validated using 20 pigs from four different breeds (Duroc, Landrace, Yorkshire, Korean native pig). Of the 33 putative cSNPs, 20 were confirmed (61%), which was similar to the frequency reported in other studies. We also identified 15 new cSNPs from the validation process, which were not detected by our in silico analysis. Our study shows that analysing genetically diverse pig breeds including the Korean native pig could serve as a useful strategy for generating a large number of cSNPs.     

Prevalence of Bartonella henselae and Bartonella clarridgeiae in cats and dogs in Korea

Blood, saliva, and nail samples were collected from 54 dogs and 151 cats and analyzed for the presence of Bartonella henselae with a novel nested polymerase chain reaction (PCR) method. Bartonella (B.) henselae was detected in feral cat blood (41.8%), saliva (44.1%), and nail (42.7%) samples. B. henselae was also detected in pet cat blood (33.3%), saliva (43.5%), and nail (29.5%) samples and in pet dog blood (16.6%), saliva (18.5%), and nail (29.6%) samples. Nine samples were infected with B. clarridgeiae and 2 were co-infected with B. henselae and B. clarridgeiae of blood samples of dogs. This report is the first to investigate the prevalence of B. henselae and B. clarridgeiae in dogs and cats in Korea, and suggests that dogs and cats may serve as potential Bartonella reservoirs.     

Estimation of instantaneous and daily net radiation from MODIS data under clear sky conditions: a case study in East Asia

The Moderate Resolution Imaging Spectroradiometer-based net radiation (R _N) model was built and applied in East Asia in 2005. Because there have hardly been simple parameterization schemes developed over a large area using remote-sensing technology, the model was aimed to present physical simplicity in complex topography at multiple spatiotemporal scales. The model successfully reproduced the instantaneous R _N values obtained at four flux tower sites having individually different ecohydrology. The diurnal cycle of R _N was contextually simulated using a simple sine curve to determine the daily and monthly average net radiation. The diurnal R _N estimation method was proven to be a reliable model as long as accurate boundary conditions, sunrise and sunset times, for example, were obtained. The monthly average net radiation (MANR) was estimated using the diurnal patterns of the instantaneous R _N. Distribution of the monthly R _N demonstrated that elevation and latitude were the primary factors affecting the MANR. The proposed R _N algorithm turned out to be a promising method for valuable applications in various fields due to systematic simplicity and fewer input parameters.     

Soybean meal and wheat flour,proper dietary protein sources for optimal growth of snail (<Emphasis Type="Italic">Semisulcospira coreana</Emphasis>)

A 10-week feeding experiment was conducted to determine the proper dietary protein source for optimal growth performance of juvenile snails, Semisulcospira coreana. Sixteen isonitrogenous (31% crude protein) and isocaloric (4.2 kcal/g DM) experimental diets (designated as FM, F-FM, SBM, F-SBM, WF, F-WF, SSM, F-SSM, SCR, F-SCR, UP, F-UP, FWP, F-FWP, MIX and F-MIX) were formulated to contain 41% fish meal, 42% fermented fish meal, 59% soybean meal, 55% fermented soybean meal, 55% wheat flour with 26% fish meal, 55% fermented wheat flour with 29% fish meal, 50% sesame seed meal with 10% fish meal, 50% fermented sesame seed meal with 11% fish meal, 55% soya-curd residue or fermented soya curd residue with 25% fish meal, 50% Undaria powder or fermented Undaria powder with 24% fish meal, 50% freshwater plant with 29% fish meal, 50% fermented freshwater plant with 28% fish meal, a mixture of 8% soybean meal, 10% wheat flour, 8% sesame seed meal, 8% soya-curd residue, 8% Undaria powder and 8% freshwater plant with 17% fish meal, and 50% the fermented mixture with 16% fish meal, respectively, as dietary protein sources. Juvenile snails (average weight, 32 ± 0.7 mg) were randomly distributed in forty-eight 25-L aquaria (16 L water each) in a flow-through system at a density of 130 snails per aquarium. Three replicate groups of snails were fed one of the experimental diets ad libitum once per day for 10 weeks. At the end of the feeding experiment, survival of snails was above 98% and not significantly different among the dietary treatments. The best final body weight was observed in snails fed the SBM and WF diets. There were no significant differences in final body weight of snails fed the F-SBM, WF, F-WF and MIX diets, but the value was higher than that of snails fed the FM, F-FM, FWP and F-FWP diets. The lowest value was observed in snails fed the SSM, F-SSM, SCR and F-SCR diets. Whole body protein and lipid contents of snails varied with dietary protein sources. Amino acid composition of whole body was altered by dietary protein sources. The results of the present study indicate that dietary protein sources significantly influence proximate and amino acid composition, and growth performance of snails. Fermentation with ferminpan instant brown yeast does not improve nutritional quality of the protein sources, and soybean meal and wheat flour in combination with fish meal could be potential protein sources in feeds for juvenile snails.     

Identification of histones as endogenous antibiotics in fish and quantification in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) skin and gill

Antimicrobial polypeptides (AMPPs) are increasingly recognized as a critical component of innate host defense. Among the AMPPs, polypeptides related to histones have been identified from many animals. Using peptide mapping, we further confirm the identity of two histone-like proteins from fish as members of the H2B (sunshine bass) and H1 (rainbow trout) histone groups. We optimized the conditions for measuring rainbow trout HLP-1/H2B via sandwich ELISA. We used two antibodies, one to the amino terminus and one to the carboxyl terminus, of trout histone H2B, as the capture antibodies, and we used peroxidase-labeled antibody raised to calf histone H2B as the secondary antibody. Specificity of the detecting antibody was confirmed by specific reactivity with histone H2B in tissue extracts via western blotting. The test was reproducible and capable of detecting as little as 5 ng of histone H2B (0.05 μg/ml). Histone H2B levels expressed in gill tissue of juvenile, healthy rainbow trout were well within concentrations that are lethal to important fish pathogens. However, there was a significant, age (size)-dependent decline in histone H2B concentrations as fish matured, until levels became virtually undetectable in market-size fish. In contrast, levels in skin appeared to remain high and unchanged in small versus large fish. Antibacterial activity in skin and gill tissues was closely correlated with histone H2B concentration measured via ELISA, which supports our previous finding that histones are the major AMPPs in rainbow trout skin and gill.