Single-molecule speckle analysis of actin filament turnover in lamellipodia

Lamellipodia are thin, veil-like extensions at the edge of cells that contain a dynamic array of actin filaments. We describe an approach for analyzing spatial regulation of actin polymerization and depolymerization in vivo in which we tracked single molecules of actin fused to the green fluorescent protein. Polymerization and the lifetime of actin filaments in lamellipodia were measured with high spatial precision. Basal polymerization and depolymerization occurred throughout lamellipodia with largely constant kinetics, and polymerization was promoted within one micron of the lamellipodium tip. Most of the actin filaments in the lamellipodium were generated by polymerization away from the tip.     

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

Among allergenic foods, soybean is known as a food causing adverse reactions in allergenic patients. To clarify the validity of labeling, the specific and sensitive detection method for the analysis of the soybean protein would be necessary. The p34 protein, originally characterized to be p34 as an oil-body associated protein in soybean, has been identified as one of the major allergenic proteins and named Gly m Bd 30K. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of the soybean protein in processed foods was developed using polyclonal antibodies raised against p34 as a soybean marker protein and the specific extraction buffer for extract. The developed sandwich ELISA method was highly specific for the soybean protein. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed ELISA were 0.47 ng/mL (equivalent to 0.19 microg/g in foods) and 0.94 ng/mL (equivalent to 0.38 microg/g in foods), respectively. The recovery ranged from 87.7 to 98.7%, whereas the intra- and interassay coefficients of variation were less than 4.2 and 7.5%, respectively. This study showed that the developed ELISA method is a specific, precise, and reliable tool for the quantitative analysis of the soybean protein in processed foods.     

Effect of plant density on growth and yield of new soybean genotypes grown under early planting condition in southwestern Japan

Soybeans planted in early to mid-June (early) are less affected by rainfall during rainy season than those conventionally planted in early to mid-July in southwestern Japan. Also, narrow row cultivation is expected to increase soybean yield and save labor for inter-tillage and ridging. Field experiments were performed in 2014 and 2015 to test the effect of plant density (high, middle, and low) under early planting condition on growth, yield, and several agronomical traits of Sachiyutaka A1 and three new genotypes (Sakukei 155, Kanto 127 and Shikoku 15). Early planting was performed in mid- to late June, even though rainy season started in early June. Higher plant densities produced 13% greater yield than low plant density through an increase in biomass accumulation, especially at R5. Among yield components, only pods m^?2 was significantly and positively correlated with yield, indicating that an increase in pods m^?2 led to a greater yield with higher plant densities. The yields of Sachiyutaka A1 were relatively stable for two years, but the lodging resistance should be further improved. Shikoku 15 had greater yield potential and lodging resistance, but its resistance to damping-off disease should be improved. Sakukei 155 with medium plant density produced relatively high and stable yield with less lodging. Although the yield of Kanto 127 fluctuated between experimental years, this genotype showed higher yield potential in higher plant densities with less lodging in 2015. Thus, Sakukei 155 and Kanto 127 with high or medium density may be suitable for early planting in this region.     

Control mechanisms for producing antimicrobial factors in ruminant mammary gland

Mastitis, a symptom of inflammation in mammary tissue by infection with various kinds of bacteria, causes huge economic losses in the milk industry. One of the popular methods for treatment of mastitis is antibiotics, although this prohibits milk shipping and sometimes causes resistant microbes. Therefore, a new strategy to treat mastitis without antibiotics is eagerly required around the world. Antimicrobial factors belong to innate immunity and can start their function extremely early after bacterial stimulation. These factors have antimicrobial activity for a broad spectrum of bacteria. Elucidation of causal mechanisms and functions of antimicrobial factors in the mammary gland is thought to result in suitable methods for prevention and treatment of mastitis. Therefore, this review introduces traits of some antimicrobial factors and the mechanisms for expressing, producing and secreting them in the mammary gland. For antimicrobial factors, lingual antimicrobial peptide (LAP), S100A7, cathelicidin and lactoferrin are controlled in different sites and different time courses, suggesting that antimicrobial factors play different roles for local defense against bacterial infection in the mammary gland. These findings will contribute to the development of prevention and treatment methods for mastitis.