Comparative analysis of neonicotinoid binding to insect membranes: I. A structure-activity study of the mode of [3H]imidacloprid displacement in Myzus persicae and Aphis craccivora

Neonicotinoids bind selectively to insect nicotinic acetylcholine receptors with nanomolar affinity to act as potent insecticides. While the members of the neonicotinoid class have many structural features in common, it is not known whether they also share the same mode of binding to the target receptor. Previous competition studies with [3H]imidacloprid, the first commercialised neonicotinoid, indicated that thiamethoxam, representing a novel structural sub-class, may bind in a different way from that of other neonicotinoids. In the present work we analysed the mode of [3H]imidacloprid displacement by established neonicotinoids and newly synthesized analogues in the aphids Myzus persicae Sulzer and Aphis craccivora Koch. We found two classes of neonicotinoids with distinct modes of interference with [3H]imidacloprid, described as direct competitive inhibition and non-competitive inhibition, respectively. Competitive neonicotinoids were acetamiprid, nitenpyram, thiacloprid, clothianidin and nithiazine, whereas thiamethoxam and the N-methyl analogues of imidacloprid and clothianidin showed non-competitive inhibition. The chloropyridine or chlorothiazole heterocycles, the polar pharmacophore parts, such as nitroimino, cyanoimino and nitromethylene, and the cyclic or acyclic structure of the pharmacophore were not relevant for the mode of inhibition. Consensus structural features of the neonicotinoids were defined for the two mechanisms of interaction with [3H]imidacloprid binding. Furthermore, two sub-classes of non-competitive inhibitors can be discriminated on the basis of their Hill coefficients for imidacloprid displacement. We conclude from the present data that the direct competitors share the binding site with imidacloprid, whereas non-competitive compounds, like thiamethoxam, bind to a different site or in a different mode.     

Genetic differentiation of indigenous (Quercus robur L.) and late flushing oak stands (Q. robur L. subsp. slavonica (Gáyer) Mátyás) in western Germany (North Rhine-Westphalia)

European Journal of Forest Research - Slavonian oaks (Quercus robur subsp. slavonica (Gáyer) Mátyás) originating from Croatia have been cultivated in Germany mainly in the...     

Detection and distribution of apoptotic cell death in normal and diseased canine cranial cruciate ligaments

One of the possible initiating factors in canine cranial cruciate ligament (CCL) rupture could be an abnormal pattern of ligament cell death. This study compared apoptotic cell death in sections of ruptured CCLs and normal controls, and examined nitric oxide (NO) production in joint tissues and correlated this to apoptosis. CCLs and cartilage from the lateral femoral condyle were harvested from 10 healthy dogs and 15 dogs with CCL rupture and ligaments were further processed to detect cleaved caspase-3 and to determine supernatant NO production in explant cultures. Apoptotic activity was greater in ruptured ligaments compared to controls. NO in ligaments showed a moderate but significant positive correlation with caspase-positive cells. The results suggest that increased apoptosis has a role in CCL rupture and that apoptosis may be influenced by local NO production.     

Osteopontin: a leading candidate adhesion molecule for implantation in pigs and sheep

Osteopontin(OPN; also known as Secreted Phosphoprotein 1, SPP1) is a secreted extra-cellular matrix(ECM) protein that binds to a variety of cell surface integrins to stimulate cell-cell and cell-ECM adhesion and communication. It is generally accepted that OPN interacts with apically expressed integrin receptors on the uterine luminal epithelium(LE)and conceptus trophectoderm to attach the conceptus to the uterus for implantation. Research conducted with pigs and sheep has significantly advanced understanding of the role(s) of OPN during implantation through exploitation of the prolonged peri-implantation period of pregnancy when elongating conceptuses are free within the uterine lumen requiring extensive paracrine signaling between conceptus and endometrium. This is followed by a protracted and incremental attachment cascade of trophectoderm to uterine LE during implantation, and development of a true epitheliochorial or synepitheliochorial placenta exhibited by pigs and sheep, respectively. In pigs, implanting conceptuses secrete estrogens which induce the synthesis and secretion of OPN in adjacent uterine LE. OPN then binds to αvβ6 integrin receptors on trophectoderm, and the αvβ3 integrin receptors on uterine LE to bridge conceptus attachment to uterine LE for implantation. In sheep, implanting conceptuses secrete interferon tau that prolongs the lifespan of CL. Progesterone released by CL then induces OPN synthesis and secretion from the endometrial GE into the uterine lumen where OPN binds integrins expressed on trophectoderm(αvβ3) and uterine LE(identity of specific integrins unknown) to adhere the conceptus to the uterus for implantation. OPN binding to the αvβ3 integrin receptor on ovine trophectoderm cells induces in vitro focal adhesion assembly, a prerequisite for adhesion and migration of trophectoderm, through activation of: 1) P70S6 K via crosstalk between FRAP1/MTOR and MAPK pathways; 2) MTOR,PI3 K, MAPK3/MAPK1(Erk1/2) and MAPK14(p38) signaling to stimulate trohectoderm cell migration; and 3) focal adhesion assembly and myosin II motor activity to induce migration of trophectoderm cells. Further large in vivo focal adhesions assemble at the uterine-placental interface of both pigs and sheep and identify the involvement of sizable mechanical forces at this interface during discrete periods of trophoblast migration, attachment and placentation in both species.     

Organic carbon enables the biotic engineering of beneficial soil structure in Profundihumic and Haplic Ferralsols

We investigated how organic matter may, directly and indirectly, modify the porosity of Ferralsols, that is, deeply weathered soils of the tropics and subtropics. Although empirical and anecdotal evidence suggests that organic matter accumulation may increase porosity, a mechanistic understanding of the processes underlying this beneficial effect is lacking, especially so for Ferralsols. To achieve our end, we leveraged the fact that the Profundihumic qualifier of Ferralsols (PF) is distinguished from Haplic Ferralsols (HF) by both a much larger average carbon content in the first 1 m of soil depth (19 kg C m⁻³ in PF vs. 10 kg C m⁻³ in HF) and a significantly lower bulk density (1.05 ± 0.08 kg L⁻¹ in PF vs. 1.21 ± 0.05 kg L⁻¹ in HF). Through exhaustive modelling of carbon – bulk density relationships, we demonstrate that the lower bulk density of PF cannot be satisfactorily explained by a simple dilution effect. Rather, we found that bulk density correlated with carbon content when combined with carbon: nitrogen ratio (r² = 0.51), black carbon content (r² = 0.75), and Δ¹⁴C (r² = 0.81). Total pore space was greater in PF (61 ± 3%) than in HF (55 ± 2%), but x-ray computed tomography revealed that pore space inside soil aggregates of 4–5 mm diameter does not vary between the studied Ferralsols. We further observed nearly twice as many roots and burrows in PF compared with HF. We thus infer that the mechanism responsible for the increase in porosity is most likely an enhancement of resource availability (e.g., energy, carbon, and nutrients) for the organisms (earthworms, ants, termites, etc.) that physically displace soil particles and promote soil aggregation. As a result of increased resource availability, soil organisms can create especially the mesoscale structural soil features necessary for unrestricted water flow and rapid gas exchange. This insight paves the way for the development of land management technologies to optimize the physical shape and capacity of the soil bioreactor.     

Tracing erosion rates in loess landscape of the Trzebnica Hills (Poland) over time using fallout and cosmogenic nuclides

Journal of Soils and Sediments - Loess landscapes are highly susceptible to soil erosion, which affects soil stability and productivity. Erosion is non-linear in time and space and determines...     

Rapid neural coding in the retina with relative spike latencies

Natural vision is a highly dynamic process. Frequent body, head, and eye movements constantly bring new images onto the retina for brief periods, challenging our understanding of the neural code for vision. We report that certain retinal ganglion cells encode the spatial structure of a briefly presented image in the relative timing of their first spikes. This code is found to be largely invariant to stimulus contrast and robust to noisy fluctuations in response latencies. Mechanistically, the observed response characteristics result from different kinetics in two retinal pathways ("ON" and "OFF") that converge onto ganglion cells. This mechanism allows the retina to rapidly and reliably transmit new spatial information with the very first spikes emitted by a neural population.     

High harmonic generation from multiple orbitals in N2

Molecular electronic states energetically below the highest occupied molecular orbital (HOMO) should contribute to laser-driven high harmonic generation (HHG), but this behavior has not been observed previously. Our measurements of the HHG spectrum of N2 molecules aligned perpendicular to the laser polarization showed a maximum at the rotational half-revival. This feature indicates the influence of electrons occupying the orbital just below the N2 HOMO, referred to as the HOMO-1. Such observations of lower-lying orbitals are essential to understanding subfemtosecond/subangstrom electronic motion in laser-excited molecules.     

Structure of precursor-bound NifEN: a nitrogenase FeMo cofactor maturase/insertase

NifEN plays an essential role in the biosynthesis of the nitrogenase iron-molybdenum (FeMo) cofactor (M cluster). It is an α(2)β(2) tetramer that is homologous to the catalytic molybdenum-iron (MoFe) protein (NifDK) component of nitrogenase. NifEN serves as a scaffold for the conversion of an iron-only precursor to a matured form of the M cluster before delivering the latter to its target location within NifDK. Here, we present the structure of the precursor-bound NifEN of Azotobacter vinelandii at 2.6 angstrom resolution. From a structural comparison of NifEN with des-M-cluster NifDK and holo NifDK, we propose similar pathways of cluster insertion for the homologous NifEN and NifDK proteins.