RESUMO
MECP2 duplication syndrome (MDS) is a neurodevelopmental disorder caused by tandem duplication of the MECP2 locus and its surrounding genes, including IRAK1. Current MDS mouse models involve transgenic expression of MECP2 only, limiting their applicability to the study of the disease. Herein, we show that an efficient and precise CRISPR/Cas9 fusion proximity-based approach can be utilized to generate an Irak1-Mecp2 tandem duplication mouse model ('Mecp2 Dup'). The Mecp2 Dup mouse model recapitulates the genomic landscape of human MDS by harboring a 160â kb tandem duplication encompassing Mecp2 and Irak1, representing the minimal disease-causing duplication, and the neighboring genes Opn1mw and Tex28. The Mecp2 Dup model exhibits neuro-behavioral abnormalities, and an abnormal immune response to infection not previously observed in other mouse models, possibly owing to Irak1 overexpression. The Mecp2 Dup model thus provides a tool to investigate MDS disease mechanisms and develop potential therapies applicable to patients.
Assuntos
Modelos Animais de Doenças , Duplicação Gênica , Quinases Associadas a Receptores de Interleucina-1 , Deficiência Intelectual Ligada ao Cromossomo X , Proteína 2 de Ligação a Metil-CpG , Animais , Proteína 2 de Ligação a Metil-CpG/genética , Proteína 2 de Ligação a Metil-CpG/metabolismo , Quinases Associadas a Receptores de Interleucina-1/genética , Quinases Associadas a Receptores de Interleucina-1/metabolismo , Deficiência Intelectual Ligada ao Cromossomo X/genética , Deficiência Intelectual Ligada ao Cromossomo X/patologia , Humanos , Camundongos Endogâmicos C57BL , Camundongos , Sistemas CRISPR-Cas/genética , Comportamento Animal , MasculinoRESUMO
Aerated concrete (AAC) or lightweight concrete is primarily used for non-load bearing structures in construction work. Generally, it is produced with cement as a main binding ingredient, and the production of cement is blamed to contribute 7 to 8% of CO2 emission in the environment. In addition, the dumping of industrial wastes is also a great environmental concern. This research is an attempt to produce low-cost and sustainable aerated concrete utilizing silica fume and fly ash as partial substitution to cement without compromising the fundamental properties of aerated concrete. The current study was divided into two phases: in the first phase, the silica fume was substituted up to 20% with a variation of 5% in each mix. In the second phase, the fly ash was replaced with cement in three variations, i.e., 10%, 20%, and 30% containing an optimum proportion of silica fume obtained in phase 1. The aluminum powder was added at 0.4% by weight of binder to introduce aeration in concrete. Before testing, samples of aerated concrete were cured with steam in an autoclaving machine for 9 h at a pressure and temperature of 1.5 bars and 127 °C respectively and oven-dried at a temperature of 105 °C for 24 h after steam curing. From the experimental results, the highest compressive and split tensile strength of AAC was recorded when 15% of the cement was replaced with silica fume and 30% of the cement was replaced with fly ash combined. At this proportion the least density was also recorded which showed the lightweight of AAC without compromising the strength characteristics. In addition, the reduction of 42.64% and 32.4% of embodied carbon and cost was observed respectively.
Assuntos
Cinza de Carvão , Dióxido de Silício , Cinza de Carvão/química , Carbono , Vapor , Materiais de ConstruçãoRESUMO
The production of cement releases an enormous amount of CO2 into the environment. Besides, industrial wastes like silica fume and fly ash need effective utilization to reduce their impacts on the environment. This research aims to explore the influence of silica fume (SF) and fly ash (FA) individually and combine them as binary cementitious material (BCM) on the hardened properties and embodied carbon of roller compacted concrete (RCC). A total of ten mixes were prepared with 1:2:4 mix ratio at the different water-cement ratios to keep the zero slump of roller compacted concrete. However, the replacement proportions for SF were 5%-15%, and FA were 5%-15% by the weight of cement individually and combine in roller compacted concrete for determining the hardened properties and embodied carbon. In this regard, several numbers of concrete specimens (cubes and cylinders) were cast and cured for 7 and 28 days correspondingly. It was observed that the compressive strength of RCC is boosted by 33.6 MPa and 30.6 MPa while using 10% of cement replaced with SF and FA individually at 28 days, respectively. Similarly, the splitting tensile strength of RCC is enhanced by 3.5 MPa at 10% cement replaced with SF and FA on 28 days, respectively. The compressive and splitting tensile strength of RCC is increased by 34.2 MPa and 3.8 MPa at SF7.5FA7.5 as BCM after 28 days consistently. In addition, the water absorption of RCC decreased while using SF and FA as cementitious material individually and together at 28 days. Besides, the embodied carbon of RCC decreased with increasing the replacement level of SF and FA by the mass of cement individually and combined.
Assuntos
Cinza de Carvão , Materiais de Construção , Carbono , Força Compressiva , Dióxido de SilícioRESUMO
BACKGROUND: This international study aimed to characterise the impact of acute SARS-CoV-2 infection in people with cystic fibrosis and investigate factors associated with severe outcomes. Methods Data from 22 countries prior to 13th December 2020 and the introduction of vaccines were included. It was de-identified and included patient demographics, clinical characteristics, treatments, outcomes and sequalae following SARS-CoV-2 infection. Multivariable logistic regression was used to investigate factors associated with clinical progression to severe COVID-19, using the primary outcome of hospitalisation with supplemental oxygen. RESULTS: SARS-CoV-2 was reported in 1555 people with CF, 1452 were included in the analysis. One third were aged <18 years, and 9.4% were solid-organ transplant recipients. 74.5% were symptomatic and 22% were admitted to hospital. In the non-transplanted cohort, 39.5% of patients with ppFEV1<40% were hospitalised with oxygen verses 3.2% with ppFEV >70%: a 17-fold increase in odds. Worse outcomes were independently associated with older age, non-white race, underweight body mass index, and CF-related diabetes. Prescription of highly effective CFTR modulator therapies was associated with a significantly reduced odds of being hospitalised with oxygen (AOR 0.43 95%CI 0.31-0.60 p<0.001). Transplanted patients were hospitalised with supplemental oxygen therapy (21.9%) more often than non-transplanted (8.8%) and was independently associated with the primary outcome (Adjusted OR 2.45 95%CI 1.27-4.71 p=0.007). CONCLUSIONS: This is the first study to show that there is a protective effect from the use of CFTR modulator therapy and that people with CF from an ethnic minority are at more risk of severe infection with SARS-CoV-2.
Assuntos
COVID-19 , Fibrose Cística , COVID-19/epidemiologia , COVID-19/terapia , Fibrose Cística/complicações , Fibrose Cística/epidemiologia , Fibrose Cística/terapia , Regulador de Condutância Transmembrana em Fibrose Cística , Etnicidade , Humanos , Grupos Minoritários , Oxigênio , SARS-CoV-2RESUMO
Tandem duplication mutations are increasingly found to be the direct cause of many rare heritable diseases, accounting for up to 10% of cases. Unfortunately, animal models recapitulating such mutations are scarce, limiting our ability to study them and develop genome editing therapies. Here, we describe the generation of a novel duplication mouse model, harboring a multi-exonic tandem duplication in the Dmd gene which recapitulates a human mutation. Duplication correction of this mouse was achieved by implementing a single-guide RNA (sgRNA) CRISPR/Cas9 approach. This strategy precisely removed a duplication mutation in vivo, restored full-length dystrophin expression, and was accompanied by improvements in both histopathological and clinical phenotypes. We conclude that CRISPR/Cas9 represents a powerful tool to accurately model and treat tandem duplication mutations. Our findings will open new avenues of research for exploring the study and therapeutics of duplication disorders.
Assuntos
Distrofina , Distrofia Muscular de Duchenne , Animais , Sistemas CRISPR-Cas , Distrofina/genética , Edição de Genes , Camundongos , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/terapia , RNA Guia de CinetoplastídeosRESUMO
BACKGROUND: The presence of co-morbidities, including underlying respiratory problems, has been identified as a risk factor for severe COVID-19 disease. Information on the clinical course of SARS-CoV-2 infection in children with cystic fibrosis (CF) is limited, yet vital to provide accurate advice for children with CF, their families, caregivers and clinical teams. METHODS: Cases of SARS-CoV-2 infection in children with CF aged less than 18 years were collated by the CF Registry Global Harmonization Group across 13 countries between 1 February and 7 August 2020. RESULTS: Data on 105 children were collated and analysed. Median age of cases was ten years (interquartile range 6-15), 54% were male and median percentage predicted forced expiratory volume in one second was 94% (interquartile range 79-104). The majority (71%) of children were managed in the community during their COVID-19 illness. Out of 24 children admitted to hospital, six required supplementary oxygen and two non-invasive ventilation. Around half were prescribed antibiotics, five children received antiviral treatments, four azithromycin and one additional corticosteroids. Children that were hospitalised had lower lung function and reduced body mass index Z-scores. One child died six weeks after testing positive for SARS-CoV-2 following a deterioration that was not attributed to COVID-19 disease. CONCLUSIONS: SARS-CoV-2 infection in children with CF is usually associated with a mild illness in those who do not have pre-existing severe lung disease.
Assuntos
COVID-19/complicações , COVID-19/terapia , Fibrose Cística/complicações , Fibrose Cística/terapia , Adolescente , COVID-19/epidemiologia , Criança , Fibrose Cística/epidemiologia , Progressão da Doença , Feminino , Humanos , Masculino , Prognóstico , Fatores de Risco , SARS-CoV-2RESUMO
With the growing SARS-CoV-2 pandemic, we need to better understand its impact in specific patient groups like those with Cystic Fibrosis (CF). We report on 181 people with CF (32 post-transplant) from 19 countries diagnosed with SARS-CoV-2 prior to 13 June 2020. Infection with SARS-CoV-2 appears to exhibit a similar spectrum of outcomes to that seen in the general population, with 11 people admitted to intensive care (7 post-transplant), and 7 deaths (3 post-transplant). A more severe clinical course may be associated with older age, CF-related diabetes, lower lung function in the year prior to infection, and having received an organ transplant. Whilst outcomes in this large cohort are better than initially feared overall, possibly due to a protective effect of the relatively younger age of the CF population compared to other chronic conditions, SARS-CoV-2 is not a benign disease for all people in this patient group.
Assuntos
COVID-19 , Fibrose Cística , Hospitalização/estatística & dados numéricos , Transplante de Pulmão/estatística & dados numéricos , SARS-CoV-2/isolamento & purificação , Adulto , Fatores Etários , Idoso , COVID-19/epidemiologia , COVID-19/prevenção & controle , COVID-19/terapia , Teste para COVID-19/métodos , Comorbidade , Fibrose Cística/epidemiologia , Fibrose Cística/cirurgia , Feminino , Saúde Global , Humanos , Pulmão/diagnóstico por imagem , Masculino , Mortalidade , Avaliação de Resultados em Cuidados de Saúde , Sistema de Registros/estatística & dados numéricos , Testes de Função Respiratória/métodos , Fatores de Risco , Fatores Sexuais , Tomografia Computadorizada por Raios X/métodosRESUMO
Single mutations in the ATP-binding cassette transporter (ABCC6) gene (OMIM 603234) are known to cause the rare autosomal recessive disease pseudoxanthoma elasticum (PXE). Recently, we have found that copy number variations (CNVs) in pseudogenes of the ABCC6 gene are quite common. The aim of this study was to investigate the frequency and possible contribution of CNV in ABCC6 and its pseudogenes in PXE. Genomic DNA from 212 PXE individuals were examined for copy number by pyrosequencing and quantitative polymerase chain reaction (PCR) and compared with healthy individuals. The frequency of PXE individuals with any CNV was higher than in healthy individuals. The majority of variation comprised known and possibly new deletions in the ABCC6 gene and duplications of the ABCC6P1 and ABCC6P2 genes. ABCC6 deletions and ABCC6P2 duplications were not observed in 142 healthy individuals. In conclusion, by pyrosequencing and quantitative PCR, we were able to detect known and possibly new deletions in the ABCC6 gene that may have caused the PXE phenotype. Pyrosequencing may be used in PXE patients who have obtained incomplete genotype from conventional techniques. The frequency of ABCC6P2 pseudogene duplication was more common in PXE patients than healthy individuals and may affect the PXE phenotype.