Genomic variability among globally distributed isolates of equine arteritis virus.

Equine arteritis virus (EAV), a non-arthropod borne togavirus, has been shown to have a global distribution. To date, no major antigenic variation has been demonstrated between EAV isolates from different geographic origins. In this study, the genomic RNA of EAV isolates obtained from horses of different breeds in various countries around the world was oligonucleotide fingerprinted. Comparisons of these fingerprints were used to determine the extent of genomic variation among such isolates. Comparisons among isolates from North American horses revealed, for the most part, oligonucleotide homologies of less than 60%. Only 29 of the 98 comparisons revealed greater than 60% oligonucleotide homology. Nonetheless, several comparisons indicated a close epidemiologic relationship between isolates from horses of different breeds located in different states. Though all European isolates were of Standardbred origin and were from horses located in northern European countries, the majority had oligonucleotide homologies of less than 60%. Where oligonucleotide homology was apparent, it was, with one exception, greater than 70%. The two isolates from New Zealand had 93.2% oligonucleotide homology. This is indicative of an extremely close epidemiologic relationship. Comparisons between EAV isolates from around the world revealed oligonucleotide homologies between viruses from North America, Europe and New Zealand. In several instances, this homology was greater than 70% and in one case greater than 80%. No oligonucleotide homology was evident in comparisons involving the virus from South Africa. The high level of genomic conservation between certain EAV isolates of disparate geographic origins may reflect dissemination of the virus associated with the international movement of horses. The extent of genomic variation demonstrated between most of the EAV isolates used in this study confirms the need for further investigation of genomic heterogeneity among strains of this virus before techniques that rely upon nucleic acid hybridization can be effectively applied as diagnostic procedures.     

Long-term application of animal slurries to grassland alters soil cation balance

Abstract. Soil samples from a 32-year grassland field experiment were taken from 0–5, 5–10, and 10–15 cm soil depths in February 2002. Plots received annual treatments of unamended control, mineral fertilizer, three rates of pig slurry and three rates of cow slurry, each with six replicates. Samples were analysed for cation exchange capacity (CEC), exchangeable cations (Na⁺, K⁺, Ca²⁺, Mg²⁺), pH and Olsen P. Exchangeable sodium percentage (ESP) was calculated as a sodicity indicator. Mean ESP was generally greater for slurry treatments than the control, with a trend of increasing ESP with application rate. This was particularly marked for cow slurry. At 0–5 cm depth ESP increased from 1.18 in the control to 1.75 at the highest rate of pig slurry and 5.60 at the highest rate of cow slurry. Similar trends were shown for CEC, exchangeable Na⁺, K⁺ and Mg²⁺, Ca²⁺ and Olsen P. The build-up of soil P due to slurry applications, together with this combination of physical and chemical factors, may increase the risk of P loss to surface waters, particularly from soils receiving high rates of cow slurry.     

Coagulopathic Effects and Therapy of Brodifacoum Toxicosis in Dogs

The clinical signs and laboratory changes of brodifacoum (BDF) intoxicated dogs and their response to vitamin K1 treatment were examined. Brodifacoum, a second-generation anticoagulant rodenticide, was fed to four dogs for 3 consecutive days producing a cumulative dose of 1.1 mg BDF/kg body weight. Clinical observations of the animals were made daily throughout the study. Monitored laboratory parameters included: one-stage prothrombin time (OSPT), activated partial thromboplastin time (APTT), activated coagulation time (ACT), complete blood counts, thrombocyte counts, and serum chemistry values. Response to vitamin K1 therapy was evaluated clinically and by laboratory tests. Serum BDF concentrations were monitored. Inappetence and hemorrhagic tendencies were exhibited by day 5 postrodenticide exposure. One-stage prothrombin time, APTT, and ACT were 25% greater than time zero values at 24, 24, and 72 hours postdosing, respectively. All laboratory parameters returned to normal within 48 hours of initiating vitamin K1 therapy (0.83 mg/kg orally, TID for 5 days). Serum brodifacoum concentrations were highest (1065-1215 ng/mL) during the 3 days after BDF dosing and were detectable (3.0-7.5 ng/mL) until day 24 postexposure. A mean BDF elimination half-life of 6 +/- 4 days was observed.     

Soluble organic nitrogen in agricultural soils

 The existence of soluble organic forms of N in rain and drainage waters has been known for many years, but these have not been generally regarded as significant pools of N in agricultural soils. We review the size and function of both soluble organic N extracted from soils (SON) and dissolved organic N present in soil solution and drainage waters (DON) in arable agricultural soils. SON is of the same order of magnitude as mineral N and of equal size in many cases; 20–30 kg SON-N ha^–1 is present in a wide range of arable agricultural soils from England. Its dynamics are affected by mineralisation, immobilisation, leaching and plant uptake in the same way as those of mineral N, but its pool size is more constant than that of mineral N. DON can be sampled from soil solution using suction cups and collected in drainage waters. Significant amounts of DON are leached, but this comprises only about one-tenth of the SON extracted from the same soil. Leached DON may take with it nutrients, chelated or complexed metals and pesticides. SON/DON is clearly an important pool in N transformations and plant uptake, but there are still many gaps in our understanding. Received: 10 June 1999     

Short-term effects of nitrogen on methane oxidation in soils

 The short-term effects of N addition on CH₄ oxidation were studied in two soils. Both sites are unfertilized, one has been under long-term arable rotation, the other is a grassland that has been cut for hay for the past 125 years. The sites showed clear differences in their capacity to oxidise CH₄, the arable soil oxidised CH₄ at a rate of 0.013 μg CH₄ kg^–1 h^–1 and the grassland soil approximately an order of magnitude quicker. In both sites the addition of (NH₄)₂SO₄ caused an immediate reduction in the rate of atmospheric CH₄ oxidation approximately in inverse proportion to the amount of NH₄ ⁺ added. The addition of KNO₃ caused no change in the rate of CH₄ oxidation in the arable soil, but in the grassland soil after 9 days the rate of CH₄ oxidation had decreased from 0.22 μg CH₄ kg^–1 h^–1 to 0.13 μg CH₄ kg^–1 h^–1 in soil treated with the equivalent of 192 kg N ha^–1. A ¹⁵N isotopic dilution technique was used to investigate the role of nitrifiers in regulating CH₄ oxidation. The arable soil showed a low rate of gross N mineralisation (0.67 mg N kg^–1 day^–1), but a relatively high proportion of the mineralised N was nitrified. The grassland soil had a high rate of gross N mineralisation (18.28 mg N kg^–1 day^–1), but negligible nitrification activity. It is hypothesised that since there was virtually no nitrification in the grassland soil then CH₄ oxidation at this site must be methanotroph mediated. Received: 31 October 1997     

Synthesis of Novel Thin-Film Materials by Pulsed Laser Deposition

Pulsed laser deposition (PLD) is a conceptually and experimentally simple yet highly versatile tool for thin-film and multilayer research. Its advantages for the film growth of oxides and other chemically complex materials include stoichiometric transfer, growth from an energetic beam, reactive deposition, and inherent simplicity for the growth of multilayered structures. With the use of PLD, artificially layered materials and metastable phases have been created and their properties varied by control of the layer thicknesses. In situ monitoring techniques have provided information about the role of energetic species in the formation of ultrahard phases and in the doping of semiconductors. Cluster-assembled nanocrystalline and composite films offer opportunities to control and produce new combinations of properties with PLD.     

Side-on copper-nitrosyl coordination by nitrite reductase

A copper-nitrosyl intermediate forms during the catalytic cycle of nitrite reductase, the enzyme that mediates the committed step in bacterial denitrification. The crystal structure of a type 2 copper-nitrosyl complex of nitrite reductase reveals an unprecedented side-on binding mode in which the nitrogen and oxygen atoms are nearly equidistant from the copper cofactor. Comparison of this structure with a refined nitrite-bound crystal structure explains how coordination can change between copper-oxygen and copper-nitrogen during catalysis. The side-on copper-nitrosyl in nitrite reductase expands the possibilities for nitric oxide interactions in copper proteins such as superoxide dismutase and prions.     

The elemental composition of asteroid 433 eros: results of the NEAR-shoemaker X-ray spectrometer

We report major element composition ratios for regions of the asteroid 433 Eros imaged during two solar flares and quiet sun conditions during the period of May to July 2000. Low aluminum abundances for all regions argue against global differentiation of Eros. Magnesium/silicon, aluminum/silicon, calcium/silicon, and iron/silicon ratios are best interpreted as a relatively primitive, chondritic composition. Marked depletions in sulfur and possible aluminum and calcium depletions, relative to ordinary chondrites, may represent signatures of limited partial melting or impact volatilization.     

Diazaquinomycins E–G,Novel Diaza-Anthracene Analogs from a Marine-Derived Streptomyces sp.

As part of our program to identify novel secondary metabolites that target drug-resistant ovarian cancers, a screening of our aquatic-derived actinomycete fraction library against a cisplatin-resistant ovarian cancer cell line (OVCAR5) led to the isolation of novel diaza-anthracene antibiotic diazaquinomycin E (DAQE; 1), the isomeric mixture of diazaquinomycin F (DAQF; 2) and diazaquinomycin G (DAQG; 3), and known analog diazaquinomycin A (DAQA; 4). The structures of DAQF and DAQG were solved through deconvolution of X-Ray diffraction data of their corresponding co-crystal. DAQE and DAQA exhibited moderate LC₅₀ values against OVCAR5 of 9.0 and 8.8 μM, respectively. At lethal concentrations of DAQA, evidence of DNA damage was observed via induction of apoptosis through cleaved-PARP. Herein, we will discuss the isolation, structure elucidation, and biological activity of these secondary metabolites.