Development and characterization of monoclonal antibodies against canine trypsin

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.     

Evaluation of truncated LipL32 expressed by Escherichia coli and Pichia pastoris for serodiagnosis of Leptospira infection in rodents

The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.     

Behavior of four broad-leaved tree species used to revegetate eroded granite hill slopes

Information on primary growth behavior after planting is required for mixed-plantation revegetation using broad-leaved species. To estimate primary growth, especially from the perspective of crown coverage and changing growth rates, we examined the growth and survival of four broad-leaved species that are frequently used in erosion-control plantations in Japan. The species studied were Myrica rubra Sieb. et Zucc., Alnus pendula Matsum., Quercus glauca Thunb., and Q. serrata Thunb. The survival, height, and basal diameter of planted trees were measured over a 4-year period, and crown area was calculated over a 3-year period. We found a negative relationship between relative growth rate (RGR) and survival rate, suggesting that fast growth may be fatal when resources are severely limited. The relative height growth rate (RHGR) of A. pendula was especially high during the early period of the study (1997–1999) and then drastically declined, whereas the opposite tendency was observed in Q. glauca. The results of stem allometry analyses conformed to the specific relationships between height growth and diameter growth of the four species; increases in stem thickness based on height increments were smaller in the pioneer species A. pendula. Between-species differences in coverage per planted tree (mean crown area multiplied by survival rate) were small as a result of the negative relationship between coverage area and survival rate.     

Estimating harvest costs and projecting quantities of logging residues for small-scale forestry in Nasushiobara,Tochigi Prefecture,Japan

We used GIS on a regional scale to estimate and compare supply potentials and costs of small-scale logging systems, a mini-forwarder and a 4-ton truck operated by private logging contractors, and manual logging and a light truck operated by individual forest own- ers, with the mechanized operational system of the Forest Owners＇ Asso- ciation. Total potential yields of timber and logging residues were esti- mated as 418,895 m3 and 254,962 m3, respectively. The economic bal- ances were estimated and available amounts were projected as supply potentials from profitable sub-compartments. As a result, available amounts of timber and logging residues were estimated at 375,466 m3 （89.9%） and 203,850 m3 （80.0%）, respectively. Because their transport expenses were lower than for other systems the most profitable sub-compartments were operated by private logging contractors who sold logging residues at a plant. The profitable sub-compartments operated by individual forest owners were few because the extracting distances were usually greater than 20 m. Raising logging residue prices from 3,000 yen·m-3 to 4,080 yen·m-3 or 6,800 yen·m-3, and establishing forest roads, which reduced some extracting distances to less than 20 m, increased the number and area of profitable sub＊compartments, and increased available amounts of logging residues.     

Examining the Optimal Method to Extract Logging Residues from Small-Scale Forestry in the Nasunogahara Area,Tochigi Prefecture,Japan

This study examines the optimal method to extract logging residues from small, fragmented and scattered forests separately from the mechanized operational systems used on aggregated forestry operation sites. First, small-scale logging systems operated by a private logging contractor and an individual forest owner were investigated. Regression equations for estimating felling and processing cycle times were established as a function of stem and log volumes, respectively. Equations to estimate the forwarding and transporting cycle times were established as a function of the forwarding and transporting distances using the forwarding and transporting velocities, respectively. Then, equations to estimate productivities and costs were established with the cycle times, volumes and hourly operational expenses consisting of labour and machinery expenses. Finally, costs of the small-scale systems—mini-forwarder and 4 t truck operated by a private logging contractor, and manual logging and light truck operated by an individual forest owner—were estimated and compared with the mechanized operation system operated by the Forest Owners’ Association. The mechanized operation system was found to have the highest cost and could not compete for small forestry operational sites and small stem volumes. The small-scale systems could be effective for harvesting small areas. This was especially true when conducting extracting operations without machines, as done by the individual forest owner, and transporting operations by a private logging contractor, in which the hourly income of the individual forest owner was higher than the hourly labour expenses.     

Effects of Rice Flour Blends on Bread Texture and Staling

The effects of blending rice flour with wheat flour on bread texture and staling were investigated with three rice varieties with different amylose contents. For the texture analysis of bread crumb, the compression test, puncture test, and tensile test were performed. A flour blend containing rice flour suppressed the recovery of the crumb after compression. For the puncture test, blends with rice flour increased the distance to penetrate the crumb. The rupture strain measured with the puncture test decreased with staling of the bread crumb for all samples tested. The fresh bread crumb sample containing waxy rice flour had much greater extensibility in the tensile test than the other samples tested, but it was dramatically decreased after one day of storage. Endothermic enthalpies corresponding to retrograded amylopectin, which is part of the staling process, were also measured. The enthalpy of bread crumb from the blended flour was lower than that of wheat flour bread up to three days but was higher on day 4. A blend of rice flour thus reduced amylopectin retrogradation during early storage, but it was accelerated later. Bread blended with waxy rice flour showed the lowest enthalpy during storage.     

Expression of CD4 T cell cytokine genes in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds

Inflammatory colorectal polyps (ICRPs) in miniature dachshunds are recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and a space-occupying large polyp in the colorectal area, and are thought to be a novel form of inflammatory bowel disease (IBD). In humans, specific cytokine patterns attributed to T helper (Th)1, Th17 and regulatory T cells have important roles in the pathogenesis of IBD. Thus, the aim of the present study was to assess the gene expression of cytokines of T cell subsets in the colorectal mucosa from dogs with ICRPs. Colorectal mucosal specimens from 10 dogs with ICRPs and 14 control dogs were used in this study. Interferon (IFN)-γ, interleukin (IL)-4, IL-17A and IL-10 mRNA expression was assessed using quantitative real-time PCR. IL-17A mRNA expression was significantly increased in large polyps compared to small polyps and controls. IFN-γ and IL-10 mRNA expression in large polyps were significantly higher than in controls. There was no significant difference in IL-4 mRNA expression among the three groups. IL-17A is thought to play important roles in the pathogenesis of ICRPs. IL-10 up-regulation could oppose the proinflammatory function of IL-17A.     

Effect of dietary fish oil on enhanced inflammation and disturbed lipophagy in white adipose tissue caused by a high fat diet

Fisheries Science - Fish oil containing omega-3 polyunsaturated fatty acids (PUFA) attenuates chronic inflammation found in obesity, leading to a reduction in insulin resistance. The effect of fish...