Generation and soil detection of nitrate-nonutilizing mutants of Diaporthe destruens causing foot rot of sweet potato

Journal of General Plant Pathology - Three nitrate-nonutilizing (nit) mutants of a soil-borne fungus, Diaporthe destruens, pathogen of foot rot of sweet potato, were generated with a...     

Chiral-selective aminoacylation of an RNA minihelix

Amino acids in natural proteins have a chiral, asymmetric center at the alpha carbon that is of the L-configuration. The sugar backbone of natural RNAs are also homochiral, but of the D-configuration. Because protein synthesis requires the aminoacylation of RNA, it is this step that could have provided chiral selectivity. Here we show that an RNA minihelix was aminoacylated by an aminoacyl-phosphate-D-oligonucleotide with a clear preference for L- as opposed to D-amino acids. A mirror-image RNA system showed the opposite selectivity. These results suggest the possibility that the selection of L-amino acids for proteins was determined by the stereochemistry of RNA.     

Dependence of self-tolerance on TRAF6-directed development of thymic stroma

The microenvironments of the thymus are generated by thymic epithelial cells (TECs) and are essential for inducing immune self-tolerance or developing T cells. However, the molecular mechanisms that underlie the differentiation of TECs and thymic compartmentalization are not fully understood. Here we show that deficiency in the tumor necrosis factor receptor-associated factor (TRAF) 6 results in disorganized distribution of medullary TECs (mTECs) and the absence of mature mTECs. Engraftment of thymic stroma of TRAF6(-/-) embryos into athymic nude mice induced autoimmunity. Thus, TRAF6 directs the development of thymic stroma and represents a critical point of regulation for self-tolerance and autoimmunity.     

Silage preparation and nutritive value of Chinese yam by-product

The objective of this experiment was to determine the ensiling characteristics, nutrient composition and nutritive value of Chinese yam by-product (CYBP) silage treated without additive (Control), with lactic acid bacteria (LY), with dried beet pulp (BY), or with mixtures of lactic acid bacteria and dried beet pulp (LBY) for a fermentation period of 150 days. The nutritive value of CYBP silage was studied in an in vivo digestibility trial using wethers in a 4 × 4 Latin square design. Addition of lactic acid bacteria and/or dried beet pulp greatly improved fermentation quality of CYBP silage. The pH was reduced to below 4.0 and lactic acid was produced at above 4.6% on dry matter basis. There were no statistical differences among treatments in the digestibilities of dry matter, organic matter, crude protein and energy in the silages, but neutral detergent fiber and acid detergent fiber digestibility were higher ( P  < 0.001) in wethers fed BY and LBY silages compared to the control and LY silages. It was estimated that total digestible nutrient and digestible energy in the CYBP silage averaged 79.1% and 14.3 MJ/kg on dry matter basis. The study suggests that CYBP can be well preserved by making a silage and it is a good potential energy source for ruminant diets.     

Simultaneous determination of neonicotinoid insecticides in agricultural samples by solid-phase extraction cleanup and liquid chromatography equipped with diode-array detection

Effective sample pretreatment procedures based on solid-phase extraction (SPE) for multiresidue determination of seven neonicotinoid insecticides in agricultural products were investigated. After extraction with acetone and concentration, the insecticides in aqueous sample extracts were transferred into organic solvent phases with a Chem Elut SPE cartridge. Finally, the eluate from the cartridge was cleaned up with a SPE cartridge packed with graphitized carbon black and aminopropyl silica gel, which showed a higher cleanup efficiency than the classical silica gel SPE cartridge. Seven insecticides were separated on a reversed-phase C18 column and a gradient system of methanol and phosphate solution based on high-performance liquid chromatography. The established multiresidue determination has been applied to several artificially spiked agricultural samples, with the result that the average recoveries were excellent, with the exception of nitenpyram. The limit of detection of the method ranged from 0.01 to 0.03 mg/kg for the insecticides.     

Effects of stratification and misspecification of covariates on species distribution models for abundance estimation from virtual line transect survey data

The aim of this study was to examine the effects of stratification of the survey region on the performance of species distribution models (SDMs) described by generalized linear models or generalized additive models when estimating school abundance by using a line transect survey. True covariates that define spatial school distribution are not always obtainable explanatory variables. When the true covariates differ from explanatory variables in the model, the explanatory variables are determined to be misspecified. We evaluated the performance of SDMs in abundance estimation with misspecified covariates by using dummy datasets for which the true abundance was known. Simulated replicates of spatial distributions of a whale school and sighting data were generated from possible scenarios motivated by the spatial school distribution of Antarctic minke whales Balaenoptera bonaerensis. This distribution was obtained from the Japanese Whale Research Program under Special Permit in the Antarctic. Our results showed that the relative bias of the abundance estimators was large when covariates were misspecified and a survey region was stratified. Although stratification of the survey region is intended to produce a conventional line transect estimator with a smaller variance than that of non-stratified survey region, it also acts to increase the bias of the abundance estimate obtained from SDMs.     

Decreasing dietary lipids improves larval survival and growth of Japanese eel Anguilla japonica

Shark eggs-based diet is the only diet by which eel larvae can grow to glass eels in captivity. However, the high level of lipids in the diet is suggested to negatively affect eel larvae. This paper examines the effect of defatted shark eggs (DSE) and hen egg yolk (HY) on growth and survival of larvae of Japanese eel Anguilla japonica. Lyophilized shark egg and commercial HY were defatted with n-hexane, and four experimental diets were prepared using both defatted and untreated shark eggs and HY. Larvae were reared for 3 weeks by feeding the experimental diets. The highest survival rate was observed in the larvae fed DSE, and larvae fed HY showed the lowest survival rate. The best growth was found in larvae fed DSE, followed by shark eggs and defatted HY, and the worst growth was in HY-fed larvae. These results show that decreasing dietary lipids improves the nutritional value of both shark eggs and HY for eel larvae. Regulation of the dietary lipid level may positively affect the larval performance of eels by combination of ingredients with a low lipid content.     

Automated acoustic method for counting and sizing farmed fish during transfer using DIDSON

Counting and sizing large farmed fish such as tuna is often performed during their transfer from one net cage to another. Dual-frequency identification sonar (DIDSON) provides an automated fish counting and sizing tool. However, its counter and sizer are not suitable for measuring farmed fish because of net movements due to currents and subsequent frequent image breakups. This paper presents a fully automated acoustic method to count and size farmed fish during fish transfer by using DIDSON imaging. The background is subtracted from the image after being stabilized by an image phase-only correlation method. The segmentation of the fish is obtained by tracing the edges with a contour tracing method. To prevent recounting the same fish, a Kalman filter algorithm was designed and adapted to predict fish movements. Automated counting was performed by analyzing the spatiotemporal trajectory of the track. The separated fish images were searched for and body length was obtained by summing down the centerline segments from the head to the tail of the fish. The proposed system was verified using farmed yellowtail, Seriola quinqueradiata (mean total length 83.1 cm) to obtain a sizing error of mean total length within 2.4 cm.     

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.