Alcohol dependence potentiates substance P/neurokinin-1 receptor signaling in the rat central nucleus of amygdala.

Behavioral and clinical studies suggest a critical role of substance P (SP)/neurokinin-1 receptor (NK-1R) signaling in alcohol dependence. Here, we examined regulation of GABA transmission in the medial subdivision of the central amygdala (CeM) by the SP/NK-1R system, and its neuroadaptation following chronic alcohol exposure. In naïve rats, SP increased action potential-dependent GABA release, and the selective NK-1R antagonist L822429 decreased it, demonstrating SP regulation of CeM activity under basal conditions. SP induced a larger GABA release in alcohol-dependent rats accompanied by decreased NK-1R expression compared to naïve controls, suggesting NK-1R hypersensitivity which persisted during protracted alcohol withdrawal. The NK-1R antagonist blocked acute alcohol-induced GABA release in alcohol-dependent and withdrawn but not in naïve rats, indicating that dependence engages the SP/NK-1R system to mediate acute effects of alcohol. Collectively, we report long-lasting CeA NK-1R hypersensitivity corroborating that NK-1Rs are promising targets for the treatment of alcohol use disorder

Computers from plants we never made. Speculations

We discuss possible designs and prototypes of computing systems that could be based on morphological development of roots, interaction of roots, and analog electrical computation with plants, and plant-derived electronic components. In morphological plant processors data are represented by initial configuration of roots and configurations of sources of attractants and repellents; results of computation are represented by topology of the roots' network. Computation is implemented by the roots following gradients of attractants and repellents, as well as interacting with each other. Problems solvable by plant roots, in principle, include shortest-path, minimum spanning tree, Voronoi diagram, $\alpha$-shapes, convex subdivision of concave polygons. Electrical properties of plants can be modified by loading the plants with functional nanoparticles or coating parts of plants of conductive polymers. Thus, we are in position to make living variable resistors, capacitors, operational amplifiers, multipliers, potentiometers and fixed-function generators. The electrically modified plants can implement summation, integration with respect to time, inversion, multiplication, exponentiation, logarithm, division. Mathematical and engineering problems to be solved can be represented in plant root networks of resistive or reaction elements. Developments in plant-based computing architectures will trigger emergence of a unique community of biologists, electronic engineering and computer scientists working together to produce living electronic devices which future green computers will be made of.Comment: The chapter will be published in "Inspired by Nature. Computing inspired by physics, chemistry and biology. Essays presented to Julian Miller on the occasion of his 60th birthday", Editors: Susan Stepney and Andrew Adamatzky (Springer, 2017

Narrow genetic base in forest restoration with holm oak (Quercus ilex L.) in Sicily

In order to empirically assess the effect of actual seed sampling strategy on genetic diversity of holm oak (Quercus ilex) forestations in Sicily, we have analysed the genetic composition of two seedling lots (nursery stock and plantation) and their known natural seed origin stand by means of six nuclear microsatellite loci. Significant reduction in genetic diversity and significant difference in genetic composition of the seedling lots compared to the seed origin stand were detected. The female and the total effective number of parents were quantified by means of maternity assignment of seedlings and temporal changes in allele frequencies. Extremely low effective maternity numbers were estimated (Nfe $\approx$ 2-4) and estimates accounting for both seed and pollen donors gave also low values (Ne $\approx$ 35-50). These values can be explained by an inappropriate forestry seed harvest strategy limited to a small number of spatially close trees

Interactions of Cathinone NPS with Human Transporters and Receptors in Transfected Cells

Pharmacological assays carried out in transfected cells have been very useful for describing the mechanism of action of cathinone new psychoactive substances (NPS). These in vitro characterizations provide fast and reliable information on psychoactive substances soon after they emerge for recreational use. Well-investigated comparator compounds, such as methamphetamine, 3,4-methylenedioxymethamphetamine, cocaine, and lysergic acid diethylamide, should always be included in the characterization to enhance the translation of the in vitro data into clinically useful information. We classified cathinone NPS according to their pharmacology at monoamine transporters and receptors. Cathinone NPS are monoamine uptake inhibitors and most induce transporter-mediated monoamine efflux with weak to no activity at pre- or postsynaptic receptors. Cathinones with a nitrogen-containing pyrrolidine ring emerged as NPS that are extremely potent transporter inhibitors but not monoamine releasers. Cathinones exhibit clinically relevant differences in relative potencies at serotonin vs. dopamine transporters. Additionally, cathinone NPS have more dopaminergic vs. serotonergic properties compared with their non-β-keto amphetamine analogs, suggesting more stimulant and reinforcing properties. In conclusion, in vitro pharmacological assays in heterologous expression systems help to predict the psychoactive and toxicological effects of NPS

Carboxylation of phenols and asymmetric nucleophile addition across C=C bond

The regioselective carboxylation of electron-rich (hetero)aromatics employing decarboxylases in the redox-neutral (reverse) carboxylation reaction using bicarbonate or CO2(g) is currently exploited for the biocatalytic synthesis of carboxylic acids.1 Three enzyme classes exert complementary regioselectivities through diverse mechanisms: (i) Whereas the o-carboxylation of phenols (an equivalent to the Kolbe-Schmitt reaction) is mediated by Zn2+-dependent o-benzoic acid (de)carboxylases,2 (ii) the -carboxylation of hydroxystyrenes is catalysed by phenolic/ferulic acid (de)carboxylases acting via a pair of Tyr-Arg residues.3 (iii) Surpringly, these enzymes also exhibit a catalytic promiscuity for the nucleophile addition of H2O,4 NH2-OMe, cyanide and n-Pr-SH across the vinyl C=C bond via a quinone-methide intermediate, which yields the corresponding (S)-configurated adducts in up to 91% e.e.5 (iv) In search of ATP-independent regio-complementary p-benzoic acid (de)carboxylases, we discovered that 3,4-dihydroxybenzoic acid decarboxylase from Enterobacter cloacae6 (DHBDC_Ec) surprisingly depends on prenylated FMN7 as cofactor. In an attempt to propose a mechanism for the carboxylation of catechol by DHBDC_Ec, QM calculations revealed that the transient formation of a 1,3-dipolar cycloaddition product (as suggested for the decarboxylation of cinnamic acid with ferulic acid decarboxylase from S. cerevisiae8) was highly disfavored (\u3e30 kcal/M). As an alternative, we propose a mono-covalent nucleophile adduct involving a prFMN iminium electrophile (~14 kcal/M). Please click Additional Files below to see the full abstract

Natural selection on cork oak: allele frequency reveals divergent selection in cork oak populations along a temperature cline

A recent study of population divergence at neutral markers and adaptive traits in cork oak has observed an association between genetic distances at locus QpZAG46 and genetic distances for leaf size and growth. In that study it was proposed that certain loci could be linked to genes encoding for adaptive traits in cork oak and, thus, could be used in adaptation studies. In order to investigate this hypothesis, here we (1) looked for associations between molecular markers and a set of adaptive traits in cork oak, and (2) explored the effects of the climate on among-population patterns in adaptive traits and molecular markers. For this purpose, we chose 9-year-old plants originating from thirteen populations spanning a broad range of climatic conditions. Plants established in a common garden site were genotyped at six nuclear microsatellites and phenotypically characterized for six functional traits potentially related to plant performance. Our results supported the proposed linkage between locus QpZAG46 and genes encoding for leaf size and growth. Temperature caused adaptive population divergence in leaf size and growth, which was expressed as differences in the frequencies of the alleles at locus QpZAG46

A fungal ascorbate oxidase with unexpected laccase activity

Ascorbate oxidases are an enzyme group that has not been explored to a large extent. So far, mainly ascorbate oxidases from plants and only a few from fungi have been described. Although ascorbate oxidases belong to the well-studied enzyme family of multi-copper oxidases, their function is still unclear. In this study, Af_AO1, an enzyme from the fungus Aspergillus flavus, was characterized. Sequence analyses and copper content determination demonstrated Af_AO1 to belong to the multi-copper oxidase family. Biochemical characterization and 3D-modeling revealed a similarity to ascorbate oxidases, but also to laccases. Af_AO1 had a 10-fold higher affinity to ascorbic acid (KM = 0.16 ± 0.03 mM) than to ABTS (KM = 1.89 ± 0.12 mM). Furthermore, the best fitting 3D-model was based on the ascorbate oxidase from Cucurbita pepo var. melopepo. The laccase-like activity of Af_AO1 on ABTS (Vmax = 11.56 ± 0.15 µM/min/mg) was, however, not negligible. On the other hand, other typical laccase substrates, such as syringaldezine and guaiacol, were not oxidized by Af_AO1. According to the biochemical and structural characterization, Af_AO1 was classified as ascorbate oxidase with unusual, laccase-like activityPeer ReviewedPostprint (published version

Pronounced turnover of vascular plant species in Central European arable fields over 90 years

We studied changes in vascular plant species occurring in Central European (Germany, Czech Republic, Slovakia, Switzerland, Liechtenstein, Austria, Hungary, Northern Italy, Slovenia and Croatia) arable fields and their edges from 1930 to 2019. To correct for bias in the data, we used occupancy modeling to analyze changes in the occupancy, i.e., distribution ranges sizes, of the 359 most common species in the AgriWeedClim database. We used ecological indicator values, native versus alien (archaeophyte, neophyte) status, and species affinity to arable habitats to assess changes in the occupancy of species with different environmental preferences and biogeographic origins. We found only a small decline in overall species occupancy over time, with a median occupancy change of −0.1 %, possibly due to the exclusion of rare species from modeling. Species turnover was more pronounced, with 72 species decreasing to less than half of their initial occupancy and 77 species more than doubling their initial occupancy. Species with environmental preferences for nutrient-rich sites with neutral pH increased in occupancy whereas species typical for arable fields decreased. No response to climate change (i.e., increased occupancy of thermophilous or drought-tolerant species) was detected. Archaeophytes and native species decreased whereas neophytes increased in occupancy. Taken together, results suggest that the biodiversity of arable fields is changing largely in response to anthropogenic habitat changes