Intracellular calcium and protein tyrosine phosphorylation during the release of bovine sperm adhering to the fallopian tube epithelium in vitro

In mammals, sperm adhesion to the epithelial cells lining the oviductal isthmus plays a key role in the maintenance of motility and in the selection of superior quality subpopulations. In the bovine species, heparin and other sulfated glycoconjugates powerfully induce the synchronous release of sperm adhering to tubal epithelium in vitro and may represent the signal which triggers release at ovulation in vivo. Sperm detachment may be due either to surface remodeling or to hyperactivation brought about by capacitation. In this paper, the dynamics of intracellular free Ca2+concentration ([Ca2+]i) and protein tyrosine phosphorylation in sperm during and after heparin-induced release from in vitro cultured oviductal monolayers were assessed to determine whether this event is due to capacitation. Moreover, Ca2+-ionophore A23187, thapsigargin, thimerosal and caffeine were used to determine whether [Ca2+]i increase and/or hyperactivation can induce sperm release. Results showed that: 1. heparin-released sperm have significantly higher [Ca2+]i than adhering sperm; 2. heparin induces a [Ca2+]i elevation in the sperm head followed by detachment from the monolayers; 3. external Ca2+is not required for heparin-induced release; 4. [Ca2+]i increase and/or hyperactivation are unable to release sperm; and 5. heparin-released sperm have an increased level of tyrosine phosphorylated proteins compared with adhering sperm. In conclusion, although heparin is considered a long-lasting capacitation agent, it quickly modulates the capacitation of bovine sperm adhering to the fallopian epithelium, probably leading to surface remodeling and therefore to the release of sperm selected and stored within the oviduct through adhesion.     

Variability within the Candida rugosa Upases family

Several fungi secrete lipase isozymes differing in biochemicalproperties and in some cases in substrate specificity. In theyeast Candida rugosa, a family of related genes encodes formultiple lipase proteins, highly homologous in sequence butpartially different in the regions interacting with the substratemolecule. Analysis of these substitutions performed on the basisof multiple alignments and using a 3-D model of the enzyme,allows identification of a restricted number of amino adds possiblyinvolved in substrate specificity of Candida lipases.     

Comprehension of cause€ffect relations in a tool-using task by chimpanzees (Pan troglodytes).

Five chimpanzees (Pan troglodytes) were tested to assess their understanding of causality in a tool task. The task consisted of a transparent tube with a trap-hole drilled in its middle. A reward was randomly placed on either side of the hole. Depending on which side the chimpanzee inserted the stick into, the candy was either pushed out of the tube or into the trap. In Experiment 1, the success rate of 2 chimpanzees rose highly above chance, but that of the other subjects did not. Results show that the 2 successful chimpanzees selected the correct side for insertion beforehand. Experiment 2 ruled out the possibility that their success was due to a distance-based associative rule, and the results favor an alternative hypothesis that relates success to an understanding of the causal relation between the tool-using action and its outcome. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Performance in a tool-using task by common chimpanzees (Pan troglodytes), bonobos (Pan paniscus), an orangutan (Pongo pygmaeus), and capuchin monkeys (Cebus apella).

Performance by individual animals of three species of great apes (Pan troglodytes, Pan paniscus, and Pongo pygmaeus) and capuchin monkeys (Cebus apella) was assessed by presenting a food treat inside a clear tube. The subjects readily used a straight stick to obtain the food. When sticks were bundled together, the apes immediately unwrapped the bundle to obtain an individual stick, whereas capuchins attempted to insert the bundled sticks. When a misshapen stick was provided, apes, but not capuchins, showed an improvement in terms of modifying the misshapen stick before insertion. Our results indicate that all these species can solve these tasks. However, only the performance of apes is consistent with emerging comprehension of the causal relations required for the avoidance of errors in the more complex tasks. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Quantitative Systems Pharmacology and Biased Agonism at Opioid Receptors: A Potential Avenue for Improved Analgesics

Chronic pain is debilitating and represents a significant burden in terms of personal and socio-economic costs. Although opioid analgesics are widely used in chronic pain treatment, many patients report inadequate pain relief or relevant adverse effects, highlighting the need to develop analgesics with improved efficacy/safety. Multiple evidence suggests that G protein-dependent signaling triggers opioid-induced antinociception, whereas arrestin-mediated pathways are credited with modulating different opioid adverse effects, thus spurring extensive research for G protein-biased opioid agonists as analgesic candidates with improved pharmacology. Despite the increasing expectations of functional selectivity, translating G protein-biased opioid agonists into improved therapeutics is far from being fully achieved, due to the complex, multidimensional pharmacology of opioid receptors. The multifaceted network of signaling events and molecular processes underlying therapeutic and adverse effects induced by opioids is more complex than the mere dichotomy between G protein and arrestin and requires more comprehensive, integrated, network-centric approaches to be fully dissected. Quantitative Systems Pharmacology (QSP) models employing multidimensional assays associated with computational tools able to analyze large datasets may provide an intriguing approach to go beyond the greater complexity of opioid receptor pharmacology and the current limitations entailing the development of biased opioid agonists as improved analgesics.     

Making Connections: Mesenchymal Stem Cells Manifold Ways to Interact with Neurons

Mesenchymal Stem Cells (MSCs) are adult multipotent cells able to increase sensory neuron survival: direct co-culture of MSCs with neurons is pivotal to observe a neuronal survival increase. Despite the identification of some mechanisms of action, little is known about how MSCs physically interact with neurons. The aim of this paper was to investigate and characterize the main mechanisms of interaction between MSCs and neurons. Morphological analysis showed the presence of gap junctions and tunneling nanotubes between MSCs and neurons only in direct co-cultures. Using a diffusible dye, we observed a flow from MSCs to neurons and further analysis demonstrated that MSCs donated mitochondria to neurons. Treatment of co-cultures with the gap junction blocker Carbenoxolone decreased neuronal survival, thus demonstrating the importance of gap junctions and, more in general, of cell communication for the MSC positive effect. We also investigated the role of extracellular vesicles; administration of direct co-cultures-derived vesicles was able to increase neuronal survival. In conclusion, our study demonstrates the presence and the importance of multiple routes of communication between MSCs and neurons. Such knowledge will allow a better understanding of the potential of MSCs and how to maximize their positive effect, with the final aim to provide the best protective treatment.     

Non-thermal atmospheric gas plasma device for surface decontamination of shell eggs

A resistive barrier discharge (RBD) prototype able to generate gas plasma at atmospheric conditions was set up. The discharge was electrically characterized and the plasma glow was analysed by optical emission spectroscopy. The decontamination power of the device was assessed on samples of shell eggs experimentally inoculated with Salmonella Enteritidis and Salmonella Typhimurium (5.5–6.5 Log CFU/eggshell) and placed in the treatment chamber. Different decontamination times (10, 20, 30, 45, 60 and 90 min) and relative humidity values (RH) of the gas mixture in the chamber (i.e. 35% and 65%, at 25 °C) were considered. All samples were treated in the plasma after-glow chamber where the measured temperature was not much higher than the room temperature, minimizing the risk of egg quality alterations.     

Imaging and elasticity measurements of the sarcolemma of fully differentiated skeletal muscle fibres

This study aimed to describe the three-dimensional structure and the elastic properties of the sarcolemma of adult, fully differentiated, skeletal muscle fibres combining Atomic Force Microscopy (AFM) and optical microscopy. Single fibres were enzymatically dissociated from Flexor Digitorum Brevis of adult mice and were maintained in culture up to 3 weeks. On the sixth day after dissociation, the upper surface of intact fibres, either alive in solution or fixed and kept in solution or fixed and exposed in air, was analysed with AFM. The most prominent features in AFM images were periodic transversal foldings with an interval that corresponded to the sarcomere length. More detailed analysis of the topography profile showed that the depth in the folding decreased with increasing sarcomere length and that the crests of the foldings corresponded to the Z-lines. Minor periodic structures could be detected in the valleys between the major foldings. AFM images also showed deep depressions on the sarcolemma likely corresponding to openings of T tubules and caveolae. Two-dimensional elasticity maps were obtained using AFM as an indenter and showed that the crests of the transversal foldings correspond to higher stiffness regions. This study provides the first complete three-dimensional topography and mechanical characterization of intact, living skeletal muscle fibres and might form the basis for further investigations aimed to compare healthy and dystrophic muscles.     

Investigation of the composition of historical and modern Italian papers by energy dispersive X-ray fluorescence (EDXRF), X-ray diffraction (XRD), and scanning electron microscopy energy dispersive spectrometry (SEM-EDS)

In this work, a study concerning the composition of Italian papers from the seventeenth to the twentieth centuries was carried out using energy dispersive X-ray fluorescence spectrometry (EDXRF), X-ray diffraction (XRD), and scanning electron microscopy coupled to energy dispersive spectrometry (SEM-EDS). The analyzed samples consisted of papers employed for drawing, writing, printing, and absorbance. Observations carried out by SEM magnified the typical paper morphology. EDXRF in combination with XRD and SEM-EDS allowed the determination of calcite, gypsum, kaolin, talc, magnesite, and dolomite, used as fillers in the production of the papers studied herein. The inks present in the handwritten and printed papers, investigated by SEM-EDS and μ-EDXRF, were synthetic, Fe based, and iron gall inks.     

Design of renewable poly(amidoamine)/hemicellulose hydrogels for heavy metal adsorption

Renewable poly(amidoamine)/hemicellulose hydrogels were prepared from O‐acetylated galactoglucomannan (AcGGM)‐rich biomass and shown to display a significantly high adsorption capacity for Cu²⁺, Cd²⁺, Pb^2+, Zn²⁺, Ni²⁺, Co²⁺, and . Two different acrylamido end‐capped poly(amidoamine) oligomers (PAA) were prepared and covalently immobilized onto an in situ formed polysaccharide network via water‐based free radical graft copolymerization and cross‐linking. The synthetic approach was shown to be viable when using a highly purified AcGGM or a crude spruce hydrolysate, an AcGGM and lignin containing biomass fraction as a reactant. Homogeneous reaction mixtures were obtained in both cases with polysaccharide contents up to 20% by weight. Oscillatory shear measurements indicated a predominantly solid‐like behavior of the hydrogels with an increase in shear storage modulus with increasing cross‐link density. The mechanical integrity of the PAA/hemicellulose hydrogels showed higher water swelling capacity and less fragility than the parent PAA hydrogels and they retained the heavy metal ion absorption ability of the PAA component, even in the presence of the least purified hemicellulose fraction. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41695.