Crispr/cas9 editing for gaucher disease modelling

Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the acid \u3b2-glucosidase gene (GBA1). Besides causing GD, GBA1 mutations constitute the main genetic risk factor for developing Parkinson\u2019s disease. The molecular basis of neurological manifestations in GD remain elusive. However, neuroinflammation has been proposed as a key player in this process. We exploited CRISPR/Cas9 technology to edit GBA1 in the human monocytic THP-1 cell line to develop an isogenic GD model of monocytes and in glioblastoma U87 cell lines to generate an isogenic GD model of glial cells. Both edited (GBA1 mutant) cell lines presented low levels of mutant acid \u3b2-glucosidase expression, less than 1% of residual activity and massive accumulation of substrate. Moreover, U87 GBA1 mutant cells showed that the mutant enzyme was retained in the ER and subjected to proteasomal degradation, triggering unfolded protein response (UPR). U87 GBA1 mutant cells displayed an increased production of interleukin-1\u3b2, both with and without inflammosome activation, \u3b1-syn accumulation and a higher rate of cell death in comparison with wild-type cells. In conclusion, we developed reliable, isogenic, and easy-to-handle cellular models of GD obtained from commercially accessible cells to be employed in GD pathophysiology studies and high-throughput drug screenings

Challenges for ecolabeling growth: lessons from the EU Ecolabel in Spain

Purpose The European Ecolabel (EU Flower) has the mission to encourage cleaner production and influence consumers to promote Europe's transition to a circular economy. Nonetheless, little is known about EU Ecolabel evolution; it is not clear what the drivers that encourage its implementation are. Thus, this study aims to assess the growing acceptance of the EU Ecolabel in the European Union, and Spain more specifically, by examining product and service categories and geographical regions. Methods The methodological approach taken in this study is a mixed methodology based on the triangulation method by consulting the EU Ecolabel scheme database, EU Ecolabel delegates from some autonomous regions, and the academic literature. Also, a geographic analysis was run in the ArcGIS Software with data about the accumulation of licenses assigned in 2016. Results and discussion The analysis shows that most products in Spain that have been awarded the EU Ecolabel belong to the following categories: Do-It-Yourself Products (paint and varnish), Paper Products, Cleaning Up Products, and Electronic Equipment. At the same time, the study showed that this ecolabel faces significant obstacles in its diffusion, such as the competition with environmental labels launched previously in Europe and other regional labels. Conclusions The results of this study indicate the existence of five drivers that may encourage the implementation of EU Flower in a region: (1) public management, (2) communication strategy, (3) sustainable public procurement criteria, (4) local income per capita, and (5) international trade incentives. Finally, this study provides essential recommendations for policymakers to trigger ecolabeling practices such as the need to improve the understanding of the EU ecolabel impact in different levels of activity, which means countries, regions, industrial clusters, firms, and consumers. Also, this investigation identifies areas for further research, and it expresses the need to develop business case studies about ecolabeling with the objective to visualize this phenomenon as an eco-innovation process

Increasing Engineering Students’ Involvement in Circular Economy Practices.

The circular economyhas become a topic of intense interest for policymakers, scholars and business managers because it has proven to bea new paradigm to achieve the sustainability of our society. However, the main efforts made in thecircular economy cannot be limited tothe actions ofprofessional or experts. We believe that if we intend to meet current needs without compromising the ability of future generations to meet their own needs, we mustteach present generations the principles for achievingeconomic, social and environmentalsustainability in the short, mediumand long-term. This paper highlights the use of participatory guided activitiesinstead of traditional coursesto teach and engage engineering students with circular economy practices

Evaluation of CSF neurotransmitters and folate in 25 patients with Rett disorder and effects of treatment

Background: Rett disorder (RD) is a progressive neurodevelopmental entity caused by mutations in the MECP2 gene. It has been postulated that there are alterations in the levels of certain neurotransmitters and folate in the pathogenesis of this disease. Here we re-evaluated this hypothesis. Patients and methods: We evaluated CSF folate, biogenic amines and pterines in 25 RD patients. Treatment with oral folinic acid was started in those cases with low folate. Patients were clinically evaluated and videotaped up to 6 months after therapy. Results: CSF folate was below the reference values in 32% of the patients. Six months after treatment no clinical improvement was observed. Three of the four patients with the R294X mutation had increased levels of a dopamine metabolite associated to a particular phenotype. Three patients had low levels of a serotonin metabolite. Two of them were treated with fluoxetine and one showed clinical improvement. No association was observed between CSF folate and these metabolites, after adjusting for the patients age and neopterin levels. Conclusion: Our results support that folinic acid supplementation has no significant effects on the course of the disease. We report discrete and novel neurotransmitter abnormalities that may contribute to the pathogenesis of RD highlighting the need for further studies on CSF neurotransmitters in clinically and genetically well characterized patients.Research in Rett syndrome is supported by FSE/FEDER and Fundação para a Ciência e Tecnologia (FCT, Portugal), Grant No. POCTI 41416/2001

Usefulness of analysis of cerebrospinal fluid for the diagnosis of neurotransmitters and pterin defects and glucose and folate transport deficiencies across blood brain barrier

Med Clin (Barc). 2006 Jun 17;127(3):81-5. [Usefulness of analysis of cerebrospinal fluid for the diagnosis of neurotransmitters and pterin defects and glucose and folate transport deficiencies across blood brain barrier] [Article in Spanish] Ormazabal A, García Cazorla A, Pérez Dueñas B, Pineda M, Ruiz A, López Laso E, García Silva M, Carilho I, Barbot C, Cormand B, Ribases M, Moller L, Fernández Alvarez E, Campistol J, Artuch R. Hospital Sant Joan de Déu, Esplugues, Barcelona, España. Abstract BACKGROUND AND OBJECTIVE: In the last few years, it has been described inborn errors of neurotransmitter and pterin metabolism and defects in folate and glucose transport across blood brain barrier. All these defects are classified as rare diseases and needs cerebrospinal fluid (CSF) sample analysis for diagnosis. Our aim was to evaluate the results of the application of a CSF analysis protocol in a pediatric population from Spain and Portugal presenting with neurological disorders of unknown origin. PATIENTS AND METHOD: We studied CSF samples from and 283 patients with neurological disorders of unknown origin and 127 controls. Neurotransmitters were analysed by HPLC with electrochemical detection, and pterins and 5-methyltetrahydrofolate were determined by HPLC with fluorescence detection. RESULTS: We diagnosed 3 patients with tyrosine hidroxylase deficiency, 2 with dopa responsive dystonia, 14 with GTP-ciclohydrolase deficiency, 2 with glucose transport deficiency and 43 with cerebral folate deficiency. CONCLUSIONS: This study allowed us to diagnose new patients, and more importantly, the establishment in all of them of a pharmacological or nutritional treatment. The most frequent defect found was CSF 5-methyltetrahydrofolate deficiency, which was present in different groups of patients. PMID: 16827996 [PubMed - indexed for MEDLINE

Surfactant-mediated variation of band-edge emission in CdS nanocomposites

The optical-structural characteristics of the direct optical band-gap semiconducting series of surfactant template-mediated laminar (CdS)x(CdCl2)y(CnH2n+4N)z nanocomposites are reported. X-ray diffraction measurements of the nanocomposites exhibited interlaminar distances in the range 2.9-3.6 nm with observations of eighth order {0 0 l} diffraction planes indicative of a high degree of laminarity and crystallographic order. Diffuse reflectance measurements have determined that the profile of their emission spectrum is that of a direct band-gap with absorption edges in the range 2.11-2.40 eV, depending on the CdS mole fraction in the nanocomposite. Photoluminescence (PL) excitation and time-resolved PL spectroscopies give an estimate of the maximum relative absorbance of the nanocomposites at ∼420 nm while the minimum was observed at ∼560 nm. The main emission was observed at ∼700 nm with emission from doubly ionized sulphur vacancies observed at ∼615 nm at room temperature. The CdS-containing nanocomposite is thus a surfactant-mediated modular system with variable band-gap energy emission

Origen y evolución del lago Yehuin (isla Grande de Tierra del Fuego, Argentina): resultados de un relevamiento geofísico

El lago Yehuin, una cuenca elongada de rumbo ONO-ESE localizada en la faja plegada y corrida externa de los Andes Fueguinos, ocupa una depresión compartimentada originada a lo largo de un segmento del sistema de fallas sinistrales del lago Deseado. Este trabajo describe un primer relevamiento geofísico llevado a cabo en el lago. Los datos de sísmica monocanal de alta resolución, integrados con información geológica de los alrededores del lago Yehuin, permitieron: (i) producir un mapa de la batimetría completa del lago, (ii) reconstruir la superficie del basamento del lago, y (iii) analizar la geometría, distribución y espesor del relleno sedimentario. Se reconocieron dos subcuencas dentro del lago Yehuin: una subcuenca oeste de 7,5 km de largo, con una profundidad máxima de 118 m; una subcuenca este de 7,2 km de largo y una profundidad máxima de 80 m. Ambas subcuencas están limitadas por un conjunto de fallas normales que cortan a una serie de corrimientos de vergencia NE. Se identificaron tres unidades sismo-estratigráficas en el registro sísmico: (1) una unidad inferior con geometría acuñada interpretada como depósitos de remoción en masa; (2) una unidad intermedia gruesa (de hasta 120 m) de origen glaciolacustre e irregularmente distribuida en la cuenca del lago; (3) una unidad superior lacustre delgada (<10 m) que cubre la cuenca entera. El lago Yehuin se considera una cuenca neógena e origen tectónico que fue luego afectada por depositación glaciaria y glaciolacustre. Se han interpretado morenas sumergidas dentro del lago Yehuin que se correlacionan con los arcos morrénicos en tierra y permiten completar el camino recesivo de los lóbulos de hielo Ewan y Fuego. Se propone un fuerte control estructural no solo para la formación del lago Yehuin, sino también para las rutas generales de los brazos norte del Paleoglaciar Fagnano.Lago Yehuin, a WNW-ESE elongated basin located in the outer fold-and-thrust belt of the Fuegian Andes, occupies a compartmented structural depression originated along a segment of the left-lateral Lago Deseado fault system. This paper describes the first geophysical survey performed within the lake. New acquired high-resolution single-channel seismic data, integrated with geological information in the surroundings of the Lago Yehuin, allowed to: (i) produce a complete bathymetric map of the lake, (ii) reconstruct the basement surface of the lake, and (iii) analyze the geometry, distribution, and thickness of the sedimentary infill. Two sub-basins were recognized within Lago Yehuin: A western sub-basin, 7.5 km long, with a maximum depth of 118 m; an eastern sub-basin, 7.2 km long with a maximum depth of 80 m. Both sub-basins are limited by a set of normal faults which overprint NE-verging thrusts. Three seismo-stratigraphic units have been identified in the seismic records: (1) a lower unit with wedged geometry interpreted as a mass flow deposits; (2) a thick (up to 120 m) intermediate unit of glacio-lacustrine nature and irregularly distributed in the Yehuin basin; (3) a thin (generally <10 m) upper lacustrine unit which drapes the entire basin. Lago Yehuin is considered a Neogene basin generated by strike-slip tectonics that was later affected by glacial and glacio-lacustrine deposition. Interpreted submerged ridge moraines within Lago Yehuin are correlated with onland moraine arcs built by the complete recessional paths of Fuego and Ewan ice lobes. A significant structural control is proposed not only for the formation of Lago Yehuin, but also for the general paths of the northern arms of the Fagnano palaeo-glacier.Fil: Lozano, Jorge Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Tassone, Alejandro Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Lodolo, Emanuele. Istituto Nazionale Di Oceanografia E Di Geofisica Sperimentale;Fil: Menichetti, Marco. Universita Degli Studi Di Urbino Carlo Bo;Fil: Cerredo, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Bran, Donaldo Mauricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Esteban, Federico Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Ormazabal, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Baradello, Luca. Instituto Nazionale di Oceanografia e di Geofísica Sperimentale; ItaliaFil: Vilas, Juan Francisco A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; Argentin

Genotypic and phenotypic spectrum of pyridoxine-dependent epilepsy (ALDH7A1 deficiency)

Pyridoxine-dependent epilepsy was recently shown to be due to mutations in the ALDH7A1 gene, which encodes antiquitin, an enzyme that catalyses the nicotinamide adenine dinucleotide-dependent dehydrogenation of L-{alpha}-aminoadipic semialdehyde/L-{Delta}1-piperideine 6-carboxylate. However, whilst this is a highly treatable disorder, there is general uncertainty about when to consider this diagnosis and how to test for it. This study aimed to evaluate the use of measurement of urine L-{alpha}-aminoadipic semialdehyde/creatinine ratio and mutation analysis of ALDH7A1 (antiquitin) in investigation of patients with suspected or clinically proven pyridoxine-dependent epilepsy and to characterize further the phenotypic spectrum of antiquitin deficiency. Urinary L-{alpha}-aminoadipic semialdehyde concentration was determined by liquid chromatography tandem mass spectrometry. When this was above the normal range, DNA sequencing of the ALDH7A1 gene was performed. Clinicians were asked to complete questionnaires on clinical, biochemical, magnetic resonance imaging and electroencephalography features of patients. The clinical spectrum of antiquitin deficiency extended from ventriculomegaly detected on foetal ultrasound, through abnormal foetal movements and a multisystem neonatal disorder, to the onset of seizures and autistic features after the first year of life. Our relatively large series suggested that clinical diagnosis of pyridoxine dependent epilepsy can be challenging because: (i) there may be some response to antiepileptic drugs; (ii) in infants with multisystem pathology, the response to pyridoxine may not be instant and obvious; and (iii) structural brain abnormalities may co-exist and be considered sufficient cause of epilepsy, whereas the fits may be a consequence of antiquitin deficiency and are then responsive to pyridoxine. These findings support the use of biochemical and DNA tests for antiquitin deficiency and a clinical trial of pyridoxine in infants and children with epilepsy across a broad range of clinical scenarios