RESUMO
E6 and E7 oncogenes are pivotal in the carcinogenic transformation in HPV infections and efficient diagnostic methods can ensure the detection and differentiation of HPV genotype. This study describes the development and validation of an electrochemical, label-free genosensor coupled with a microfluidic system for detecting the E6 and E7 oncogenes in cervical scraping samples. The nanostructuring employed was based on a cysteine and graphene quantum dots layer that provides functional groups, surface area, and interesting electrochemical properties. Biorecognition tests with cervical scraping samples showed differentiation in the voltammetric response. Low-risk HPV exhibited a lower biorecognition response, reflected in ΔI% values of 82.33 % ± 0.29 for HPV06 and 80.65 % ± 0.68 for HPV11 at a dilution of 1:100. Meanwhile, high-risk, HPV16 and HPV18, demonstrated ΔI% values of 96.65 % ± 1.27 and 93 % ± 0.026, respectively, at the same dilution. Therefore, the biorecognition intensity followed the order: HPV16 >HPV18 >HPV06 >HPV11. The limit of detection and the limit of quantification of E6E7 microfluidic LOC-Genosensor was 26 fM, and 79.6 fM. Consequently, the E6E7 biosensor is a valuable alternative for clinical HPV diagnosis, capable of detecting the potential for oncogenic progression even in the early stages of infection.
Assuntos
Técnicas Biossensoriais , Proteínas Oncogênicas Virais , Técnicas Biossensoriais/métodos , Humanos , Proteínas Oncogênicas Virais/genética , Feminino , Limite de Detecção , Proteínas E7 de Papillomavirus/genética , Colo do Útero/virologia , Grafite/química , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Técnicas Eletroquímicas/métodos , Proteínas Repressoras/genética , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/instrumentação , Pontos Quânticos/química , Dispositivos Lab-On-A-Chip , Papillomaviridae/genética , Papillomaviridae/isolamento & purificaçãoRESUMO
In recent years, emerging environmental pollutants have increasingly endangered the health of freshwater organisms. The gut microbiota exhibits sensitivity to medications, dietary factors and environmental pollutants, rendering it a novel target for toxicological studies. The gut microbiota can be a potential exposure route affecting the host's health. Herein, we review the current knowledge on two different but concurrent pollutants, microplastics and pesticides, regarding their impact on the gut microbiota, which includes alterations in microbial composition, gene expression, function, and health effects in the hosts. Moreover, synergetic interactions between microplastics and pesticides can exacerbate dysbiosis and health risks. We discuss health-related implications of gut microbial changes based on the consequences in metabolism, immunity, and physiology function. Further research is needed to discover the mechanisms underlying these effects and develop strategies for mitigating their harmful impacts on freshwater animals.
Assuntos
Água Doce , Microbioma Gastrointestinal , Microplásticos , Praguicidas , Poluentes Químicos da Água , Microbioma Gastrointestinal/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Microplásticos/toxicidade , Animais , Praguicidas/toxicidadeRESUMO
The differential diagnosis of supratentorial brain tumours in children can be challenging, especially considering the recent changes to the WHO classification of CNS tumours published in 2021. Many new tumour types have been proposed which frequently present in children and young adults and their imaging features are currently being described by the neuroradiology community. The purpose of this article is to provide guidance to residents and fellows new to the field of paediatric neuroradiology on how to evaluate an MRI of a patient with a newly diagnosed supratentorial tumour. Six different approaches are discussed including: 1. Tumour types, briefly discussing the main changes to the recent WHO classification of CNS tumours, 2. Patient age and its influence on incidence rates of specific tumour types, 3. Growth patterns, 4. Tumour location and how defining the correct location helps in narrowing down the differential diagnoses and 5. Imaging features of the tumour on DWI, SWI, FLAIR and post contrast sequences.
Assuntos
Imageamento por Ressonância Magnética , Neoplasias Supratentoriais , Humanos , Neoplasias Supratentoriais/diagnóstico por imagem , Neoplasias Supratentoriais/patologia , Criança , Imageamento por Ressonância Magnética/métodos , Diagnóstico Diferencial , Fatores Etários , Meios de ContrasteRESUMO
The cefazolin inoculum effect (CzIE) has been associated with poor clinical outcomes in patients with MSSA infections. We aimed to investigate the point prevalence of the CzIE among nasal colonizing MSSA isolates from ICU patients in a multicenter study in Colombia (2019-2023). Patients underwent nasal swabs to assess for S. aureus colonization on admission to the ICU and some individuals had follow-up swabs. We performed cefazolin MIC by broth-microdilution using standard and high-inoculum and developed a modified nitrocefin-based rapid test to detect the CzIE. Whole genome sequencing was carried out to characterize BlaZ types and allotypes, phylogenomics and Agr-typing. All swabs were subjected to 16S-rRNA metabarcoding sequencing to evaluate microbiome characteristics associated with the CzIE. A total of 352 patients were included; 46/352 (13%) patients were colonized with S. aureus; 22% (10/46) and 78% (36/46) with MRSA and MSSA, respectively. Among 36 patients that contributed with 43 MSSA colonizing isolates, 21/36 (58%) had MSSA exhibiting the CzIE. BlaZ type A and BlaZ-2 were the predominant type and allotype in 56% and 52%, respectively. MSSA belonging to CC30 were highly associated with the CzIE and SNP analyses supported transmission of MSSA exhibiting the CzIE among some patients of the same unit. The modified nitrocefin rapid test had 100%, 94.4% and 97.7% sensitivity, specificity and accuracy, respectively. We found a high prevalence point prevalence of the CzIE in MSSA colonizing the nares of critically-ill patients in Colombia. A modified rapid test was highly accurate in detecting the CzIE in this patient population.
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Bioactive and biodegradable scaffolds that mimic the natural extracellular matrix of bone serve as temporary structures to guide new bone tissue growth. In this study, 3D-printed scaffolds composed of poly (lactic acid) (PLA)-tricalcium phosphate (TCP) (90-10 wt.%) were modified with 1%, 5%, and 10 wt.% of ZnO to enhance bone tissue regeneration. A commercial chain extender named Joncryl was incorporated alongside ZnO to ensure the printability of the composites. Filaments were manufactured using a twin-screw extruder and subsequently used to print 3D scaffolds via fused filament fabrication (FFF). The scaffolds exhibited a homogeneous distribution of ZnO and TCP particles, a reproducible structure with 300 µm pores, and mechanical properties suitable for bone tissue engineering, with an elastic modulus around 100 MPa. The addition of ZnO resulted in enhanced surface roughness on the scaffolds, particularly for ZnO microparticles, achieving values up to 241 nm. This rougher topography was responsible for enhancing protein adsorption on the scaffolds, with an increase of up to 85% compared to the PLA-TCP matrix. Biological analyses demonstrated that the presence of ZnO promotes mesenchymal stem cell (MSC) proliferation and differentiation into osteoblasts. Alkaline phosphatase (ALP) activity, an important indicator of early osteogenic differentiation, increased up to 29%. The PLA-TCP composite containing 5% ZnO microparticles exhibited an optimized degradation rate and enhanced bioactivity, indicating its promising potential for bone repair applications.
Assuntos
Materiais Biocompatíveis , Regeneração Óssea , Fosfatos de Cálcio , Diferenciação Celular , Proliferação de Células , Células-Tronco Mesenquimais , Osteoblastos , Poliésteres , Impressão Tridimensional , Engenharia Tecidual , Alicerces Teciduais , Óxido de Zinco , Alicerces Teciduais/química , Fosfatos de Cálcio/química , Poliésteres/química , Regeneração Óssea/efeitos dos fármacos , Engenharia Tecidual/métodos , Células-Tronco Mesenquimais/citologia , Óxido de Zinco/química , Materiais Biocompatíveis/química , Diferenciação Celular/efeitos dos fármacos , Osteoblastos/citologia , Osteogênese/efeitos dos fármacos , Teste de Materiais , Osso e Ossos , Regeneração Tecidual Guiada/métodos , Humanos , Animais , Fosfatase Alcalina/metabolismo , Módulo de Elasticidade , Porosidade , Propriedades de SuperfícieRESUMO
Rocky shore communities are shaped by complex interactions among environmental drivers and a range of biological processes. Here, we investigated the importance of abiotic and biotic drivers on the population structure of key rocky intertidal species at 62 sites, spanning â¼50% of the Brazilian rocky shoreline (i.e., â¼500 km). Large-scale population patterns were generally explained by differences in ocean temperature and wave exposure. For the gastropod species Lottia subrugosa, differences at smaller scales (i.e., 0.1-1 km) were better explained by other abiotic influences such as freshwater discharge and substrate roughness. Based on the general population patterns of intertidal species identified, three main oceanographic groups were observed: a cold-oligotrophic grouping at northern sites (Lakes sub-region), a eutrophic group associated with large estuaries and urban zones (Santos and Guanabara bays); and a transitional warm-water group found between the two more productive areas. Larger individuals of Stramonita brasiliensis, L. subrugosa and Echinolittorina lineolata were generally found in the cold-oligotrophic system (i.e., upwelling region), while small suspension feeders dominate the warm-eutrophic systems. Evidence of bottom-up regulation was not observed, and top-down regulation effects were only observed between the whelk S. brasiliensis and its mussel prey Pernaperna. Environmental drivers as compared to biotic interactions, therefore, play a key role determining the population structure of multiple intertidal species, across a range of spatial scales along the SW Atlantic shores.
Assuntos
Ecossistema , Brasil , Animais , Monitoramento Ambiental , Gastrópodes/fisiologia , Dinâmica Populacional , Biodiversidade , TemperaturaRESUMO
Although Antarctica is the most isolated continent on Earth, its remote location does not protect it from the impacts of human activities. Antarctic metazoans such as filter-feeding invertebrates are a crucial component of the Antarctic benthos. They play a key role in the benthic-pelagic carbon flux in coastal areas by filtering particles and planktonic organisms from the sediment-water interface. Due to their peculiar ecological niche, these organisms can be considered a wasp-waist in the ecosystem, making them highly sensitive to marine pollution. Recently, anthropogenic particles such as micro-nanoplastics and manufactured nanoparticles (MNP) have been classified as contaminants of emerging concern (CEC) due to their small size range, which also overlaps with the preferred particle size ingested by aquatic metazoans. Indeed, it has been demonstrated that some species such as Antarctic krill can ingest, transform, and release MNPs, making them newly bioavailable for other Antarctic filter-feeding organisms. Similarly, the production and use of anthropogenic MNP are rapidly increasing, leading to a growing presence of materials, such as nano-sized metal-oxides, in the environment. For these reasons, it is important to provide evidence of the adverse effects of such emerging contaminants at sub-lethal concentrations in environmental risk assessments. These contaminants may cause cascade effects with consequences not only on individuals but also at the community and ecosystem levels. In this review, we discuss the state-of-the-art knowledge on the physiological and molecular effects of anthropogenic MNP in Antarctic aquatic metazoans. We further highlight the importance of identifying early biomarkers using sessile metazoans as sentinels of environmental health.
Assuntos
Organismos Aquáticos , Poluentes Químicos da Água , Regiões Antárticas , Animais , Poluentes Químicos da Água/análise , Monitoramento Ambiental , Nanopartículas , EcossistemaRESUMO
The lack of synthesized information regarding biodiversity is a major problem among researchers, leading to a pervasive cycle where ecologists make field campaigns to collect information that already exists and yet has not been made available for a broader audience. This problem leads to long-lasting effects in public policies such as spending money multiple times to conduct similar studies in the same area. We aim to identify this knowledge gap by synthesizing information available regarding two Brazilian long-term biodiversity programs and the metadata generated by them. Using a unique dataset containing 1904 metadata, we identified patterns of metadata distribution and intensity of research conducted in Brazil, as well as where we should concentrate research efforts in the next decades. We found that the majority of metadata were about vertebrates, followed by plants, invertebrates, and fungi. Caatinga was the biome with least metadata, and that there's still a lack of information regarding all biomes in Brazil, with none of them being sufficiently sampled. We hope that these results will have implications for broader conservation and management guiding, as well as to funding allocation programs.
Assuntos
Biodiversidade , Conservação dos Recursos Naturais , Brasil , Conservação dos Recursos Naturais/métodos , Ecologia , Monitoramento Ambiental/métodosRESUMO
The human FoxP transcription factors dimerize via three-dimensional domain swapping, a unique feature among the human Fox family, as result of evolutionary sequence adaptations in the forkhead domain. This is the case for the conserved glycine and proline residues in the wing 1 region, which are absent in FoxP proteins but present in most of the Fox family. In this work, we engineered both glycine (G) and proline-glycine (PG) insertion mutants to evaluate the deletion events in FoxP proteins in their dimerization, stability, flexibility, and DNA-binding ability. We show that the PG insertion only increases protein stability, whereas the single glycine insertion decreases the association rate and protein stability and promotes affinity to the DNA ligand.