Spectrometer for X-ray emission experiments at FERMI free-electron-laser

A portable and compact photon spectrometer to be used for photon in-photon out experiments, in particular x-ray emission spectroscopy, is presented. The instrument operates in the 25\u2013800 eV energy range to cover the full emissions of the FEL1 and FEL2 stages of FERMI. The optical design consists of two interchangeable spherical varied-lined-spaced gratings and a CCD detector. Different input sections can be accommodated, with/without an entrance slit and with/without an additional relay mirror, that allow to mount the spectrometer in different end-stations and at variable distances from the target area both at synchrotron and at free-electron-laser beamlines. The characterization on the Gas Phase beamline at ELETTRA Synchrotron (Italy) is presented

Estado actual de institucionalización y regulación de la evaluación y gestión ambiental de las obras de transporte en Argentina

La incorporación de la componente ambiental en las obras de transporte en Argentina se ha dado en las últimas décadas del siglo XX, con un desarrollo heterogéneo según el medio del que se trate. En ese sentido se destaca el estudio de las relaciones causales (planteadas generalmente en términos de "impactos") del transporte sobre el ambiente. Su ejemplo más concreto está dado por las Evaluaciones de Impacto Ambiental, entendidas como herramientas jurídico-administrativas basadas en informes técnicos, como los Estudios de Impacto Ambiental. Se presenta el estado de situación de la consideración de la dimensión ambiental en la evaluación y gestión de proyectos y construcción de obras de infraestructura para los distintos medios de transporte en la Argentina, a través del marco institucional y normativo ambiental vinculado a cada medio de transporte, acompañado por un conjunto de estudios de caso

Estado actual de institucionalización y regulación de la evaluación y gestión ambiental de las obras de transporte en Argentina

La incorporación de la componente ambiental en las obras de transporte en Argentina se ha dado en las últimas décadas del siglo XX, con un desarrollo heterogéneo según el medio del que se trate. En ese sentido se destaca el estudio de las relaciones causales (planteadas generalmente en términos de "impactos") del transporte sobre el ambiente. Su ejemplo más concreto está dado por las Evaluaciones de Impacto Ambiental, entendidas como herramientas jurídico-administrativas basadas en informes técnicos, como los Estudios de Impacto Ambiental. Se presenta el estado de situación de la consideración de la dimensión ambiental en la evaluación y gestión de proyectos y construcción de obras de infraestructura para los distintos medios de transporte en la Argentina, a través del marco institucional y normativo ambiental vinculado a cada medio de transporte, acompañado por un conjunto de estudios de caso.Fil: Daniele, Claudio. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Departamento e Instituto de Geografía; Argentina.Fil: Mereb, Juan Francisco. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Departamento de Geografía; Argentina.Fil: Frassetto, Andrea. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Departamento e Instituto de Geografía; Argentina.Fil: Pérez, Jimena. Aves Argentinas. Carrera de Naturalista de Campo e Intérprete de la Naturaleza; Argentina

Macrophage Inhibitor Clodronate Enhances Liver Transduction of Lentiviral but Not Adeno-Associated Viral Vectors or mRNA Lipid Nanoparticles in Neonatal and Juvenile Mice

Recently approved adeno-associated viral (AAV) vectors for liver monogenic diseases haemophilia A and B are exemplifying the success of liver-directed viral gene therapy. In parallel, additional gene therapy strategies are rapidly emerging to overcome some inherent AAV limitations, such as the non-persistence of the episomal transgene in the rapidly growing liver and immune response. Viral integrating vectors such as in vivo lentiviral gene therapy and non-viral vectors such as lipid nanoparticles encapsulating mRNA (LNP-mRNA) are rapidly being developed, currently at the preclinical and clinical stages, respectively. Macrophages are the first effector cells of the innate immune response triggered by gene therapy vectors. Macrophage uptake and activation following administration of viral gene therapy and LNP have been reported. In this study, we assessed the biodistribution of AAV, lentiviral, and LNP-mRNA gene therapy following the depletion of tissue macrophages by clodronate pre-treatment in neonatal and juvenile mice. Both neonatal and adult clodronate-treated mice showed a significant increase in lentiviral-transduced hepatocytes. In contrast, clodronate pre-treatment did not modify hepatocyte transduction mediated by hepatotropic AAV8 but reduced LNP-mRNA transfection in neonatal and juvenile animals. These results highlight the importance of age-specific responses in the liver and will have translational applications for gene therapy programs

mRNA therapy corrects defective glutathione metabolism and restores ureagenesis in preclinical argininosuccinic aciduria

The urea cycle enzyme argininosuccinate lyase (ASL) enables the clearance of neurotoxic ammonia and the biosynthesis of arginine. Patients with ASL deficiency present with argininosuccinic aciduria, an inherited metabolic disease with hyperammonemia and a systemic phenotype coinciding with neurocognitive impairment and chronic liver disease. Here, we describe the dysregulation of glutathione biosynthesis and upstream cysteine utilization in ASL-deficient patients and mice using targeted metabolomics and in vivo positron emission tomography (PET) imaging using ( S)-4-(3-18F-fluoropropyl)-l-glutamate ([18F]FSPG). Up-regulation of cysteine metabolism contrasted with glutathione depletion and down-regulated antioxidant pathways. To assess hepatic glutathione dysregulation and liver disease, we present [18F]FSPG PET as a noninvasive diagnostic tool to monitor therapeutic response in argininosuccinic aciduria. Human hASL mRNA encapsulated in lipid nanoparticles improved glutathione metabolism and chronic liver disease. In addition, hASL mRNA therapy corrected and rescued the neonatal and adult Asl-deficient mouse phenotypes, respectively, enhancing ureagenesis. These findings provide mechanistic insights in liver glutathione metabolism and support clinical translation of mRNA therapy for argininosuccinic aciduria. </p

The incidence of movement disorder increases with age and contrasts with subtle and limited neuroimaging abnormalities in argininosuccinic aciduria

Argininosuccinate lyase (ASL) is integral to the urea cycle detoxifying neurotoxic ammonia and the nitric oxide (NO) biosynthesis cycle. Inherited ASL deficiency causes argininosuccinic aciduria (ASA), a rare disease with hyperammonemia and NO deficiency. Patients present with developmental delay, epilepsy and movement disorder, associated with NO-mediated downregulation of central catecholamine biosynthesis. A neurodegenerative phenotype has been proposed in ASA. To better characterise this neurodegenerative phenotype in ASA, we conducted a retrospective study in six paediatric and adult metabolic centres in the UK in 2022. We identified 60 patients and specifically looked for neurodegeneration-related symptoms: movement disorder such as ataxia, tremor and dystonia, hypotonia/fatigue and abnormal behaviour. We analysed neuroimaging with diffusion tensor imaging (DTI) magnetic resonance imaging (MRI) in an individual with ASA with movement disorders. We assessed conventional and DTI MRI alongside single photon emission computer tomography (SPECT) with dopamine analogue radionuclide 123 I-ioflupane, in Asl-deficient mice treated by hASL mRNA with normalised ureagenesis. Movement disorders in ASA appear in the second and third decades of life, becoming more prevalent with ageing and independent from the age of onset of hyperammonemia. Neuroimaging can show abnormal DTI features affecting both grey and white matter, preferentially basal ganglia. ASA mouse model with normalised ureagenesis did not recapitulate these DTI findings and showed normal 123 I-ioflupane SPECT and cerebral dopamine metabolomics. Altogether these findings support the pathophysiology of a late-onset movement disorder with cell-autonomous functional central catecholamine dysregulation but without or limited neurodegeneration of dopaminergic neurons, making these symptoms amenable to targeted therapy

Development of an Integrated Countermeasure Device for Use in Long-Duration Space Flight

Prolonged weightlessness is associated with declines in musculoskeletal, cardiovascular, and sensorimotor health. Consequently, in-flight countermeasures are required to preserve astronaut health. We developed and tested a novel exercise countermeasure device (CCD) for use in spaceflight with the aim of preserving musculoskeletal and cardiovascular health along with an incorporated balance-training component. Additionally, the CCD features a compact footprint, and a low power requirement. Methods: After design and development of the CCD, we carried out a training study to test its ability to improve cardiovascular and muscular fitness in healthy volunteers. Fourteen male and female subjects (41.4+/-9.0 years, 69.5+/-15.4Kg) completed 12 weeks (3 sessions per week) of concurrent strength and endurance training on the CCD. Subjects were tested at baseline and after 12 weeks for 1-repetition max leg press strength (1RM), peak oxygen consumption (VO2peak), and isokinetic joint torque (ISO) at the hip, knee, and ankle. Additionally, we evaluated subjects after 6 weeks of training for changes in VO2peak and 1RM. Results: VO2peak and 1RM improved after 6-weeks, with additional improvements after 12 weeks (1.95+/-0.5, 2.28+/-0.5, 2.47+/-0.6 LY/min and 131.2+/-63.9,182.8+/-75.0, 207.0+/-75.0 Kg) for baseline, 6 weeks, and 12 weeks respectively. ISO for hip adduction, adduction, and ankle plantar flexion improved after 12 weeks of training (70.3+/-39.5, 76.8+/-39.2 and 55.7+/-21.7 N-m vs. 86.1+/-37.3, 85.1+/-34.3 and 62.1+/-26.4 N-m respectively). No changes were observed for ISO during hip flexion, knee extension, or knee flexion. Conclusions: The CCD is effective at improving cardiovascular fitness and isotonic leg strength in healthy adults. Further, the improvement in hip adductor and abductor torque provides support that the CCD may provide additional protection for the preservation of bone health at the hip

The Comet Interceptor Mission

Here we describe the novel, multi-point Comet Interceptor mission. It is dedicated to the exploration of a little-processed long-period comet, possibly entering the inner Solar System for the first time, or to encounter an interstellar object originating at another star. The objectives of the mission are to address the following questions: What are the surface composition, shape, morphology, and structure of the target object? What is the composition of the gas and dust in the coma, its connection to the nucleus, and the nature of its interaction with the solar wind? The mission was proposed to the European Space Agency in 2018, and formally adopted by the agency in June 2022, for launch in 2029 together with the Ariel mission. Comet Interceptor will take advantage of the opportunity presented by ESA's F-Class call for fast, flexible, low-cost missions to which it was proposed. The call required a launch to a halo orbit around the Sun-Earth L2 point. The mission can take advantage of this placement to wait for the discovery of a suitable comet reachable with its minimum ΔV capability of 600 ms-1. Comet Interceptor will be unique in encountering and studying, at a nominal closest approach distance of 1000 km, a comet that represents a near-pristine sample of material from the formation of the Solar System. It will also add a capability that no previous cometary mission has had, which is to deploy two sub-probes - B1, provided by the Japanese space agency, JAXA, and B2 - that will follow different trajectories through the coma. While the main probe passes at a nominal 1000 km distance, probes B1 and B2 will follow different chords through the coma at distances of 850 km and 400 km, respectively. The result will be unique, simultaneous, spatially resolved information of the 3-dimensional properties of the target comet and its interaction with the space environment. We present the mission's science background leading to these objectives, as well as an overview of the scientific instruments, mission design, and schedule

Nanostructural analysis of the adhesive interface in dentistry

2012/2013The subject of this thesis is the stability of the adhesive interface in dentistry. Success in adhesive dentistry means long lasting restorations. However, there is substantial evidence that this ideal objective is not achieved. Current research in this field aims at increasing the resin-dentin bond durability. This doctoral research examines the fundamental processes responsible for the aging mechanisms involved in the degradation of resin-bonded interfaces, as well as some potential approaches to prevent and counteract this degradation. Resin-dentin bond degradation is a complex process that is not completely understood, involving the hydrolysis of both the resin and the collagen component of the hybrid layer. The hydrophilic and acidic characteristics of current dentin adhesives have made hybrid layers highly prone to water sorption, which causes polymer degradation and results in decreased resin-dentin bond strength over time. These unstable polymers inside the hybrid layer may result in an incomplete encapsulation of collagen fibers, which become vulnerable to mechanical and hydrolytical fatigue, as well as degradation by host-derived proteases with collagenolytic activity. These enzymes, such as matrix metalloproteinases (MMPs) and cysteine cathepsins, have a crucial role in the degradation of type I collagen, the organic component of the hybrid layer. The first part of this thesis aims to review the current knowledge regarding adhesion to the tooth substrate (Chapter 1), focusing on the fundamental processes that are responsible for the degradation of the adhesive interface (Chapter 2). Since the permeability of adhesives to water is particularly evident in simplified adhesive formulations, the research activity was focused on self-etch and universal adhesive systems’ behavior. Thus, the research study reported in Chapter 3 showed that the bond strength and nanoleakage expression of two-step and one-step self-etch tested bonding systems were affected by storage for 6 month and 1 year in artificial saliva. Although it is generally accepted that the permeability of adhesives to water is particularly evident in simplified adhesive formulations, the stability over time was not related to the number of steps of bonding systems, but to their chemical formulations. The performance of a new universal (or multi-mode) adhesive system through storage in artificial saliva was also investigated. The original results presented in Chapter 4 found that improved bonding effectiveness of the tested universal adhesive system on dentin was obtained when the adhesive was applied with the self-etch approach. Indeed, the etch-and-rinse approaches tested (both on wet and dry dentin) resulted in immediate bond strength comparable to the self-etch mode but expedited long-term aging resulted in reduced bond strength and increased nanoleakage expression, irrespective of dentin wetness. Moreover, the results of the zymographic analysis showed evident changes in dentinal MMP-2 and -9 enzyme activities after the application of the tested adhesives, revealing differences in the extent of enzyme activation. These findings exhibit that the activation of endogenous MMPs is not related to the adhesive system or the strategy employed. Thus, regardless of the approach and the material used in bonding procedures, a stable and durable bond is not achieved. Therefore, experimental strategies that aim to enhance the adhesive interface, particularly improving the durability of the resin-dentin bond strength by inhibiting intrinsic collagenolytic activity and increasing the resistance of dentin collagen matrix to enzymatic degradation are needed. The last part of the thesis is focused on both the strategies to inhibit the proteolytic and collagenolytic activity of the endogenous proteases and the methods to increase the mechanical strength of collagen network and its resistance to enzymatic degradation (Chapter 5). Chlorhexidine (CHX) has been used as a non-specific MMP inhibitor to prevent degradation of hybrid layers. However, CHX is water-soluble and may leach out of hybrid layers, compromising its long-term anti-MMP effectiveness. An entirely different approach is to treat the acid-etched dentin containing activated matrix-bound MMPs with cross-linking agents that inactivate the catalytic site of proteases. In particular, the ability of a cross-linker agent, 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (EDC), to prevent collagen degradation was evaluated under occlusal cycle loading. Previous research successfully utilized EDC to increase the durability of resin-dentin bonds by increasing the mechanical properties of the collagen matrix; however, the 1 to 4 hrs required for that procedure was clinically unacceptable. For this reason, the purpose of the last part of the research, presented in Chapter 6, was to evaluate the ability of 0.5 M EDC short-time (1 min) pre-treatment to improve the stability of demineralized dentin collagen matrices by quantifying the release of telopeptide fragments over time. The results showed that EDC application for 1 min may be a clinically relevant and effective means for stabilizing the collagen network not only by strengthening the fibrils, but also by reducing the enzymatic degradation rate. Thus, dentin collagen reinforcement and strengthening through EDC cross-linking might be of importance to improve the bond strength and structural integrity of the resin-dentin interface over time against the enzymatic and hydrolytic degradation.La tesi qui presentata riguarda la stabilità dell'interfaccia adesiva in odontoiatria. Il successo delle moderne terapie conservative è rappresentato dalla longevità dei restauri adesivi. Tuttavia, vi è una sostanziale evidenza che questo obiettivo ideale non sia raggiunto. La stabilità dell’interfaccia adesiva dipende dalla formazione di uno strato ibrido, compatto e omogeneo, durante l’impregnazione del substrato dentinale da parte dei monomeri adesivi. Poiché lo strato ibrido rappresenta un’entità complessa, in cui interagiscono componenti biologiche diverse (matrice dentinale collagenica e cristalli d’idrossiapatite residui) e non (monomeri resinosi e solventi), i fenomeni d’invecchiamento interessano in maniera sinergica sia la porzione resinosa che quella dentale. L’articolato processo che porta alla degradazione dell’interfaccia adesiva coinvolge infatti la componente resinosa, attraverso l’idrolisi della resina negli spazi interfibrillari, e quella organica, attraverso la disorganizzazione delle fibre collagene dovuta ad un incompleto incapsulamento delle stesse, nonché alla degradazione da parte di proteasi intrinseche con attività collagenolitica. È stato dimostrato come questi enzimi, le metalloproteinasi della matrice (MMP) e le catepsine, abbiano un ruolo cruciale nella degradazione del collagene di tipo I, la principale componente organica dello strato ibrido. Inoltre le caratteristiche idrofile e acide degli attuali sistemi adesivi dentinali hanno reso lo strato ibrido molto suscettibile all'assorbimento di acqua, comportando, attraverso l’idrolisi, la degradazione dello stesso e andando così a contribuire ad una diminuzione della forza di legame nel tempo. Attualmente l’interesse della comunità scientifica mira ad aumentare la durata del legame adesivo con il substrato dentinale. Dopo un’attenta analisi delle attuali conoscenze riguardanti adesione al substrato dentale (Capitolo 1), la prima parte della tesi si propone di valutare i processi fondamentali che sono responsabili della degradazione dell'interfaccia adesiva (Capitolo 2). Poiché la permeabilità all’acqua degli adesivi è particolarmente evidente nelle formulazioni semplificate, l'attività di ricerca si è concentrata sull’analisi del comportamento dei sistemi adesivi self-etch e dei recenti sistemi adesivi universali. I risultati riportati nel Capitolo 3 ha dimostrato come la forza di legame e l’espressione del nanoleakage dei sistemi adesivi self-etch two-step e one-step testati sia negativamente influenzata dall’invecchiamento in saliva artificiale per 6 mesi e 1 anno. Sebbene sia generalmente accettato che la permeabilità degli adesivi all'acqua è particolarmente evidente in formulazioni di adesivi semplificati, la stabilità nel tempo non è stata correlata al numero di passaggi dei sistemi adesivi, bensì alle loro composizioni chimiche. Sono state in seguito analizzate anche le prestazioni di un nuovo sistema adesivo universale (o multimodale). I risultati presentati nel Capitolo 4 hanno stabilito una migliore efficienza adesiva del sistema universale, testato sul substrato dentinale, quando l'adesivo è stato applicato con l'approccio self-etch. Infatti, la tecnica etch-and-rinse, testata sia su dentina umida che secca, ha comportato una forza di adesione immediata paragonabile alla modalità self-etch, ma a tempi di invecchiamento incrementali si è evidenziata una diminuzione della forza di legame e una maggiore espressione del nanoleakage, a prescindere dalla condizione di umidità dentinale. Inoltre, i risultati dell'analisi zimografica hanno mostrato evidenti variazioni dell’attività enzimatica delle metalloproteinasi MMP-2 e -9 dopo l'applicazione degli adesivi testati. Questi risultati dimostrano come l'attivazione delle MMP endogene non sia correlata al sistema adesivo o alla strategia adottata. Ne evince che, indipendentemente dal metodo e dal materiale utilizzato nelle procedure adesive, non si è in grado di stabilire un legame affidabile e duraturo. Pertanto si avverte l’esigenza di strategie sperimentali che mirino a migliorare la stabilità dell’interfaccia adesiva, in particolare incrementando la durata della forza di legame in dentina inibendo l'attività collagenolitica intrinseca e aumentando la resistenza del collagene alla degradazione enzimatica. L'ultima parte della tesi è focalizzata quindi sulle strategie per inibire l'attività proteolitica e collagenolitica delle proteasi endogene e sui metodi per aumentare la resistenza meccanica del collagene alla degradazione enzimatica (Capitolo 5). Un potente agente antibatterico, la clorexidina (CHX), è stato usato come inibitore non specifico delle MMP al fine di impedire la degradazione dello strato ibrido. Tuttavia la CHX, essendo solubile in acqua, può dissolversi nello strato ibrido, compromettendo la sua efficacia anti-MMP a lungo termine. Un approccio completamente diverso è quello di trattare la dentina mordenzata con agenti cross-linker. In particolare, simulando il carico occlusale, è stata valutata la capacità di un agente cross-linker, l’1-etil-3-(3-dimetilammino-propil) carbodiimmide (EDC), per prevenire la degradazione del collagene. Precedenti ricerche hanno utilizzato con successo l’EDC con lo scopo di aumentare la durata dell’interfaccia adesiva, aumentando le proprietà meccaniche della matrice di collagene; tuttavia, il tempo necessario (da 1 a 4 ore) richiesto per tali procedure è clinicamente inaccettabile. Per questo motivo, lo scopo dell’ultima parte della ricerca, presentata nel Capitolo 6, è stato quello di valutare la capacità di 0,5 M EDC nel breve periodo di pretrattamento (1 min), andando a quantificare il rilascio di frammenti di telopeptidi di collagene nel corso del tempo. I risultati hanno dimostrato che l'applicazione di EDC per 1 min può essere un approccio clinicamente rilevante ed efficace nello stabilizzare il collagene, non solo rafforzando le fibrille, ma anche riducendo la velocità di degradazione enzimatica. Di conseguenza, l’utilizzo di questo cross-linker può garantire una valida strategia per migliorare la forza di legame e l'integrità strutturale dell'interfaccia adesiva nel tempo contro l’attività enzimatica intrinseca del collagene e la degradazione idrolitica.XXVI Ciclo198

Ultrafast charge dynamics in an amino acid induced by attosecond pulses

In the past few years, attosecond techniques have been implemented for the investigation of ultrafast dynamics in molecules. The generation of isolated attosecond pulses characterized by a relatively high photon flux has opened up new possibilities in the study of molecular dynamics. In this paper, we report on experimental and theoretical results of ultrafast charge dynamics in a biochemically relevant molecule, namely, the amino acid phenylalanine. The data represent the first experimental demonstration of the generation and observation of a charge migration process in a complexmolecule, where electron dynamics precede nuclear motion. The application of attosecond technology to the investigation of electron dynamics in biologically relevant molecules represents a multidisciplinary work, which can open new research frontiers: those in which few-femtosecond and even subfemtosecond electron processes determine the fate of biomolecules. It can also open new perspectives for the development of new technologies, for example, in molecular electronics, where electron processes on an ultrafast temporal scale are essential to trigger and control the electron current on the scale of the molecule