A Burkholderia pseudomallei toxin inhibits helicase activity of translation factor eIF4A

The structure of BPSL1549, a protein of unknown function from Burkholderia pseudomallei, reveals a similarity to Escherichia coli cytotoxic necrotizing factor 1. We found that BPSL1549 acted as a potent cytotoxin against eukaryotic cells and was lethal when administered to mice. Expression levels of bpsl1549 correlate with conditions expected to promote or suppress pathogenicity. BPSL1549 promotes deamidation of glutamine-339 of the translation initiation factor eIF4A, abolishing its helicase activity and inhibiting translation. We propose to name BPSL1549 Burkholderia lethal factor 1.     

Wnt3a-mediated formation of phosphatidylinositol 4,5-bisphosphate regulates LRP6 phosphorylation

The canonical Wnt-beta-catenin signaling pathway is initiated by inducing phosphorylation of one of the Wnt receptors, low-density lipoprotein receptor-related protein 6 (LRP6), at threonine residue 1479 (Thr1479) and serine residue 1490 (Ser1490). By screening a human kinase small interfering RNA library, we identified phosphatidylinositol 4-kinase type II alpha and phosphatidylinositol-4-phosphate 5-kinase type I (PIP5KI) as required for Wnt3a-induced LRP6 phosphorylation at Ser1490 in mammalian cells and confirmed that these kinases are important for Wnt signaling in Xenopus embryos. Wnt3a stimulates the formation of phosphatidylinositol 4,5-bisphosphates [PtdIns (4,5)P2] through frizzled and dishevelled, the latter of which directly interacted with and activated PIP5KI. In turn, PtdIns (4,5)P2 regulated phosphorylation of LRP6 at Thr1479 and Ser1490. Therefore, our study reveals a signaling mechanism for Wnt to regulate LRP6 phosphorylation.     

Carotenogenesis up-regulation in Scenedesmus sp. using a targeted metabolomics approach by liquid chromatography-high-resolution mass spectrometry

Carotenoids have potent antioxidant activity as well as therapeutic value, and their formation has been seen to be induced in algae by stress, including high-salt culture conditions. A differential profiling of carotenoids was conducted using a targeted metabolomics approach with accurate mass data generated by liquid chromatography-electrospray-time-of-flight (LC-ESI-TOF) mass spectrometry followed by postacquisition filtering based on isotope patterns and mass defects to detect carotenoids up-regulated in Scenedesmus sp. exposed to high-salt conditions. Algal cultures treated with high concentrations of sodium acetate or sodium chloride were found to cause an increase in various carotenoids. On the basis of differential analysis, astaxanthin and canthaxanthin increased upon salt treatment. Astaxanthin, in its free form and as fatty acid esters, was seen to increase in Scenedesmus sp. using accurate mass MS. A few other carotenoid compounds increased upon salt treatment, including echinenone and adonirubin, involved in the pathway of astaxanthin biosynthesis from β-carotene, as well as isomers of astaxanthin and canthaxanthin. A time course study of acetate treatment was done to observe the time-dependent up-regulation of carotenogenesis.     

Optimized denitrification bioreactor treatment through simulated drainage containment

In the design of wood-based, enhanced-denitrification bioreactors to treat nitrate in agricultural drainage, the consideration of the highly variable flow rates and nitrate concentrations inherent to many drainage systems is important. For optimized mitigation of these nitrate loads, it may be best to contain drainage water prior to treatment in order to facilitate longer, more constant retention times rather than to allow cycles of flushing and dry periods in the denitrification bioreactor. Simulated containment prior to bioreactor treatment compared to passing drainage directly through a bioreactor was investigated with the use of six pilot-scale denitrification bioreactors constructed with plywood and filled with Pinus radiata woodchips at Massey University No. 4 Dairy Farm (Palmerston North, New Zealand). Initial bromide tracer tests were followed with a series of five simulated drainage events each at successively declining inflow nitrate concentrations. During each drainage event, three pilot bioreactors received a simulated hydrograph lasting 1.5 days (Non-Containment treatment) and three pilot bioreactors received the same total drainage volume treated over 4 days at a constant flow rate (i.e. constant retention time; Containment treatment). Results showed significantly different total mass removal efficiencies of 14.0% vs. 36.9% and significantly different removal rates of 2.1 g N m⁻³ day⁻¹ vs. 6.7 g N m⁻³ day⁻¹ for the Non-Containment and Containment treatments, respectively, which indicated that treating drainage at constant retention times provided more optimized nitrate removal. While this work was done to evaluate treatment under New Zealand drainage conditions, it also provides valuable information for optimizing agricultural drainage denitrification bioreactor performance in general.     

A comparison of the relative toxicity of bone meal and other P sources used as remedial treatments to the earthworm Eisenia fetida

It has been suggested that sources of P could be used to remediate metal-contaminated soil. The toxicity of four potential P sources, potassium hydrogen phosphate (PHP), triple superphosphate (TSP), rock phosphate (RP) and raw bone meal (RBM) to Eisenia fetida was determined. The concentration of P that is statistically likely to kill 50% of the population (LC50) for PHP, TSP and RBM was determined in OECD acute toxicity tests. 14 day LC50s expressed as bulk P concentration lay in the range 3319–4272 mg kg^?1 for PHP, 3107–3590 mg kg^?1 for TSP and 1782–2196 mg kg^?1 for RBM (ranges present the 95% confidence intervals). For PHP and TSP mortality was significantly impacted by the electrical conductivity of the treated soils. No consistent relationship existed between mortality and electrical conductivity, soil pH and available (Olsen) P across the PHP, TSP and RBM amendment types. In RP toxicity tests mortality was low and it was not possible to determine a LC50 value. Incineration of bone meal at temperatures between 200 and 300 °C, pre-washing the bone meal, co-amendment with 5% green waste compost and delaying introduction of earthworms after bone meal amendments by 21 days or more led to significant reductions in the bone meal toxicity. These results are consistent with the toxicity being associated with the release and/or degradation of a soluble organic component present in raw bone meal. Bone meal can be used as an earthworm-friendly remedial amendment in metal-contaminated soils but initial additions may have a negative effect on any earthworms surviving in the contaminated soil before the organic component in the bone meal degrades in the soil.     

Development of a novel diagnostic test for detection of bovine viral diarrhea persistently infected animals using hair

The purpose of this study was to determine whether manually plucked hairs might serve as an alternative sample for a quantitative real time polymerase chain reaction (qRT-PCR) testing. Twenty three, 1~3 week old, non-bovine viral diarrhea virus (BVDV) vaccinated calves, found to be positive for BVDV by immunohistochemical staining, were selected and hairs were manually plucked from the ear. qRT-PCR was performed on samples consisting of more than 30 hairs (30~100) and whole blood. All 23 animals were positive for the virus by qRT-PCR performed on the whole blood and when samples of more than 30 hairs were assayed. Additionally, qRT-PCR was performed on groups of 10 and 20 hairs harvested from 7 out of 23 immunohistochemical staining-positive calves. When groups of 20 and 10 hairs were tested, 6 and 4 animals, respectively, were positive for the virus.     

The Effect of Lake-Water Infiltration on the Acidity and Base Cation Status of Forest Soil

In this article, the effects of the artificialrecharging of groundwater by infiltrating surface water throughforest soil, i.e. sprinkling infiltration, on the acidity andbase cation status of the soil are described. The study wascarried out in the Ahvenisto esker area, Hämeenlinna,southern Finland, during 1996–1998. The sample plots werelocated in a 110- to 160-yr-old Scots pine (Pinussylvestris L.) stand. The site was classified as the Oxalis-Maianthemum type. The soil consisted of a mixture oftill and glaciofluvial sediments. The pH of the organic layerincreased from 4.7 to >6.5 soon after the start of irrigationon the infiltration plot. The pH of the 0–10 cm mineral soillayer also increased from 4.9 to 6.4 as a result ofinfiltration. Sprinkling infiltration increased theexchangeable Ca and Mg concentrations in the organic anduppermost mineral soil layers. The output of Ca and Mg inpercolation water from the 0–100 cm thick layer was lower thanthe input to the soil surface via irrigation in 1996. Theretention of Ca and Mg on cation exchange sites took placewithin a relatively short period of time, since retention wasobserved only in 1996 but no longer in 1997 or 1998 indicatingsaturation of the cation exchange sites by base cations. Lakewater infiltration leads to the neutralisation of forest soilacidity, and increases the capacity of the soil to withstandacidic inputs by increasing the concentrations of exchangeablebase cations on cation exchange sites in the soil.     

Production of cod (Gadus morhua) muscle hydrolysates. Influence of combinations of commercial enzyme preparations on hydrolysate peptide size range

The complement of enzyme activities of a selection of commercial protease preparations were determined using fluorogenic substrates. Alcalase was used in combination with other commercial enzyme preparations to produce cod muscle (Gadus morhua) hydrolysates. Each muscle hydrolysate was characterized with respect to the percentage degree of hydrolysis (DH %), peptide molecular weight range, and free amino acid content. The enzyme preparations containing predominantly protease or endopeptidase activities achieved high DH % and produced significant amounts of peptides below a molecular weight of 3000. Alcalase combined with exopeptidase-rich preparations produced hydrolysates rich in low-molecular-weight peptides. Selecting combinations of enzyme preparations with complementary activity profiles could be used to manipulate the peptide molecular weight profile of hydrolysates.     

Fecal streptococci recoveries in different marine areas

The usual media and procedures were followed to measure the concentration of fecal streptococci (MPN on Azide Dextrose and Ethyl Violet Azide broths, membrane filtration on m-Enterococcus, KF and Pfizer Selective Enterococcus agars and according to the mE procedure) in samples collected along two different marine areas. The results were evaluated on the basis of three parameters: total concentrations, number of enterococci-like colonies (namely colonies gram positive, catalase negative, coccus shaped) and rate of strictly named fecal streptococci. From the results it appears that the various media and procedures employed gave different yields and their capacity to measure fecal streptococci varies according to the origin of samples. The accompanying bacterial flora may play an important role on the selectivity of each technique to measure the fecal streptococci.     

Modified method for the analysis of free fatty acids in fish

The objective of this study was the development of a method for the quantification of free fatty acids (FFA) using less aggressive reactants against the handler and the environment than those used in the classic method of Lowry and Tinsley. The modified procedure is a variation of the Lowry and Tinsley method employing cyclohexane in place of benzene. The use of benzene is prohibited in certain work processes and laboratories, and the competent authority in each country is actively promoting research into harmless or less harmful products that could replace benzene. A comparison with the traditional AOCS titration method for oil analysis was performed. FFA content in mackerel frozen at -10 degrees C was measured according to the three methods over a 12 month period. The results showed similar values, and good correlations were obtained.