AAV-Mediated Gene Therapy for CRB1-Hereditary Retinopathies

Variations in the Crumbs homolog-1 (CRB1) gene lead to autosomal recessive retinal dystrophies such as early-onset retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA). No treatment is yet available for these patients. Adeno-associated virus (AAV) mediated gene therapy for hereditary retinal diseases holds great promise proven by the large number of active clinical trials. We here summarized the knowledge about the localization and function of CRB1 in the retina and the main pathological features resulting from loss of CRB1 function in humans and in rodents. This know-how is being applied to design and develop AAV gene therapy vectors for the treatment of CRB1-Hereditary retinopathies. Knowing which cell types express the CRB proteins, the possible redundancy of function between CRB1 and CRB2, and the AAV tropism in the human retina, will allow us to rationalize about the AAV capsid, promoter and route of administration that should be used in the AAV vector in order to efficiently and specifically deliver CRB1 or CRB2 into the human retina

Animal Models of Bone Loss in Inflammatory Arthritis: from Cytokines in the Bench to Novel Treatments for Bone Loss in the Bedside—a Comprehensive Review

Throughout life, bone is continuously remodelled. Bone is formed by osteoblasts, from mesenchymal origin, while osteoclasts induce bone resorption. This process is tightly regulated. During inflammation, several growth factors and cytokines are increased inducing osteoclast differentiation and activation, and chronic inflammation is a condition that initiates systemic bone loss. Rheumatoid arthritis (RA) is a chronic inflammatory auto-immune disease that is characterised by active synovitis and is associated with early peri-articular bone loss. Peri-articular bone loss precedes focal bone erosions, which may progress to bone destruction and disability. The incidence of generalised osteoporosis is associated with the severity of arthritis in RA and increased osteoporotic vertebral and hip fracture risk. In this review, we will give an overview of different animal models of inflammatory arthritis related to RA with focus on bone erosion and involvement of pro-inflammatory cytokines. In addition, a humanised endochondral ossification model will be discussed, which can be used in a translational approach to answer osteoimmunological questions

Pulsed FCAW of Martensitic Stainless Clads onto Mild Steel: Microstructure, Hardness, and Residual Stresses

The low carbon martensitic stainless AWS 410NiMo steel has in its chemical composition 13% chromium, 4% nickel, and 0.4% molybdenum (wt.%) and is used in turbine recovery, rotors, and high-pressure steam pump housings due to its resistance to impact at low temperatures, as well as to corrosion and cavitation. Those applications of the AWS 410NiMo steel frequently demand repair, which is performed by welding or cladding. Arc welding is a well-established technique for joining materials and presents several parameters that influence the mechanical performance of the weld bead. Although numerous welding processes exist, optimizing welding parameters for specific applications and materials is always challenging. The present work deals with a systematic study to verify the correlation between the pulsed fluxed core arc welding (FCAW) parameters, namely pulse current and frequency, welding speed, and contact tip work distance (CTWD), and the bead morphology, microstructure formation, residual stress, and hardness of the martensitic clad. The substrate used was the AISI 1020 steel, and the AWS 410NiMo steel was the filler metal for clad deposition. From the initial nine (9) samples, three (3) were selected for in-depth characterization. Lower heat input resulted in lower dilution, more elevated hardness, and lower compressive residual stresses. Therefore, the results highlight the need for selecting the proper heat input, even when using a pulsed FCAW procedure, to achieve the desired performance of the clad. In the present case, a higher heat input appears to be more advantageous owing to the lower convexity index, smooth hardness transition between fusion and heat-affected zones in addition to more elevated compressive stresses.Peer ReviewedPostprint (published version

Microarray and morphological analysis of early postnatal CRB2 mutant retinas on a pure C57BL/6J genetic background

In humans, the Crumbs homologue-1 (CRB1) gene is mutated in progressive types of autosomal recessive retinitis pigmentosa and Leber congenital amaurosis. Th

Dendritic cell-specific deletion of β-catenin results in fewer regulatory T-cells without exacerbating autoimmune collagen-induced arthritis

Dendritic cells (DCs) are professional antigen presenting cells that have the dual ability to stimulate immunity and maintain tolerance. However, the signalling pathways mediating tolerogenic DC function in vivo remain largely unknown. The β-catenin pathway has been suggested to promote a regulatory DC phenotype. The aim of this study was to unravel the role of β-catenin signalling to control DC function in the autoimmune collagen-induced arthritis model (CIA). Deletion of β-catenin specifically in DCs was achieved by crossing conditional knockout mice with a CD11c-Cre transgen

Genetic diversity and structure of two species of Enyalius (Squamata: Leiosauridae) from neotropical biodiversity hotspots

Diversidade e estrutura genética de duas espécies de Enyalius (Squamata: Leiosauridae) em hotspots neotropicais de biodiversidade. Os lagartos do gênero Enyalius são endêmicos da América do Sul, sendo predominantemente encontrados no Cerrado e em fragmentos da Floresta Atlântica. Este é um gênero pouco estudado, e não foram encontrados dados relacionados à diversidade e à estrutura genética das espécies do gênero. Neste trabalho, investigamos a diversidade genética de populações de E. bilineatus (N = 20) e E. perditus (N = 28), usando um fragmento de 234-pb do citocromo b, e comparamos as sequências geradas com outras publicadas. Dezenove haplótipos distintos foram encontrados (11 de E. perditus e oito de E. bilineatus), sendo oito deles novos registros. Os valores de diversidade haplotípica foram muito similares para as duas espécies (0.684 para E. perditus e 0.647 para E. bilineatus). A distância genética entre as duas espécies foi de 20.3%, e as distâncias intraespecíficas foram 2.0% para E. perditus e 5.6% para E. bilineatus. Nossos dados sugerem que as populações de E. bilineatus são altamente divergentes e que a espécie deve apresentar diversidade críptica. Este é o primeiro estudo medindo a diversidade genética de espécies do gênero Enyalius oriundas de regiões consideradas hotspots da biodiversidade neotropical e apresenta dados relevantes para um melhor entendimento das relações inter e intrapopulacionais, assim como a distribuição das linhagens genéticas desse gênero endêmico.Genetic diversity and structure of two species of Enyalius (Squamata: Leiosauridae) from neotropical biodiversity hotspots. Enyalius, a lizard genus endemic to South America, is mostly distributed in the remains of the Atlantic Forest and in the Cerrado. The genus has been the topic of a few studies but none has quantified the genetic diversity and structure within and among populations of Enyalius. The genetic diversity and structure of populations of E. bilineatus (N = 20) and E. perditus (N = 28) are examined using a 234-bp fragment of the cytochrome b gene and compared with the sequences reported in other published data. Nineteen distinct haplotypes (eleven for E. perditus and eight for E. bilineatus) were found, eight of which were recorded for the first time. The haplotype diversities are highly similar for both species (0.684 for E. perditus and 0.647 for E. bilineatus). The genetic distance between the two species is 20.3% and the distance within species is 2.0% and 5.6% for E. perditus and E. bilineatus, respectively. Our data suggest that populations of E. bilineatus are genetically divergent and may reveal cryptic diversity. This is the first study to quantify the genetic diversity of species of Enyalius from Neotropical biodiversity hotspots. These data facilitate a better understanding of both within and among population variation, and highlight the distribution of genetic lineages of an endemic and poorly studied genus

Sustainable plant polyesters as substrates for optical gas sensors

UIDB/04378/2020 PTDC/BII-BIO/28878/2017 PTDC/SAU-SER/30388/2017 SFRH-BD-110467-2015The fast and non-invasive detection of odors and volatile organic compounds (VOCs) by gas sensors and electronic noses is a growing field of interest, mostly due to a large scope of potential applications. Additional drivers for the expansion of the field include the development of alternative and sustainable sensing materials. The discovery that isolated cross-linked polymeric structures of suberin spontaneously self-assemble as a film inspired us to develop new sensing composite materials consisting of suberin and a liquid crystal (LC). Due to their stimuli-responsive and optically active nature, liquid crystals are interesting probes in gas sensing. Herein, we report the isolation and the chemical characterization of two suberin types (from cork and from potato peels) resorting to analyses of gas chromatography–mass spectrometry (GC-MS), solution nuclear magnetic resonance (NMR), and X-ray photoelectron spectroscopy (XPS). The collected data highlighted their compositional and structural differences. Cork suberin showed a higher proportion of longer aliphatic constituents and is more esterified than potato suberin. Accordingly, when casted it formed films with larger surface irregularities and a higher C/O ratio. When either type of suberin was combined with the liquid crystal 5CB, the ensuing hybrid materials showed distinctive morphological and sensing properties towards a set of 12 VOCs (comprising heptane, hexane, chloroform, toluene, dichlormethane, diethylether, ethyl acetate, acetonitrile, acetone, ethanol, methanol, and acetic acid). The optical responses generated by the materials are reversible and reproducible, showing stability for 3 weeks. The individual VOC-sensing responses of the two hybrid materials are discussed taking as basis the chemistry of each suberin type. A support vector machines (SVM) algorithm based on the features of the optical responses was implemented to assess the VOC identification ability of the materials, revealing that the two distinct suberin-based sensors complement each other, since they selectively identify distinct VOCs or VOC groups. It is expected that such new environmentally-friendly gas sensing materials derived from natural diversity can be combined in arrays to enlarge selectivity and sensing capacity.publishersversionpublishe