Synthesis of Phospho-Amino Acid Analogues as Tissue Adhesive Cement Additives

In this paper we report the synthesis of a library of phospho-amino acid analogues, via a novel single-step allyl-phosphoester protection/Pd-mediated deprotection strategy. These phosphoserine and phosphotyrosine analogues were then applied as additives to create adhesive calcium phosphate cements, allowing us to probe the chemical origins of the increased surface binding strength. We demonstrate the importance of multiple calcium binding motifs in mediating adhesion, as well as highlighting the crucial role played by substrate hydrophobicity and orientation in controlling binding strength

Task-dependent workload adjustment of female breeders in a cooperatively breeding fish

©The Author(s) 2017. Published by Oxford University Press on behalf of the International Society for Behavioral Ecology. All rights reserved. Parental investment affects the future survival and reproductive success of breeders. Therefore, breeders should optimize the amount of care they invest into the current offspring. In cooperative breeding systems, the amount of breeders' parental care is influenced by the behavior of brood-care helpers. Such workload adjustment is expected to depend on the task that needs to be fulfilled. While investment rules of breeders in respect to single tasks are well investigated in many bird and mammal species, little is known about behavioral adjustment of breeders when dealing with multiple tasks. Here, we examined the workload adjustment in multiple tasks of female breeders in the cooperatively breeding fish Neolamprologus obscurus. By combining behavioral observations with helper removal experiments in a wild population, we show that female territory defense and offspring care significantly decreased with increasing helper number. Furthermore, the workload invested in these tasks significantly increased after the removal of a helper, suggesting load-lightening effects in territory defense and offspring care. On the other hand, female territory maintenance behavior (i.e., excavating sand from the breeding shelter) did not correlate with helper number. While sand excavation significantly increased after the helper removal experiment, the size of the excavated stone area decreased after the helper removal in the recent study, suggesting that sand excavation may have additive effects for the breeders. These results demonstrate and underline the importance of task-dependent workload adjustment in cooperative breeders

The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro

Huntingtin-interacting protein 1 related (Hip1R) is a novel component of clathrin-coated pits and vesicles and is a mammalian homologue of Sla2p, an actin-binding protein important for both actin organization and endocytosis in yeast. Here, we demonstrate that Hip1R binds via its putative central coiled-coil domain to clathrin, and provide evidence that Hip1R and clathrin are associated in vivo at sites of endocytosis. First, real-time analysis of Hip1R–YFP and DsRed–clathrin light chain (LC) in live cells revealed that these proteins show almost identical temporal and spatial regulation at the cell cortex. Second, at the ultrastructure level, immunogold labeling of ‘unroofed’ cells showed that Hip1R localizes to clathrin-coated pits. Third, overexpression of Hip1R affected the subcellular distribution of clathrin LC. Consistent with a functional role for Hip1R in endocytosis, we also demonstrated that it promotes clathrin cage assembly in vitro. Finally, we showed that Hip1R is a rod-shaped apparent dimer with globular heads at either end, and that it can assemble clathrin-coated vesicles and F-actin into higher order structures. In total, Hip1R's properties suggest an early endocytic function at the interface between clathrin, F-actin, and lipids

Directed Evolution of Gloeobacter violaceus Rhodopsin Spectral Properties

Proton-pumping rhodopsins (PPRs) are photoactive retinal-binding proteins that transport ions across biological membranes in response to light. These proteins are interesting for light-harvesting applications in bioenergy production, in optogenetics applications in neuroscience, and as fluorescent sensors of membrane potential. Little is known, however, about how the protein sequence determines the considerable variation in spectral properties of PPRs from different biological niches or how to engineer these properties in a given PPR. Here we report a comprehensive study of amino acid substitutions in the retinal binding pocket of Gloeobacter violacaeus rhodopsin (GR) that tune its spectral properties. Directed evolution generated 70 GR variants with absorption maxima shifted by up to +/- 80 nm, extending the protein’s light absorption significantly beyond the range of known natural PPRs. While proton pumping activity was disrupted in many of the spectrally shifted variants, we identified single tuning mutations that incurrred blue and red shifts of 42 nm and 22 nm, respectively, that did not disrupt proton pumping. Blue-shifting mutations were distributed evenly along the retinal molecule while red-shifting mutations were clustered near the residue K257, which forms a covalent bond with retinal through a Schiff base linkage. Thirty-four of the identified tuning mutations are not found in known microbial rhodopsins. We discovered a subset of red-shifted GRs that exhibit high levels of fluorescence relative to the wild-type protein

Archaerhodopsin variants with enhanced voltage-sensitive fluorescence in mammalian and Caenorhabditis elegans neurons

Probing the neural circuit dynamics underlying behaviour would benefit greatly from improved genetically encoded voltage indicators. The proton pump ​Archaerhodopsin-3 (​Arch), an optogenetic tool commonly used for neuronal inhibition, has been shown to emit voltage-sensitive fluorescence. Here we report two ​Arch variants with enhanced radiance (Archers) that in response to 655 nm light have 3–5 times increased fluorescence and 55–99 times reduced photocurrents compared with ​Arch WT. The most fluorescent variant, Archer1, has 25–40% fluorescence change in response to action potentials while using 9 times lower light intensity compared with other ​Arch-based voltage sensors. Archer1 is capable of wavelength-specific functionality as a voltage sensor under red light and as an inhibitory actuator under green light. As a proof-of-concept for the application of ​Arch-based sensors in vivo, we show fluorescence voltage sensing in behaving Caenorhabditis elegans. Archer1’s characteristics contribute to the goal of all-optical detection and modulation of activity in neuronal networks in vivo

Distinct Dynamics of Endocytic Clathrin-Coated Pits and Coated Plaques

Here we classify endocytic structures at the adherent (bottom) surface of many cells in culture into shorter-lived coated pits and longer-lived coated plaques which internalize by different mechanisms

Child and adolescent psychiatric patients and later criminality

<p>Abstract</p> <p>Background</p> <p>Sweden has an extensive child and adolescent psychiatric (CAP) research tradition in which longitudinal methods are used to study juvenile delinquency. Up to the 1980s, results from descriptions and follow-ups of cohorts of CAP patients showed that children's behavioural disturbances or disorders and school problems, together with dysfunctional family situations, were the main reasons for families, children, and youth to seek help from CAP units. Such factors were also related to registered criminality and registered alcohol and drug abuse in former CAP patients as adults. This study investigated the risk for patients treated 1975–1990 to be registered as criminals until the end of 2003.</p> <p>Methods</p> <p>A regional sample of 1,400 former CAP patients, whose treatment occurred between 1975 and 1990, was followed to 2003, using database-record links to the Register of Persons Convicted of Offences at the National Council for Crime Prevention (NCCP).</p> <p>Results</p> <p>Every third CAP patient treated between 1975 and 1990 (every second man and every fifth woman) had entered the Register of Persons Convicted of Offences during the observation period, which is a significantly higher rate than the general population.</p> <p>Conclusion</p> <p>Results were compared to published results for CAP patients who were treated between 1953 and 1955 and followed over 20 years. Compared to the group of CAP patients from the 1950s, the results indicate that the risk for boys to enter the register for criminality has doubled and for girls, the risk seems to have increased sevenfold. The reasons for this change are discussed. Although hypothetical and perhaps speculative this higher risk of later criminality may be the result of lack of social control due to (1) rising consumption of alcohol, (2) changes in organisation of child social welfare work, (3) the school system, and (4) CAP methods that were implemented since 1970.</p

HVOF-Deposited WCCoCr as Replacement for Hard Cr in Landing Gear Actuators

WCCoCr coatings deposited by HVOF can replace hard Cr on landing gear components. Powders with two different WC particle sizes (micro and nano-) and geometries have been employed to study the effects on the coating’s properties. Moreover, coatings produced employing two sets of parameters resulting in high and low flame temperatures have been evaluated. Minor differences in microstructure and morphology were observed for the two powders employing the same spraying parameters, but the nano-sized powder exhibited a higher spraying efficiency. However, more significant microstructural changes result when the low- and high-energy spray parameters are used. Moreover, results of various tests which include adhesion, wear, salt fog corrosion resistance, liquid immersion, and axial fatigue strength, indicate that the coatings produced with high-energy flame are similar in behavior. On the other hand, the nanostructured low-energy flame coating exhibited a significantly lower salt fog corrosion resistanc

Population density and group size effects on reproductive behavior in a simultaneous hermaphrodite

<p>Abstract</p> <p>Background</p> <p>Despite growing evidence that population dynamic processes can have substantial effects on mating system evolution, little is known about their effect on mating rates in simultaneous hermaphrodites. According to theory, mating rate is expected to increase with mate availability because mating activity is primarily controlled by the male sexual function. A different scenario appears plausible in the hermaphroditic opisthobranch <it>Chelidonura sandrana</it>. Here, field mating rates are close to the female fitness optimum, suggesting that mating activity remains unresponsive to variation in mate availability.</p> <p>Results</p> <p>Applying an experimental design that aims at independent experimental manipulation of density and social group size, we find substantial increases in mate encounter rate with both factors, but no statistically detectable effects on mating rate in <it>C. sandrana</it>. Instead, mating rate remained close to the earlier determined female fitness optimum.</p> <p>Conclusions</p> <p>We demonstrate that mating rate in <it>C. sandrana </it>is largely unresponsive to variation in mate availability and is maintained close to the female fitness optimum. These findings challenge the prevailing notion of male driven mating rates in simultaneous hermaphrodites and call for complementary investigations of mating rate effects on fitness through the male sexual function.</p