Mitochondrial injury induced by triclopyr in the rat liver.

The herbicide triclopyr (3,5,6-trichloro-2-pyridinyloxyacetic acid) is already considered an environmental problem due to damage caused by incorrect disposal, leaching, and aerial dispersion, which may pose risks to the environment and human health. Studies have evaluated metabolism, absorption, excretion, and active transport but there is no clear information about its mode of action (MoA) and its cytotoxic action potential remains unknown. In this context, mitochondria have been used to assess the toxicity of xenobiotics, for this reason, to identify the toxic mechanism of triclopyr, hepatic mitochondria from Wistar rats were exposed in vitro to different concentrations of triclopyr (0.5-500 µM). There was neither formation/accumulation of reactive oxygen and nitrogen species, nor lipid peroxidation or changes in the mitochondrial antioxidant system, in addition to proper functioning of oxidative phosphorylation and ATP production. Changes were found in NAD(P)H oxidation, membrane potential dissipation and mitochondrial calcium gradient. These results demonstrate that mitochondria suffer damage related to their bioenergetics and redox status but not to their structure when exposed to concentrations of triclopyr considered higher than those described as found in the environment so far.HighlightsTriclopyr has a low mitochondrial uncoupling potential.The damage caused to the bioenergetics and redox state of the mitochondria is related to concentrations considered higher than those found in the environment.Even at high concentrations, triclopyr was not able to change the structure of the organelle after exposure.Oxidative phosphorylation and ATP production were not impaired after exposure.NAD(P)H oxidation resulted in potential membrane dissipation and mitochondrial calcium gradient dissipation.Triclopyr does not have RONS-forming properties, as well as it does not peroxide membrane lipids, it preserves membrane sulfhydryl groups and maintains the normality of the GSH/GSSG ratio.

Historical Pollution of an Urban Watershed Based in Geochemical, Geoacumulation, and EROD Activity in PLHC-1 Analyses in Sediment Cores.

In the present study, the environmental changes over the past 100 years in the Cambé River watershed in Southern Brazil were investigated. For this, a sediment core was collected to examine the distribution of metals, polycyclic aromatic hydrocarbons (PAHs), and ecotoxicological markers. The core corresponds from 1914 to 2012, which was obtained by the decay of 210Pb. Changes in the study area also were examined by: Geoaccumulation index (Igeo) and enrichment factor (EF), sediment quality guidelines (SGQs), and ethoxyresorufin-O-deethylase (EROD) activity in Poeciliopsis lucida hepatocellular carcinoma (PLHC-1) cells line. The Σ16 PAHs ranged from 242.6 to 40,775 ng g-1, with pyrogenic source at the beginning of the core, which likely corresponds to the burning of forests to establish the city and the later use of fossil fuels from 1960 to 2012. In the 1930s, most of metals presented a concentration below the background. After the 1930s, high concentrations can be associated with the intense use of fertilizers (Cd, Cu, Zn, Hg) and increases in urban traffic (Zn and Pb). Igeo showed that the distribution of Cu is considered moderately to strongly polluted, and the EF of Pb was considered as moderately polluted. Statistical analyses showed a strong relation between the presence of the ANP, FLU, PHE, BaP, IND, and Bghi compounds and the induction of EROD activity, and no correlation with heavy metals. A prediction model for the linear regression was obtained between the ANP and BaP concentrations and the EROD activity, with an accuracy of 99%.

Climate change impacts on marine water quality: The case study of the Northern Adriatic sea.

Climate change is posing additional pressures on coastal ecosystems due to variations in water biogeochemical and physico-chemical parameters (e.g., pH, salinity) leading to aquatic ecosystem degradation. With the main aim of analyzing the potential impacts of climate change on marine water quality, a Regional Risk Assessment methodology was developed and applied to coastal marine waters of the North Adriatic. It integrates the outputs of regional biogeochemical and physico-chemical models considering future climate change scenarios (i.e., years 2070 and 2100) with site-specific environmental and socio-economic indicators. Results showed that salinity and temperature will be the main drivers of changes, together with macronutrients, especially in the area of the Po' river delta. The final outputs are exposure, susceptibility and risk maps supporting the communication of the potential consequences of climate change on water quality to decision makers and stakeholders and provide a basis for the definition of adaptation and management strategies.

Regional Risk Assessment for climate change impacts on coastal aquifers.

Coastal aquifers have been identified as particularly vulnerable to impacts on water quantity and quality due to the high density of socio-economic activities and human assets in coastal regions and to the projected rising sea levels, contributing to the process of saltwater intrusion. This paper proposes a Regional Risk Assessment (RRA) methodology integrated with a chain of numerical models to evaluate potential climate change-related impacts on coastal aquifers and linked natural and human systems (i.e., wells, river, agricultural areas, lakes, forests and semi-natural environments). The RRA methodology employs Multi Criteria Decision Analysis methods and Geographic Information Systems functionalities to integrate heterogeneous spatial data on hazard, susceptibility and risk for saltwater intrusion and groundwater level variation. The proposed approach was applied on the Esino River basin (Italy) using future climate hazard scenarios based on a chain of climate, hydrological, hydraulic and groundwater system models running at different spatial scales. Models were forced with the IPCC SRES A1B emission scenario for the period 2071-2100 over four seasons (i.e., winter, spring, summer and autumn). Results indicate that in future seasons, climate change will cause few impacts on the lower Esino River valley. Groundwater level decrease will have limited effects: agricultural areas, forests and semi-natural environments will be at risk only in a region close to the coastline which covers less than 5% of the total surface of the considered receptors; less than 3.5% of the wells will be exposed in the worst scenario. Saltwater intrusion impact in future scenarios will be restricted to a narrow region close to the coastline (only few hundred meters), and thus it is expected to have very limited effects on the Esino coastal aquifer with no consequences on the considered natural and human systems.

Autochthonous case of dengue in France, October 2013.

In October 2013, autochthonous dengue fever was diagnosed in a laboratory technician in Bouches-du-Rhone, southern France, a department colonised by Aedes albopictus since 2010. After ruling out occupational contamination, we identified the likely chain of local vector-borne transmission from which the autochthonous case arose. Though limited, this second occurrence of autochthonous dengue transmission in France highlights that efforts should be continued to rapidly detect dengue virus introduction and prevent its further dissemination in France.

X-ray phase contrast imaging and noise evaluation using a single phase grating interferometer.

In this paper we present some quantitative measurements of X-ray phase contrast images and noise evaluation obtained with a recent grating based X-ray phase contrast interferometer. This device is built using a single phase grating and a large broadband X-ray source. It was calibrated using a reference sample and finally used to perform measurements of a biological fossil: a mosquito trapped in amber. As phase images, noise was evaluated from the measured interferograms.

Climate change impact assessment in Veneto and Friuli Plain groundwater. Part II: a spatially resolved regional risk assessment.

Climate change impact assessment on water resources has received high international attention over the last two decades, due to the observed global warming and its consequences at the global to local scale. In particular, climate-related risks for groundwater and related ecosystems pose a great concern to scientists and water authorities involved in the protection of these valuable resources. The close link of global warming with water cycle alterations encourages research to deepen current knowledge on relationships between climate trends and status of water systems, and to develop predictive tools for their sustainable management, copying with key principles of EU water policy. Within the European project Life+ TRUST (Tool for Regional-scale assessment of groundwater Storage improvement in adaptation to climaTe change), a Regional Risk Assessment (RRA) methodology was developed in order to identify impacts from climate change on groundwater and associated ecosystems (e.g. surface waters, agricultural areas, natural environments) and to rank areas and receptors at risk in the high and middle Veneto and Friuli Plain (Italy). Based on an integrated analysis of impacts, vulnerability and risks linked to climate change at the regional scale, a RRA framework complying with the Sources-Pathway-Receptor-Consequence (SPRC) approach was defined. Relevant impacts on groundwater and surface waters (i.e. groundwater level variations, changes in nitrate infiltration processes, changes in water availability for irrigation) were selected and analyzed through hazard scenario, exposure, susceptibility and risk assessment. The RRA methodology used hazard scenarios constructed through global and high resolution model simulations for the 2071-2100 period, according to IPCC A1B emission scenario in order to produce useful indications for future risk prioritization and to support the addressing of adaptation measures, primarily Managed Artificial Recharge (MAR) techniques. Relevant outcomes from the described RRA application highlighted that potential climate change impacts will occur with different extension and magnitude in the case study area. Particularly, qualitative and quantitative impacts on groundwater will occur with more severe consequences in the wettest and in the driest scenario (respectively). Moreover, such impacts will likely have little direct effects on related ecosystems - croplands, forests and natural environments - lying along the spring area (about 12% of croplands and 2% of natural environments at risk) while more severe consequences will indirectly occur on natural and anthropic systems through the reduction in quality and quantity of water availability for agricultural and other uses (about 80% of agricultural areas and 27% of groundwater bodies at risk).

Potent selective nonpeptidic inhibitors of human lung tryptase.

Human lung tryptase, a homotetrameric serine protease unique to mast cell secretory granules, has been implicated in the pathogenesis of asthma. A hypothesis that tethered symmetrical inhibitors might bridge two adjacent active sites was explored via a rationally designed series of bisbenzamidines. These compounds demonstrated a remarkable distanced-defined structure-activity relationship against human tryptase with one series possessing subnanomolar potencies. Additional evidence supporting the concept of active-site bridging is also presented.

UK-78,282, a novel piperidine compound that potently blocks the Kv1.3 voltage-gated potassium channel and inhibits human T cell activation.

1. UK-78,282, a novel piperidine blocker of the T lymphocyte voltage-gated K+ channel, Kv1.3, was discovered by screening a large compound file using a high-throughput 86Rb efflux assay. This compound blocks Kv1.3 with a IC50 of approximately 200 nM and 1:1 stoichiometry. A closely related compound, CP-190,325, containing a benzyl moiety in place of the benzhydryl in UK-78,282, is significantly less potent. 2 Three lines of evidence indicate that UK-78,282 inhibits Kv1.3 in a use-dependent manner by preferentially blocking and binding to the C-type inactivated state of the channel. Increasing the fraction of inactivated channels by holding the membrane potential at - 50 mV enhances the channel's sensitivity to UK-78,282. Decreasing the number of inactivated channels by exposure to approximately 160 mM external K+ decreases the sensitivity to UK-78,282. Mutations that alter the rate of C-type inactivation also change the channel's sensitivity to UK-78,282 and there is a direct correlation between tau(h) and IC50 values. 3. Competition experiments suggest that UK-78,282 binds to residues at the inner surface of the channel overlapping the site of action of verapamil. Internal tetraethylammonium and external charybdotoxin do not compete UK-78,282's action on the channel. 4. UK-78,282 displays marked selectivity for Kv1.3 over several other closely related K+ channels, the only exception being the rapidly inactivating voltage-gated K+ channel, Kv1.4. 5. UK-78,282 effectively suppresses human T-lymphocyte activation.

Inhibition of MMP-1 and MMP-13 with phosphinic acids that exploit binding in the S2 pocket.

Through the use of empirical and computational methods, phosphinate-based inhibitors of MMP-1 and MMP-13 that bind into the S2 pocket of these enzymes were designed. The synthesis and testing of 2 suggested that binding was occurring as hypothesized. Structure determination of a co-crystal of 2 bound to the catalytic domain of MMP-1 confirmed the binding mode. Substituents binding into S2, S1', S2' and S3', were optimized yielding compounds with low double-digit nM IC50's against these enzymes.

The signature sequence of voltage-gated potassium channels projects into the external vestibule.

A highly conserved motif, GYGD, contributes to the formation of the ion selectivity filter in voltage-gated K+ channels and is thought to interact with the scorpion toxin residue, Lys27. By probing the pore of the Kv1.3 channel with synthetic kaliotoxin-Lys27 mutants, each containing a non-natural lysine analog of a different length, and using mutant cycle analysis, we determined the spatial locations of Tyr400 and Asp402 in the GYGD motif, relative to His404 located at the base of the outer vestibule. Our data indicate that the terminal amines of the shorter Lys27 analogs lie close to His404 and to Asp402, while Lys27 itself interacts with Tyr400. Based on these data, we developed a molecular model of this region of the channel. The junction between the outer vestibule and the pore is defined by a ring ( approximately 8-9-A diameter) formed from alternating Asp402 and His404 residues. Tyr400 lies 4-6 A deeper into the pore, and its interaction with kaliotoxin-Lys27 is in competition with K+ ions. Studies with dimeric Kv1.3 constructs suggest that two Tyr400 residues in the tetramer are sufficient to bind K+ ions. Thus, at least part of the K+ channel signature sequence extends into a shallow trough at the center of a wide external vestibule.

Topology of the pore-region of a K+ channel revealed by the NMR-derived structures of scorpion toxins.

The architecture of the pore-region of a voltage-gated K+ channel, Kv1.3, was probed using four high affinity scorpion toxins as molecular calipers. We established the structural relatedness of these toxins by solving the structures of kaliotoxin and margatoxin and comparing them with the published structure of charybdotoxin; a homology model of noxiustoxin was then developed. Complementary mutagenesis of Kv1.3 and these toxins, combined with electrostatic compliance and thermodynamic mutant cycle analyses, allowed us to identify multiple toxin-channel interactions. Our analyses reveal the existence of a shallow vestibule at the external entrance to the pore. This vestibule is approximately 28-32 A wide at its outer margin, approximately 28-34 A wide at its base, and approximately 4-8 A deep. The pore is 9-14 A wide at its external entrance and tapers to a width of 4-5 A at a depth of approximately 5-7 A from the vestibule. This structural information should directly aid in developing topological models of the pores of related ion channels and facilitate therapeutic drug design.