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Discrete- and continuous-time approaches are frequently used to model the role of heterogeneity on dynamical interacting agents on the top of complex networks. While, on the one hand, one does not expect drastic differences between these approaches, and the choice is usually based on one's expertise or methodological convenience, on the other hand, a detailed analysis of the differences is necessary to guide the proper choice of one or another approach. We tackle this problem by investigating both discrete- and continuous-time mean-field theories for the susceptible-infected-susceptible (SIS) epidemic model on random networks with power-law degree distributions. We compare the discrete epidemic link equations (ELE) and continuous pair quenched mean-field (PQMF) theories with the corresponding stochastic simulations, both theories that reckon pairwise interactions explicitly. We show that ELE converges to the PQMF theory when the time step goes to zero. We performed an epidemic localization analysis considering the inverse participation ratio (IPR). Both theories present the same localization dependence on network degree exponent γ: for γ<5/2 the epidemics are localized on the maximum k-core of networks with a vanishing IPR in the infinite-size limit while, for γ>5/2, the localization happens on hubs that do not form a densely connected set and leads to a finite value of the IPR. However, the IPR and epidemic threshold of ELE depend on the time-step discretization such that a larger time step leads to more localized epidemics. A remarkable difference between discrete- and continuous-time approaches is revealed in the epidemic prevalence near the epidemic threshold, in which the discrete-time stochastic simulations indicate a mean-field critical exponent θ=1 instead of the value θ=1/(3-γ) obtained rigorously and verified numerically for the continuous-time SIS on the same networks.
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Interest in measuring immunoglobulin G Subclasses (IgG Subclasses) is increasing as more information is gathered and understanding regarding conditions associated with deficiencies of each IgG Subclass grows. Different methodologies are available for the measurement of IgG Subclasses, but their specificities vary. As a result, laboratories choose the methodology that better suits their routine, but which may not necessarily align with the needs of their population. In addition, the lack of standardization for the quantification of IgG Subclasses causes diagnostic gaps when comparing results provided by different methodologies. Thus, the purpose of our research is to compare the analytical performance of The Binding Site's (TBS) Optilite® human Immunoglobulin G (IgG) and IgG Subclasses Immunoturbidimetry assay, with the Nephelometry method routinely used in our clinical laboratory, Siemens BNII®. Our results show that the Immunoturbidimetry assay appears to be the most reliable to evaluate IgG Subclasses: the sum of IgG Subclasses and Total IgG correlate better than by Nephelometry. Although these methodologies share a similar principle, the comparison of results appears to be compromised. Therefore, prior to switching methodologies, further studies should be conducted to assess which methodology could be better applied to specific populations. It is also essential to standardise IgG Subclasses assays to reduce discrepancies that arise from comparing results.
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Inmunoglobulina G , Inmunoturbidimetría , Nefelometría y Turbidimetría , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/clasificación , Inmunoglobulina G/inmunología , Nefelometría y Turbidimetría/métodos , Inmunoturbidimetría/métodos , Reproducibilidad de los Resultados , Femenino , Masculino , Adulto , Persona de Mediana Edad , AncianoRESUMEN
The presence of basal lineage characteristics signifies hyperaggressive human adenocarcinomas of the breast, bladder and pancreas. However, the biochemical mechanisms that maintain this aberrant cell state are poorly understood. Here we performed marker-based genetic screens in search of factors needed to maintain basal identity in pancreatic ductal adenocarcinoma (PDAC). This approach revealed MED12 as a powerful regulator of the basal cell state in this disease. Using biochemical reconstitution and epigenomics, we show that MED12 carries out this function by bridging the transcription factor ΔNp63, a known master regulator of the basal lineage, with the Mediator complex to activate lineage-specific enhancer elements. Consistent with this finding, the growth of basal-like PDAC is hypersensitive to MED12 loss when compared to PDAC cells lacking basal characteristics. Taken together, our genetic screens have revealed a biochemical interaction that sustains basal identity in human cancer, which could serve as a target for tumor lineage-directed therapeutics.
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Carcinoma Ductal Pancreático , Complejo Mediador , Neoplasias Pancreáticas , Factores de Transcripción , Proteínas Supresoras de Tumor , Humanos , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patología , Carcinoma Ductal Pancreático/metabolismo , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/metabolismo , Complejo Mediador/genética , Complejo Mediador/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismo , Regulación Neoplásica de la Expresión Génica , Línea Celular Tumoral , Linaje de la Célula/genética , Elementos de Facilitación GenéticosRESUMEN
Objective.Modern medical imaging plays a vital role in clinical practice, enabling non-invasive visualization of anatomical structures. Dynamic contrast enhancement (DCE) imaging is a technique that uses contrast agents to visualize blood flow dynamics in a time-resolved manner. It can be applied to different modalities, such as computed tomography (CT) and electrical impedance tomography (EIT). This study aims to develop a common theoretical and practical hemodynamic extraction basis for DCE modelling across modalities, based on the gamma-variate function.Approach.The study introduces a framework to generate time-intensity curves for multiple DCE imaging modalities from user-defined hemodynamic parameters. Thus, extensive datasets were simulated for both DCE-CT and EIT, representing different hemodynamic scenarios. Additionally, gamma-variate extensions to account for several physiological effects were detailed in a modality-agnostic manner, and three corresponding fitting strategies, namely nonlinear, linear, and a novel hybrid approach, were implemented and compared on the basis of accuracy of parameter estimation, first pass reconstruction, speed of computation, and failure rate.Main results.As a result, we found the linear method to be the most modality-dependent, exhibiting the greatest bias, variance and failure rates, although remaining the fastest alternative. The hybrid method at least matches the state-of-the-art nonlinear method's accuracy, while improving its robustness and speed by 10 times.Significance.Our research suggests that the hybrid method may bring noteworthy accuracy and efficiency improvements in handling the high-dimensionality of DCE imaging in general, being a step towards real-time processing. Moreover, our generative model presents a potential asset to produce benchmarking and data augmentation datasets across modalities.
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Medios de Contraste , Impedancia Eléctrica , Procesamiento de Imagen Asistido por Computador , Tomografía , Tomografía/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Tomografía Computarizada por Rayos X/métodos , Humanos , HemodinámicaRESUMEN
BACKGROUND: To validate pulmonary computed tomography (CT) perfusion in a porcine model by invasive monitoring of cardiac output (CO) using thermodilution method. METHODS: Animals were studied at a single center, using a Swan-Ganz catheter for invasive CO monitoring as a reference. Fifteen pigs were included. Contrast-enhanced CT perfusion of the descending aorta and right and left pulmonary artery was performed. For variation purposes, a balloon catheter was inserted to block the contralateral pulmonary vascular bed; additionally, two increased CO settings were created by intravenous administration of catecholamines. Finally, stepwise capillary occlusion was performed by intrapulmonary arterial injection of 75-µm microspheres in four stages. A semiautomatic selection of AFs and a recirculation-aware tracer-kinetics model to extract the first-pass of AFs, estimating blood flow with the Stewart-Hamilton method, was implemented. Linear mixed models (LMM) were developed to calibrate blood flow calculations accounting with individual- and cohort-level effects. RESULTS: Nine of 15 pigs had complete datasets. Strong correlations were observed between calibrated pulmonary (0.73, 95% confidence interval [CI] 0.6-0.82) and aortic blood flow measurements (0.82, 95% CI, 0.73-0.88) and the reference as well as agreements (± 2.24 L/min and ± 1.86 L/min, respectively) comparable to the state of the art, on a relatively wide range of right ventricle-CO measurements. CONCLUSIONS: CT perfusion validly measures CO using LMMs at both individual and cohort levels, as demonstrated by referencing the invasive CO. RELEVANCE STATEMENT: Possible clinical applications of CT perfusion for measuring CO could be in acute pulmonary thromboembolism or to assess right ventricular function to show impairment or mismatch to the left ventricle. KEY POINTS: ⢠CT perfusion measures flow in vessels. ⢠CT perfusion measures cumulative cardiac output in the aorta and pulmonary vessels. ⢠CT perfusion validly measures CO using LMMs at both individual and cohort levels, as demonstrated by using the invasive CO as a reference standard.
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Cateterismo de Swan-Ganz , Arteria Pulmonar , Humanos , Porcinos , Animales , Gasto Cardíaco/fisiología , Arteria Pulmonar/diagnóstico por imagen , Cateterismo de Swan-Ganz/métodos , Perfusión , Tomografía Computarizada por Rayos XRESUMEN
There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans.
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Contaminación de Alimentos , Microplásticos , Alimentos Marinos , Alimentos Marinos/análisis , Contaminación de Alimentos/análisis , Microplásticos/análisis , Animales , Alimentos en Conserva/análisisRESUMEN
BACKGROUNDING: Stayability, which may be defined as the probability of a cow remaining in the herd until a reference age or at a specific number of calvings, is usually measured late in the animal's life. Thus, if used as selection criteria, it will increase the generation interval and consequently might decrease the annual genetic gain. Measuring stayability at an earlier age could be a reasonable strategy to avoid this problem. In this sense, a better understanding of the genetic architecture of this trait at different ages and/or at different calvings is important. This study was conducted to identify possible regions with major effects on stayability measured considering different numbers of calvings in Nellore cattle as well as pathways that can be involved in its expression throughout the female's productive life. RESULTS: The top 10 most important SNP windows explained, on average, 17.60% of the genetic additive variance for stayability, varying between 13.70% (at the eighth calving) and 21% (at the fifth calving). These SNP windows were located on 17 chromosomes (1, 2, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 18, 19, 20, 27, and 28), and they harbored a total of 176 annotated genes. The functional analyses of these genes, in general, indicate that the expression of stayability from the second to the sixth calving is mainly affected by genetic factors related to reproductive performance, and nervous and immune systems. At the seventh and eighth calvings, genes and pathways related to animal health, such as density bone and cancer, might be more relevant. CONCLUSION: Our results indicate that part of the target genomic regions in selecting for stayability at earlier ages (from the 2th to the 6th calving) would be different than selecting for this trait at later ages (7th and 8th calvings). While the expression of stayability at earlier ages appeared to be more influenced by genetic factors linked to reproductive performance together with an overall health/immunity, at later ages genetic factors related to an overall animal health gain relevance. These results support that selecting for stayability at earlier ages (perhaps at the second calving) could be applied, having practical implications in breeding programs since it could drastically reduce the generation interval, accelerating the genetic progress.
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Estudio de Asociación del Genoma Completo , Genómica , Femenino , Animales , Bovinos/genética , Fenotipo , Probabilidad , Reproducción/genéticaRESUMEN
Introdução: os programas liderados por enfermeiros desempenham um papel fundamental no engajamento do indivíduo na autogestão do diabetes mellitus tipo 2 (DM2), pois impactam na melhoria da qualidade de vida. A autogestão apoiada pelo suporte social torna-se crucial, sobretudo por descrever a importância do enfermeiro na promoção da saúde da pessoa com DM2, tendo em vista que essa intervenção pode contribuir para reduzir as consequências e os agravamentos da doença. Objetivo: mapear os programas com foco no suporte social para pessoas com DM2 liderados por enfermeiros. Materiais e método: revisão de escopo que seguiu as diretrizes metodológicas do Joanna Briggs Institute. Utilizou-se como descritores de busca "autocuidado", "intervenção liderada por enfermeiro", "diabetes mellitus tipo 2", com busca nos bancos de dados Web of Science, Medline, CINAHL, Catálogo de Teses e Dissertações da Capes, Scopus, JBI Evidence Synthesis, PsycINFO e Repositórios Científicos de Acesso Aberto de Portugal, entre fevereiro e março de 2022. Os critérios de elegibilidade foram estudos publicados nos últimos 10 anos, em inglês, espanhol e português, abrangendo a experiência do indivíduo em programas com foco na autogestão por meio do suporte social liderados por enfermeiros. Resultados: 12 estudos incluídos, publicados nos Estados Unidos, na Inglaterra e no Irã, com foco no suporte social de apoio à autogestão de pessoas com DM2 e com concentração em locais de atendimento às pessoas com doenças crônicas. Observou-se que a maioria dos programas liderados por enfermeiros é implementada em países desenvolvidos e apresenta como característica principal ênfase na teoria social cognitiva. O público se caracterizou por ser maioria mulheres, faixa etária de adultos maiores de 22 anos, com baixa escolaridade. Conclusões: o estudo respondeu ao objetivo proposto ao demonstrar que, apesar do baixo custo para a implementação com alta devolutiva social e econômica, ainda é considerado escasso o desenvolvimento desses programas. Reitera-se a necessidade de estudos posteriores com ênfase nos níveis emocionais, como depressão e ansiedade.
Introducción: los programas dirigidos por enfermeras desempeñan un papel fundamental a la hora de implicar a las personas en el autocontrol de la diabetes mellitus tipo 2 (DM2), ya que repercuten en la mejora de la calidad de vida. El autocontrol respaldado por el apoyo social es crucial, especialmente porque describe la importancia de los profesionales de enfermería en la promoción de la salud de las personas con diabetes mellitus tipo 2 (DM2), dado que esta intervención puede ayudar a reducir las consecuencias y el empeoramiento de la enfermedad. Objetivo: mapear los programas centrados en el apoyo social para personas con DM2 dirigidos por enfermeros. Materiales y método: una revisión de alcance que siguió las directrices metodológicas del Instituto Joanna Briggs. Los descriptores de búsqueda utilizados fueron "autocuidado", "intervención dirigida por enfermeros", "diabetes mellitus tipo 2", con búsqueda en la Web of Science, Medline, CINAHL, Catálogo de Tesis y Disertaciones de la Capes [Coordinación de Perfeccionamiento de Personal de Nivel Superior], Scopus, JBI Evidence Synthesis, PsycINFO y Repositorios Científicos de Acceso Abierto de Portugal, entre febrero y marzo de 2022. Los criterios de elegibilidad fueron estudios publicados en los últimos 10 años, en inglés, español y portugués, que cubrieran la experiencia individual de programas dirigidos por profesionales de enfermería centrados en la autogestión a través del apoyo social. Resultados: se incluyeron 12 estudios, publicados en Estados Unidos, Inglaterra e Irán, centrados en el apoyo social para la autogestión de personas con DM2 y en entornos de atención a personas con enfermedades crónicas. Se observó que la mayoría de los programas dirigidos por enfermeros se aplican en países desarrollados y tienen como característica principal el énfasis en la teoría cognitiva social. El público se caracterizó por ser mayoritariamente femenino, mayor de 22 años y con bajo nivel de estudios. Conclusiones: el estudio respondió al objetivo propuesto demostrando que, a pesar del bajo costo de implementación con alto retorno social y económico, el desarrollo de estos programas aún es considerado escaso. Se reitera la necesidad de nuevos estudios que enfaticen niveles emocionales como la depresión y la ansiedad.
Introduction: Nurse-led programs have a determinant role in the engagement of individuals in the self-management of type 2 diabetes mellitus (T2DM), as they contribute to improving quality of life. Self-management assisted by social support is vital, especially as it describes the importance of nurses in promoting health for people with T2DM, as this intervention can help reduce the consequences and progression of the disease. Objective: To map programs focused on nurse-led social support for people with T2DM. Materials and Methods: This is a scoping review in line with the methodological guidelines of the Joanna Briggs Institute. The search descriptors used were "autocuidado" (self-care), "intervenção liderada por enfermeiro" (nurse-led intervention), and "diabetes mellitus tipo 2" (type 2 diabetes mellitus), with searches in the Web of Science, Medline, CINAHL, Capes Theses and Dissertations Catalog, Scopus, JBI Evidence Synthesis, PsycINFO, and the Portuguese Open Access Scientific Repositories databases conducted between February and March 2022. The eligibility criteria were studies published within the last 10 years, in English, Spanish, and Portuguese, covering the individual's experience in programs focused on self-management through nurse-led social support. Results: A total of 12 studies were included, all of which were published in the United States, England, and Iran, focusing on social support for self-management in people with T2DM, and centered on care facilities for people with chronic diseases. It was found that most nurse-led programs have been implemented in developed countries and that their main characteristic is their emphasis on social cognitive theory. The population was characterized by being mostly women, aged over 22, with a low level of education. Conclusions: The study met the proposed objective by demonstrating that, despite the low implementation cost with high social and economic returns, the development of these programs is still scarce. The need for further studies focusing on emotional levels, such as depression and anxiety, is highlighted.
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Apoyo Social , Diabetes Mellitus Tipo 2 , Automanejo , Programas Sociales , Atención de EnfermeríaRESUMEN
Intoxication by colchicine is rare, and its rapid recognition is crucial, as severe toxicity or death is reported in 10% of cases. Here, we present the case of a 50-year-old female admitted to the emergency department 24 hours after ingesting 10 mg of colchicine. Upon examination, she was conscious and hemodynamically stable. Analytically, she exhibited leukocytosis with neutrophilia and an elevation of lactate dehydrogenase (LDH). She was initiated on fluid therapy and transferred to the intermediate care unit of medicine. On the third day of hospitalization, she developed anterior chest pain, reduced breath sounds in the right hemithorax, and dullness on percussion. Arterial blood gas analysis showed partial respiratory failure, and chest X-rays and a computed tomography (CT) scan revealed a right-sided pleural effusion. The likely diagnosis was pleural effusion secondary to cardiac dysfunction due to colchicine intoxication. This case aims to describe the potential toxic effects of colchicine in cases of overdose and to reflect on ways to reduce its morbidity and mortality.
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BACKGROUND: Tumor budding (TB) is a dynamic process associated with the epithelial-mesenchymal transition and a well-established prognostic biomarker for colorectal cancer. As part of the tumor microenvironment, tumor buds demonstrate increased cell motility and invasiveness. Current evidence demonstrates that high levels of TB correlate with disease progression and worst outcomes across different solid tumors. Our work aims to demonstrate the clinical applicability of TB analysis and its utility as a prognostic factor for patients with early breast cancer (EBC). METHODS: Retrospective, single-center, observational study, enrolling patients with EBC diagnosed in a Portuguese hospital between 2014 and 2015. TB classification was performed according to the International Tumor Budding Conference 2016 guidelines. RESULTS: A statistically significant relation was found between higher TB score and aggressive clinicopathological features (angiolymphatic/perineural invasion-p < 0.001; tumor size-p = 0.012; nuclear grading-p < 0.001; and Ki-67 index-p = 0.011), higher number of relapses (p < 0.001), and short disease-free survival (DFS) (p < 0.001). CONCLUSION: We demonstrate that high TB correlates with shorter DFS and aggressive clinicopathological features used in daily practice to decide on the benefit of chemotherapy for EBC. TB represents a needed prognostic biomarker for EBC, comprising a new factor to be considered in the adjuvant decision-making process by identifying patients at a high risk of relapse and with higher benefit on treatment intensification. Clinical trials incorporating TB are needed to validate its prognostic impact.
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Rett Syndrome is an X-linked neurodevelopmental disorder (RTT; OMIM#312750) associated to MECP2 mutations. MeCP2 dysfunction is seen as one cause for the deficiencies found in brain-derived neurotrophic factor (BDNF) signaling, since BDNF is one of the genes under MeCP2 jurisdiction. BDNF signaling is also dependent on the proper function of the adenosinergic system. Indeed, both BDNF signaling and the adenosinergic system are altered in Mecp2-null mice (Mecp2-/y), a representative model of severe manifestation of RTT. Considering that symptoms severity largely differs among RTT patients, we set out to investigate the BDNF and ADO signaling modifications in Mecp2 heterozygous female mice (Mecp2+/-) presenting a less severe phenotype. Symptomatic Mecp2+/- mice have lower BDNF levels in the cortex and hippocampus. This is accompanied by a loss of BDNF-induced facilitation of hippocampal long-term potentiation (LTP), which could be restored upon selective activation of adenosine A2A receptors (A2AR). While no differences were observed in the amount of adenosine in the cortex and hippocampus of Mecp2+/- mice compared with healthy littermates, the density of the A1R and A2AR subtype receptors was, respectively, upregulated and downregulated in the hippocampus. Data suggest that significant changes in BDNF and adenosine signaling pathways are present in an RTT model with a milder disease phenotype: Mecp2+/- female animals. These features strengthen the theory that boosting adenosinergic activity may be a valid therapeutic strategy for RTT patients, regardless of their genetic penetrance.
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Factor Neurotrófico Derivado del Encéfalo , Síndrome de Rett , Animales , Femenino , Humanos , Ratones , Adenosina/metabolismo , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Estudios Transversales , Modelos Animales de Enfermedad , Proteína 2 de Unión a Metil-CpG/genética , Proteína 2 de Unión a Metil-CpG/metabolismo , Ratones Noqueados , Síndrome de Rett/metabolismoRESUMEN
The presence of basal lineage characteristics signifies hyper-aggressive human adenocarcinomas of the breast, bladder, and pancreas. However, the biochemical mechanisms that maintain this aberrant cell state are poorly understood. Here we performed marker-based genetic screens in search of factors needed to maintain basal identity in pancreatic ductal adenocarcinoma (PDAC). This approach revealed MED12 as a powerful regulator of the basal cell state in this disease. Using biochemical reconstitution and epigenomics, we show that MED12 carries out this function by bridging the transcription factor p63, a known master regulator of the basal lineage, with the Mediator complex to activate lineage-specific enhancer elements. Consistent with this finding, the growth of basal-like PDAC is hypersensitive to MED12 loss when compared to classical PDAC. Taken together, our comprehensive genetic screens have revealed a biochemical interaction that sustains basal identity in human cancer, which could serve as a target for tumor lineage-directed therapeutics.
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A splenic abscess (SA) following sleeve gastrectomy (SG) is a rare manifestation of a gastric leak (GL). The clinical findings include fever, abdominal pain, leukocytosis, and an elevated C-reactive protein. A computed tomography scan is diagnostic and can show signs of GL, or diffuse peritonitis. Treatment can either be non-operative (including large-spectrum antibiotics and percutaneous drainage) or surgical (including splenectomy). We present the case of a 41-year-old female patient with SA, with septic shock and diffuse peritonitis, successfully treated with a splenectomy three months post-SG.
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Introduction: Multiple sclerosis (MS) is an inflammatory and demyelinating disorder of central nervous system that can be diagnosed in pediatric age (<18 years) in 3-5% of the cases. This early onset is associated with higher relapse rates and earlier progression to neurological disability. By using NEDA-3 (No Evidence of Disease Activity-3) criteria, we aimed to identify clinical predictors associated with absence of disease activity and control of disease progression 12 months after the diagnosis, in a cohort of pediatric-onset MS (POMS) patients regularly followed-up in our center. Methods: We analyzed demographic, clinical, laboratorial and imaging variables of patients with POMS identified in our center, between 2010 and 2021, in two moments: at the diagnosis and 12 months after it. Statistical tests were applied to compare the distribution of those variables between groups defined by NEDA-3 status and by each one of its three variable components. Results: We included 27 patients in the study (18 female), with a mean age of 14.8 years (± 2.8), being all diagnosed with relapsing-remitting MS and with a median score of 1.5 at the Expanded Disability Status Scale (EDSS). The use of natalizumab (p = 0.017) and the negativity for anti-EBV IgG antibodies (p = 0.018) at diagnosis were associated with a higher achievement of NEDA-3 status 12 months after, in our cohort. Prescribed treatment was also associated with statistically significant differences in the "absence of MRI activity" component of NEDA-3 (p = 0.006): patients under treatment with natalizumab had a higher probability of achieving this status, and the opposite was observed in glatiramer acetate-treated children. Discussion and conclusion: Our exploratory results underline the pivotal importance that an early and more effective therapeutical approach may have in the control of disease activity, in POMS. Additionally, they also seem to suggest that the presence of anti-EBV antibodies is not innocent, as it can be related to a less favorable evolution of the disease, even at a very early stage. Further studies are needed to confirm the applicability of these variables as prognostic and personalized tools in this clinical setting.
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Inverse PCR is a powerful tool for the rapid introduction of desired mutations at desired positions in a circular double-stranded DNA sequence. In this technique, custom-designed mutant primers oriented in the inverse direction are used to amplify the entire circular template with incorporation of the required mutation(s). By careful primer design, it can be used to perform such diverse modifications as the introduction of point or multiple mutations, the insertion of new sequences, and even sequence deletions. Three primer formats are commonly used, nonoverlapping, partially overlapping, and fully overlapping primers, and here we describe the use of nonoverlapping primers for introduction of a point mutation. Use of such a primer setup in the PCR, with one of the primers containing the desired mismatch mutation, results in the amplification of a linear, double-stranded, mutated product. Methylated template DNA is removed from the non-methylated PCR product by DpnI digestion, and the PCR product is then phosphorylated by polynucleotide kinase treatment before being recircularized by ligation and transformed to E. coli. This relatively simple site-directed mutagenesis procedure is of major importance in biology and biotechnology where it is commonly employed for the study and engineering of DNA, RNA, and proteins.
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Mutagénesis Sitio-Dirigida , Reacción en Cadena de la Polimerasa , Mutagénesis Sitio-Dirigida/métodos , Reacción en Cadena de la Polimerasa/métodos , Mutación , Ingeniería de Proteínas , TemperaturaRESUMEN
Phoretic mites attach to different body parts of the red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier, 1790), to disperse. However, the question of how the patterns of attachment sites are formed remains intriguing. Here, we examined RPW-associated phoretic mites in four districts in Northern Portugal (macrohabitat), and investigated the patterns of mite distribution on six body parts of RPW (microhabitat). At the macrohabitat level, we detected seven phoretic mite taxa using the RPW host in each of the four studied districts, all documented for the first time in association with this invasive exotic species in Portugal. However, their relative abundance (species evenness) varied between districts, as did species diversity. All examined weevils carried mites, and the prevalence of the different taxa did not differ between districts or sex of weevils. Measured by mean abundance and degree of aggregation, Centrouropoda sp. proved to be the dominant taxon, while Acarus sp. and Curculanoetus rhynchophorus were considered common subordinate taxa and Uroovobella sp., Mesostigmata, Nenteria extremica and Dendrolaelaps sp. sparse taxa. At the microhabitat level, all taxa were present on all body parts of the RPW; the highest abundance was in a region encompassing the inner surface of the elytra and the membranous hind wings (subelytral space). Analysis of niche overlap revealed that the distribution patterns of phoretic mite taxa on the RPW were not randomly structured. In the subelytral space, interspecific coexistence of mites increased as a function of body size difference with the dominant Centrouropoda sp. We found that in the subelytral space the large dominant species Centrouropoda sp. displaced the larger species Uroobovella sp. and the similarly sized species Nenteria extremica, but coexisted with smaller taxa.
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One hundred and sixty-six cats from two animal shelters were subjected to enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence antibody test (IFAT), conventional polymerase chain reaction (cPCR), quantitative PCR (qPCR) and parasitological tests (PA) for the diagnosis of Leishmania spp. Among them, 15% (25/166), 53.6% (89/166), 3.6% (06/166) and 1.8% (03/166) were positive by ELISA, IFAT, both PCRs and PA, respectively. The sequencing of ITS-1 PCR amplicons revealed a 100% match with Leishmania infantum. After the Leishmania spp. survey, 12 cats were selected and divided into two groups for clinical, hematological, and biochemical analysis: six L. infantum positive cats (G1) and six Leishmania spp. negative cats (G2). All the cats were negative for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). A statistical analysis indicated significantly low platelet counts and significant hyperproteinemia associated with hypoalbuminemia in positive cats (p<0.05). Our results suggest that in endemic areas, cats with clinical signs of feline leishmaniosis (such as skin lesions, weight loss and/or enlarged lymph nodes) and that exhibit hematological and biochemical changes, such as low platelet counts and hyperproteinemia with hypoalbuminemia, should be tested for Leishmania spp. infection.
Asunto(s)
Enfermedades de los Gatos , Hipoalbuminemia , Virus de la Inmunodeficiencia Felina , Leishmania infantum , Leishmaniasis Visceral , Leishmaniasis , Gatos , Animales , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/veterinaria , Hipoalbuminemia/veterinaria , Leishmaniasis/diagnóstico , Leishmaniasis/veterinaria , Leishmaniasis/epidemiología , Virus de la Leucemia Felina , Enfermedades de los Gatos/diagnósticoRESUMEN
The propagation of strelitzia plants can be carried out in vitro as an alternative to combine the aseptic conditions of the culture medium with the use of strategies to promote germination and controlled abiotic conditions. However, this technique is still limited by the prolonged time and low percentage of seed germination, which is the most viable explant source, due to dormancy. Thus, the objective of this study was to evaluate the influence of chemical and physical scarification processes of seeds combined with gibberellic acid (GA3), as well as the effect of graphene oxide in the in vitro cultivation of strelitzia plants. Seeds were subjected to chemical scarification with sulfuric acid for different periods (10 to 60 min) and physical scarification (sandpaper), in addition to a control treatment without scarification. After disinfection, the seeds were inoculated in MS (Murashige and Skoog) medium with 30 g L-1 sucrose, 0.4 g L-1 PVPP (polyvinylpyrrolidone), 2.5 g L-1 Phytagel®, and GA3 at different concentrations. Growth data and antioxidant system responses were measured from the formed seedlings. In another experiment, the seeds were cultivated in vitro in the presence of graphene oxide at different concentrations. The results showed that the highest germination was observed in seeds scarified with sulfuric acid for 30 and 40 min, regardless of the addition of GA3. After 60 days of in vitro cultivation, physical scarification and scarification time with sulfuric acid promoted greater shoot and root length. The highest seedling survival was observed when the seeds were immersed for 30 min (86.66%) and 40 min (80%) in sulfuric acid without GA3. The concentration of 50 mg L-1 graphene oxide favored rhizome growth, while the concentration of 100 mg L-1 favored shoot growth. Regarding the biochemical data, the different concentrations did not influence MDA (Malondialdehyde) levels, but caused fluctuations in antioxidant enzyme activities.
RESUMEN
During the progression of pancreatic ductal adenocarcinoma (PDAC), tumor cells are known to acquire transcriptional and morphological properties of the basal (also known as squamous) epithelial lineage, which leads to more aggressive disease characteristics. Here, we show that a subset of basal-like PDAC tumors aberrantly express p73 (TA isoform), which is a known transcriptional activator of basal lineage identity, ciliogenesis, and tumor suppression in normal tissue development. Using gain- and loss- of function experiments, we show that p73 is necessary and sufficient to activate genes related to basal identity (e.g. KRT5), ciliogenesis (e.g. FOXJ1), and p53-like tumor suppression (e.g. CDKN1A) in human PDAC models. Owing to the paradoxical combination of oncogenic and tumor suppressive outputs of this transcription factor, we propose that PDAC cells express a low level of p73 that is optimal for promoting lineage plasticity without severe impairment of cell proliferation. Collectively, our study reinforces how PDAC cells exploit master regulators of the basal epithelial lineage during disease progression.