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Delivery of adeno-associated viral vectors (AAVs) to cerebrospinal fluid (CSF) has emerged as a promising approach to achieve widespread transduction of the central nervous system (CNS) and peripheral nervous system (PNS), with direct applicability to the treatment of a wide range of neurological diseases, particularly lysosomal storage diseases. Although studies in small animal models have provided proof of concept and experiments in large animals demonstrated feasibility in bigger brains, there is not much information on long-term safety or durability of the effect. Here, we report a 7-year study in healthy beagle dogs after intra-CSF delivery of a single, clinically relevant dose (2 × 1013 vg/dog) of AAV9 vectors carrying the canine sulfamidase, the enzyme deficient in mucopolysaccharidosis type IIIA. Periodic monitoring of CSF and blood, clinical and neurological evaluations, and magnetic resonance and ultrasound imaging of target organs demonstrated no toxicity related to treatment. AAV9-mediated gene transfer resulted in detection of sulfamidase activity in CSF throughout the study. Analysis at tissue level showed widespread sulfamidase expression and activity in the absence of histological findings in any region of encephalon, spinal cord, or dorsal root ganglia. Altogether, these results provide proof of durability of expression and long-term safety for intra-CSF delivery of AAV-based gene transfer vectors encoding therapeutic proteins to the CNS., Graphical abstract, Seven-year follow-up of dogs after intra-CSF administration of AAV9-sulfamidase vectors results in detection of sulfamidase activity in CSF and widespread transgene expression in CNS, PNS, and liver, in the absence of any adverse events. Proof of durability and safety of this gene therapy support its clinical translation to treat CNS diseases.

En Valentia y su entorno se registró una intensa actividad constructiva en determinados lugares y momentos entre los siglos VI y VIII. La construcción del monumental centro episcopal en la primera mitad del s. VI, iniciativa del obispo Justiniano, cambió el paisaje urbano. En sus edificios (catedral, baptisterio, mausoleo y otros) se aprecia el uso continuado, pero selectivo y diferenciado, según cada uno de los conjuntos, de gran cantidad de elementos romanos. A partir de mediados del s. VI la actividad edilicia en Valentia decayó. Lo único a destacar es que se urbanizó la arena del Circo, un amplio espacio de 70 por 350 metros. Estos hallazgos se conocen peor, por ser excavaciones de poca extensión que no permiten identificar ningún edificio entre los muros aparecidos, pero sí que atestiguan un proyecto constructivo unitario, aunque de menor entidad que en la zona episcopal. Los materiales arqueológicos presentan una importante proporción de importaciones orientales y africanas.A finales del s. VI, a 16 km. de Valentia surgió un nuevo asentamiento fortificado de cierta extensión, 5 hectáreas, València la Vella. Su construcción, ex novo, supuso un gran esfuerzo edilicio. Además del gran recinto amurallado se han localizado dos edificios públicos en la parte baja y otro en la ciudadela. Debió ser el centro del poder de la provincia Carthaginensis.El yacimiento se abandonó a finales del s. VII o inicios del VIII. Al mismo tiempo, se creó otro centro monumental, el importante centro de poder de Pla de Nadal, a unos 2 km. de València la Vella. Su elemento más notorio fue el gran palacio de patio central, pero los diversos edificios que van apareciendo a su alrededor, indican que sería un complejo aún por delimitar. An intense constructive activity was developed in certain places and times in Valentia and around between the 6th and 8th centuries. The construction of the monumental episcopal center in the first half of the 6th century changed the urban landscape. The new buildings (cathedral, baptistery, mausoleum and others) continuously and selectively used a large number of Roman elements. The Circus arena was urbanized from the second half of the 6th century until the middle of the 7th. These findings are small excavations. There are a lot of walls but they do not allow to identify any building. Anyway, they attest the continuity in the reuse of Roman materials. This area had less entity than the Episcopal area. In both cases, the supply of the reused material would be from the remains of the old Roman town. Archaeological materials have a significant proportion of Eastern and African imports.Valencia la Vella, a new and very important fortified settlement of 5 hectares, emerged at the end of the 6th century. The site is 16 km away from Valentia on the Turia river. This new city was made ex novo, so it was necessary a great building effort. In addition to the large walled enclosure two public buildings have been located in the lower part and another in the citadel. This site must have been the center of power in Carthaginensis province.Valencia la Vella was abandoned in the late 7th or early 8th centuries. Contemporarily another important power center was created in Pla de Nadal, about 2 km. from Valencia la Vella. Probably the recently disappeared site would be used as a quarry.

Background: The prevalence of multimorbidity and the number of chronic pathologies increases with age, and hospitalization rates are higher in very elderly patients. Aims: To identify predictors of mortality in patients aged 90 years or more at 6 and 12 months after admission to the internal medicine ward. Materials and methods: Prospective cohort study in patients aged 90 years or older who were admitted to the internal medicine ward of Alicante General University Hospital from November 2018 to March 2019. We collected data for patient scores on the Barthel, Lawton-Brody, and Short Portable Mental Status Questionnaire; a nutritional questionnaire; the Charlson index; PALIAR and PROFUND prognostic indexes; and mortality at 6 and 12 months. Results: In the bivariate analysis adjusted for gender and comorbidity, malnutrition was associated with mortality at 6 and 12 months (odds ratio (OR) 5.17, 95% confidence interval (C11 1.30, 20.50; OR 7.78, 95% CI 1.47, 40.9, respectively), as were moderate-severe cognitive impairment (6 months: OR 12.0, 95% CI 2.79, 52.0; 12 months: OR 10.73, 95% CI 2.40, 47.8) and high-risk scores on the PALIAR (6 months: OR 9.18, 95% CI 1.99, 42.27;12 months: OR 11.7, 95% CI 2.62, 51.60) and PROFUND (6 months: OR 10.7, 95% CI 1.68, 68.80; 12 months: OR 20.9, 95% CI 3.15, 139.0) prognostic indexes. The Barthel index was associated with mortality only at 6 months (OR 6.16; 95% CI 1.69, 22.4). Conclusions: In nonagenarian inpatients, a worse score on the comprehensive assessment and a higher score on prognostic indexes were associated with increased risk of mortality at 6 and 12 months. Copyright (C) 2021, Taiwan Society of Geriatric Emergency & Critical Care Medicine.

Neovascularization associated with diabetic retinopathy (DR) and other ocular disorders is a leading cause of visual impairment and adult-onset blindness. Currently available treatments are merely palliative and offer temporary solutions. Here, we tested the efficacy of antiangiogenic gene transfer in an animal model that mimics the chronic progression of human DR. Adeno-associated viral (AAV) vectors of serotype 2 coding for antiangiogenic Pigment Epithelium Derived Factor (PEDF) were injected in the vitreous of a 1.5 month-old transgenic model of retinopathy that develops progressive neovascularization. A single intravitreal injection led to long-term production of PEDF and to a striking inhibition of intravitreal neovascularization, normalization of retinal capillary density, and prevention of retinal detachment. This was parallel to a reduction in the intraocular levels of Vascular Endothelial Growth Factor (VEGF). Normalization of VEGF was consistent with a downregulation of downstream effectors of angiogenesis, such as the activity of Matrix Metalloproteinases (MMP) 2 and 9 and the content of Connective Tissue Growth Factor (CTGF). These results demonstrate long-term efficacy of AAV-mediated PEDF overexpression in counteracting retinal neovascularization in a relevant animal model, and provides evidence towards the use of this strategy to treat angiogenesis in DR and other chronic proliferative retinal disorders.

Presentamos los primeros resultados obtenidos en el proyecto de excavaciones arqueológicas en València la Vella y ejecutados entre los años 2016 y 2018. En estas primeras campañas hemos optado por la documentación topográfica del asentamiento y la ejecución de sondeos de verificación estratigráfica. Además se ha incidido en la documentación de los tramos de muralla más afectados por destrucciones realizadas en el siglo XX.Estas actividades han permitido corroborar la cronología visigoda de un núcleo amurallado de casi 5 ha y constatar la importancia del urbanismo intramuros, así como de las actividades económicas que se pudieron realizar en su interior. De este modo, València la Vella se ofrece a la comunidad científica como uno de los yacimientos arqueológicos visigodos más relevantes para el conocimiento del urbanismo y la sociedad de los siglos VI y VII. We present the first results obtained in the archaeological research project carried out in València la Vella between 2016 and 2018. In these first campaigns we documented the settlement topographically and dug stratigraphic verification trenches. We have also started to document the section of the wall most damaged by actions in the twentieth century. These activities have allowed us to corroborate the Visigothic chronology of a walled nucleus of almost 5 hectares and to confirm the importance of the intramural urban planning and the economic activities undertaken in it. In this way we are able to offer València la Vella to the scientific community as one of the most important Visigothic archaeological sites for contributing to our knowledge of urban planning and society in the sixth and seventh centuries.

The first historically documented pandemic caused by Yersinia pestis began as the Justinianic Plague in 541 within the Roman Empire and continued as the so-called First Pandemic until 750. Although paleogenomic studies have previously identified the causative agent as Y. pestis , little is known about the bacterium’s spread, diversity, and genetic history over the course of the pandemic. To elucidate the microevolution of the bacterium during this time period, we screened human remains from 21 sites in Austria, Britain, Germany, France, and Spain for Y. pestis DNA and reconstructed eight genomes. We present a methodological approach assessing single-nucleotide polymorphisms (SNPs) in ancient bacterial genomes, facilitating qualitative analyses of low coverage genomes from a metagenomic background. Phylogenetic analysis on the eight reconstructed genomes reveals the existence of previously undocumented Y. pestis diversity during the sixth to eighth centuries, and provides evidence for the presence of multiple distinct Y. pestis strains in Europe. We offer genetic evidence for the presence of the Justinianic Plague in the British Isles, previously only hypothesized from ambiguous documentary accounts, as well as the parallel occurrence of multiple derived strains in central and southern France, Spain, and southern Germany. Four of the reported strains form a polytomy similar to others seen across the Y. pestis phylogeny, associated with the Second and Third Pandemics. We identified a deletion of a 45-kb genomic region in the most recent First Pandemic strains affecting two virulence factors, intriguingly overlapping with a deletion found in 17th- to 18th-century genomes of the Second Pandemic.

Significance The first historically reported pandemic attributed to Yersinia pestis started with the Justinianic Plague (541–544) and continued for around 200 y as the so-called First Pandemic. To date, only one Y. pestis strain from this pandemic has been reconstructed using ancient DNA. In this study, we present eight genomes from Britain, France, Germany, and Spain, demonstrating the geographic range of plague during the First Pandemic and showing microdiversity in the Early Medieval Period. Moreover, we detect similar genome decay during the First and Second Pandemics (14th to 18th century) that includes the same two virulence factors, thus providing an example of potential convergent evolution of Y. pestis during large-scale epidemics., The first historically documented pandemic caused by Yersinia pestis began as the Justinianic Plague in 541 within the Roman Empire and continued as the so-called First Pandemic until 750. Although paleogenomic studies have previously identified the causative agent as Y. pestis, little is known about the bacterium’s spread, diversity, and genetic history over the course of the pandemic. To elucidate the microevolution of the bacterium during this time period, we screened human remains from 21 sites in Austria, Britain, Germany, France, and Spain for Y. pestis DNA and reconstructed eight genomes. We present a methodological approach assessing single-nucleotide polymorphisms (SNPs) in ancient bacterial genomes, facilitating qualitative analyses of low coverage genomes from a metagenomic background. Phylogenetic analysis on the eight reconstructed genomes reveals the existence of previously undocumented Y. pestis diversity during the sixth to eighth centuries, and provides evidence for the presence of multiple distinct Y. pestis strains in Europe. We offer genetic evidence for the presence of the Justinianic Plague in the British Isles, previously only hypothesized from ambiguous documentary accounts, as well as the parallel occurrence of multiple derived strains in central and southern France, Spain, and southern Germany. Four of the reported strains form a polytomy similar to others seen across the Y. pestis phylogeny, associated with the Second and Third Pandemics. We identified a deletion of a 45-kb genomic region in the most recent First Pandemic strains affecting two virulence factors, intriguingly overlapping with a deletion found in 17th- to 18th-century genomes of the Second Pandemic.

This contribution presents the results of the archaeometric characterization of Late Roman pottery from the city of Valencia (Spain). The ceramic samples, including common wares, amphorae, and cooking wares, have been characterized using WD-X-ray fluorescence for the chemical analysis, X-ray diffraction for the mineralogical study, and optical microscopy by thin-section analysis for the mineralogical and petrographic characterization. The results show a wide range of fabrics with cooking wares of imported origin. The most important result is, however, the identification of a possible regional ware that resembles African cooking wares but that could have been locally or regionally produced, as well as the possible production in the area of some common wares. These results provide a better insight into the production and distribution of Late Roman ceramics in the area and contribute to a better understanding of trade dynamics in the Western Mediterranean for imported wares.

Sustainable building and/or green building strategies have been founded on two basic principles: energy efficiency and environmental responsibility. Energy efficiency attempts to serve a dual purpose, the first geopolitical – to end dependency on fossil fuel supplies from other countries – and the second to reduce energy consumption. While various methods for assessing the environmental impact of residential buildings are available and a number of certifications have been created, these certifications do not actually help home owners and users, but rather are endorsing buildings that have been constructed with no attempt at sustainability, thus making the diffusion of any sustainability criteria in construction that much more difficult. This study aims to develop a simple user-friendly tool for home owners that provides the same rigor that current sustainability certificates offer the professional builder and property developer. A sustainability appraisal is carried out based on social, environmental and economic impacts, each of which is evaluated using easily understandable parameters. The integrated system is then verified through the case study of a real residential building. The subsequent results demonstrate a better building performance when pursuing green demand in housing and also indicate the changes required for improvement. In applying this new integrated tool we are able to confirm its ease of use, along with its highly comparable results with any other evaluation tools currently available.

By 2020, different European Directives on the energy performance of buildings (EPBD) aim to reduce energy consumption by 20%, reduce greenhouse gas emissions by 20% and endorse a target of a 20% share of energy from renewable sources. These Directives have led EU Member States to develop systems, following their corresponding transposition into national law, to evaluate the energy efficiency of buildings, and create different Energy Performance Certificates (EPCs). This has led to different energy efficiency measurement systems in the EU and a CO 2 emissions scale has not been included by some Member States. Just as EPC is aimed at influencing the demand for energy-efficient buildings, this study is aimed at developing a straightforward procedure to obtain the environmental impact in the use phase of a building in kgCO 2 /m 2 year in order to influence the demand for buildings with a low impact on climate change. By applying this methodology to different case studies, we can conclude that obtaining the formula drastically simplifies the calculation of CO 2 emissions in KgCO 2 /m 2 year. The results obtained from applying this methodology will serve to record CO 2 emission rates in single-family houses on a yearly basis, thus contributing to the study of applications that address climate change mitigation.

The first historically documented pandemic caused byYersinia pestisstarted as the Justinianic Plague in 541 within the Roman Empire and continued as the so-called First Pandemic until 750. Although palaeogenomic studies have previously identified the causative agent asY. pestis, little is known about the bacterium’s spread, diversity and genetic history over the course of the pandemic.To elucidate the microevolution of the bacterium during this time period, we screened human remains from 20 sites in Austria, Britain, Germany, France and Spain forY. pestisDNA and reconstructed six new genomes. We present a novel methodological approach assessing SNPs in ancient bacterial genomes, facilitating qualitative analyses of low coverage genomes from a metagenomic background. Phylogenetic analysis reveals the existence of previously undocumentedY. pestisdiversity during the 6th–7thcenturies, and provides evidence for the presence of multiple distinctY. pestisstrains in Europe. We offer genetic evidence for the presence of the Justinianic Plague in the British Isles, previously only hypothesized from ambiguous documentary accounts, as well as southern France and Spain, and that southern Germany seems to have been affected by at least two distinctY. pestisstrains. Four of the reported strains form a polytomy similar to others seen across theY. pestisphylogeny, associated with the Second and Third Pandemics. We identified a deletion of a 45 kb genomic region in the most recent First Pandemic strain affecting two virulence factors, intriguingly overlapping with a deletion found in 17th–18th-century genomes of the Second Pandemic.Significance StatementThe first historically reported pandemic attributed toYersinia pestisstarted with the Justinianic Plague (541–544) and continued for around 200 years as the so-called First Pandemic. To date, only oneY. pestisstrain from this pandemic has been reconstructed using ancient DNA. In this study, we present six new genomes from Britain, France, Germany and Spain, demonstrating the geographic range of plague during the First pandemic and showing microdiversity in the Early Medieval Period. Moreover, we detect similar genome decay during the First and Second Pandemic (17thto 18thcentury) that includes the same two virulence factors, thus providing an example of potential convergent evolution ofY. pestisduring large scale epidemics.

Many different research papers have been written about sustainable building, outlining which indicators should be considered for the construction of sustainable buildings. However, due to the wide range of European, national and local regulations, the application of these indicators is complicated by numerous incongruencies. Furthermore, there are many definitions of concepts such as bio-construction, bioclimatic architecture, ecological building, sustainable construction and others which have the same objective. This paper aims to establish a procedure for facilitating the current system of environmental certification and at the same time, help apply the measures for sustainable construction by creating a scale of CO2 emissions according to construction system. The methodology employed is based on the standardized systems of simulation and classification of energy efficiency in Spain, although they could be adapted to systems used in other countries. Applying this methodology to a case study enables the conclusion that the value obtained from the different simulations of the whole building with the different typologies of roofing proposed have a classification A and B, and when the different roof typologies are studied separately, they are classified as A, B and C. This methodology simplifies the decision-making process at an environmental level and also in terms of energy efficiency so that decision-making can be carried out at the architectural design stage and not subsequently, when energy simulations are currently carried out. The results will enable the application of the scale obtained as a new indicator in the definitions of construction systems.

Gene therapy is a promising therapeutic alternative for Lysosomal Storage Disorders (LSD), as it is not necessary to correct the genetic defect in all cells of an organ to achieve therapeutically significant levels of enzyme in body fluids, from which non-transduced cells can uptake the protein correcting their enzymatic deficiency. Animal models are instrumental in the development of new treatments for LSD. Here we report the generation of the first mouse model of the LSD Muccopolysaccharidosis Type IIID (MPSIIID), also known as Sanfilippo syndrome type D. This autosomic recessive, heparan sulphate storage disease is caused by deficiency in N-acetylglucosamine 6-sulfatase (GNS). Mice deficient in GNS showed lysosomal storage pathology and loss of lysosomal homeostasis in the CNS and peripheral tissues, chronic widespread neuroinflammation, reduced locomotor and exploratory activity and shortened lifespan, a phenotype that closely resembled human MPSIIID. Moreover, treatment of the GNS-deficient animals with GNS-encoding adeno-associated viral (AAV) vectors of serotype 9 delivered to the cerebrospinal fluid completely corrected pathological storage, improved lysosomal functionality in the CNS and somatic tissues, resolved neuroinflammation, restored normal behaviour and extended lifespan of treated mice. Hence, this work represents the first step towards the development of a treatment for MPSIIID.

For most lysosomal storage diseases (LSDs) affecting the CNS, there is currently no cure. The BBB, which limits the bioavailability of drugs administered systemically, and the short half-life of lysosomal enzymes, hamper the development of effective therapies. Mucopolysaccharidosis type IIIA (MPS IIIA) is an autosomic recessive LSD caused by a deficiency in sulfamidase, a sulfatase involved in the stepwise degradation of glycosaminoglycan (GAG) heparan sulfate. Here, we demonstrate that intracerebrospinal fluid (intra-CSF) administration of serotype 9 adenoassociated viral vectors (AAV9s) encoding sulfamidase corrects both CNS and somatic pathology in MPS IIIA mice. Following vector administration, enzymatic activity increased throughout the brain and in serum, leading to whole body correction of GAG accumulation and lysosomal pathology, normalization of behavioral deficits, and prolonged survival. To test this strategy in a larger animal, we treated beagle dogs using intracisternal or intracerebroventricular delivery. Administration of sulfamidase-encoding AAV9 resulted in transgenic expression throughout the CNS and liver and increased sulfamidase activity in CSF. High-titer serum antibodies against AAV9 only partially blocked CSF-mediated gene transfer to the brains of dogs. Consistently, anti-AAV antibody titers were lower in CSF than in serum collected from healthy and MPS IIIA-affected children. These results support the clinical translation of this approach for the treatment of MPS IIIA and other LSDs with CNS involvement.

Mucopolysaccharidosis type II (MPSII) is an X-linked lysosomal storage disease characterized by severe neurologic and somatic disease caused by deficiency of iduronate-2-sulfatase (IDS), an enzyme that catabolizes the glycosaminoglycans heparan and dermatan sulphate. Intravenous enzyme replacement therapy (ERT) currently constitutes the only approved therapeutic option for MPSII. However, the inability of recombinant IDS to efficiently cross the blood-brain barrier (BBB) limits ERT efficacy in treating neurological symptoms. Here, we report a gene therapy approach for MPSII through direct delivery of vectors to the CNS. Through a minimally invasive procedure, we administered adeno-associated virus vectors encoding IDS (AAV9-Ids) to the cerebrospinal fluid of MPSII mice with already established disease. Treated mice showed a significant increase in IDS activity throughout the encephalon, with full resolution of lysosomal storage lesions, reversal of lysosomal dysfunction, normalization of brain transcriptomic signature, and disappearance of neuroinflammation. Moreover, our vector also transduced the liver, providing a peripheral source of therapeutic protein that corrected storage pathology in visceral organs, with evidence of cross-correction of nontransduced organs by circulating enzyme. Importantly, AAV9-Ids-treated MPSII mice showed normalization of behavioral deficits and considerably prolonged survival. These results provide a strong proof of concept for the clinical translation of our approach for the treatment of Hunter syndrome patients with cognitive impairment.

Mucopolysaccharidosis type IIIC (MPSIIIC) is a severe lysosomal storage disease caused by deficiency in activity of the transmembrane enzyme heparan-α-glucosaminide N-acetyltransferase (HGSNAT) that catalyses the N-acetylation of α-glucosamine residues of heparan sulfate. Enzyme deficiency causes abnormal substrate accumulation in lysosomes, leading to progressive and severe neurodegeneration, somatic pathology and early death. There is no cure for MPSIIIC, and development of new therapies is challenging because of the unfeasibility of cross-correction. In this study, we generated a new mouse model of MPSIIIC by targeted disruption of the Hgsnat gene. Successful targeting left LacZ expression under control of the Hgsnat promoter, allowing investigation into sites of endogenous expression, which was particularly prominent in the CNS, but was also detectable in peripheral organs. Signs of CNS storage pathology, including glycosaminoglycan accumulation, lysosomal distension, lysosomal dysfunction and neuroinflammation were detected in 2-month-old animals and progressed with age. Glycosaminoglycan accumulation and ultrastructural changes were also observed in most somatic organs, but lysosomal pathology seemed most severe in liver. Furthermore, HGSNAT-deficient mice had altered locomotor and exploratory activity and shortened lifespan. Hence, this animal model recapitulates human MPSIIIC and provides a useful tool for the study of disease physiopathology and the development of new therapeutic approaches., Summary: A new animal model of the severe neurodegenerative lysosomal disorder mucopolysaccharidosis IIIC recapitulates the human disease, with progressive CNS and somatic lysosomal pathology, and shortened lifespan.

Blood-retinal barrier (BRB) breakdown is a key event in diabetic retinopathy and other ocular disorders that leads to increased retinal vascular permeability. This causes edema and tissue damage resulting in visual impairment. Insulin-like growth factor-I (IGF-I) is involved in these processes, although the relative contribution of increased systemic versus intraocular IGF-I remains controversial. Here, to elucidate the role of this factor in BRB breakdown, transgenic mice with either local or systemic elevations of IGF-I have been examined. High intraocular IGF-I, resulting from overexpression of IGF-I in the retina, increased IGF-I receptor content and signaling and led to accumulation of vascular endothelial growth factor. This was parallel to up-regulation of vascular Intercellular adhesion molecule I and retinal infiltration by bone marrow-derived microglial cells. These alterations resulted in increased vessel paracellular permeability to both low and high molecular weight compounds in IGF-I-overexpressing retinas and agreed with the loss of vascular tight junction integrity observed by electron microscopy and the altered junctional protein content. In contrast, mice with chronically elevated serum IGF-I did not show alterations in the retinal vasculature structure and permeability, indicating that circulating IGF-I cannot initiate BRB breakdown. Consistent with a key role of IGF-I signaling in retinal diseases, a strong up-regulation of the IGF-I receptor in human retinas with marked gliosis was also observed. Thus, this study demonstrates that intraocular IGF-I, but not systemic IGF-I, is sufficient to trigger processes leading to BRB breakdown and increased retinal vascular permeability. Therefore, therapeutic interventions designed to counteract local IGF-I effects may prove successful to prevent BRB disruption.

International audience; Gene therapy is an attractive tool for the treatment of monogenic disorders, in particular for lysosomal storage diseases (LSD) caused by deficiencies in secretable lysosomal enzymes in which neither full restoration of normal enzymatic activity nor transduction of all affected cells are necessary. However, some LSD such as Mucopolysaccharidosis Type IIIB (MPSIIIB) are challenging because the disease's main target organ is the brain and enzymes do not efficiently cross the blood-brain barrier even if present at very high concentration in circulation. To overcome these limitations, we delivered AAV9 vectors encoding for alpha-N-acetylglucosaminidase (NAGLU) to the Cerebrospinal Fluid (CSF) of MPSIIIB mice with the disease already detectable at biochemical, histological and functional level. Restoration of enzymatic activity in Central Nervous System (CNS) resulted in normalization of glycosaminoglycan content and lysosomal physiology, resolved neuroinflammation and restored the pattern of gene expression in brain similar to that of healthy animals. Additionally, transduction of the liver due to passage of vectors to the circulation led to whole-body disease correction. Treated animals also showed reversal of behavioural deficits and extended lifespan. Importantly, when the levels of enzymatic activity were monitored in the CSF of dogs following administration of canine NAGLU-coding vectors to animals that were either naive or had pre-existing immunity against AAV9, similar levels of activity were achieved, suggesting that CNS efficacy would not be compromised in patients seropositive for AAV9. Our studies provide a strong rationale for the clinical development of this novel therapeutic approach as the treatment for MPSIIIB.

L’équipe franco-espagnole travaillant sur les boutiques flanquant la Via degli Augustali au nord du forum a achevé son programme en 2013 en complétant la documentation sur la boutique VII 4, 26. Fig. 1 - Pompéi. Plan de la boutique. G. Chapelin (CNRS-CJB). Fig. 2 - Pompéi. Vue générale de la boutique VII 4, 26. J.-P. Brun. Ce secteur a été dégagé dans les années 1820 et la documentation sur ces fouilles est lacunaire. Réinterpréter ces boutiques et comprendre leur évolution ont été les buts...