Isolation and identification of indigestible pyroglutamyl peptides in an enzymatic hydrolysate of wheat gluten prepared on an industrial scale

An enzymatic hydrolysate of wheat gluten was further digested in vitro with porcine pepsin and pancreatin to obtain an indigestible peptide. Indigestible pyroglutamyl peptide was isolated from the digest by strong cation-exchange, size-exclusion, and reversed-phase chromatographies. The pyroglutamyl peptide was digested with pyroglutamate aminopeptidase, and the digest was reacted with phenyl isothiocyanate. The resultant phenylthiocarbamyl (PTC) peptides were purified by reversed-phase HPLC by using binary gradient elution with ammonium acetate buffer, pH 6.0, and acetonitrile. The PTC peptides were analyzed with an automatic peptide sequencer on the basis of the Edman degradation method with a modified program. Some pyroglutamyl peptides were also analyzed by fast-atom bombardment ionization mass spectrometry without the pyroglutamate amino peptidase digestion. Consequently, pyroGlu-Asn-Pro-Gln, pyroGlu-Gln-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gly-Gln-Gly-Gln, pyroGlu-Gln, pyroGlu-Gln-Pro, pyroGlu-Ile-Pro-Gln, pyroGlu-Ile-Pro, pyroGlu-Gln-Pro-Leu, pyroGlu-Gln-Phe-Pro-Gln, pyroGlu-Ser-Phe-Pro-Gln, pyroGlu-Phe-Pro-Gln, and pyroGlu-Gln-Pro-Pro-Phe-Ser were identified.     

High susceptibility to zymbal gland and intestinal carcinogenesis in diabetic Otsuka long-evans Tokushima Fatty rats

Diabetes mellitus (DM) and obesity are believed to be risk factors for colorectal cancer in humans. In experiment 1, male nondiabetic Long-Evans Tokushima Otsuka (LETO) rats and Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a model animal of type 2 DM, were whole-body X-irradiated (4 Gy) at 6 and 8 weeks of age and euthanized at 78 weeks of age (n=15, respectively). The incidences of small intestine adenocarcinoma in LETO and OLETF rats were 0% and 30%, respectively. In experiment 2, male LETO and OLETF rats (n=24, respectively) were given s.c. injections of 15 mg/kg azoxymethane (AOM) once weekly for 3 weeks and euthanized at 36 weeks of age. The incidences of Zymbal gland tumors in LETO and OLETF rats were 0% and 67%, respectively (P<0.001), whereas those of small intestine adenocarcinoma were 0% and 43% (P<0.001) and those of cecum/colon adenocarcinoma were 46% and 79% (P<0.05), respectively. Fatty change of hepatocytes was common in OLETF rats (63%) but not in LETO rats. Serum triglyceride and free fatty acid levels in OLETF rats were significantly higher than in LETO rats at sacrifice, whereas serum insulin levels in OLETF rats were very diverse. These data suggest that hyperlipidemia plays a significant role in high susceptibility to lower intestinal tract carcinogenesis in OLETF rats; this strain is susceptible to AOM-induced Zymbal gland carcinogenesis.     

A Case Report of a Malignant Fibrous Histiocytoma in a T-cell Receptor β Chain and p53 Double-knockout Mouse

A subcutaneous tumor was found in the right abdomen of a 16-week-old male TCRβ and p53 double-knockout mouse. The tumor had indistinct borders with the surrounding tissue. The cut surface after formalin fixation was pale yellowish white, partially dark red and partly white. Histologically, the tumor was composed of three distinct regions. The first region showed pleomorphic cells arranged in sheets. The second region showed spindle cells arranged in interlacing fascicles. The final region contained a mixture of the above mentioned two types of cells. Furthermore, a small amount of collagen fibers, round cells, multinucleated giant cells, and cells with eosinophilic granules were observed between these tumor cells. Immunohistochemical examination and electron microscopy identified that the pleomorphic cells and spindle cells were histiocytes and fibroblasts, respectively, and that the round cells were undifferentiated mesenchymal cells. Based on these findings, the tumor was diagnosed as a malignant fibrous histiocytoma.     

A retrospective study and gene analysis of canine sterile panniculitis

In this study, a retrospective analysis was conducted to assess the current aspects and predisposing factors of canine sterile panniculitis. Miniature dachshund, neutered, and younger dogs appeared to be predisposed. In addition, histories of previous surgery and injection were associated in 46.5% of the cases, with several types of surgical suture materials used. About 88% of the dogs had multifocal lesions, frequently with signs of systemic illnesses. Whereas systemic immunosuppressive therapy was effective in most dogs, surgical excision of lesions was rarely curative. In order to prevent recurrences, over 65% of the cases required prolonged immunosuppressive therapy. Polymorphism of canine alpha(1)AT gene was investigated as a candidate gene for sterile panniculitis. Eight polymorphisms were discovered in seven Miniature dachshunds by direct nucleotide sequencing, which included a 12-bp deletion, three non-synonymous single nucleotide polymorphisms, and four silent substitutions. Genotyping of the two polymorphisms, c.109_120del12 and c.483A>C, which identified at high incidence in the dachshunds, was conducted in 83 dogs of 6 popular breeds. The frequencies of neither polymorphism differed between Miniature dachshunds and other breeds, suggesting that neither is responsible for developing panniculitis.     

Methods for determining the consistency limits and constants of soil

The physical analysis of soil tilth has long been one of the important problems in relationship to the solution of soil productivity. Various methods have been proposed to measure soil tilth and most of them are based upon granulation, porosity, penetration and compressibility of soils. In our laboratory here studies³,⁴⁾ for the physical improvement of soil are being continued to find a suitable method of reclamation for heavy clayey polder soils of extremely bad structure in Kojima Bay. In this paper several methods for measuring the consistency limits, and consistency of dry and wet soil are reported, and the value of these methods as a measure of soil tilth is discussed.     

Biodegradation kinetics of monosaccharides and their contribution to basal respiration in tropical forest soils

Low molecular weight (LMW) organic compounds in soil solution are easily biodegradable and could fuel respiration by soil microorganisms. Our main aim was to study the mineralization kinetics of monosaccharides using ¹⁴C-radiolabelled glucose. Based on these data and the soil solution concentrations of monosaccharides, we evaluated the contribution of monosaccharides to basal respiration for a variety of tropical forest soils. Further, the factors controlling the mineralization kinetics of monosaccharides were examined by comparing tropical and temperate forest soils. Monosaccharides comprised on average 5.2 to 47.7% of dissolved organic carbon in soil solution. Their kinetic parameters (V _max and K_M ), which were described by a single Michaelis-Menten equation, varied widely from 11 to 152?nmol?g^?1?h^?1 and 198 to 1294?µmol?L^?1 for tropical soils, and from 182 to 400?nmol?g^?1?h^?1 and 1277 to 3150?µmol?L^?1 for temperate soils, respectively. The values of V _max increased with increasing microbial biomass-C in tropical and temperate soils, while the K_M values had no correlations with soil biological or physicochemical properties. The positive correlation between V _max values and microbial biomass-C indicates that microbial biomass-C is an essential factor to regulate the V _max values in tropical and temperate forest soils. The biodegradation kinetics of monosaccharides indicate that the microbial capacity of monosaccharide mineralization far exceeds its rate at soil solution concentration. Monosaccharides in soil solution are rapidly mineralized, and their mean residence times in this study were very short (0.4–1.9?h) in tropical forests. The rates of monosaccharide mineralization at actual soil solution concentrations made up 22–118% of basal respiration. Probably because of the rapid and continuous production and consumption of monosaccharides, monosaccharide mineralization is shown to be a dominant fraction of basal respiration in tropical forest soils, as well as in temperate and boreal forest soils.     

Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

Programmed death-1 (PD-1) is a known immunoinhibitory receptor that contributes to immune evasion of various tumor cells and pathogens causing chronic infection, such as bovine leukemia virus (BLV) infection. First, in this study, to establish a method for the expression and functional analysis of bovine PD-1, hybridomas producing monoclonal antibodies (mAb) specific for bovine PD-1 were established. Treatment with these anti-PD-1 mAb enhanced interferon-gamma (IFN-γ) production of bovine peripheral blood mononuclear cells (PBMC). Next, to examine whether PD-1 blockade by anti-PD-1 mAb could upregulate the immune reaction during chronic infection, the expression and functional analysis of PD-1 in PBMC isolated from BLV-infected cattle with or without lymphoma were performed using anti-PD-1 mAb. The frequencies of both PD-1⁺ CD4⁺ T cells in blood and lymph node and PD-1⁺ CD8⁺ T cells in lymph node were higher in BLV-infected cattle with lymphoma than those without lymphoma or control uninfected cattle. PD-1 blockade enhanced IFN-γ production and proliferation and reduced BLV-gp51 expression and B-cell activation in PBMC from BLV-infected cattle in response to BLV-gp51 peptide mixture. These data show that anti-bovine PD-1 mAb could provide a new therapy to control BLV infection via upregulation of immune response.     

A refined hydro-environmental watershed model with field-plot-scale resolution

A distributed hydro-environmental model is developed that achieves detailed analysis of the movement of water at a field-plot-scale resolution in a mesoscale watershed including lowland areas where, especially for agricultures, it is an essential need to get rid of redundant groundwater by drainage facilities such as rivers, canals and/or underdrains. For this, the problem geometry is meshed with unstructured cells of triangular shape. Profile of a column cell is zoned into two: surface zone and groundwater zone in which water movement is represented by combined tank and soil moisture sub-models, and well-defined two-dimensional unconfined shallow groundwater flow sub-model, respectively. The top-two sub-models serve to evaluate evapotranspiration, infiltration, soil water content, lateral surface water flow, and vertical percolation. The vertical percolation so evaluated is given as longitudinal recharge to the bottom sub-model for computing groundwater flow. Surface water–groundwater interactions through beds and stream-banks of perennial and ephemeral canals are considered by treating the canal courses as internal boundaries in the groundwater flow model. The finite volume method (FVM) that allows of unstructured mesh and produces conservative solutions is employed for groundwater flow computation. The model developed is applied to an actual watershed which includes a low-lying paddy area to quantify the hydrological impact of land-use management practices over a period of 29 years in which the farmland consolidation project was implemented and part of the paddy fields were converted to upland crop fields and housing lands. From the results obtained, it is concluded that the model presently developed lends itself to water—as well as land-use management practices.     

A hydro-environmental watershed model improved in canal-aquifer water exchange process

Long-term simulation using the distributed hydro-environmental watershed model is efficacious for assessing irrigation impacts on hydrological cycle in detail and for implementing watershed management successfully. In this article, the previously developed hydro-environmental watershed model (HEWM-1) is improved in the water exchange process caused by surface water-groundwater interaction via drainage canals and/or underdrains. The time-varying stream flow in canals is described by the complete one-dimensional shallow water equations in a newly introduced submodel, the open channel flow submodel. This submodel coordinates with the other submodels: the tank, soil moisture and groundwater flow submodels which are interlinked in a cascade manner. The improved model (HEWM-2) is applied to an agricultural watershed covering an area from an alluvial fan onto a nearly level alluvial plain, to be validated. The simulation by HEWM-2 is informative for identifying whether any drainage canal is gaining or losing water in relation to groundwater level. It could thus provide useful information for conserving a complex network of drainage canals which also functions as a passage for aquatic animals like fishes.