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The Association for the Study of Reproductive Biology (ASEBIR) Interest Group in Embryology (in Spanish 'Grupo de Interés de Embriología') reviewed key morphokinetic parameters to assess the contribution of time-lapse technology (TLT) to the ASEBIR grading system. Embryo grading based on morphological characteristics is the most widely used method in human assisted reproduction laboratories. The introduction and implementation of TLT has provided a large amount of information that can be used as a complementary tool for morphological embryo evaluation and selection. As part of IVF treatments, embryologists grade embryos to decide which embryos to transfer or freeze. At the present, the embryo grading system developed by ASEBIR does not consider dynamic events observed through TLT. Laboratories that are using TLT consider those parameters as complementary data for embryo selection. The aim of this review was to evaluate review time-specific morphological changes during embryo development that are not included in the ASEBIR scoring system, and to consider them as candidates to add to the scoring system.
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Embrião de Mamíferos , Desenvolvimento Embrionário , Humanos , Imagem com Lapso de Tempo/métodos , Transferência Embrionária/métodos , Biologia , Técnicas de Cultura Embrionária , Implantação do Embrião , Fertilização in vitro/métodos , BlastocistoRESUMO
The purpose of the study was to investigate to which extent a healthy lifestyle in female healthcare workers with chronic pain contributes to reducing the risk of disability pension. We conducted a prospective cohort study with an 11-year registry follow-up. Overall, 2386 Danish female healthcare workers with chronic pain completed a questionnaire about work and lifestyle (leisure-time physical activity, smoking, and body mass index (BMI)). Data on disability benefit payments were obtained from the Danish Register for Evaluation of Marginalization. Two models (minimally and fully adjusted for different potential confounders) were tested using the Cox proportional hazards model. During the follow-up period, 17.9% of the healthcare workers obtained disability pension. Low levels of leisure time physical activity (reference: moderate level) increased the risk of disability pension in the minimally (Hazard Ratio: 1.38 (95% CI: 1.14-1.69)) and fully adjusted models (Hazard Ratio: 1.27 (95% CI: 1.04-1.56)). Being highly physically active, as opposed to being moderately active, did not confer additional protection. Additionally, a positive association was observed between smoking and disability pension in the minimally adjusted model (Hazard Ratio: 1.27 (95% CI: 1.05-1.54)). BMI was not an influential factor. In female healthcare workers with chronic pain, at least moderate levels of physical activity is a protective factor for disability pension. Effective promotion strategies should be designed for both workplace and non-workplace settings.
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Dor Crônica , Pessoas com Deficiência , Humanos , Feminino , Estudos Prospectivos , Seguimentos , Dor Crônica/epidemiologia , Pensões , Inquéritos e Questionários , Estilo de Vida Saudável , Fatores de Risco , Modelos de Riscos ProporcionaisRESUMO
INTRODUCTION: The assessment of quality of life (QoL) should be one of the main objectives in paediatric clinical trials. Even though researchers, regulators and advocates support the use of patient-reported outcomes (PROs), this has not been fully implemented. The aim of this study is to assess the measurement of QoL and the usage of PROs, palatability assessments and medication diaries in early-phase clinical trials for childhood and adolescent cancer. METHODS: Early-phase clinical trials for children and adolescents with cancer opened between 2005 and 2022 at the Royal Marsden Hospital (London, UK) and Vall d'Hebron University Hospital (Barcelona, Spain) were interrogated for trial characteristics and the use of QoL questionnaires, PROs, palatability assessments and medication diaries. RESULTS: Overall, 72 clinical trials were analysed: 12 (16.7%) evaluated QoL and eight (11.1%) evaluated PROs. Palatability was tested in 21/40 (52.5%) trials of oral drugs and 23/72 (31.9%) incorporated medication diaries. No studies mentioned patient involvement in the trial protocol. Use of PROs increased from one of 36 (2.8%) to seven of 36 (19.4%) between the first period (2005-2016) and the second period (2017-2022) (p = .02). Implementation of medication diaries increased from seven of 36 (19.4%) to 16/36 (44.4%) in each period, respectively (p = .02). CONCLUSION: Only a minor proportion of the international/multicentric early-phase trials evaluated included QoL/PROs and medication diaries or palatability questionnaires to help assess these, although this trend seems to be increasing over recent years. Greater implementation of QoL/PROs has the potential to improve the patient's wellbeing and facilitate symptom control, to enhance patient/parent involvement in future trial designs and to provide information for drug prioritisation.
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Neoplasias , Medidas de Resultados Relatados pelo Paciente , Qualidade de Vida , Humanos , Adolescente , Criança , Neoplasias/psicologia , Neoplasias/tratamento farmacológico , Neoplasias/terapia , Feminino , Masculino , Ensaios Clínicos como Assunto , Inquéritos e Questionários , Pré-EscolarRESUMO
Mobile genetic elements control their life cycles by the expression of a master repressor, whose function must be disabled to allow the spread of these elements in nature. Here, we describe an unprecedented repression-derepression mechanism involved in the transfer of Staphylococcus aureus pathogenicity islands (SaPIs). Contrary to the classical phage and SaPI repressors, which are dimers, the SaPI1 repressor StlSaPI1 presents a unique tetrameric conformation never seen before. Importantly, not just one but two tetramers are required for SaPI1 repression, which increases the novelty of the system. To derepress SaPI1, the phage-encoded protein Sri binds to and induces a conformational change in the DNA binding domains of StlSaPI1, preventing the binding of the repressor to its cognate StlSaPI1 sites. Finally, our findings demonstrate that this system is not exclusive to SaPI1 but widespread in nature. Overall, our results characterize a novel repression-induction system involved in the transfer of MGE-encoded virulence factors in nature.
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Ilhas Genômicas , Fagos de Staphylococcus , Ilhas Genômicas/genética , Fagos de Staphylococcus/genética , Staphylococcus aureus/genéticaRESUMO
We describe a pertussis outbreak in the Vallès region of Catalonia, from September 2023 to April 2024. Incidence was high in children aged 10-14â¯years compared with previous outbreaks. Limited impact in newborns could be explained by the high vaccination coverage during pregnancy and at 11 months of age in 2022, at 85% and 94.1 %, respectively. A third booster vaccine dose during preadolescence should be considered and vaccination coverage in pregnant women be improved to control future outbreaks.
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Surtos de Doenças , Vacina contra Coqueluche , Coqueluche , Humanos , Coqueluche/epidemiologia , Coqueluche/prevenção & controle , Coqueluche/diagnóstico , Espanha/epidemiologia , Feminino , Adolescente , Criança , Incidência , Lactente , Vacina contra Coqueluche/administração & dosagem , Gravidez , Pré-Escolar , Masculino , Recém-Nascido , Vacinação/estatística & dados numéricos , Adulto , Cobertura Vacinal/estatística & dados numéricos , Imunização Secundária , Adulto Jovem , Bordetella pertussis/isolamento & purificação , Distribuição por Idade , Vigilância da PopulaçãoRESUMO
Psychopathic traits in childhood have been revealed as potential identifiers of risk, being predictive of later forms of behavioral maladjustment. Yet, it is still under debate how psychopathic traits in children should be best conceptualized and which are the core dimensions for construct definition and prediction. The present study aims to examine the structure of psychopathic traits in childhood, and its predictive value, by using a combination of traditional factor analysis and more recent network-based methods. Data on psychopathic traits, as measured by the Child Problematic Traits Inventory (CPTI), were collected in a large sample of children (n = 2454; 48.2% girls), aged 3 to 6 at the onset of the study (Mage = 4.26; SD = 0.91), who were followed-up one and two years later using parent- and teacher-reports. Results showed that psychopathic traits measured via CPTI are best conceptualized as five latent factors encompassing grandiosity, deceitfulness, callousness, impulsivity and need of stimulation, a result that converged across informants and time. Callousness and grandiosity emerged as central traits using network analysis of parent-reports, while deceitfulness was most central using teacher-reports. Finally, callousness, impulsivity and deceitfulness emerged as the best predictors of concurrent, prospective and stable conduct problems. These results provide a refined structure of psychopathic traits in children that better accounts for the core elements of the construct. Additional theoretical and practical implications will be discussed in terms of assessment, diagnostic classification and tailored prevention/intervention.
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Breast cancer is the leading malignancy in women worldwide, both in terms of incidence and mortality. Triple-negative breast cancer (TNBC) is the type with the worst clinical outcomes and with fewer therapeutic options than other types of breast cancer. GK-1 is a peptide that in the experimental model of the metastatic 4T1 breast cancer has demonstrated anti-tumor and anti-metastatic properties. Herein, GK-1 (5 mg/kg, i.v.) weekly administrated not only decreases tumor growth and the number of lung macro-metastases but also lung and lymph nodes micro-metastases. Histological analysis reveals that GK-1 reduced 57% of the intra-tumor vascular areas, diminished the leukemoid reaction's progression, and the spleens' weight and length. A significant reduction in VEGF-C, SDF-1, angiopoietin-2, and endothelin-1 angiogenic factors was induced. Moreover, GK-1 prevents T cell exhaustion in the tumor-infiltrating lymphocytes (TILs) decreasing PD-1 expression. It also increased IFN-γ and granzyme-B expression and the cytotoxic activity of CD8+ TILs cells against tumor cells. All these features were found to be associated with a better antitumor response and prognosis. Altogether, these results reinforce the potential of GK-1 to improve the clinical outcome of triple-negative breast cancer immunotherapy. Translation research is ongoing towards its evaluation in humans.
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Antineoplásicos , Neoplasias de Mama Triplo Negativas , Humanos , Feminino , Animais , Camundongos , Neoplasias de Mama Triplo Negativas/patologia , Exaustão das Células T , Linfócitos do Interstício Tumoral/metabolismo , Prognóstico , Antineoplásicos/uso terapêutico , Linfócitos T CD8-Positivos/metabolismoRESUMO
BACKGROUND AND AIMS: Computer-aided detection (CADe) has been shown to improve polyp detection in clinical trials. Limited data exist on the impact, utilization, and attitudes toward artificial intelligence (AI)-assisted colonoscopy in daily clinical practice. We aimed to evaluate the effectiveness of the first U.S. Food and Drug Administration-approved CADe device for polyp detection in the United States and the attitudes toward its implementation. METHODS: We performed a retrospective analysis of a prospectively maintained database of patients undergoing colonoscopy at a tertiary center in the United States before and after a real-time CADe system was made available. The decision to activate the CADe system was at the discretion of the endoscopist. An anonymous survey was circulated among endoscopy physicians and staff at the beginning and conclusion of the study period regarding their attitudes toward AI-assisted colonoscopy. RESULTS: CADe was activated in 52.1% of cases. Compared with historical control subjects, there was no statistically significant difference in adenomas detected per colonoscopy (1.08 vs 1.04, P = .65), even after excluding diagnostic and therapeutic indications and cases where CADe was not activated (1.27 vs 1.17, P = .45). In addition, there was no statistically significant difference in adenoma detection rate (ADR), median procedure, and withdrawal times. Survey results demonstrated mixed attitudes toward AI-assisted colonoscopy, of which main concerns were high number of false-positive signals (82.4%), high level of distraction (58.8%), and impression it prolonged procedure time (47.1%). CONCLUSIONS: CADe did not improve adenoma detection in daily practice among endoscopists with high baseline ADRs. Despite its availability, AI-assisted colonoscopy was only activated in half of the cases, and multiple concerns were raised by staff and endoscopists. Future studies will help elucidate the patients and endoscopists that would benefit most from AI-assisted colonoscopy.
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Adenoma , Pólipos do Colo , Neoplasias Colorretais , Humanos , Pólipos do Colo/diagnóstico , Estudos Retrospectivos , Inteligência Artificial , Centros de Atenção Terciária , Colonoscopia/métodos , Computadores , Adenoma/diagnóstico , Neoplasias Colorretais/diagnósticoRESUMO
Staphylococcus aureus infections can lead to diseases that range from localized skin abscess to life-threatening toxic shock syndrome. The SrrAB two-component system (TCS) is a global regulator of S. aureus virulence and critical for survival under environmental conditions such as hypoxic, oxidative, and nitrosative stress found at sites of infection. Despite the critical role of SrrAB in S. aureus pathogenicity, the mechanism by which the SrrAB TCS senses and responds to these environmental signals remains unknown. Bioinformatics analysis showed that the SrrB histidine kinase contains several domains, including an extracellular Cache domain and a cytoplasmic HAMP-PAS-DHp-CA region. Here, we show that the PAS domain regulates both kinase and phosphatase enzyme activity of SrrB and present the structure of the DHp-CA catalytic core. Importantly, this structure shows a unique intramolecular cysteine disulfide bond in the ATP-binding domain that significantly affects autophosphorylation kinetics. In vitro data show that the redox state of the disulfide bond affects S. aureus biofilm formation and toxic shock syndrome toxin-1 production. Moreover, with the use of the rabbit infective endocarditis model, we demonstrate that the disulfide bond is a critical regulatory element of SrrB function during S. aureus infection. Our data support a model whereby the disulfide bond and PAS domain of SrrB sense and respond to the cellular redox environment to regulate S. aureus survival and pathogenesis.
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Proteínas de Bactérias/metabolismo , Cisteína/metabolismo , Proteínas Repressoras/metabolismo , Staphylococcus aureus/metabolismo , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Toxinas Bacterianas , Sequência de Bases , Biofilmes , Domínio Catalítico , Modelos Animais de Doenças , Endocardite , Enterotoxinas , Feminino , Regulação Bacteriana da Expressão Gênica , Histidina Quinase/metabolismo , Masculino , Modelos Moleculares , Mutação , Oxirredução , Domínios Proteicos , Coelhos , Proteínas Repressoras/química , Proteínas Repressoras/genética , Sepse , Infecções Estafilocócicas/metabolismo , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidade , Superantígenos , Thermotoga maritima , Virulência/genética , Virulência/fisiologiaRESUMO
The present study aims to further examine the four-factor model of psychopathy in adolescence with a new alternate model for the assessment of psychopathic traits and conduct disorder (CD): The Proposed Specifiers for Conduct Disorder-Short version (PSCD-SV). Data were collected in a sample of 414 adolescents (49.2% females) aged 12-15 at the first assessment who were then followed-up 2 years later. Results supported the usefulness of the PSCD-SV to assess the broader construct of psychopathy showing good psychometric properties, including adequate reliability and validity, while accounting for all its dimensions. In addition, the study showed close associations between psychopathic traits and adolescent behavioral, emotional and psychosocial maladjustment. Finally, the findings elucidated the PSCD's connection to parental support and psychological control, and reinforced the potential role of parenting practices as predictors that can act as mechanisms of change in the development of psychopathy. Overall, current findings shed light on conceptual and developmental models of psychopathy that may have implications for assessment, diagnostic classification, prevention, and intervention.
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Transtorno da Conduta , Adolescente , Feminino , Humanos , Masculino , Transtorno da Conduta/diagnóstico , Transtorno da Conduta/psicologia , Poder Familiar , Psicometria , Reprodutibilidade dos Testes , Transtorno da Personalidade Antissocial/diagnóstico , Transtorno da Personalidade Antissocial/psicologiaRESUMO
BACKGROUND: Impaired insight into illness is a common feature of schizophrenia. Improved insight is associated with better treatment adherence and clinical outcomes. At the same time, improving insight has been suggested to increase depressive symptoms and diminish quality of life. The aim of this study was to examine the associations between impaired insight and degree of subjective happiness, perceived level of success, and life satisfaction in patients with schizophrenia spectrum disorders. METHODS: A total of 108 participants with schizophrenia or schizoaffective disorder were included. Data for this study were obtained from our group's previous investigation that examined the relationship between impaired insight and visuospatial attention. Insight into illness was measured by the VAGUS scale, which assesses general illness awareness, accurate symptom attribution, awareness of the need for treatment, and awareness of the negative consequences attributable to the illness. RESULTS: Our results revealed no association among the VAGUS average and subscale scores and degree of subjective happiness, perceived level of success, and life satisfaction. CONCLUSIONS: Our study suggests that insight into illness is not related to subjective happiness, life satisfaction, or perceived level of success in patients with schizophrenia, which is in contrast to previous reports that demonstrate an association between insight into illness and depression.
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Esquizofrenia , Humanos , Esquizofrenia/complicações , Psicologia do Esquizofrênico , Felicidade , Qualidade de Vida , Satisfação PessoalRESUMO
RegulonDB, first published 20 years ago, is a comprehensive electronic resource about regulation of transcription initiation of Escherichia coli K-12 with decades of knowledge from classic molecular biology experiments, and recently also from high-throughput genomic methodologies. We curated the literature to keep RegulonDB up to date, and initiated curation of ChIP and gSELEX experiments. We estimate that current knowledge describes between 10% and 30% of the expected total number of transcription factor- gene regulatory interactions in E. coli. RegulonDB provides datasets for interactions for which there is no evidence that they affect expression, as well as expression datasets. We developed a proof of concept pipeline to merge binding and expression evidence to identify regulatory interactions. These datasets can be visualized in the RegulonDB JBrowse. We developed the Microbial Conditions Ontology with a controlled vocabulary for the minimal properties to reproduce an experiment, which contributes to integrate data from high throughput and classic literature. At a higher level of integration, we report Genetic Sensory-Response Units for 200 transcription factors, including their regulation at the metabolic level, and include summaries for 70 of them. Finally, we summarize our research with Natural language processing strategies to enhance our biocuration work.
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Biologia Computacional/métodos , Escherichia coli K12/genética , Regulação Bacteriana da Expressão Gênica , Genômica , Ontologia Genética , Redes Reguladoras de Genes , Genômica/métodos , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
The precise determination of de novo genetic variants has enormous implications across different fields of biology and medicine, particularly personalized medicine. Currently, de novo variations are identified by mapping sample reads from a parent-offspring trio to a reference genome, allowing for a certain degree of differences. While widely used, this approach often introduces false-positive (FP) results due to misaligned reads and mischaracterized sequencing errors. In a previous study, we developed an alternative approach to accurately identify single nucleotide variants (SNVs) using only perfect matches. However, this approach could be applied only to haploid regions of the genome and was computationally intensive. In this study, we present a unique approach, coverage-based single nucleotide variant identification (COBASI), which allows the exploration of the entire genome using second-generation short sequence reads without extensive computing requirements. COBASI identifies SNVs using changes in coverage of exactly matching unique substrings, and is particularly suited for pinpointing de novo SNVs. Unlike other approaches that require population frequencies across hundreds of samples to filter out any methodological biases, COBASI can be applied to detect de novo SNVs within isolated families. We demonstrate this capability through extensive simulation studies and by studying a parent-offspring trio we sequenced using short reads. Experimental validation of all 58 candidate de novo SNVs and a selection of non-de novo SNVs found in the trio confirmed zero FP calls. COBASI is available as open source at https://github.com/Laura-Gomez/COBASI for any researcher to use.
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Variações do Número de Cópias de DNA , Genoma Humano , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Pais , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA/métodos , Software , Algoritmos , Criança , HumanosRESUMO
BACKGROUND: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic is a current public health concern. Rapid diagnosis is crucial, and reverse transcription polymerase chain reaction (RT-PCR) is presently the reference standard for SARS-CoV-2 detection. OBJECTIVE: Automated RT-PCR analysis (ARPA) is a software designed to analyze RT-PCR data for SARSCoV-2 detection. ARPA loads the RT-PCR data, classifies each sample by assessing its amplification curve behavior, evaluates the experiment's quality, and generates reports. METHODS: ARPA was implemented in the R language and deployed as a Shiny application. We evaluated the performance of ARPA in 140 samples. The samples were manually classified and automatically analyzed using ARPA. RESULTS: ARPA had a true-positive rate = 1, true-negative rate = 0.98, positive-predictive value = 0.95, and negative-predictive value = 1, with 36 samples correctly classified as positive, 100 samples correctly classified as negative, and two samples classified as positive even when labeled as negative by manual inspection. Two samples were labeled as invalid by ARPA and were not considered in the performance metrics calculation. CONCLUSIONS: ARPA is a sensitive and specific software that facilitates the analysis of RT-PCR data, and its implementation can reduce the time required in the diagnostic pipeline.
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COVID-19/diagnóstico , Diagnóstico por Computador , SARS-CoV-2/isolamento & purificação , Software , Teste para COVID-19 , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saliva/virologiaRESUMO
Human T cell leukemia virus type 1 (HTLV-1) is the ethological agent of adult T cell leukemia/lymphoma (ATLL) and a number of lymphocyte-mediated inflammatory conditions, including HTLV-1-associated myelopathy/tropical spastic paraparesis. HTLV-1 orf-I encodes two proteins, p8 and p12, whose functions in humans are to counteract innate and adaptive responses and to support viral transmission. However, the in vivo requirements for orf-I expression vary in different animal models. In macaques, the ablation of orf-I expression by mutation of its ATG initiation codon abolishes the infectivity of the molecular clone HTLV-1p12KO In rabbits, HTLV-1p12KO is infective and persists efficiently. We used humanized mouse models to assess the infectivity of both wild-type HTLV-1 (HTLV-1WT) and HTLV-1p12KO We found that NOD/SCID/γC-/- c-kit+ mice engrafted with human tissues 1 day after birth (designated NSG-1d mice) were highly susceptible to infection by HTLV-1WT, with a syndrome characterized by the rapid polyclonal proliferation and infiltration of CD4+ CD25+ T cells into vital organs, weight loss, and death. HTLV-1 clonality studies revealed the presence of multiple clones of low abundance, confirming the polyclonal expansion of HTLV-1-infected cells in vivo HTLV-1p12KO infection in a bone marrow-liver-thymus (BLT) mouse model prone to graft-versus-host disease occurred only following reversion of the orf-I initiation codon mutation within weeks after exposure and was associated with high levels of HTLV-1 DNA in blood and the expansion of CD4+ CD25+ T cells. Thus, the incomplete reconstitution of the human immune system in BLT mice may provide a window of opportunity for HTLV-1 replication and the selection of viral variants with greater fitness.IMPORTANCE Humanized mice constitute a useful model for studying the HTLV-1-associated polyclonal proliferation of CD4+ T cells and viral integration sites in the human genome. The rapid death of infected animals, however, appears to preclude the clonal selection typically observed in human ATLL, which normally develops in 2 to 5% of individuals infected with HTLV-1. Nevertheless, the expansion of multiple clones of low abundance in these humanized mice mirrors the early phase of HTLV-1 infection in humans, providing a useful model to investigate approaches to inhibit virus-induced CD4+ T cell proliferation.
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Linfócitos T CD4-Positivos/virologia , Proliferação de Células , Infecções por HTLV-I/patologia , Infecções por HTLV-I/virologia , Interações Hospedeiro-Patógeno , Vírus Linfotrópico T Tipo 1 Humano/crescimento & desenvolvimento , Proteínas Virais Reguladoras e Acessórias/metabolismo , Animais , Modelos Animais de Doenças , Transmissão de Doença Infecciosa , Camundongos , Camundongos Knockout , Camundongos SCID , Proteínas Virais Reguladoras e Acessórias/deficiênciaRESUMO
The RcsCDB phosphorelay system controls an extremely large regulon in Enterobacteriaceae that involves processes such as biofilm formation, flagella production, synthesis of extracellular capsules and cell division. Therefore, fine-tuning of this system is essential for virulence in pathogenic microorganisms of this group. The final master effector of the RcsCDB system is the response regulator (RR) RcsB, which activates or represses multiple genes by binding to different promoter regions. This regulatory activity of RcsB can be done alone or in combination with additional transcriptional factors in phosphorylated or dephosphorylated states. The capacity of RcsB to interact with multiple promoters and partners, either dephosphorylated or phosphorylated, suggests an extremely conformational dynamism for this RR. To shed light on the activation mechanism of RcsB and its implication on promoter recognition, we solved the crystal structure of full-length RcsB from Salmonella enterica serovar Typhimurium in the presence and absence of a phosphomimetic molecule BeF3-. These two novel structures have guided an extensive site-directed mutagenesis study at the structural and functional level that confirms RcsB conformational plasticity and dynamism. Our data allowed us to propose a ß5-T switch mechanism where phosphorylation is coupled to alternative DNA binding ways and which highlights the conformational dynamism of RcsB to be so pleiotropic.
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Proteínas de Bactérias/química , DNA/química , Conformação de Ácido Nucleico , Domínios Proteicos , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cristalografia por Raios X , DNA/genética , DNA/metabolismo , Modelos Moleculares , Fosforilação , Ligação Proteica , Salmonella typhimurium/genética , Salmonella typhimurium/metabolismo , Homologia de Sequência de AminoácidosRESUMO
BACKGROUND: The relationship between community-acquired respiratory viruses (CARVs) and chronic lung allograft dysfunction (CLAD) in lung transplant recipients is still controversial. METHODS: We performed a prospective cohort study (2009-2014) in all consecutive adult patients (≥18 years) undergoing lung transplantation in the Hospital Universitari Vall d'Hebron (Barcelona, Spain). We systematically collected nasopharyngeal swabs from asymptomatic patients during seasonal changes, from patients with upper respiratory tract infectious disease, lower respiratory tract infectious disease (LRTID), or acute rejection. Nasopharyngeal swabs were analyzed by multiplex polymerase chain reaction. Primary outcome was to evaluate the potential association of CARVs and development of CLAD. Time-dependent Cox regression models were performed to identify the independent risk factors for CLAD. RESULTS: Overall, 98 patients (67 bilateral lung transplant recipients; 63.3% male; mean age, 49.9 years) were included. Mean postoperative follow-up was 3.4 years (interquartile range [IQR], 2.5-4.0 years). Thirty-eight lung transplant recipients (38.8%) developed CLAD, in a median time of 20.4 months (IQR, 12-30.4 months). In time-controlled multivariate analysis, CARV-LRTID (hazard ratio [HR], 3.00 [95% confidence interval {CI}, 1.52-5.91]; P = .002), acute rejection (HR, 2.97 [95% CI, 1.51-5.83]; P = .002), and cytomegalovirus pneumonitis (HR, 3.76 [95% CI, 1.23-11.49]; P = .02) were independent risk factors associated with developing CLAD. CONCLUSIONS: Lung transplant recipients with CARVs in the lower respiratory tract are at increased risk to develop CLAD.
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Infecções Comunitárias Adquiridas/etiologia , Infecções Comunitárias Adquiridas/fisiopatologia , Pneumopatias/etiologia , Pneumopatias/fisiopatologia , Transplante de Pulmão/efeitos adversos , Pneumonia Viral/etiologia , Pneumonia Viral/fisiopatologia , Adulto , Aloenxertos , Infecções Comunitárias Adquiridas/epidemiologia , Feminino , Seguimentos , Humanos , Pneumopatias/diagnóstico , Pneumopatias/epidemiologia , Masculino , Pessoa de Meia-Idade , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Modelos de Riscos Proporcionais , Estudos Prospectivos , Testes de Função Respiratória , Fatores de Risco , TransplantadosRESUMO
Perforin-2 (P-2) is a recently described antimicrobial protein with unique properties to kill intracellular bacteria. We investigated P-2 expression pattern and cellular distribution in human skin and its importance in restoration of barrier function during wound healing process and infection with the common wound pathogen Staphylococcus aureus. We describe a novel approach for the measurement of P-2 mRNA within individual skin cells using an amplified fluorescence in situ hybridization (FISH) technique. The unique aspect of this approach is simultaneous detection of P-2 mRNA in combination with immune-phenotyping for cell surface proteins using fluorochrome-conjugated antibodies. We detected P-2 transcript in both hematopoietic (CD45+ ) and non-hematopoietic (CD45- ) cutaneous cell populations, confirming the P-2 expression in both professional and non-professional phagocytes. Furthermore, we found an induction of P-2 during wound healing. P-2 overexpression resulted in a reduction of intracellular S. aureus, while infection of human wounds by this pathogen resulted in P-2 suppression, revealing a novel mechanism by which S. aureus may escape cutaneous immunity to cause persistent wound infections.
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Proteínas Citotóxicas Formadoras de Poros/metabolismo , Análise de Célula Única/métodos , Pele/metabolismo , Infecções Estafilocócicas/metabolismo , Cicatrização , Animais , Membrana Celular/metabolismo , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Fibroblastos/metabolismo , Células HEK293 , Humanos , Imunofenotipagem , Hibridização in Situ Fluorescente , Linfócitos Intraepiteliais/imunologia , Linfócitos Intraepiteliais/metabolismo , Queratinócitos/imunologia , Queratinócitos/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Camundongos , Pele/microbiologia , Staphylococcus aureusRESUMO
Francisellosis is a disease caused by different species of the bacterial genus Francisella and has been diagnosed in a wide variety of animals, including fish. Francisellosis in fish is characterized by the development of non-specific clinical signs as well as the presence of numerous granulomas in several organs (mainly spleen and kidney). Ten neon jewel cichlids Hemichromis bimaculatus were submitted for diagnosis from a farm located in Morelos, Mexico. Gross examination, wet preparations, cytology, histopathology and PCR were performed. Affected fish showed lethargy, erratic swimming, imbalance and gasping. At the post mortem examination, multiple granulomas were observed in the kidney and spleen. Microscopically, granulomatous inflammation was observed in several organs. Species-specific PCR assay using DNA from the affected tissues of H. bimaculatus as a template demonstrated the presence of F. noatunensis subsp. orientalis (Fno) by amplifying a hypothetical protein gene of the Fno species. The end diagnosis of francisellosis is important for Mexican ornamental aquaculture, since it is necessary to implement measures for treatment, prevention, control and diagnosis. This is the first report of francisellosis in the neon jewel cichlid.
Assuntos
Ciclídeos , Doenças dos Peixes , Francisella , Infecções por Bactérias Gram-Negativas , Animais , Surtos de Doenças , MéxicoRESUMO
Seborrhoeic Dermatitis (SD) is a common inflammatory skin disorder, but its molecular pathogenesis remains elusive. Previously, we have established the Mpzl3 knockout (-/-) mice as a model for SD. In this study, we focused on early phases of skin inflammation and determined the cytokine profiles and identified immune cell types in the lesional skin in the Mpzl3 -/- mice. Using flow cytometry, we detected a significant increase of CD45+ leucocytes, CD3+ T lymphocytes and especially γδ T cells but not αß T cells in the lesional skin compared to control. We also detected high levels of IL-17 and determined that the γδ T cells were a major contributing source. CD3+ and γδ T cell localization in the skin was verified by indirect immunofluorescent staining. Since neither γδ T cells nor IL-17 had been implicated in SD, our study provides novel insights into the role of MPZL3 in the pathogenesis of SD-like skin inflammation.