Optimization of Immobilized Gallium (III) Ion Affinity Chromatography for Selective Binding and Recovery of Phosphopeptides from Protein Digests

Although widely used in proteomics research for the selective enrichment of phosphopeptides from protein digests, immobilized metal-ion affinity chromatography (IMAC) often suffers from low specificity and differential recovery of peptides carrying different numbers of phosphate groups. By systematically evaluating and optimizing different loading, washing, and elution conditions, we have developed an efficient and highly selective procedure for the enrichment of phosphopeptides using a commercially available gallium(III)-IMAC column (PhosphoProfile, Sigma). Phosphopeptide enrichment using the reagents supplied with the column is incomplete and biased toward the recovery and/or detection of smaller, singly phosphorylated peptides. In contrast, elution with base (0.4 M ammonium hydroxide) gives efficient and balanced recovery of both singly and multiply phosphorylated peptides, while loading peptides in a strong acidic solution (1% trifluoracetic acid) further increases selectivity toward phosphopeptides, with minimal carryover of nonphosphorylated peptides. 2,5-Dihydroxybenzoic acid, a matrix commonly used when analyzing phosphopeptides by matrix-assisted laser desorption/ionization mass spectrometry was also evaluated as an additive in loading and eluting solvents. Elution with 50% acetonitrile containing 20 mg/mL dihydroxybenzoic acid and 1% phosphoric acid gave results similar to those obtained using ammonium hydroxide as the eluent, although the latter showed the highest specificity for phosphorylated peptides.     

Olfactory responses of adult Potato Tuber Moth,Phthorimaea operculella (Zeller) measured by attraction relative to the tomato leaf volatiles

The potato tuber moth (PTM) is an oligophagous herbivore and a severe pest of solanaceous crops in many countries of the world. Previously, we reported host expansion and damage potential of PTM on tomato, a congeneric crop of potato. Here we tested adult olfactory behaviour of PTM to leaves of five different cultivated tomato varieties including Moneymaker, Campari, Ailsa craig, LA3475 and E6203, and one wild species, Solanum pimpinellifolium. Tomato leaf hydro-distilled oils of Moneymaker, Campari, Ailsa craig, S. pimpinellifolium and E6203 showed strong attractiveness and LA3475 exhibited repulsiveness for adult PTM of both sexes in two-armed bioassays. Volatiles of Moneymaker, Campari, Ailsa Craig, S. pimpinellifolium and E6203 showed attractiveness for mated adult PTM of both sexes (> 70%). The extracted oil of all tomato leaves contains monoterpenes, sesquiterpenes, diterpenes and C₆ alcohols as well as other components. Phytol is one of the common and major compounds in all tomato varieties. Phytol showed weak attractiveness: 60.8–63.6% and 57.6–60.6% for male and female PTM, respectively when tested at concentrations of 0.1–10 mg/mL. Therefore, the attractiveness of tomato leaf volatiles might be due to synergistic effects of plant volatile mixtures. Identification PTM attractant volatile in tomato leaf is important in its control in environmentally friendly manner.     

A potential threat to tomato,a congener crop to potato from invaded potato tuber moth,Phthorimaea operculella (Zeller)

Potato tuber moth (PTM) is a primary pest of potato which had invaded into Korea in 1968 and its distribution was restricted in the southern part of the peninsula. Possibly due to the climate change, its distribution expanded toward the Northern part of South Korea including Gyounggi and Gangwon province where tomato production is high. Host expansion and damage potential of PTM was evaluated on tomato, a congeneric crop of potato. Three commercial varieties of tomato (Dafnis, Bangul and Jujube) and a potato (Superior-Sumi) were subjected to PTM larvae. The results showed that host suitability of tomato to PTM was lower than potato, but PTM could completely develop and reproduce on tomato leaves or fruits. Development time was varied relative to tomato varieties and plant age. PTM showed lower performance on the leaves and fruits of cherry type tomatoes. Higher survival was observed in the large sized fruit of tomato varieties. This study showed substantial evidence that the invaded PTM could damage the tomato cultivation.     

Fine Wood Decomposition Rates Decline with the Age of Tropical Successional Forests in Southern Mexico: Implications to Ecosystem Carbon Storage

Ecosystems - Wood decomposition in tropical forests is strongly linked to the terrestrial carbon cycle. Our understanding of the successional changes in wood decomposition in diverse tropical...     

Antigenic property of pediocin AcH produced by Pediococcus acidilactici H

Pediocin AcH, a bacteriocin of Pediococcus acidilactici H, inhibits the growth of several food spoilage and pathogenic bacteria. The antigenic property of partially purified pediocin AcH was tested by immunizing mice and a rabbit. Pediocin AcH was not immunogenic in these animals as determined by immunoblotting even after conjugation to bovine serum albumin. The non-immunogenic nature of pediocin AcH, its non-toxicity to laboratory animals and its hydrolysis by gastric proteolytic enzymes may be considered favourably in its possible use as a food preservative.     

Diet and macronutrient niche of Asiatic black bear (Ursus thibetanus) in two regions of Nepal during summer and autumn

Relatively little is known about the nutritional ecology of omnivorous Asiatic black bears (Ursus thibetanus) in Nepal. We characterized the diet of black bears in two seasons (June–July, “summer”; and October–November “autumn”) and two study areas (Dhorpatan Hunting Reserve [DHR]; and Kailash Sacred Landscape [KSL]). We then conducted nutritional analysis of species consumed by black bears in each study area, in combination with nutritional estimates from the literature, to estimate the proportions of macronutrients (i.e., protein [P], lipid [L], and carbohydrate [C]) in the seasonal bear foods and diets, as well as their macronutrient niche breadth. We found that bamboo (Arundinaria spp.) had the highest relative frequency in both study areas and seasons. Ants and termites were found in DHR diets, but not KSL diets. One anthropogenic crop was found in DHR summer diets (Zea mays) and two were found in KSL summer diets (Z. mays; and Kodo millet [Paspalum scrobiculatum]). Other than insects, no animal prey was found in either diet. The proportions of macronutrients in diets (i.e., realized macronutrient niches) were relatively high in carbohydrate for both study areas and seasons: DHR_summer 24.1P:8.7L:67.2C; KSL_summer 16.7P:8.2L:75.1C; DHR_autumn 21.1P:10.5L:68.4C; KSH_autumn 19.0P:11.0L:70.0C. Macronutrient niche breadth was 3.1 × greater in the DHR than KSL during summer, and 4.0 × greater in the autumn, primarily due to the higher proportion of lipid in ants and termites relative to plant foods. Within‐study area differences in niche breadth were greater during summer than autumn; in the KSH the macronutrient breadth was 1.4 × greater in summer, while in the DHR it was 1.1 × greater in summer. Similarity in dietary macronutrient proportions despite differences in foods consumed and niche breadth are suggestive of foraging to reach a preferred macronutrient balance.     

PHYTOACCUMULATION OF ANTIMICROBIALS BY HYDROPONIC CUCURBITA PEPO

Consumer use of antimicrobial-containing products continuously introduces triclocarban and triclosan into the environment. Triclocarban and triclosan adversely affect plants and animals and have the potential to affect human health. Research examined the phytoaccumulation of triclocarban and triclosan by pumpkin (Cucurbita pepo cultivar Howden) and zucchini (Cucurbita pepo cultivar Gold Rush) grown hydroponically. Pumpkin and zucchini were grown in nutrient solution spiked with 0.315 μg/mL triclocarban and 0.289 μg/mL triclosan for two months. Concentrations of triclocarban and triclosan in nutrient solutions were monitored weekly. At the end of the trial, roots and shoots were analyzed for triclocarban and triclosan. Research demonstrated that pumpkin and zucchini accumulated triclocarban and triclosan. Root accumulation factors were 1.78 and 0.64 and translocation factors were 0.001 and 0.082 for triclocarban and triclosan, respectively. The results of this experiment were compared with a previous soil column study that represented environmentally relevant exposure of antimicrobials from biosolids and had similar root mass. Plants were not as efficient in removing triclocarban and triclosan in hydroponic systems as in soil systems. Shoot concentrations of antimicrobials were the same or lower in hydroponic systems than in soil columns, indicating that hydroponic system does not overpredict the concentrations of antimicrobials.     

Predictive habitat modelling to estimate petrel breeding colony sizes: Sooty shearwaters (Puffinus griseus) and mottled petrels (Pterodroma inexpectata) on Whenua Hou Island

Abstract

Between 2001 and 2006, we systematically sampled the entire coast of Whenua Hou, a rugged offshore island in southern New Zealand, to estimate the population densities of sooty shearwaters (Puffinus griseus) and mottled petrels (Pterodroma inexpectata) by counting the entrances of breeding burrows. A two‐step regression modelling process using binomial errors was used to predict the presence of a colony, and a normal general linear model was used to predict the density of entrances within colonies. Aerial photography, GIS and a Digital Elevation Model were used to extract relevant habitat and location variables, and a combination of both regression models was used to predict the density of breeding burrows within each 5.32 m² pixel on the island. This complex GIS and habitat prediction modelling approach gave population estimates very similar to a more traditional simple area extrapolation method and gave no improvement in precision. However, correction for the slope of the land increased our simple area estimates of population size by 11%. We estimate populations of sooty shearwater and mottled petrel breeding pairs at 173 000 (162 000–190 000) and 160 000 (123 000–197 000) respectively. Based on this number of breeding pairs, we calculate that Whenua Hou supports a total population of 868 000 (554 000–1 270 000) sooty shearwaters. Our estimate of the total mottled petrel population 202 000 pairs (162 000–242 000) is comparable with the only published estimate, but could be an underestimate because mottled petrels are sometimes found in large burrows. More research for robust estimation of population trends is needed to assess the conservation status of mottled petrels.