Reliable enzyme-linked immunosorbent assay for the determination of walnut proteins in processed foods

Among food allergens of tree nuts, walnuts are a frequent cause of adverse food reactions in allergic patients. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of walnut soluble proteins in processed foods was developed. The sandwich ELISA is highly specific for walnut soluble proteins. The recovery ranged from 83.4 to 123%, whereas the intra- and interassay coefficients of variation were less than 8.8 and 7.2%, respectively. This study showed that the proposed method is a reliable tool for detection in the presence of hidden walnut proteins in processed foods.     

Yield and growth characteristics of erect panicle type rice (Oryza sativa L.) cultivar,Shennong265 under various crop management practices in Western Japan

Erect panicle rice cultivars utilize solar energy effectively and have improved ecological growing conditions. Among such cultivars, Shennong265 has been grown successfully throughout Northern China. Nevertheless, no studies have yet examined the relationships between crop dry matter productivity, weather conditions, and nitrogen uptake of the erect panicle type rice cultivar in Japan. The objective of our study was to evaluate the productivity of erect panicle rice Shennong265 in Western Japan under varied conditions. Three rice cultivars, Shennong265, Nipponbare, and Takanari were grown in the field under different fertilizer and plant density conditions in Western Japan; using this information, we compared yield and growth characteristics of Shennong265 with those of Nipponbare and Takanari. Although Shennong265 had radiation use efficiency similar to that of the high yielding cultivar (Takanari) and much higher leaf nitrogen content than Takanari and Nipponbare, the average grain yield of Shennong265 grown under normal fertilizer and plant density conditions was approximately 6.9 t ha^?1 as against 6.2 t ha^?1 for Nipponbare and 9.6 t ha^?1 for Takanari. These results suggest that, while Shennong265 has a high yield potential, the environmental conditions including climate, fertilizer, and planting period provided in this study were not suitable for achieving its maximum yield. The reduced performance of Shennong265 may be caused by insufficient fertilizer after heading and by shorter growth periods, as well as by the climate of Western Japan. Additional fertilizer application during the heading stage and earlier transplanting may be needed to obtain higher Shennong265 yields in Western Japan.     

Long-term shifts in the growth and maturation size of Miyabe charr <Emphasis Type="Italic">Salvelinus malma miyabei</Emphasis>

Overfishing can affect life history traits, resulting in population collapse and oftentimes a decrease in length-at-age and maturation size in fish populations. However, little is known about the recovery mechanisms and time scales of these traits in exploited wild populations. In the study reported here, we documented long-term shifts in growth and mature size in Miyabe charr Salvelinus malma miyabei associated with a history of recreational fishing in Lake Shikaribetsu for approximately 80 years. Downsizing in the charr was observed when the charr population collapsed due to intensive recreational fishing. However, subsequent moratoriums and the introduction of fishing regulations, especially the implementation of a catch-and-release policy, during the following 10–30 years facilitated the recovery of population size, length-at-age and mature fish size. This study provides important insights into the biological changes and required recovery time scales of a heavily harvested population and supports management and conservation strategies.     

Investigation of serum concentrations and immunohistochemical localization of α1-acid glycoprotein in tumor dogs

The purpose of this study was to measure the dynamics of serum α1-acid glycoprotein (AGP) concentration in dogs with various tumors, and to investigate the localization of AGP in some tissues using immunohistochemical staining. Sera were obtained from 171 dogs bearing tumors of various types. Serum AGP concentration was measured by single radial immunodiffusion. Tumors occurring in the liver and spleen were also investigated immunohistochemically using anti-canine AGP antibody. Mean serum AGP levels were 749 ± 602 mg/L in dogs with carcinoma (n = 39), 1,014 ± 971 mg/L with sarcoma (n = 18), and 887 ± 935 mg/L with round cell tumors (n = 46), all significantly higher than serum AGP level in healthy dogs (n = 137, 364 ± 106 mg/L). Mean serum AGP levels were significantly higher than in healthy dogs in complex mammary gland carcinoma (n = 5, 876 ± 721 mg/L), malignant melanoma (n = 7, 1,010 ± 821 mg/L), and hepatocellular carcinoma (n = 5, 936 ± 741 mg/L) among carcinomas, hemangiosarcoma (n = 5, 1,740 ± 1,323 mg/L) among sarcomas, and lymphoma (n = 19, 1,072 ± 965 mg/L) and histiocytic tumor (n = 6, 1,800 ± 1,387 mg/L) among round cell tumors. In an immunohistochemical investigation of AGP localization, both weak and strong staining for anti-AGP antibody were seen in hepatic tissue in dogs with primary non-tumorous lesions originating in the spleen (hematoma) and elevated serum AGP, but all tumor tissue in the spleen was negative. Among dogs with primary tumor lesions of the spleen (hemangiosarcoma) and elevated serum AGP levels, both weak, moderate and strong staining for anti-AGP antibody were seen in hepatic tissue, while strong positive staining was apparent in all tumorous tissue from the spleen. In primary tumor lesions in the liver (hepatocellular carcinoma), both moderate and strong staining for anti-AGP antibody were seen in normal hepatic tissue, and both weak, moderate and strong staining were seen in tumor tissues of the liver. AGP levels thus appear to be elevated in dogs with carcinomas, sarcomas, and round cell tumors. With some of these malignant tumors, localization of AGP in tumor tissue was seen.     

Effects of mycoplasmal pneumonia of swine (MPS) lung lesion‐selected Landrace pigs on MPS resistance and immune competence in three‐way crossbred pigs

To clarify the genetic influence of mycoplasmal pneumonia of swine (MPS) lesion‐selected Landrace (La) on MPS resistance and immune characteristics in three‐way crossbred pigs (LaWaDa), the LaWaDa pigs were compared with the non‐selected crossbred (LbWbDb) and purebred (La) pigs. The MPS lesion score in the three lines was as follows: La line < LaWaDa line < LbWbDb line, with significant differences among the lines. The proportions of myeloid cells and T cells were lower and higher, respectively, in the LaWaDa pigs compared with those in the other two lines. Messenger RNA (mRNA) expression of interleukin (IL)‐6, IL‐10, transforming growth factor‐β, and interferon‐γ in peripheral blood was significantly increased after vaccination in the La and LaWaDa lines. IL‐4 mRNA expression in the LaWaDa line was intermediate to the La and LbWbDb lines. Furthermore, principal component analysis for immune traits and MPS lesions was executed to clarify the characteristics of each pig line. These findings suggest that the immune responses in the three pig lines are genetically distinct and that MPS resistance and some immunity characteristics from the La line were transmitted to the three‐way crossbred pigs.     

Studies on the immune status of calves with chronic inflammation and thymus atrophy

The thymus is a primary lymphoid organ where the primary T cell repertoire is generated. Thymus atrophy is induced by various conditions, including infectious diseases, glucocorticoid treatment, and poor breeding management. Cattle with thymus atrophy tend to exhibit weak calf syndrome, a condition in which approximately half of neonates die shortly after birth. Calves with thymus atrophy that survive the first month typically contract chronic inflammatory diseases. In this study, we analyzed the populations of the peripheral blood mononuclear cells and thymocytes in calves with thymus atrophy. In addition, we evaluated polarization of master gene and cytokine mRNA expression in peripheral blood CD4⁺ cells in the calves. The population of CD4⁺CD8⁺ cells in thymus of the calves with thymus atrophy was lower than that of control calves. IL10 mRNA expression in peripheral blood CD4⁺ cells of calves with thymus atrophy was significantly lower than that of control calves. TBX21 mRNA expression in peripheral CD4⁺ cells of thymus atrophy calves was tended to be higher than that of the control group. In addition, FOXP3 mRNA expression in peripheral CD4⁺ cells of the thymus atrophy calves was tended to be lower than that of the control calves. Thymus atrophy calves exhibited chronic inflammatory disease leading, in severe situations, to conditions such as pneumonia with caseous necrosis. These severe inflammatory responses likely are due to decreases in IL10 mRNA expression, impairing control of macrophages, one of the main cell fractions of natural immunity.     

Structural insights into the interaction between gabazine (SR-95531) and Laodelphax striatellus GABA receptors

γ-Aminobutyric acid receptors (GABARs) mediate fast inhibitory neurotransmission and are targets for insecticides. GABARs are composed of five subunits, the composition of which dictates the pharmacological characteristics of GABARs. Both competitive and noncompetitive GABAR antagonists can be used as insecticides. Gabazine is a potent competitive antagonist of mammalian α1β2γ2 GABARs; however, it is less potent against insect GABARs. To explore how gabazine interacts with GABARs, we examined whether the sensitivity of the small brown planthopper (Laodelphax striatellus) RDL GABAR (LsRDLR) to gabazine is increased when its amino acid residues are substituted with α1β2γ2 GABAR residues. In the results, two of the generated mutants showed enhanced gabazine sensitivity. Docking simulations of gabazine using LsRDLR homology models and an α1β2γ2 GABAR cryo-EM structure revealed that the accommodation of gabazine into the “aromatic box” in the orthosteric site lowered the binding energy. This information may help in designing GABAR-targeting insecticides with novel modes of action.     

Evaluation of bovine uterine gland functions in 2D and 3D culture system

In ruminants, uterine glands play key roles in the establishment of pregnancy by secreting various factors into the uterine lumen. Although a three-dimensional (3D) culture system has been used for investigating cellular functions in vitro, the detailed functions of uterine gland have not been fully elucidated. In this study, we examined the benefits of 3D culture system to examine the innate functions of bovine uterine glands. Isolated bovine uterine glands were cultured on Matrigel (2D) or in Matrigel (3D), respectively, and the mRNA levels of secreted proteins (SERPINA14, MEP1B, APOA1, ARSA, CTGF, and SPP1) were measured in isolated and cultured uterine glands. The protein expression of estrogen receptor β (ERβ) and progesterone receptor (PR) and the establishment of apico-basal polarity were examined. In isolated uterine glands, the mRNA levels of secreted proteins changed during the estrous cycle. Although uterine glands cultured in both 2D and 3D expressed ERβ and PR, progesterone did not affect SERPINA14 mRNA expression. The expression of APOA1 mRNA in 2D cultured uterine glands did not respond to estrogen and progesterone. Additionally, the mRNA levels of secreted proteins in the 3D culture system were significantly higher than those in the 2D culture system, which might be attributed to the different cellular morphology between them. The locations of ZO-1 and β-catenin in 2D cultured uterine glands were disordered compared with 3D cultured uterine glands. These results showed that the hormonal responsiveness of secreted factor expression and cellular morphology were different between 2D and 3D cultured bovine uterine glands.