Development and evaluation of an indirect enzyme-linked immunosorbent assay for detection of foot-and-mouth disease virus nonstructural protein antibody using a chemically synthesized 2B peptide as antigen.

Forty peptides were synthesized corresponding to hydrophilic clusters of amino acids within the sequences of foot-and-mouth disease virus (FMDV) nonstructural proteins (NSP). Six peptides were studied in more detail and the most promising, a 2B peptide, was evaluated in enzyme-linked immunosorbent assay (ELISA) using sera from naive, vaccinated, and vaccinated-and-challenged cattle as well as bovine sera from field outbreaks. The performance of the new NSP peptide ELISA was compared to that of 4 commercial NSP ELISA kits. Antibody to 2B was detectable from the end of the first week to the second week after infection in most of the nonvaccinated animals and by the second to third week in vaccinated-and-challenged animals. The sensitivity of the 2B peptide ELISA was comparable to the 3ABC Ceditest (Ceditest FMDV-NS, Cedi Diagnostics B.V.; Chung et al., 2002). With some modification and further validation, this 2B test could be useful as a screening or conformational NSP test in postvaccination surveillance for FMD.     

Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice

We previously induced protective immune response by oral immunization with yeast expressing the ApxIIA antigen. The ApxI antigen is also an important factor in the protection against Actinobacillus pleuropneumoniae serotype 5 infection; therefore, the protective immunity in mice following oral immunization with Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) was compared with that in two control groups (group A and B). The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p < 0.01). The highest systemic (IgG) and local (IgA) humoral immune responses to ApxIA and ApxIIA were detected in group E after the third immunization (p < 0.05). The levels of IL-1β and IL-6 after challenge with an A. pleuropneumoniae field isolate did not change significantly in the vaccinated groups. The level of TNF-α increased in a time-dependent manner in group E but was not significantly different after the challenge. After the challenge, the mice in group E had a significantly lower infectious burden and a higher level of protection than the mice in the other groups (p < 0.05). The survival rate in each group was closely correlated to the immune response and histopathological observations in the lung following the challenge. These results suggested that immunity to the ApxIA antigen is required for optimal protection.     

Guided bone regeneration with beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

This study was performed to evaluate the effect of beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/PLGC group. However, the new cortical bone in the TCP/PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.     

Nutrient-flow wick culture system for potted plant production: System characteristics and plant growth

To compliment the current subirrigation systems used for production of potted plants, a nutrient-flow wick culture (NFW) system was developed and compared with other subirrigation systems, such as an ebb and flow culture (EBB) system and a nutrient-stagnant wick culture (NSW) system in relation to their system characteristics and plant growth. Kalanchoe (Kalanchoe blossfeldiana cv. New Alter) was cultivated in a 6 cm pot for 10 weeks in each subirrigation system. The water-absorption pattern of the medium, water content of the medium, water loss, algal growth, salt-buildup and plant growth under various culture systems were observed. The water contents of medium under the NFW and EBB systems showed fluctuations from 30 to 40% and from 50 to 60% (by volume), respectively, whereas the water content under the NSW system gradually increased to over 40% without fluctuation. Relative to other systems, the water loss in the NFW system was 50–70% due to the reduction in the evaporation from the surfaces of the trough and medium. Algae appeared in the NSW system because the nutrient solution was always stagnant in the trough, while it was not observed under the NFW system. The dissolved oxygen in the nutrient solution was the highest during the irrigation period and the salinity in the medium was the lowest in the NFW system. With regard to system characteristics, the NFW system was simple, water-saving and efficient. In addition, the growth of kalanchoes in the NFW system was similar to those in the NSW and EBB systems at an irrigation frequency of five times a day.     

Long- and short-term flooding effects on survival and sink-source relationships of swamp-adapted tree species

About 95% of swamp tupelo (Nyssa sylvatica var. biflora (Walt.) Sarg.) and sweetgum (Liquidambar styraciflua L.) seedlings survived continuous root flooding for more than two years, whereas none of the swamp chestnut oak (Quercus michauxii Nutt.) and cherrybark oak (Q. falcata var. pagodifolia Ell.) seedlings survived one year of flooding. Death of oak seedlings occurred in phases associated with periods of major vegetative growth, e.g., after bud burst in spring, after summer stem elongation, and during the winter deciduous stage, suggesting that stored reserves and sources were inadequate to maintain the seedlings when vegetative sinks were forming. Additional evidence that flooding induced a source deficiency in oak was that leaves of flooded oak were 65 to 75% smaller than leaves of nonflooded oak. Flooded swamp tupelo seedlings had a normal leaf size and patchy stomatal opening compared with nonflooded seedlings. Flooding caused increases in alcohol dehydrogenase (ADH) specific activity in taproot cambial tissues and increases in starch concentrations of swamp tupelo seedlings that were reversed when seedlings were removed from flooding. Flooding had little effect on soluble sugar concentrations in swamp tupelo or sweetgum. In the long-term flood-dry-flood treatment, in which all species had survivors, upper canopy leaf photosynthetic rates were higher in all species during the dry period than in nonflooded controls, whereas their starch and soluble sugars concentrations were similar to those of nonflooded controls. Based on seedling survival and the sink-source relationships, the order of flood tolerance was: swamp tupelo > sweetgum > swamp chestnut oak > cherrybark oak.     

Characterization of proanthocyanidin in hot water extract isolated from <Emphasis Type="Italic">Pinus radiata</Emphasis> bark

Proanthocyanidins (PAs) in the hot water extract prepared from Pinus radiata bark were isolated by a Sephadex LH-20 chromatography. The PAs were characterized using UV/Visible, FT-IR and ¹³C nuclear magnetic resonance (NMR) spectroscopic techniques, matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS), and gel permeation chromatography (GPC) analysis. Bate-Smith acidic alcoholysis and FT-IR spectrum of the PAs indicated the predominant presence of the procyanidin (PC) structure. The ¹³C NMR spectrum revealed that the PAs comprise PC (94%) together with a small amount of prodelphinidin (PD, 6%), and also that they contain the higher trans configuration rather than the cis configuration of C2–C3. The MALDI-TOF MS results obviously proved the presence of PD units, but the maximum degree of polymerization (DP) was limited to octamer. The average molar mass (Mw, Mn), polydispersity (Mw/Mn), and DP of the pine bark PAs were 3,800 (Mw), 1,200 (Mn), 3.2, and 13, respectively.     

Prediction of bending properties for structural glulam using optimized distributions of knot characteristics and laminar MOE

This study established a prediction model for bending properties of glued-laminated timber (glulam) using optimized knot and modulus of elasticity (MOE) distributions of lumber laminate as the main input variables. For this purpose, knot and MOE data were investigated for all pieces of lumber that were prepared for glulam manufacturing, and statistical distributions of knot size, knot number in one lumber, and MOE of each laminate were optimized as distribution functions. These knot and MOE data were used as input variables in the prediction model for bending properties, and were also used in generating virtual glulam using the inverse transform method. Prediction of bending properties for glulam was carried out using the transformed section method, which is partially provided in ASTM D 3737 (Annex A4). Predicted values were compared with those from full-scale four-point bending tests for 60 six-layered glulams with 10 different laminar combinations. Finally, the allowable bending properties of glulam for each specific laminate combination were determined by calculating the fifth percentile of the modulus of rupture and the average modulus of elasticity from virtual test results of more than 1000 virtual glulams. From the results of this study, predicted bending properties for glulam and their distributions could be used for structural design in both allowable stress design and limit state design.     

Determination of reference intervals for metabolic profile of Hanwoo cows at early,middle and late gestation periods

Background

Metabolic profile was initially designed as a presymptomatic diagnostic aid based on statistical analyses of blood metabolites to provide an early warning of certain types of metabolic disorder. However, there is little metabolic profile data available about Korean Hanwoo cows. Therefore, this study aimed to determine the reference intervals of metabolic profile for Korean Hanwoo cows.

Methods

Healthy animals (2,205) were selected and divided into early (day 1 to 95), middle (day 96 to 190) and late (day 191 to 285) period according to their gestating period. Metabolic profile including total protein (TP), albumin (Alb), urea (UREA), glucose (Glu), total cholesterol (T-Cho), long-chain fatty acid (LCFA), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), creatinine (Crea), calcium (Ca), inorganic phosphorous (iP) and magnesium (Mg) were analyzed using a TBA-40FR automatic biochemical analyzer. The data of Korean Hanwoo cows were then compared to those of the Japanese Wagyu cows.

Results

Most of the data of the Korean Hanwoo cows were relatively higher than those of Japanese Wagyu cows, with the exception of Glu and GGT. This may indicate that the nutritional level of feed for the Korean Hanwoo cows was higher than that of the Japanese Wagyu cows because of the different feeding system. In particular, relatively higher levels of UREA and LCFA were observed in the Korean Hanwoo cows, and this may also contribute to the low reproduction efficiency.

Conclusions

These findings may provide some theoretical basis for understanding the reproductive and feeding situation of Korean Hanwoo cows.     

Antioxidant and antimelanogenic activities of polyamine conjugates from corn bran and related hydroxycinnamic acids

The antioxidant activity of three major polyamine conjugates, N,N'-dicoumaroyl-putrescine (DCP), N-p-coumaroyl-N'-feruloylputrescine (CFP), and N,N'-diferuloyl-putrescine (DFP) isolated from corn bran, and their related hydroxycinnamic acids, p-coumaric acid and ferulic acid, were evaluated by three antioxidant in vitro assay systems, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide and hydroxyl radicals generated by enzymatic and nonenzymatic reactions. Additionally, five phenolic compounds were evaluated for melanogenesis inhibitory activity using mushroom tyrosinase and B16 melanoma cells. Most of the phenolic compounds significantly scavenged DPPH, superoxide, and hydroxyl radicals in a dose-dependent manner. Particularly, DFP showed potent DPPH (IC50 = 38.46 microM) and superoxide (IC50 = 291.62 microM) radical scavenging activities, while DCP exhibited the strongest hydroxyl radical scavenging activity (IC50 = 120.55 microM). CFP also exerted moderate DPPH, superoxide, and hydroxyl radical scavenging activities. Meanwhile, DCP (IC50 = 181.73 microM) showed potent tyrosinase inhibitory activity toward l-tyrosine as the substrate, whereas DFP (IC50 = 733.64 microM) significantly inhibited melanin synthesis in B16 melanoma cells. These current results indicate that these three polyamine conjugates from corn bran may be useful potential sources of natural antioxidants and skin-whitening agents.