RESUMO
The hallmark of epidermolysis bullosa (EB) is fragile attachment of epithelia due to genetic variants in cell adhesion genes. We describe 16 EB patients treated in the ear, nose, and throat department of a tertiary pediatric hospital linked to the United Kingdom's national EB unit between 1992 and 2023. Patients suffered a high degree of morbidity and mortality from laryngotracheal stenosis. Variants in laminin subunit alpha-3 (LAMA3) were found in 10/15 patients where genotype was available. LAMA3 encodes a subunit of the laminin-332 heterotrimeric extracellular matrix protein complex and is expressed by airway epithelial basal stem cells. We investigated the benefit of restoring wild-type LAMA3 expression in primary EB patient-derived basal cell cultures. EB basal cells demonstrated weak adhesion to cell culture substrates, but could otherwise be expanded similarly to non-EB basal cells. In vitro lentiviral overexpression of LAMA3A in EB basal cells enabled them to differentiate in air-liquid interface cultures, producing cilia with normal ciliary beat frequency. Moreover, transduction restored cell adhesion to levels comparable to a non-EB donor culture. These data provide proof of concept for a combined cell and gene therapy approach to treat airway disease in LAMA3-affected EB.
Assuntos
Adesão Celular , Epidermólise Bolhosa , Laminina , Lentivirus , Humanos , Laminina/metabolismo , Laminina/genética , Epidermólise Bolhosa/genética , Epidermólise Bolhosa/metabolismo , Epidermólise Bolhosa/terapia , Epidermólise Bolhosa/patologia , Criança , Lentivirus/genética , Masculino , Feminino , Pré-Escolar , Terapia Genética/métodos , Vetores Genéticos/genética , Células Epiteliais/metabolismo , Células Cultivadas , Expressão Gênica , Adolescente , LactenteRESUMO
Epidemiologic studies in the general population show that the level of dietary salt intake is associated with increases in blood pressure (BP), cardiovascular events, and mortality. According to trial data, reducing salt intake lowers the incidence of these 3 outcomes. On the basis of this evidence, the World Health Organization and other bodies recommend restricting salt intake. The association of salt intake with BP and cardiovascular disease has also been seen in chronic kidney disease (CKD), and trials of salt reduction in CKD have shown benefit, reflected by reduced BP and a lower rate of cardiovascular events. However, these trials have typically used resourceintensive approaches to dietary salt reduction that are not suitable for routine clinical care, and salt intake typically remains high in people with CKD. The OxSalt care bundle is a lowcost intervention that was demonstrated in the OxCKD1 trial to help people with CKD lower their salt intake, and could be applied in routine clinical practice.
Assuntos
Insuficiência Renal Crônica , Cloreto de Sódio na Dieta , Humanos , Insuficiência Renal Crônica/dietoterapia , Cloreto de Sódio na Dieta/efeitos adversos , Cloreto de Sódio na Dieta/administração & dosagem , Hipertensão/dietoterapia , Dieta Hipossódica , Doenças Cardiovasculares/prevenção & controle , Doenças Cardiovasculares/etiologia , Pressão Sanguínea/efeitos dos fármacos , Feminino , MasculinoRESUMO
A performance evaluation was conducted on the new General Electric (GE) digital Omni Legend PET-CT system with 32 cm extended field of view. The first commercially available clinical digital bismuth germanate system. The system does not use time of flight (ToF). Testing was performed in accordance with the NEMA NU2-2018 standard. A comparison was made between two other commercial GE scanners with extended fields of view; the Discovery MI - 6 ring (ToF enabled) and the Discovery IQ (non-ToF). A genetic evolutionary algorithm was developed to optimize image reconstruction parameters from image quality assessments. The Omni demonstrated average spatial resolutions at 1 cm radial offset as 3.9 mm FWHM. The total system sensitivity at the center was 44.36 cps/kBq. The peak NECR was measured as 501 kcps at 17.8 kBq ml-1with a 35.48% scatter fraction. The maximum count-rate error below NECR peak was 5.5%. Using standard iterative reconstructions, sphere contrast recovery coefficients were from 52.7 ± 3.2% (10 mm) to 92.5 ± 2.4% (37 mm). The PET-CT co-registration accuracy was 2.4 mm. In place of ToF, the Omni employs software corrections through a pre-trained neural network (PDL) (trained on non-ToF to ToF) that takes Bayesian penalized likelihood reconstruction (Q.Clear) images as input. The optimum parameters for image reconstruction, determined using the genetic algorithm were a Q.Clear parameter,ß, of 350 and a 'medium' PDL setting. Using standard iterative reconstructions, the Omni initially showed increased background variability compared to the Discovery MI. With optimized PDL reconstruction parameters selected using the genetic algorithm the performance of the Omni surpassed that of the Discovery MI on all NEMA tests. The genetic algorithm's demonstrated ability to enhance image quality in PET-CT imaging underscores the importance of algorithm driven optimization and underscores the requirement to validate its use in the clinical setting.
Assuntos
Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Tomografia Computadorizada por Raios X , Teorema de Bayes , Imagens de Fantasmas , AlgoritmosRESUMO
INTRODUCTION: Although contact days-days with health-care contact outside home-are increasingly adopted as a measure of time toxicity and treatment burden, they could also serve as a surrogate of treatment-related harm. We sought to assess the association between contact days and patient-reported outcomes and the prognostic ability of contact days. METHODS: We conducted a secondary analysis of CO.17 that evaluated cetuximab vs supportive care in patients with advanced colorectal cancer. CO.17 collected European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire C30 instrument data. We assessed the association between number of contact days in a window and changes in physical function and global health status and the association between number of contact days in the first 4 weeks with overall survival. RESULTS: There was a negative association between the number of contact days and change in physical function (per each additional contact day: at 4 weeks, 1.50-point decrease; 8 weeks, 1.06-point decrease; P < .0001 for both) but not with global health status. This negative association was seen in patients receiving cetuximab but not supportive care. More contact days in the first 4 weeks was associated with worse overall survival for all participants and patients receiving cetuximab (per each additional contact day: all participants, adjusted hazard ratio [HR] = 1.07, 95% confidence interval [CI] = 1.05 to 1.10; and cetuximab, adjusted HR = 1.08, 95% CI = 1.05 to 1.11; P < .0001 for both). CONCLUSIONS: In this secondary analysis of a clinical trial, more contact days early in the course were associated with declines in physical function and worse survival in all participants and in participants receiving cancer-directed treatment. TRIAL REGISTRATION: ClinicalTrials.gov number, NCT00079066.
Assuntos
Cetuximab , Neoplasias Colorretais , Qualidade de Vida , Humanos , Neoplasias Colorretais/mortalidade , Neoplasias Colorretais/tratamento farmacológico , Masculino , Feminino , Cetuximab/efeitos adversos , Cetuximab/uso terapêutico , Cetuximab/administração & dosagem , Pessoa de Meia-Idade , Idoso , Prognóstico , Medidas de Resultados Relatados pelo Paciente , Inquéritos e Questionários , Fatores de Tempo , Antineoplásicos Imunológicos/uso terapêutico , Antineoplásicos Imunológicos/efeitos adversos , Nível de Saúde , Desempenho Físico FuncionalRESUMO
Isolation of tissue-specific fetal stem cells and derivation of primary organoids is limited to samples obtained from termination of pregnancies, hampering prenatal investigation of fetal development and congenital diseases. Therefore, new patient-specific in vitro models are needed. To this aim, isolation and expansion of fetal stem cells during pregnancy, without the need for tissue samples or reprogramming, would be advantageous. Amniotic fluid (AF) is a source of cells from multiple developing organs. Using single-cell analysis, we characterized the cellular identities present in human AF. We identified and isolated viable epithelial stem/progenitor cells of fetal gastrointestinal, renal and pulmonary origin. Upon culture, these cells formed clonal epithelial organoids, manifesting small intestine, kidney tubule and lung identity. AF organoids exhibit transcriptomic, protein expression and functional features of their tissue of origin. With relevance for prenatal disease modeling, we derived lung organoids from AF and tracheal fluid cells of congenital diaphragmatic hernia fetuses, recapitulating some features of the disease. AF organoids are derived in a timeline compatible with prenatal intervention, potentially allowing investigation of therapeutic tools and regenerative medicine strategies personalized to the fetus at clinically relevant developmental stages.