Autophagy in health and disease: a double-edged sword

Autophagy, the process by which cells recycle cytoplasm and dispose of excess or defective organelles, has entered the research spotlight largely owing to the discovery of the protein components that drive this process. Identifying the autophagy genes in yeast and finding orthologs in other organisms reveals the conservation of the mechanism of autophagy in eukaryotes and allows the use of molecular genetics and biology in different model systems to study this process. By mostly morphological studies, autophagy has been linked to disease processes. Whether autophagy protects from or causes disease is unclear. Here, we summarize current knowledge about the role of autophagy in disease and health.     

Split luciferase complementation assay to detect regulated protein-protein interactions in rice protoplasts in a large-scale format

Background

The rice interactome, in which a network of protein-protein interactions has been elucidated in rice, is a useful resource to identify functional modules of rice signal transduction pathways. Protein-protein interactions occur in cells in two ways, constitutive and regulative. While a yeast-based high-throughput method has been widely used to identify the constitutive interactions, a method to detect the regulated interactions is rarely developed for a large-scale analysis.

Results

A split luciferase complementation assay was applied to detect the regulated interactions in rice. A transformation method of rice protoplasts in a 96-well plate was first established for a large-scale analysis. In addition, an antibody that specifically recognizes a carboxyl-terminal fragment of Renilla luciferase was newly developed. A pair of antibodies that recognize amino- and carboxyl- terminal fragments of Renilla luciferase, respectively, was then used to monitor quality and quantity of interacting recombinant-proteins accumulated in the cells. For a proof-of-concept, the method was applied to detect the gibberellin-dependent interaction between GIBBERELLIN INSENSITIVE DWARF1 and SLENDER RICE 1.

Conclusions

A method to detect regulated protein-protein interactions was developed towards establishment of the rice interactome.     

Molecular structures of gellan gum imaged with atomic force microscopy in relation to the rheological behavior in aqueous systems in the presence or absence of various cations

Aqueous solutions of gellan gum with comparable molecular masses but with different acyl contents were investigated by atomic force microscopy and rheological measurements in the presence or absence of various cations. For a high-acyl sample, no continuous network structures were identified microscopically, except in the presence of Ca (2+), where structural inhomogeneity was the highest in terms of the height distribution of molecular assemblies. Rheological thermal hysteresis between sol-gel transitions was detected in the presence of K (+) and Ca (2+), particularly Ca (2+). The storage modulus at 20 degrees C was larger in the order Na (+) < Ca (2+) < K (+). For a low-acyl sample, continuous network structures were identified in the presence of each cation, involving greater thermal hysteresis than the corresponding data for the high-acyl sample. Structural homogeneity was the highest in the presence of K (+). Thermal hysteresis and elasticity of the system were discussed in terms of continuousness and homogeneity of network structures.     

Effect of mitotane on pituitary corticotrophs in clinically normal dogs

OBJECTIVE: To evaluate the effects of mitotane administration on the function and morphology of pituitary corticotrophs in clinically normal dogs. ANIMALS: 12 clinically normal adult Beagles. PROCEDURES: Dogs were randomly assigned to the control group or the mitotane treatment group. In mitotane treatment group dogs, mitotane was administered for 1 month. In both groups, ACTH stimulation testing and corticotrophin-releasing hormone (CRH) stimulation testing were performed. Magnetic resonance imaging (MRI) of the pituitary gland and brain was performed in mitotane treatment group dogs before and after administration of mitotane. After CRH stimulation testing and MRI, dogs were euthanatized and the pituitary gland and adrenal glands were excised for gross and histologic examination. RESULTS: ACTH concentrations in mitotane treatment group dogs were significantly higher than in the control group dogs following CRH stimulation. Magnetic resonance imaging revealed that pituitary glands were significantly larger in treatment group dogs after administration of mitotane, compared with before administration. On gross and histologic examinations, the adrenal cortex was markedly atrophied. Immunohistochemistry revealed hypertrophy of corticotrophs in pituitary glands of mitotane treatment group dogs. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicate that inhibition of the adrenal cortex by continuous administration of mitotane leads to functional amplification and morphologic enhancement of corticotrophs in clinically normal dogs. In instances of corticotroph adenoma, hypertrophy of individual corticotrophs induced by mitotane may greatly facilitate enlargement of the pituitary gland and increases in ACTH secretion.     

Geographic variability in taxonomic composition,standing stock,and productivity of the mesozooplankton community around the Kuroshio Current in the East China Sea

Despite the low productivity that has been thought to characterize plankton communities in the western boundary current of the North Pacific Subtropical Gyre, many migratory fishes risk encountering low food availability during crucial life history stages by reproducing and recruiting in the Kuroshio region (i.e., the Kuroshio Paradox). Here, we report on geographic variability in taxonomic composition, biomass, and productivity of the mesozooplankton community in the Kuroshio Current and neighboring waters in the East China Sea. Calanoid copepods were the most abundant mesozooplankton taxon throughout our sampling stations. Small copepods, which include nauplii and poecilostomatoids, and gelatinous metazoans were the next most abundant. Seasonal variability in mesozooplankton standing stock (i.e., abundance and biomass) and productivity (i.e., production rate and protein synthetase activity) exceeded spatial variability across the stations and regions. The mesozooplankton community was characterized by high biomass and production rates in the summer, as well as high abundance and protein synthetase activity in the fall. No significant differences were found for mesozooplankton standing stock and productivity in the Kuroshio Path relative to those on the continental shelf or on the outside of the Kuroshio Path. Our results indicate that the standing stock and productivity of the mesozooplankton community in the Kuroshio Path are equivalent to those on the continental shelf, and that these communities are supported by small copepods and gelatinous zooplankton. We suggest that the mesozooplankton standing stocks and productivity provide adequate food availability for migratory fishes in the Kuroshio and neighboring waters in the East China Sea.     

Estimation of nutrient excretion factors of broiler and layer chickens in Japan

We estimated the nitrogen (N), phosphorus (P), potassium (K) and magnesium (Mg) excretion factors of broiler and layer chickens in Japan, using two approaches and the latest data available. In the top‐down approach, we determined the nutrient amounts in the feeds and those in the products (i.e. the liveweight gain, eggs), and the national nutrient excretions were determined as the difference between these amounts. We then calculated the nutrient excretion factors by dividing the national excretions by the number of animals. In the bottom‐up approach, we calculated the amounts of nutrients in the feed and product per head using productivity parameters (feed conversion ratio, etc.). The differences between these amounts were considered the nutrient excretion factors. The average nutrient excretion factors of broilers (g/day/head) estimated using the top‐down and bottom‐up approaches were: N, 1.40 and 1.87; P, 0.36, 0.50; K, 0.54, 0.77; Mg, 0.13, 0.18, respectively. The excretion factors obtained by the top‐down approach can be used to calculate the national/regional excretions. The two approaches resulted in almost the same excretion factors for layers, and the average nutrient excretion factors of layers (g/day/head) estimated were: N, 2.20; P, 0.55; K, 0.68; Mg, 0.23. The estimated excretion factors for N (only) are smaller than the reported factors.     

Relationship Between Physicochemical Properties of Starches and White Salted Noodle Quality in Japanese Wheat Flours

This study describes the effect of starch properties of Japanese wheat flours on the quality of white salted noodles (WSN). Starch was isolated from 24 flours of 17 Japanese cultivars and amylose content was determined along with pasting properties by Rapid Visco Analyser (RVA), thermal properties by differential scanning calorimetry (DSC), and the distribution of amylopectin chain length by high‐performance anion exchange chromatography (HPAEC). Twenty flours were used to prepare WSN. As expected, 5–6% lower amylose content was associated with good WSN quality (higher scores in softness, elasticity, and smoothness). RVA analysis indicated that the pasting temperature had the greatest influence on WSN quality, while breakdown and setback showed slight effects on WSN quality. DSC results showed that lower endothermal enthalpy (ΔH) in the amylose‐lipid complex was associated with good WSN quality. Chainlength distribution of amylopectin by HPAEC was not an important factor in relation to WSN quality.     

Properties of Starches from Several Low‐Amylose Rice Cultivars

The starch properties of five low‐amylose rice cultivars, Yawarakomachi, Soft 158, Hanabusa, Aya, and Snow Pearl, were compared with those of two normal amylose rice cultivars, Nipponbare and Hinohikari. There were no large differences in the distributions of the amylopectin chain length determined by high‐performance anion‐exchange chromatography, and the starch gelatinization properties determined by differential scanning calorimetry, between normal and low‐amylose rice cultivars. Results obtained using rapid viscosity analysis indicated that low‐amylose rice starches had lower peak viscosity, breakdown, and setback values than normal amylose rice starches. Starch granules from low‐amylose rice cultivars had a higher susceptibility to glucoamylase than those from normal amylose rice cultivars. The results of this study showed some differences between normal and low‐amylose rice starches in pasting properties and enzymatic digestibility.