Substrate-specificity and toxicological significance of pyrethroid-hydrolyzing esterases of mouse liver microsomes

An esterase or esterases in acetone powder preparations of mouse liver microsomes hydrolyze the cyclopropanecarboxylate ester linkage of pyrethroid insecticide chemicals derived from primary alcohols. The rate of cleavage of (+)-trans-chrysanthemates with various alcohol moieties decreases in the following order: 5-propargyl-2-furylmethyl; 5-benzyl-3-furylmethyl (bioresmethrin); 3-phenoxybenzyl; tetrahydrophthalimidomethyl esters. The hydrolysis rate of benzylfurylmethyl esters with various acid moieties decreases in the order: (+)- or (?)-trans-chrysanthemate; (+)-trans-ethanochrysanthemate; tetramethylcyclopropanecarboxylate; (+)- or (?)-cis-chrysanthemate or (+)-cis-ethanochrysanthemate. The trans-isomers of chrysanthemates and ethanochrysanthemates are hydrolyzed from 2.6- to more than 50-fold more rapidly than the corresponding cis-isomers. This enzyme system does not hydrolyze secondary alcohol esters, i.e., allethronyl (+)-trans- and (+)-cis-chrysanthemates.On intraperitoneal administration to mice, the (+)-trans-chrysanthemate and -ethanochrysanthemate of benzylfurylmethanol are of very low toxicity relative to the corresponding (+)-cis-isomers and the tetramethylcyclopropanecarboxylate. S,S,S-tributyl phosphorotrithioate (DEF) pretreatment increases the toxicity of these five compounds by 2.6- to more than 188-fold, with the exception of bioresmethrin whose toxicity is not altered. When the toxicity is increased, it is probably the result of esterase inhibition since DEF strongly inhibits the esterase activity of fresh liver microsomes while the mixed-function oxidase system remains active. The oxidase system metabolizes the chrysanthemates more rapidly than the ethanochrysanthemates of benzylfuryl-methanol. Depending upon the pyrethroid involved, the esterase or the mixed-function oxidase system, or both may be responsible for limiting the toxicity of these pyrethroids to mice.     

Pesticide interactions: Effects of organophosphorus pesticides on the metabolism,toxicity, and persistence of selected pyrethroid insecticides

The organophosphorus pesticides profenofos, sulprofos, O-ethyl O-(4-nitrophenyl) phenylphosphonothioate (EPN), and S,S,S,-tributyl phosphorotrithioate (DEF) administered intraperitoneally to mice at 0.5 to 5 mg/kg strongly inhibit the liver microsomal esterase(s) hydrolyzing trans-permethrin. Profenofos, EPN, and DEF at 25 mg/kg increase the intraperitoneal toxicity of fenvalerate > 25-fold and of malathion > 100-fold. Topically applied profenofos, sulprofos, and DEF significantly synergize the toxicity of cis-cypermethrin to cabbage looper larvae and house fly adults but these phosphorus compounds are much less effective in synergizing the toxicity of trans-permethrin. The magnitude of synergism appears to depend on the species, organophosphorus compound, and pyrethroid involved. Profenofos, sulprofos, and EPN do not significantly alter the persistence of trans-permethrin on bean foliage.     

Evolution of the composition of a selected bitter Camembert cheese during ripening: release and migration of taste-active compounds.

The aim of this study was to add to the understanding of changes in taste that occur during the ripening of a bitter Camembert cheese by the evolution of its composition. Physicochemical analyses were performed on rind, under-rind, and center portions of a Camembert cheese selected for its intense bitterness. At each of the six steps of ripening studied organic acids, sugars, total nitrogen, soluble nitrogen, phosphotungstic acid soluble nitrogen, non-protein nitrogen, Na, K, Ca, Mg, Pi, Cl, and biogenic amines were quantified in each portion. Changes in cheese composition seemed to mainly result from the development of Penicillium camemberti on the cheese outer layer. Migration phenomena and the release of potentially taste-active compounds allowed for the evolution of saltiness, sourness, and bitterness throughout ripening to be better understood. Apart from taste-active compounds, the impact of the cheese matrix on its taste development is discussed.     

Mass spectrometry analysis of volatile compounds in raw meat for the authentication of the feeding background of farm animals

The authentication of the conditions of animal production, based on the analysis of meat commercial cuts, is a major challenge on both societal and analytical grounds. The aim of the present work was to propose a method for the extraction of the volatile compounds from ruminant raw muscles trimmed of fat and to assess by mass spectrometry-based techniques the relevance of these compounds for the authentication of the type of feeding offered to the animals. The first step of the study consisted of validating conditions of dynamic headspace (DH) extraction of volatile compounds that enabled us to minimize the appearance of heat-induced artifacts and to maximize the richness of the DH-gas chromatography-mass spectrometry profile (DH-GC-MS) of raw lamb muscle. An extraction temperature of 35 degrees C (vs 60 and 90 degrees C) and a sample mass of 6.25 g (vs 12.5, 25, and 50 g) were shown to be suitable. The second step aimed at identifying volatile compounds enabling us to discriminate muscle samples from 16 experimental lambs fed either concentrate (n = 8) or pasture (n = 8). Before, to carefully explore the information given by the DH-GC-MS signal, the MS spectra acquired along the chromatogram were summed and then converted in a virtual-DH-MS spectral fingerprint to have a quick overview of the discriminative potential of the volatile fraction. According to univariate (analysis of variance) and to multivariate (principal component analysis) data treatments performed on virtual-DH-MS fingerprints, the meat volatile fraction was relevant to reveal the type of feeding of the living animal. The detailed examination of the information given by the GC dimension showed that 33 volatile compounds among the 204 detected in the muscle by DH-GC-MS enabled us to discriminate the type of feeding of the lambs. The relevance of these results is discussed in light of previous studies performed on adipose tissues.     

Metabolite profiling of maize kernels--genetic modification versus environmental influence

A metabolite profiling approach based on gas chromatography-mass spectrometry (GC-MS) was applied to investigate the metabolite profiles of genetically modified (GM) Bt-maize (DKC78-15B, TXP 138F) and Roundup Ready-maize (DKC78-35R). For the comparative investigation of the impact of genetic modification versus environmental influence on the metabolite profiles, GM maize was grown together with the non-GM near-isogenic comparators under different environmental conditions, including several growing locations and seasons in Germany and South Africa. Analyses of variance (ANOVA) revealed significant differences between GM and non-GM maize grown in Germany and South Africa. For the factor genotype, 4 and 3%, respectively, of the total number of peaks detected by GC-MS showed statistically significant differences (p < 0.01) in peak heights as compared to the respective isogenic lines. However, ANOVA for the factor environment (growing location, season) revealed higher numbers of significant differences (p < 0.01) between the GM and the non-GM maize grown in Germany (42%) and South Africa (10%), respectively. This indicates that the majority of differences observed are related to natural variability rather than to the genetic modifications. In addition, multivariate data assessment by means of principal component analysis revealed that environmental factors, that is, growing locations and seasons, were dominant parameters driving the variability of the maize metabolite profiles.     

Factors influencing variability of proteolytic genes and activities in arable soils

Microorganisms, capable of proteolysis, are widely distributed in soil but almost nothing is known about the abundance of genes related to protein degradation and the regulation of their activity in terrestrial ecosystems. Therefore, the aim of this study was: (1) to quantify two bacterial genes involved in protein degradation, (2) to investigate factors affecting the abundance of these genes, and (3) to relate this data to potential proteolytic activities. For this purpose, an arable field in southern Germany under integrated management was studied. The uniformly managed field showed pronounced soil heterogeneity with four different soil types. In April, July and October 2003, soil samples were taken from the four soil types at three different depths. We applied a real-time PCR assay for quantification of subtilisin (sub) and neutral metalloprotease (npr) genes, both encoding for extracellular proteases, as well as the 16S rRNA gene representing a rough estimate of the size of the bacterial populations. Potential proteolytic activity was measured using casein as a substrate. Both soil type and time of sampling influenced the size and activity of the bacterial protease genes under investigation. Total nitrogen and carbon availability was, beside soil texture, the main factor responsible for the observed changes in the abundance of proteolytic genes and potential proteolytic activity. Whereas a positive relationship was found between sub and npr gene copy numbers and the number of 16S rRNA gene copies in all cases, a positive relationship between sub and npr coding genes and potential proteolytic activity was only found for sandy soils. This indicates that sandy soils cannot stabilize proteolytic enzymes and the activity of npr and sub genes is strictly dependent on the presence of the corresponding genes. In contrast, in clay soils proteolytic activity was not correlated with the abundance of the genes analyzed, probably due to the stabilization of the proteolytic enzymes.     

Effects of climate change on paddy water use efficiency with temporal change in the transplanting and growing season in South Korea

The aim of this study was to assess the effects of climate change on green and blue water use with temporal change in the transplanting and growing season for paddy fields in South Korea using the representative concentration pathway (RCP) 4.5 and 8.5 scenarios in 2025 (2011–2040), 2055 (2041–2070), and 2085 (2071–2100). The optimal transplant date determined by an accumulated temperature analysis, between 27 April and 21 May in 1995, was delayed until 29 June in 2085, and the average growing period was decreased by about 20 days in 2085. Changes in the transplanting season and growing period could influence the efficiency of future green water use and blue water savings. Approximately 810.5 Mm³ of green water was available during the 1995 transplanting season, but the predicted use of green water was 1524.7 Mm³ in future for the optimal transplanting season. In addition, 923.0 Mm³ of blue water savings can be expected in 2025. This study showed that appropriate change of transplanting season could mitigate the water requirements in paddy fields by optimizing phenology, improving green water efficiency, and blue water savings.     

Histochemical and functional correlations in anterior horn neurons of the cat spinal cord

The histochemical reaction for phosphorylase is completely lost from anterior horn neurons rich in phosphorylase within 72 hours after proximal or distal axonal section. Using this new type of axonal reaction as a marking technique in the anterior horn of the seventh lumbar spinal cord segment of the cat, we demonstrated that (i) alpha motor neurons of slow twitch motor units, like those of fast twitch motor units, are rich in phosphorylase and poor in succinate dehydrogenase, and (ii) interneurons and Renshaw neurons are rich in succinate dehydrogenase and poor in phosphorylase. Gamma motor neurons, because of their small size, are considered to be rich in succinate dehydrogenase and poor in phosphorylase. Thus, anterior horn neurons capable of higher firing frequencies (Renshaw neurons, interneurons, and gamma motor neurons) are richer in mitochondrial oxidative enzyme activity as marked by succinate dehydrogenase. Those firing at lower frequencies (both types of alpha motor neurons) are richer in phosphorylase activity and glycogen content and, thus, apparently better equipped for anaerobic glycolysis.     

Effects of Three Different Liquid Additives Mixed with Whole Oats or Rolled Oats on the Generation of Airborne Particles from an Experimental Simulating Horse Feeding

This study was designed to analyze the reduction potential of three different liquid additives (water, rapeseed oil, and molasses) in three different concentrations mixed with cleaned whole or rolled oats on the generation of airborne particles under standardized laboratory conditions. In addition, the influence of two different drop heights (20 and 40 cm) was analyzed. Airborne particle concentrations were detected online with the gravimetrically measuring analyzer TEOM 1400a that was equipped successively with different inlets to measure the particle fractions PM₂₀, PM₁₀, and PM_2.5. The mixing of oats with just 1% (w/w) of each of the liquid additives led to a significant reduction in airborne particle generation in all three particle fractions with respect to the samples without additives (P < .0001). By the addition of 1% oil, a reduction in the PM₂₀ fraction of 90.6% (120.3 μg/m³) could be achieved. The same dosage of water or molasses only resulted in a reduction of 60.4% (504.6 μg/m³) or 69.1% (393.2 μg/m³), respectively. In general, the highest significant reduction in the PM₂₀ fraction was achieved using 3% rapeseed oil: 96.5% as compared with 75.6% for 3% water and 81.9% for 3% molasses. The drop height only had a significant effect on the generation of the PM₁₀ fraction, with it being lower at 20 cm than at 40 cm (P = .0261).     

Modulation of neuronal interactions through neuronal synchronization

Brain processing depends on the interactions between neuronal groups. Those interactions are governed by the pattern of anatomical connections and by yet unknown mechanisms that modulate the effective strength of a given connection. We found that the mutual influence among neuronal groups depends on the phase relation between rhythmic activities within the groups. Phase relations supporting interactions between the groups preceded those interactions by a few milliseconds, consistent with a mechanistic role. These effects were specific in time, frequency, and space, and we therefore propose that the pattern of synchronization flexibly determines the pattern of neuronal interactions.