A Novel GUCA1A Variant Associated with Cone Dystrophy Alters cGMP Signaling in Photoreceptors by Strongly Interacting with and Hyperactivating Retinal Guanylate Cyclase

Guanylate cyclase-activating protein 1 (GCAP1), encoded by the GUCA1A gene, is a neuronal calcium sensor protein involved in shaping the photoresponse kinetics in cones and rods. GCAP1 accelerates or slows the cGMP synthesis operated by retinal guanylate cyclase (GC) based on the light-dependent levels of intracellular Ca²⁺, thereby ensuring a timely regulation of the phototransduction cascade. We found a novel variant of GUCA1A in a patient affected by autosomal dominant cone dystrophy (adCOD), leading to the Asn104His (N104H) amino acid substitution at the protein level. While biochemical analysis of the recombinant protein showed impaired Ca²⁺ sensitivity of the variant, structural properties investigated by circular dichroism and limited proteolysis excluded major structural rearrangements induced by the mutation. Analytical gel filtration profiles and dynamic light scattering were compatible with a dimeric protein both in the presence of Mg²⁺ alone and Mg²⁺ and Ca²⁺. Enzymatic assays showed that N104H-GCAP1 strongly interacts with the GC, with an affinity that doubles that of the WT. The doubled IC₅₀ value of the novel variant (520 nM for N104H vs. 260 nM for the WT) is compatible with a constitutive activity of GC at physiological levels of Ca²⁺. The structural region at the interface with the GC may acquire enhanced flexibility under high Ca²⁺ conditions, as suggested by 2 μs molecular dynamics simulations. The altered interaction with GC would cause hyper-activity of the enzyme at both low and high Ca²⁺ levels, which would ultimately lead to toxic accumulation of cGMP and Ca²⁺ in the photoreceptor outer segment, thus triggering cell death.     

High and low surface area NiO–MgO and CoO–MgO solid solutions: a study of XPS surface composition and CO oxidation activity

Oxide solid solutions NiO–MgO of high surface area were studied by XPS. The surface Ni²⁺ concentration was found to be equal, within experimental errors, to the bulk concentration. The result is analogous to that found previously for the low surface area NiO–MgO system and for both the high and low surface area systems of CoO–MgO. The catalytic oxidation of CO by O₂, on high and low surface area NiO–MgO and CoO–MgO materials, was investigated with the aim of relating the catalytic activity with transition metal ion nature and concentration. Turnover frequency data (CO₂ molecules produced per second per surface atom) show that the activity is due primarily to the transition metal ions and is not subject to the ions being in special configurations (dimers or trimers) or in special positions (edges, corners). The activity of CoO–MgO is higher than that of NiO–MgO solid solution. This revised version was published online in June 2006 with corrections to the Cover Date.     

The Role of the Inflammatory Response in Mediating Functional Recovery Following Composite Tissue Injuries

Composite tissue injuries (CTI) are common among US Military Service members during combat operations, and carry a high potential of morbidity. Furthermore, CTI are often complicated due to an altered wound healing response, resulting in part from a dysregulation of the innate and adaptive immune responses. Unlike normal wound healing, in CTI, disruptions occur in innate immune responses, altering neutrophil functions, macrophage activation and polarization, further impacting the functions of T regulatory cells. Additionally, the biological underpinnings of these unfavorable wound healing conditions are multifactorial, including various processes, such as: ischemia, hypoxia, low nutrient levels, and altered cell metabolic pathways, among others, all of which are thought to trigger anergy in immune cells and destabilize adaptive immune responses. As a result, impaired wound healing is common in CTI. Herein, we review the altered innate and adaptive immune cells and their metabolic status and responses following CTI, and discuss the role a multi-pronged immunomodulatory approach may play in facilitating improved outcomes for afflicted patients.     

Microstructural Study of the Intermetallic Bonding Between AI Foam and Low Carbon Steel

Bonding between a metal foam core and a metallic skin is a pre requisite for the technological application of aluminum foam as filling reinforcement material to improve energy absorption and vibration damping of hollow components. This work is a preliminary study for the microstructural characterization of the interface layer formed between a commercial powder metallurgy （PM） precursor and a steel mould during foaming. The microstructure of the intermetallic layer was characterized by scanning electron microscopy, electron probe microanalysis and nanohardness measurements on the cross section. X-ray diffraction measurements, performed on the foam/substrate surface after stepwise material removal, allow the identification of the intermetallic phases. Two intermetallic layers, identified as Fe2Als and FeAI3, characterize the low Si foam/substrate while the AISiI0 foam/substrate interface evidences the presence of three Fe（Si, AI） intermetallic layers with different composition. Two and three different phases of increasing hardness could be distinguished going from the foam to the steel substrate for AIMglSi0.6 and AISiI0 precursors respectively. The results suggest the importance of elemental diffusion from steel substrate in the molten aluminum matrix （foam）. The possibility to control and tailor the microstructural properties of the interface between foam and steel skin is of fundamental importance in the technological process of foam filled structures manufacturing.     

On a scenario assessment tool for the operation of the JT-60SA superconducting tokamak

JT-60SA is a superconducting tokamak to be assembled and operated at the JAEA laboratories in Naka (Japan). The tokamak is designed, manufactured and operated under the funding of the Broader Approach Agreement (between the government of Japan and the European Commission) and of the Japan Fusion National Programme; JT-60SA aims to prepare, support and complement the ITER experimental programme. The European contribution to the JT-60SA is, for a large fraction, procured by France, Germany, Italy, Spain and Belgium.This paper summarizes the activities carried out at F4E to develop a user-friendly software tool able to assess in real-time if an operational scenario could be structurally withstood by the magnet system of JT-60SA. Such tool is based on a theoretical formulation which is supported by a series of dedicated finite element method (FEM) calculations, and is able to provide a comparative assessment of any candidate scenario with respect to the baseline scenarios, and a quantitative assessment of all electro magnetic (EM) forces acting on the magnet system at any time during the candidate scenario. The tool as it is presented is specifically designed to be used for the JT-60SA tokamak, though it is designed so to that its usage could be extended easily to any other tokamak.     

Upgrading the data acquisition and control systems of the European Breeding Blanket Test Facility

The EBBTF (European Breeding Blanket Test Facility) experimental plant is a key component for the development of the breeding blankets (TBMs test blanket modules, HCLL helium cooled lithium lead and HCPB helium cooled pebble bed types) used by ITER. EBBTF is an experimental plant which provides the double breeding/cooling loops (liquid metal and gas) required for HCLL testing. EBBTF is composed of four subsystems (TBM, IELLLO integrated European lead lithium loop, HE-FUS3 helium fusion loop, version 3 and helium compressor build by ATEKO) with dedicated control systems realized with hardware/software combinations covering 15 years (1995–2010) time span. At the end of 2010 we began to upgrade the HE-FUS3 data acquisition control systems (DACS) replacing the obsolete PLC Siemens S5 with National Instruments Compact FieldPoint and LabVIEW. The control room has been completely reorganized using high resolution monitors and workstations linked with standard Ethernet interfaces. The data acquisition, control, safety and SCADA software has been completely developed in ENEA using LabVIEW. In this paper we are going to discuss the technical difficulties and the solutions that we have used to accomplish the upgrade.     

Serine Hydroxymethyltransferase from the Cold Adapted Microorganism Psychromonas ingrahamii: A Low Temperature Active Enzyme with Broad Substrate Specificity

Serine hydroxymethyltransferase from the psychrophilic microorganism Psychromonas ingrahamii was expressed in Escherichia coli and purified as a His-tag fusion protein. The enzyme was characterized with respect to its spectroscopic, catalytic, and thermodynamic properties. The properties of the psychrophilic enzyme have been contrasted with the characteristics of the homologous counterpart from E. coli, which has been structurally and functionally characterized in depth and with which it shares 75% sequence identity. Spectroscopic measures confirmed that the psychrophilic enzyme displays structural properties almost identical to those of the mesophilic counterpart. At variance, the P. ingrahamii enzyme showed decreased thermostability and high specific activity at low temperature, both of which are typical features of cold adapted enzymes. Furthermore, it was a more efficient biocatalyst compared to E. coli serine hydroxymethyltransferase (SHMT) particularly for side reactions. Many β-hydroxy-α-amino acids are SHMT substrates and represent important compounds in the synthesis of pharmaceuticals, agrochemicals and food additives. Thanks to these attractive properties, this enzyme could have a significant potential for biotechnological applications.     

Advances in cultivar choice,hazelnut orchard management,and nut storage to enhance product quality and safety: an overview

European hazelnut (Corylus avellana L.) is a major species of interest for nutritional use within the Betulaceae family and its nuts are widely used throughout the world in the chocolate, confectionery, and bakery industries. Recently its cultivation has been expanded in traditional producer countries and established in new places in the southern hemisphere, including Chile, South Africa, and Australia. Introducing hazelnut in new environments could reduce its productivity, lead the trees to experience eco‐physiological disorders, and expose the crop to high pressure from common and new pests and diseases. Thus, new approaches in cultivar choice guidance, in the sustainable orchard management and even in nut storage and kernel quality evaluation are urgently required to improve the hazelnut production and processing chain. The main objective of this study was to systematize the published information regarding recent findings about the cultural operations that directly influence nut and kernel quality, support varietal choice for new plantations, and list the recent advances in nut storage and in quality and safety evaluation. © 2020 Society of Chemical Industry     

Multiphoton molecular photorelease in click-chemistry-functionalized gold nanoparticles

Yellow-green controlled photorelease: probes click-linked to peptide-coated gold nanospheres by a triazole ring can be released in living cells under a focused 561 nm laser at low power. Photocleaving follows a three-photon event stimulated by the excitation of the localized surface plasmon resonance.