Domestic hens succeed at serial reversal learning and perceptual concept generalisation using a new automated touchscreen device

Improving the welfare of farm animals depends on our knowledge on how they perceive and interpret their environment; the latter depends on their cognitive abilities. Hence, limited knowledge of the range of cognitive abilities of farm animals is a major concern. An effective approach to explore the cognitive range of a species is to apply automated testing devices, which are still underdeveloped in farm animals. In screen-like studies, the uses of automated devices are few in domestic hens. We developed an original fully automated touchscreen device using digital computer-drawn colour pictures and independent sensible cells adapted for cognitive testing in domestic hens, enabling a wide range of test types from low to high complexity. This study aimed to test the efficiency of our device using two cognitive tests. We focused on tasks related to adaptive capacities to environmental variability, such as flexibility and generalisation capacities as this is a good start to approach more complex cognitive capacities. We implemented a serial reversal learning task, categorised as a simple cognitive test, and a delayed matching-to-sample (dMTS) task on an identity concept, followed by a generalisation test, categorised as more complex. In the serial reversal learning task, the hens performed equally for the two changing reward contingencies in only three reversal stages. In the dMTS task, the hens increased their performance rapidly throughout the training sessions. Moreover, to the best of our knowledge, we present the first positive result of identity concept generalisation in a dMTS task in domestic hens. Our results provide additional information on the behavioural flexibility and concept understanding of domestic hens. They also support the idea that fully automated devices would improve knowledge of farm animals’ cognition.     

Evaluation of the prevalence and economic burden of adverse drug reactions presenting to the medical emergency department of a tertiary referral centre: a prospective study

Background  

Adverse drug reactions (ADRs) are now recognized as an important cause of hospital admissions, with a proportion ranging from 0.9–7.9%. They also constitute a significant economic burden. We thus aimed at determining the prevalence and the economic burden of ADRs presenting to Medical Emergency Department (ED) of a tertiary referral center in India     

A novel mechanism of autophagic cell death in dystrophic muscle regulated by P2RX7 receptor large-pore formation and HSP90

P2RX7 is an ATP-gated ion channel, which can also exhibit an open state with a considerably wider permeation. However, the functional significance of the movement of molecules through the large pore (LP) and the intracellular signaling events involved are not known. Here, analyzing the consequences of P2RX7 activation in primary myoblasts and myotubes from the Dmd^mdx mouse model of Duchenne muscular dystrophy, we found ATP-induced P2RX7-dependent autophagic flux, leading to CASP3-CASP7-independent cell death. P2RX7-evoked autophagy was triggered by LP formation but not Ca²⁺ influx or MAPK1-MAPK3 phosphorylation, 2 canonical P2RX7-evoked signals. Phosphoproteomics, protein expression inference and signaling pathway prediction analysis of P2RX7 signaling mediators pointed to HSPA2 and HSP90 proteins. Indeed, specific HSP90 inhibitors prevented LP formation, LC3-II accumulation, and cell death in myoblasts and myotubes but not in macrophages. Pharmacological blockade or genetic ablation of p2rx7 also proved protective against ATP-induced death of muscle cells, as did inhibition of autophagy with 3-MA. The functional significance of the P2RX7 LP is one of the great unknowns of purinergic signaling. Our data demonstrate a novel outcome—autophagy—and show that molecules entering through the LP can be targeted to phagophores. Moreover, we show that in muscles but not in macrophages, autophagy is needed for the formation of this LP. Given that P2RX7-dependent LP and HSP90 are critically interacting in the ATP-evoked autophagic death of dystrophic muscles, treatments targeting this axis could be of therapeutic benefit in this debilitating and incurable form of muscular dystrophy.     

Reconstitution of allogeneic hemopoietic stem cells: the essential role of FcR gamma and the TCR beta-chain-FCp33 complex

Transplantation of purified allogeneic hemopoietic stem cells (SC) alone is characterized by a decreased risk of graft-vs-host disease but increased incidence of engraftment failure. It has been established that the facilitating cell (FC) promotes allogeneic SC reconstitution and results in donor-specific transplantation tolerance across MHC disparities, without graft-vs-host disease. Although the requirements for this facilitating function are not well-characterized, it is known that facilitation is dependent on FC expression of a unique heterodimer consisting of the TCR beta-chain (TCRbeta) and a 33-kDa protein, FCp33. The current study confirms that CD3epsilon and TCRbeta expression are present on the FC at the time of transplantation and demonstrates that the majority of cells in the FC population express the TCR signaling molecule, FcRgamma, rather than the more conventional CD3zeta receptor. Of particular significance, we have now demonstrated that FC-mediated allogeneic SC reconstitution is critically dependent on FcRgamma expression and that FcRgamma coprecipitates with the TCRbeta-FCp33 heterodimer. The mandatory requirement of TCRbeta and FcRgamma for FC function provides the first evidence of a previously undescribed role for FcRgamma in the facilitation of allogeneic SC reconstitution and establishes that FcRgamma is part of the TCRbeta-FCp33 complex uniquely expressed on FC.     

Elevated CO2 lowers relative and absolute herbivore density across all species of a scrub-oak forest

The unabated increase in global atmospheric CO(2) is expected to induce physiological changes in plants, including reduced foliar nitrogen, which are likely to affect herbivore densities. This study employs a field-based CO(2 )enrichment experiment at Kennedy Space Center, Florida, to examine plant-herbivore (insect) interactions inside eight open-topped chambers with elevated CO(2) (710 ppm) and eight control chambers with ambient CO(2). In elevated CO(2) we found decreased herbivore densities per 100 leaves, especially of leaf miners, across all five plant species we examined: the oak trees Quercus myrtifolia, Q. geminata, and Q. chapmanii, the nitrogen-fixing vine Galactia elliottii and the shrub Vaccinium myrsinites. Both direct and indirect effects of lowered plant nitrogen may influence this decrease in herbivore densities. Direct effects of lowered nitrogen resulted in increased host-plant related death and an increase in compensatory feeding: per capita herbivore leaf consumption in elevated CO(2) was higher than in ambient CO(2). Indirectly, compensatory feeding may have prolonged herbivore development and increased exposure to natural enemies. For all leaf miners we examined, mortality from natural enemies increased in elevated CO(2). These increases in host-plant induced mortality and in attack rates by natural enemies decreased leaf miner survivorship, causing a reduction in leaf miner density per 100 leaves. Despite increased leaf production in elevated CO(2) from the carbon fertilization effect, absolute herbivore abundance per chamber was also reduced in elevated CO(2). Because insects cause premature leaf abscission, we also thought that leaf abscission would be decreased in elevated CO(2). However, for all plant species, leaf abscission was increased in elevated CO(2), suggesting a direct effect of CO(2) on leaf abscission that outweighs the indirect effects of reduced insect densities on leaf abscission.     

Selection and gene flow between microenvironments: the case of Drosophila at Lower Nahal Oren,Mount Carmel,Israel

Drosophila melanogaster and D. simulans from contrasted microenvironments (south- and north-facing slopes of Lower Nahal Oren Canyon in Israel) were tested for genetic differentiation at microsatellite loci, which might be linked to differential adaptation to local ecological factors. No overall genetic differentiation was observed in either species. This indicates that the contrasted selective pressures on the two sides of the valley are not strong enough to cause population subdivision in highly mobile species such as Drosophila. Significant differences in allele frequencies were observed at two microsatellite loci, but on the whole the level of divergence we observed is far lower than has been reported previously.     

Genetic analysis and overexpression of lipolytic activity in Bacillus subtilis.

The previously cloned Bacillus subtilis lipase gene (lip) was mapped on the chromosome and used in the construction of a B. subtilis derivative totally devoid of any lip sequence. Homologous overexpression was performed in this strain by subcloning the lip open reading frame on a multicopy plasmid under the control of a strong gram-positive promoter. A 100-fold overproducing strain was obtained, which should facilitate purification of the secreted protein. Furthermore, the delta lip strain BCL1050 constitutes an ideal host for the cloning of heterologous lipase genes.     

Methylated amino acids in ribosomal proteins from Escherichia coli treated with ethionine and from a mutant lacking methylation of protein L11.

In the present study, the nature, proportions and distribution of methylated amino acids in ribosomal proteins from Escherichia coli grown in the presence of ethionine and from mutant prm 1 were studied. The undermethylated ribosomes had been labeled by addition in vitro or in vivo of radioactive methyl groups from S-adenosylmethionine or from methionine. The following compounds were identified : N alpha-mono-, di- and trimethylalanines, N epsilon-mono-, di- and trimethyllysines, methylamine and N alpha-trimethylalanyllysine. Except for the latter compound and N-alpha-dimethylalanine, all other derivatives had been previously identified in the literature. It is shown that the dipeptide had been in the past mistaken for N epsilon-monomethyllysine, and arises through incomplete hydrolysis in 24 hrs of the N-terminal peptide bond of protein L11. The results of the present study are discussed in the light of previous work on ribosomal protein methylation by the authors and other workers in the field.     

In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold

In situ hybridization has become a standard method for localizing DNA or RNA sequences in cytological preparations. We developed two methods to extend this technique to the transmission electron microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope in situ hybridization. Radioactively labeled complementary RNA (cRNA) is hybridized to metaphase chromosomes deposited on electron microscope grids and fixed in 70 percent ethanol vapor; hybridixation site are detected by autoradiography. Specific and intense labeling of chromosomal centromeric regions is observed even after relatively short exposure times. Inerphase nuclei present in some of the metaphase chromosome preparations also show defined paatterms of satellite DNA labeling which suggests that satellite-containing regions are associate with each other during interphase. The sensitivity of this method is estimated to at least as good as that at the light microscope level while the resolution is improved at least threefold. The second method, which circumvents the use of autoradiogrphic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction is improved at least threefold. The second method, which circumvents the use of autoradiographic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction with an antibody against biotin and secondary antibody adsorbed to the surface of over centromeric heterochromatin and along the associated peripheral fibers. Labeling is on average ten times that of background binding. This method is rapid and possesses the potential to allow precise ultrastructual localization of DNA sequences in chromosomes and chromatin.