Preparation of phytate-removed deamidated soybean globulins by ion exchangers and characterization of their calcium-binding ability.

Phytate-removed deamidated soybean globulins were prepared using ion-exchange resins to provide them with a functional property to enhance calcium absorption in the body. The phosphorus level was reduced from 45 to 20 micromol/g using 0.05 g/mL of AE-4, an anion-exchange resin with a (2-hydroxyethyl)dimethylammonium group, in 0.2% soybean globulin solution for 1 h at 4 degrees C, and 90-92% of the phosphorus in defatted soybeans could be removed. As for deamidation, CE-4, a cation-exchange resin of the carboxylate type, showed a much higher deamidation activity than CE-1 and CE-2, cation-exchange resins of the sulfonate type. No peptide bond hydrolysis was observed for any cation-exchange resin treated at 4 degrees C. There was no significant difference in the amount of acid amide deamidated at temperatures between 4 and 50 degrees C. The deamidation level was able to increase to 73% using 0.10 g/mL of CE-4 in a 0.2% soybean globulin solution for 6 h at 4 degrees C. The amount of calcium bound to the soybean globulins decreased with removal of the phytate but increased with deamidation.     

Growth, Net Photosynthetic Rate, Nutrient Status and Secondary Xylem Anatomical Characteristics of Fagus Crenata Seedlings Grown in Brown Forest Soil Acidified with H2SO4 Solution

Dry matter production, net photosynthetic rate, leaf nutrient status and trunk anatomical characteristics of Fagus crenata seedlings grown in brown forest soil acidified by adding H₂SO₄ solution were investigated. The soil acidification leaded to decreased (Ca+Mg+K)/Al molar ratio in the soil solution. Dry mass per plant of the seedlings grown in the soil treated with H⁺ at 120 mg·L^?1 was significantly reduced compared with the control value at 0 mg·L^?1. When net photosynthetic rate was reduced in the seedlings grown in the soil treated with H⁺ at 120 mg·L^?1, the carboxylation efficiency and maximum net photosynthetic rate at saturated CO₂-concentration were lower than the control values. The addition of H⁺ to the soil at 120 mg·L^?1iinduced a reduction in the concentration of Ca in the leaf. By contrast, the concentration of Al in the leaf was increased with increasing the amount of H⁺ added to the soil. The annual ring formed in the seedlings grown in the soil treated with H⁺ at 120 mg·L^?1 was significantly narrower than that at 0 (control), 10, 30, 60 or 90 mg·L^?1. Based on the results obtained in the present study, we conclude that Fagus crenata is relatively sensitive to a reduction in the (Ca+Mg+K)/Al molar ratio of soil solution compared with Picea abies.     

Lymphocyte blastogenic responses to inciting food allergens in dogs with food hypersensitivity

Lymphocyte blastogenic responses against food allergens in dogs with food hypersensitivity were evaluated in this study. Eleven dogs with food hypersensitivity, based on food elimination and oral food provocation tests and allergic responses to food allergens, were examined by various tests such as intradermal testing, antigen-specific IgE testing, and lymphocyte blastogenic responses. The number and kinds of food allergens identified as positive by these tests were compared with the offending food allergens that were found in an oral food provocation test. In 9 (82%) of the 11 dogs with food hypersensitivity, there was close agreement for positive allergens between the results of lymphocyte blastogenic responses and oral food provocation test; however, there was little agreement for intradermal and IgE testing of the positive allergens with those of the oral food provocation test (11% and 31%, respectively). In the 9 dogs, the stimulation indices of lymphocyte blastogenic responses increased to 2.0-10.1 upon food provocation but decreased significantly to 0.7-1.4 upon feeding the elimination diet until clinical signs disappeared. These results indicate that lymphocyte blastogenic responses may fluctuate because of exposure to offending food allergens in dogs with food hypersensitivity. Lymphocytes reactive to food allergens may play an important role in the pathogenesis of food hypersensitivity in dogs.     

Cathepsin gene expression in mouse placenta during the latter half of pregnancy

Gene expressions and their interaction are complex and have not been definitely clarified in the placenta. To identify interactions of gene products previously not studied, we applied cDNA subtraction analyses to the placenta between days 12 and 16, days 12 and 14, days 14 and 16 of pregnancy. Among subtracted cDNAs cathepsin M, Q and R in PECs were specifically identified on days 14 and 16 pregnancy. All of these gene expressions exhibited a similar pattern to the mPL-II gene expression determined by northern blot and RT-PCR analyses. By means of in situ hybridization, these mRNAs were localized in the basal and labyrinth zones of the placenta on day 16 of pregnancy. Double staining studies of cathepsin Q or cathepsin R mRNA by in situ hybridization followed by immunohistochemical staining of mPL-II in the same section revealed that signals for cathepsin Q and cathepsin R mRNAs were colocalized in mPL-II immunopositive trophoblast cells in the basal and labyrinth zones of the placenta on day 16 of pregnancy. Possible association of cathepsins with mPL-II may play important roles in placental functions during the latter half of pregnancy in mice.     

Identification of Card15/Nod2 mRNA in intestinal tissue of experimentally induced colitis in rats

Card15/Nod2 has been suggested to be an intracellular pathogen-associated molecular pattern (PAMPs) recognition molecule, which contains a leucine-rich repeat region similar to the Toll-like receptors (TLRs). Card15/Nod2 gene variants play an important role in the susceptibility to Crohn's disease. In this study, we examined the kinetics of Card15/Nod2 expression in intestinal tissue during inflammation in the 2, 4, 6-trinitrobenzenesulfonic acid (TNBS)-treated rat experimental colitis model. At 2 and 4 days after TNBS administration, the mononuclear cells remarkably infiltrated the mucosal layer and tunica muscularis, which was followed by a gradual decrease to resting levels at 14 days after TNBS administration. Card15/Nod2 mRNA expression increased and peaked at 4 days after the TNBS administration, followed by a gradual decrease in accordance with the amelioration of the inflammatory response. Expressions of Tlr2, Tlr4 and Myd88 were also upregulated in the inflamed colonic region, and in an in situ hybridization study, a positive signal for Card15/Nod2 was observed in the crypt of the epithelial cell layer and in the infiltrated cells of the submucosal and myenteric regions. These results suggest that in addition to the TLR recognition systems, Card15/Nod2 may contribute to the inflammatory process not only in the epithelial and submucosal layers but also in the tunica muscularis.     

Mastitis caused by Mycobacterium kansasii infection in a dog

A 2‐year, 7‐month‐old female Chihuahua was admitted for a mammary mass measuring one cm in diameter. The dog had a history of demodicosis for 4 months and showed signs of pseudopregnancy at the time of the visit. Cytologic examination of an aspirate of the mass revealed a large number of macrophages containing nonstaining bacterial rods, which were acid‐fast in a Ziehl–Neelsen stain, suggesting mycobacterial infection. Histologic examination of the mass revealed a pyogranulomatous mastitis characterized by an infiltration with macrophages containing acid‐fast bacteria. Mycobacterium kansasii was subsequently cultured and identified by PCR. Surgical excision of the mass resulted in the growth of other dermal masses, but antimycobacterial treatment with rifampin and clarithromycin resolved these masses within 1 month. Three months after discontinuation of the treatment, similar organisms were found in aspirates of the enlarged bilateral inguinal lymph nodes by cytologic examination. Despite antimycobacterial treatment for another 4 months, there was no improvement and demodicosis also recurred. The dog eventually died of lymphoma 5 months after the relapse of mycobacterial infection. Although M kansasii is considered an important pathogen for pulmonary and cutaneous disease in people, there is only one report in a dog with an infection in a pleural effusion. As both adult‐onset demodicosis in dogs as well as mycobacterial infection in people have been associated with T‐lymphocyte deficiency, the M kansasii infection in this dog may have been associated with a condition of immune compromise.