Electrophysiological identification of site-insensitive mechanisms in knockdown-resistant strains (kdr,super-kdr) of the housefly larva (Musca domestica)

The effects of pyrethroids were studied upon isolated segmental nerves and neuromuscular junctions in both susceptible (Cooper) and knockdown-resistant (kdr; super-kdr) strains of housefly larvae (Musca domestica L.). Isolated segmental nerves contained neither cell bodies nor synaptic contacts; thus, any effects of pyrethroids were attributed solely to their actions upon voltage-dependent Na⁺ channels. Threshold concentrations of the type II pyrethroid, deltamethrin, required to elevate the spontaneous firing rate of these nerves were determined. Both resistant strains were about ten times less sensitive to deltamethrin than the susceptible strain, but insensitivity of super-kdr nerves was no greater than in the less resistant kdr strain. At neuromuscular junctions, the minimum concentrations of pyrethroids needed to trigger massive increases in the frequency of miniature excitatory postsynaptic potentials (mEPSPs) were determined for deltamethrin and the type I pyrethroid, fenfluthrin. With fenfluthrin there was no detectable difference between the junctions of kdr and super-kdr strains, which were both about ten-fold less sensitive than Cooper junctions. With deltamethrin, kdr junctions were about 30 times less sensitive than those of Cooper; super-kdr junctions were dramatically insensitive to deltamethrin, being some 10000- and 300-fold less sensitive than those of Cooper and kdr respectively. Thus, in the synaptic assay, super-kdr conferred an extension in resistance over kdr only against the type II pyrethroid, it being ineffective against fenfluthrin. We suggest that kdr resistance comprises at least two site-insensitive areas within the nervous system. One involves insensitivity of the Na⁺ channel and has similar efficacy in both kdr and super-kdr strains against type I and II pyrethroids; the other is associated with the presynaptic terminal and is particularly effective in super-kdr resistance against type II pyrethroids. The latter could be associated with Ca²⁺-activated phosphorylation of proteins involved with neurotransmitter release. Such phosphorylation reactions are known to be perturbed by pyrethroids, especially by type II compounds.     

Grain composition of Virginia winter barley and implications for use in feed,food, and biofuels production

Grain compositional components impacting barley (Hordeum vulgare L.) use in food, feed and fuel products, must be combined with improved agronomic traits to produce a commercially viable barley cultivar. Little current information is available on grain composition and variability among winter barley genotypes. This study was conducted to determine the variability among modern hulled and hulless winter barley genotypes in grain composition. Barley types varied significantly in grain and kernel weight, starch, beta-glucan, oil and ash content, but not in protein concentration. Hulless barley had significantly higher grain test weight and starch concentration than hulled and malting types, and significantly higher beta-glucan than malting barley. Hulless barley had significantly lower kernel weights, oil, and ash concentrations than hulled and malting types. Higher starch and lower fiber and ash in grain of hulless barley versus hulled feed or malting type barley are characteristics that increase hulless barley desirability. Selection for high starch concentration among all barley types is feasible and will facilitate development of barley cultivars better suited for use in feed, malt, and ethanol production.     

Cardiovascular and autonomic nervous effects of edrophonium and atropine combinations during neuromuscular blockade antagonism in sheep

ObjectiveTo study heart rate (HR), arterial blood pressure (BP) and autonomic nervous (AN) effects of edrophonium–atropine combinations during neuromuscular blockade (NMB) antagonism in sheep.Experimental design Randomized, prospective and experimental study.AnimalsSeventy-eight Scottish blackface ewes; mean age: 4.5 years; mean body mass: 54 kg.MethodsAfter induction with IV etomidate (0.5 mg kg⁻¹) and midazolam (0.5 mg kg⁻¹), anaesthesia was maintained with halothane and NMB produced with atracurium or mivacurium. In the first study (n = 53), the electrocardiographic (ECG), HR, BP and AN effects of low (40 μg kg⁻¹) and high (80 μg kg⁻¹) atropine doses combined with either of two edrophonium doses (0.5 or 1.0 mg kg⁻¹) were investigated. These variables were also measured in a second study when edrophonium (1.0 mg kg⁻¹) was administered 5 minutes before atropine (80 μg kg⁻¹) and vice versa. Data were analysed using one-way within-subjects and repeated measures anova.ResultsIn the first study, all combinations reversed NMB but significantly (p < 0.001) increased HR and BP within 2 minutes without arrhythmias. In the second study, edrophonium (1.0 mg kg⁻¹) significantly increased HR and BP, saliva flow (n = 1) and lung sounds (n = 3) and caused ECG changes (n = 1). Cardiovascular changes were partially reversed by atropine (80 μg kg⁻¹) administered 5 minutes later. Administered first, atropine (80 μg kg⁻¹) significantly decreased HR and BP effects which were fully (HR) and partially (BP) reversed by edrophonium (1 mg kg⁻¹) administered 5 minutes later.Conclusion and clinical relevance The cardiovascular effects of edrophonium and atropine were opposite to those reported in humans and dogs. Edrophonium (0.5 mg kg⁻¹) and atropine (80 μg kg⁻¹) caused the mildest HR changes without ECG and noncardiac AN disturbances, and is recommended for the antagonism of NMB in sheep.     

Development and evaluation of pyramiding lines carrying early or late heading QTLs in the indica rice cultivar ‘IR64’

The heading date is an important trait for determining regional and climatic adaptability in rice. To expand the adaptability of the indica rice cultivar ‘IR64’, we pyramided multiple early or late heading quantitative trait locus (QTLs) in the ‘IR64’ genetic background by crossing previously developed near-isogenic lines (NILs) with a single QTL for early or late heading. The effects of pyramiding QTLs were observed in three different climatic zones of the Philippines, Madagascar, and Japan. The early heading pyramiding lines (PYLs) headed 6.2 to 12.8 days earlier than ‘IR64’ while the late heading PYLs headed 18.8 to 27.1 days later than ‘IR64’. The PYLs tended to produce low grain yield compared to ‘IR64’. The low yield was not improved by combining SPIKE, which is a QTL that increases the number of spikelets per panicle. Conversely, ‘IR64-PYL(7+10)’ carrying Hd5 and Hd1 headed earlier, produced more tillers, and more panicles per m² than ‘IR64’, and mitigated the yield decrease in early heading. These results suggest that the effects of pyramided QTLs on heading date were consistent across various environments and PYLs could be used to enhance the adaptation of ‘IR64’ in other rice growing environments.     

New molecular markers linked to qualitative and quantitative powdery mildew and scald resistance genes in barley for dry areas

A partial genetic linkage map was constructed on 71 doubled-haploid lines derived from a cross between the barley lines Tadmor and WI2291 with 181 molecular markers. The segregating population was used to detect markers linked to the gene Mlg conferring resistance to powdery mildew (Erysiphe graminis f. sp. hordei) and to genes for quantitative resistance to scald (Rhynchosporium secalis). The gene Mlg on chromosome 4H was flanked by two AFLP markers at a distance of 2.0 and 2.4 cM, respectively. QTLs for resistance to scald were detected on chromosomes 2H and 3H. This association of molecular markers with qualitative and quantitative disease resistance loci represents a valuable starting-point for marker-assisted selection. This revised version was published online in July 2006 with corrections to the Cover Date.     

Recycling endosomes supply AMPA receptors for LTP

Long-term potentiation (LTP) of synaptic strength, the most established cellular model of information storage in the brain, is expressed by an increase in the number of postsynaptic AMPA receptors. However, the source of AMPA receptors mobilized during LTP is unknown. We report that AMPA receptors are transported from recycling endosomes to the plasma membrane for LTP. Stimuli that triggered LTP promoted not only AMPA receptor insertion but also generalized recycling of cargo and membrane from endocytic compartments. Thus, recycling endosomes supply AMPA receptors for LTP and provide a mechanistic link between synaptic potentiation and membrane remodeling during synapse modification.     

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.     

Chondrules with peculiar REE patterns: implications for solar nebular condensation at high C/O

Rare earth element (REE) data from two ordinary chondrite chondrules show distinct negative chondrite-normalized concentration anomalies of samarium, europium, and ytterbium. The peculiar patterns may be the result of REE gas/solid fractionation at an oxygen fugacity lower than has been assumed for the canonical solar nebula. We suggest that the two ordinary chondrite chondrules acquired the fractionated REE patterns by incorporation of highly reduced, ultrarefractory condensates in their precursors. This interpretation implies that high-temperature condensation processes occurred in nebular environments with a strong deficit in oxygen, such as regions with an enhanced carbon/oxygen ratio.