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BACKGROUND: Recent studies have revealed atypical features in the plastomes of the family Cactaceae, the largest lineage of succulent species adapted to arid and semi-arid regions. Most plastomes sequenced to date are from short-globose and cylindrical cacti, while little is known about plastomes of epiphytic cacti. Published cactus plastomes reveal reduction and complete loss of IRs, loss of genes, pseudogenization, and even degeneration of tRNA structures. Aiming to contribute with new insights into the plastid evolution of Cactaceae, particularly within the tribe Rhipsalideae, we de novo assembled and analyzed the plastomes of Lepismium cruciforme and Schlumbergera truncata, two South American epiphytic cacti. METHODS AND RESULTS: Our data reveal many gene losses in both plastomes and the first loss of functionality of the trnT-GGU gene in Cactaceae. The trnT-GGU is a pseudogene in L. cruciforme plastome and appears to be degenerating in the tribe Rhipsalideae. Although the plastome structure is conserved among the species of the tribe Rhipsalideae, with tribe-specific rearrangements, we mapped around 200 simple sequence repeats and identified nine nucleotide polymorphism hotspots, useful to improve the phylogenetic resolutions of the Rhipsalideae. Furthermore, our analysis indicated high gene divergence and rapid evolution of RNA editing sites in plastid protein-coding genes in Cactaceae. CONCLUSIONS: Our findings show that some characteristics of the Rhipsalideae tribe are conserved, such as plastome structure with IRs containing only the ycf2 and two tRNA genes, structural degeneration of the trnT-GGU gene and ndh complex, and lastly, pseudogenization of rpl33 and rpl23 genes, both plastid translation-related genes.
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Cactaceae , Filogenia , Plastídeos , Cactaceae/genética , Plastídeos/genética , Evolução Molecular , Genes de Plantas/genética , Pseudogenes/genética , Genomas de Plastídeos/genética , RNA de Transferência/genética , Rearranjo Gênico/genéticaRESUMO
The Amazon rivers constitute the largest river basin in the world, with a high level of biodiversity. The Tocantins River is one of the most important rivers in this region, which has been impacted by different land uses. The objective of this study was to carry out a multi-evidence analysis focusing on the water quality of the Tocantins River, close to the municipality of Marabá-PA. We analyzed forest cover and water quality and, using the model organism Danio rerio, performed toxicity tests for histopathological effects, as well as the habitat selection approach by exposing fish to different river water samples in a multi-compartment device. The results showed that the studied area has already lost almost 30% of its forests in recent decades. Regarding water quality, the upstream (C1) and downstream (C5) points are the least impacted. On the other hand, the other points (C2-C4), closer to the city, greater input of pollutants was detected. Fish exposed to water samples from the most impacted sites showed several oedemas and hyperplastic cells in the gills. Regarding habitat selection behavior, there was a marked avoidance by samples with the highest contamination load. The results of this study lead to the understanding of the potential negative effects of human activities on local Amazonian biodiversity, since the potential toxicity of the environment, in conjunction with changes in the habitat selection process, could lead to a decline in populations of aquatic organisms, altering the environmental balance.
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Ecossistema , Monitoramento Ambiental , Rios , Poluentes Químicos da Água , Qualidade da Água , Peixe-Zebra , Animais , Rios/química , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade , Biodiversidade , Brasil , Florestas , PeixesRESUMO
The Brazil nut shell was used as a precursor material for preparing activated carbon by chemical activation with potassium hydroxide. The obtained material (BNSAC) was characterized, and the adsorptive features of phenol were investigated. The characterization showed that the activated carbon presented several rounded cavities along the surface, with a specific surface area of 332 m2 g-1. Concerning phenol adsorption, it was favored using an adsorbent dosage of 0.75 g L-1 and pH 6. The kinetic investigation revealed that the system approached the equilibrium in around 180 min, and the Elovich model represented the kinetic curves. The Sips model well represented the equilibrium isotherms. In addition, the increase in temperature from 25 to 55 °C favored the phenol adsorption, increasing the maximum adsorption capacity value (qs) from 83 to 99 mg g-1. According to the estimated thermodynamic parameters, the adsorption was spontaneous, favorable, endothermic, and governed by physical interactions. Therefore, the Brazil nut shell proved a good precursor material for preparing efficient activated carbon for phenol removal.
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Bertholletia , Poluentes Químicos da Água , Fenol/química , Carvão Vegetal/química , Concentração de Íons de Hidrogênio , Fenóis , Termodinâmica , Adsorção , Água , Cinética , Poluentes Químicos da Água/análise , SoluçõesRESUMO
Sapelli wood sawdust-derived magnetic activated carbon (SWSMAC) was produced by single-step pyrolysis using KOH and NiCl2 as activating and magnetization agents. SWSMAC was characterized by several techniques (SEM/EDS, N2 adsorption/desorption isotherms, FTIR, XRD, VSM, and pHPZC) and applied in the brilliant blue FCF dye adsorption from an aqueous medium. The obtained SWSMAC was a mesoporous material and showed good textural properties. Metallic nanostructured Ni particles were observed. Also, SWSMAC exhibited ferromagnetic properties. In the adsorption experiments, adequate conditions were an adsorbent dosage of 0.75 g L-1 and a solution pH of 4. The adsorption was fast, and the pseudo-second-order demonstrated greater suitability to the kinetic data. The Sips model fitted the equilibrium data well, and the maximum adsorption capacity predicted by this model was 105.88 mg g-1 (at 55 °C). The thermodynamic study revealed that the adsorption was spontaneous, favorable, and endothermic. Besides, the mechanistic elucidation suggested that electrostatic interactions, hydrogen bonding, π-π interactions, and n-π interactions were involved in the brilliant blue FCF dye adsorption onto SWSMAC. In summary, an advanced adsorbent material was developed from waste by single-step pyrolysis, and this material effectively adsorbs brilliant blue FCF dye.
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Carvão Vegetal , Poluentes Químicos da Água , Adsorção , Carvão Vegetal/química , Madeira , Poluentes Químicos da Água/química , Termodinâmica , Cinética , Fenômenos Magnéticos , Concentração de Íons de Hidrogênio , Azul de Metileno/químicaRESUMO
The cotton boll weevil (CBW) (Anthonomus grandis) is one of the major insect pests of cotton in Brazil. Currently, CBW control is mainly achieved by insecticide application, which is costly and insufficient to ensure effective crop protection. RNA interference (RNAi) has been used in gene function analysis and the development of insect control methods. However, some insect species respond poorly to RNAi, limiting the widespread application of this approach. Therefore, nanoparticles have been explored as an option to increase RNAi efficiency in recalcitrant insects. Herein, we investigated the potential of chitosan-tripolyphosphate (CS-TPP) and polyethylenimine (PEI) nanoparticles as a dsRNA carrier system to improve RNAi efficiency in the CBW. Different formulations of the nanoparticles with dsRNAs targeting genes associated with juvenile hormone metabolism, such as juvenile hormone diol kinase (JHDK), juvenile hormone epoxide hydrolase (JHEH), and methyl farnesoate hydrolase (MFE), were tested. The formulations were delivered to CBW larvae through injection (0.05-2 µg), and the expression of the target genes was evaluated using RT-qPCR. PEI nanoparticles increased targeted gene silencing compared with naked dsRNAs (up to 80%), whereas CS-TPP-dsRNA nanoparticles decreased gene silencing (0%-20%) or led to the same level of gene silencing as the naked dsRNAs (up to 50%). We next evaluated the effects of targeting a single gene or simultaneously targeting two genes via the injection of naked dsRNAs or dsRNAs complexed with PEI (500 ng) on CBW survival and phenotypes. Overall, the gene expression analysis showed that the treatments with PEI targeting either a single gene or multiple genes induced greater gene silencing than naked dsRNA (â¼60%). In addition, the injection of dsJHEH/JHDK, either naked or complexed with PEI, significantly affected CBW survival (18% for PEI nanoparticles and 47% for naked dsRNA) and metamorphosis. Phenotypic alterations, such as uncompleted pupation or malformed pupae, suggested that JHEH and JHDK are involved in developmental regulation. Moreover, CBW larvae treated with dsJHEH/JHDK + PEI (1,000 ng/g) exhibited significantly lower survival rate (55%) than those that were fed the same combination of naked dsRNAs (30%). Our findings demonstrated that PEI nanoparticles can be used as an effective tool for evaluating the biological role of target genes in the CBW as they increase the RNAi response.
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Water pollution by dyes has been a major environmental problem to be tackled, and magnetic adsorbents appear as promising alternatives to solve it. Herein, magnetic activated carbons were prepared by the single-step method from Sapelli wood sawdust, properly characterized, and applied as adsorbents for brilliant blue dye removal. In particular, two magnetic activated carbons, MAC1105 and MAC111, were prepared using the proportion of biomass KOH of 1:1 and varying the proportion of NiCl2 of 0.5 and 1. The characterization results demonstrated that the different proportions of NiCl2 mainly influenced the textural characteristics of the adsorbents. An increase in the surface area from 260.0 to 331.5 m2 g-1 and in the total pore volume from 0.075 to 0.095 cm3 g-1 was observed with the weight ratio of NiCl2. Both adsorbents exhibit ferromagnetic properties and the presence of nanostructured Ni particles. The different properties of the materials influenced the adsorption kinetics and equilibrium of brilliant blue dye. MAC111 showed faster kinetics, reaching the equilibrium in around 10 min, while for MAC1105, it took 60 min for the equilibrium to be reached. In addition, based on the Sips isotherm, the maximum adsorption capacity was 98.12 mg g-1 for MAC111, while for MAC1105, it was 60.73 mg g-1. Furthermore, MAC111 presented the potential to be reused in more adsorption cycles than MAC1105, and the use of the adsorbents in the treatment of a simulated effluent exhibited high effectiveness, with removal efficiencies of up to 90%.
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Carvão Vegetal , Poluentes Químicos da Água , Adsorção , Corantes , Fenômenos Magnéticos , Cinética , Azul de Metileno , Concentração de Íons de HidrogênioRESUMO
Cotton is the most important crop for fiber production worldwide. However, the cotton boll weevil (CBW) is an insect pest that causes significant economic losses in infested areas. Current control methods are costly, inefficient, and environmentally hazardous. Herein, we generated transgenic cotton lines expressing double-stranded RNA (dsRNA) molecules to trigger RNA interference-mediated gene silencing in CBW. Thus, we targeted three essential genes coding for chitin synthase 2, vitellogenin, and ecdysis-triggering hormone receptor. The stability of expressed dsRNAs was improved by designing a structured RNA based on a viroid genome architecture. We transformed cotton embryos by inserting a promoter-driven expression cassette that overexpressed the dsRNA into flower buds. The transgenic cotton plants were characterized, and positive PCR transformed events were detected with an average heritability of 80%. Expression of dsRNAs was confirmed in floral buds by RT-qPCR, and the T1 cotton plant generation was challenged with fertilized CBW females. After 30 days, data showed high mortality (around 70%) in oviposited yolks. In adult insects fed on transgenic lines, chitin synthase II and vitellogenin showed reduced expression in larvae and adults, respectively. Developmental delays and abnormalities were also observed in these individuals. Our data remark on the potential of transgenic cotton based on a viroid-structured dsRNA to control CBW.
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Gorgulhos , Humanos , Animais , Gorgulhos/genética , RNA de Cadeia Dupla/genética , RNA de Cadeia Dupla/metabolismo , Gossypium/genética , Gossypium/metabolismo , Vitelogeninas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismoRESUMO
Breast cancer is a type of cancer with the highest incidence worldwide in 2021, with early diagnosis and rapid treatment intervention being the reasons for the decreasing mortality rate associated with the disease. The major challenge faced by clinicians and pathologists is the lack of accuracy in the histopathological analysis of biopsy or resection samples, leading to classification misdiagnosis and compromising the prognosis of patients. Spectral histopathology has provided great advances regarding cancer diagnosis, especially through the use of FTIR spectroscopy, proving to be a valuable complement to histopathological analyses. In this study unstained formalin-fixed paraffin embedded breast cancer tissue samples, collected from patients undergoing surgery and mounted on glass slides, were probed through FTIR and Raman microspectrocopy. Two classification models were constructed using the AdaBoost algorithm, both achieving >90% accuracy and successfully discriminating invasive breast carcinoma from surrounding normal tissue. Chemical maps from the interfaces of invasive breast carcinoma-surrounding normal tissue were also generated. This study showed the potential of spectral histopathology, in particular FTIR, for daily use in pathology laboratories, introducing few disruptions to the routine workflow while increasing the sensitivity, specificity and accuracy of the diagnoses.
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Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Formaldeído/química , Análise Espectral Raman/métodosRESUMO
Chest radiographs allow for the meticulous examination of a patient's chest but demands specialized training for proper interpretation. Automated analysis of medical imaging has become increasingly accessible with the advent of machine learning (ML) algorithms. Large labeled datasets are key elements for training and validation of these ML solutions. In this paper we describe the Brazilian labeled chest x-ray dataset, BRAX: an automatically labeled dataset designed to assist researchers in the validation of ML models. The dataset contains 24,959 chest radiography studies from patients presenting to a large general Brazilian hospital. A total of 40,967 images are available in the BRAX dataset. All images have been verified by trained radiologists and de-identified to protect patient privacy. Fourteen labels were derived from free-text radiology reports written in Brazilian Portuguese using Natural Language Processing.
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Algoritmos , Processamento de Linguagem Natural , Radiografia Torácica , Brasil , Humanos , Raios XRESUMO
Infectious diseases of different etiologies have been associated with acute and long-term neurological consequences. The primary cause of these consequences appears to be an inflammatory process characterized primarily by a pro-inflammatory microglial state. Microglial cells, the local effectors' cells of innate immunity, once faced by a stimulus, alter their morphology, and become a primary source of inflammatory cytokines that increase the inflammatory process of the brain. This inflammatory scenario exerts a critical role in the pathogenesis of neurodegenerative diseases. In recent years, several studies have shown the involvement of the microglial inflammatory response caused by infections in the development of neurodegenerative diseases. This has been associated with a transitory microglial state subsequent to an inflammatory response, known as microglial priming, in which these cells are more responsive to stimuli. Thus, systemic inflammation and infections induce a transitory state in microglia that may lead to changes in their state and function, making priming them for subsequent immune challenges. However, considering that microglia are long-lived cells and are repeatedly exposed to infections during a lifetime, microglial priming may not be beneficial. In this review, we discuss the relationship between infections and neurodegenerative diseases and how this may rely on microglial priming.
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MOTIVATION: Time-lapse microscopy is a powerful technique that relies on images of live cells cultured ex vivo that are captured at regular intervals of time to describe and quantify their behavior under certain experimental conditions. This imaging method has great potential in advancing the field of precision oncology by quantifying the response of cancer cells to various therapies and identifying the most efficacious treatment for a given patient. Digital image processing algorithms developed so far require high-resolution images involving very few cells originating from homogeneous cell line populations. We propose a novel framework that tracks cancer cells to capture their behavior and quantify cell viability to inform clinical decisions in a high-throughput manner. RESULTS: The brightfield microscopy images a large number of patient-derived cells in an ex vivo reconstruction of the tumor microenvironment treated with 31 drugs for up to 6 days. We developed a robust and user-friendly pipeline CancerCellTracker that detects cells in co-culture, tracks these cells across time and identifies cell death events using changes in cell attributes. We validated our computational pipeline by comparing the timing of cell death estimates by CancerCellTracker from brightfield images and a fluorescent channel featuring ethidium homodimer. We benchmarked our results using a state-of-the-art algorithm implemented in ImageJ and previously published in the literature. We highlighted CancerCellTracker's efficiency in estimating the percentage of live cells in the presence of bone marrow stromal cells. AVAILABILITY AND IMPLEMENTATION: https://github.com/compbiolabucf/CancerCellTracker. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
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Antineoplásicos , Neoplasias , Humanos , Microscopia/métodos , Imagem com Lapso de Tempo , Software , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Medicina de Precisão , Algoritmos , Microambiente TumoralRESUMO
Bacillus thuringiensis produces several virulence factors, the main ones being the Cry and Cyt toxins, present in the parasporal body produced during sporulation. The Cyt toxins have mechanisms specific for mosquitoes and Cyt1Aa, the most studied cytolytic toxin, is effective for mosquito control by acting in synergism with Cry toxins. The goal of the present work was to study the frequency of the codifying gene for Cyt1Aa in B. thuringiensis native isolates acquired from samples of soil, insect and water, as well as to verify any possible genetic polymorphism. 1,448 B thuringiensis strains were used for DNA extraction and PCR technique, all with the use of a primer that amplifies a fragment of 300 pairs of the cyt1Aa gene. The strains that showed amplification in the PCR reaction were sequenced and compared to each other and to the sequences available at Genbank. 32 (2.3%) strains of B. thuringiensis showed positive amplification for the cyt1Aa gene. The highest frequency of isolates with cyt1Aa gene was acquired from samples coming from the Cerrado biome, both isolates from soil and from insects, equally with 3.4%. The cyt1Aa gene sequencing highlighted that, for that 300 bp region, the gene is conserved and there is no single-base polymorphism.
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Bacillus thuringiensis , Culicidae , Animais , Bacillus thuringiensis/genética , Proteínas de Bactérias/genética , Frequência do Gene , Solo , VirulênciaRESUMO
MAIN CONCLUSION: The plastome of Melocactus glaucescens shows unique rearrangements, IR expansion, and unprecedented gene losses in Cactaceae. Our data indicate tRNA import from the cytosol to the plastids in this species. Cactaceae represents one of the richest families in keystone species of arid and semiarid biomes. This family shows various specific features comprehending morphology, anatomy, and metabolism, which allow them to grow under unfavorable environmental conditions. The subfamily Cactoideae contains the most divergence of species, which are highly variable in growth habit and morphology. This subfamily includes the endangered species Melocactus glaucescens (tribe Cereeae), which is a cactus endemic to the biome Caatinga in Brazil. Aiming to analyze the plastid evolution and develop molecular markers, we sequenced and analyzed in detail the plastome of M. glaucescens. Our analyses revealed that the M. glaucescens plastome is the most divergent among the species of the family Cactaceae sequenced so far. We characterized here unique rearrangements, expanded IRs containing an unusual set of genes, and several gene losses. Some genes related to the ndh complex were lost during the plastome evolution, while others have lost their functionality. Additionally, the loss of three tRNA genes (trnA-UGC, trnV-UAC, and trnV-GAC) suggests tRNA import from the cytosol to the plastids in M. glaucescens. Moreover, we identified high gene divergence, several putative positive signatures, and possible unique RNA-editing sites. Furthermore, we mapped 169 SSRs in the plastome of M. glaucescens, which are helpful to access the genetic diversity of natural populations and conservation strategies. Finally, our data provide new insights into the evolution of plastids in Cactaceae, which is an outstanding lineage adapted to extreme environmental conditions and a notorious example of the atypical evolution of plastomes.
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Cactaceae , Evolução Molecular , Cactaceae/genética , Filogenia , Plastídeos/genética , RNA de Transferência/genéticaRESUMO
Knowledge of the mechanisms that trigger infection-related encephalopathies is still very limited and cell therapies are one of the most promising alternatives for neurodegenerative diseases, and its application in infectious diseases can be of great relevance. Mesenchymal stromal cells are cells with great immunomodulatory potential; therefore, their use in clinical and preclinical studies in a variety of diseases, including central nervous system diseases, increased in the last decade. Mesenchymal stromal cells can exert their beneficial effects through several mechanisms, such as direct cell contact, through surface receptors, and also through paracrine or endocrine mechanisms. The paracrine mechanism is widely accepted by the scientific community and involves the release of soluble factors, which include cytokines, chemokines and trophic factors, and extracellular vesicles. This mini review discusses mesenchymal stromal cells mechanisms of action in neurological disorders, the neuroinflammatory process that takes place in the brain as a result of peripheral inflammation and changes in the brain's cellular scenario as a common factor in central nervous system diseases, and mesenchymal stromal cells therapy in encephalopathies. Mesenchymal stromal cells have been shown to act in neuroinflammation processes, leading to improved survival and mitigating behavioral damage. More recently, these cells have been tested in preclinical models of infectious diseases-associated encephalopathies (e.g., cerebral malaria and sepsis associated encephalopathy) and have shown satisfactory results.
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MAIN CONCLUSION: Host-derived suppression of nematode essential genes decreases reproduction of Meloidogyne incognita in cotton. Root-knot nematodes (RKN) represent one of the most damaging plant-parasitic nematode genera worldwide. RNAi-mediated suppression of essential nematode genes provides a novel biotechnological strategy for the development of sustainable pest-control methods. Here, we used a Host Induced Gene Silencing (HIGS) approach by stacking dsRNA sequences into a T-DNA construct to target three essential RKN genes: cysteine protease (Mi-cpl), isocitrate lyase (Mi-icl), and splicing factor (Mi-sf), called dsMinc1, driven by the pUceS8.3 constitutive soybean promoter. Transgenic dsMinc1-T4 plants infected with Meloidogyne incognita showed a significant reduction in gall formation (57-64%) and egg masses production (58-67%), as well as in the estimated reproduction factor (60-78%), compared with the susceptible non-transgenic cultivar. Galls of the RNAi lines are smaller than the wild-type (WT) plants, whose root systems exhibited multiple well-developed root swellings. Transcript levels of the three RKN-targeted genes decreased 13- to 40-fold in nematodes from transgenic cotton galls, compared with those from control WT galls. Finally, the development of non-feeding males in transgenic plants was 2-6 times higher than in WT plants, indicating a stressful environment for nematode development after RKN gene silencing. Data strongly support that HIGS of essential RKN genes is an effective strategy to improve cotton plant tolerance. This study presents the first application of dsRNA sequences to target multiple genes to promote M. incognita tolerance in cotton without phenotypic penalty in transgenic plants.
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Gossypium , Tylenchoidea , Animais , Gossypium/genética , Doenças das Plantas/genética , Plantas Geneticamente Modificadas/genética , RNA de Cadeia Dupla , Tylenchoidea/genéticaRESUMO
In 2020, approximately 10 million people died of cancer, rendering this disease the second leading cause of death worldwide. Detecting cancer in its early stages is paramount for patients' prognosis and survival. Hence, the scientific and medical communities are engaged in improving both therapeutic strategies and diagnostic methodologies, beyond prevention. Optical vibrational spectroscopy has been shown to be an ideal diagnostic method for early cancer diagnosis and surgical margins assessment, as a complement to histopathological analysis. Being highly sensitive, non-invasive and capable of real-time molecular imaging, Raman and Fourier transform infrared (FTIR) spectroscopies give information on the biochemical profile of the tissue under analysis, detecting the metabolic differences between healthy and cancerous portions of the same sample. This constitutes tremendous progress in the field, since the cancer-prompted morphological alterations often occur after the biochemical imbalances in the oncogenic process. Therefore, the early cancer-associated metabolic changes are unnoticed by the histopathologist. Additionally, Raman and FTIR spectroscopies significantly reduce the subjectivity linked to cancer diagnosis. This review focuses on breast and head and neck cancers, their clinical needs and the progress made to date using vibrational spectroscopy as a diagnostic technique prior to surgical intervention and intraoperative margin assessment.
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Infectious diseases may affect brain function and cause encephalopathy even when the pathogen does not directly infect the central nervous system, known as infectious disease-associated encephalopathy. The systemic inflammatory process may result in neuroinflammation, with glial cell activation and increased levels of cytokines, reduced neurotrophic factors, blood-brain barrier dysfunction, neurotransmitter metabolism imbalances, and neurotoxicity, and behavioral and cognitive impairments often occur in the late course. Even though infectious disease-associated encephalopathies may cause devastating neurologic and cognitive deficits, the concept of infectious disease-associated encephalopathies is still under-investigated; knowledge of the underlying mechanisms, which may be distinct from those of encephalopathies of non-infectious cause, is still limited. In this review, we focus on the pathophysiology of encephalopathies associated with peripheral (sepsis, malaria, influenza, and COVID-19), emerging therapeutic strategies, and the role of neuroinflammation.
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Encefalopatias/imunologia , COVID-19/complicações , Citocinas/imunologia , Influenza Humana/complicações , Malária/complicações , Sepse/complicações , Barreira Hematoencefálica/imunologia , Encefalopatias/prevenção & controle , COVID-19/imunologia , Humanos , Influenza Humana/imunologia , Malária/imunologia , Sepse/imunologiaRESUMO
BACKGROUND: The hemolymph and insect gut together have an essential role in the immune defense against microorganisms, including the production of antimicrobial peptides (AMP). AMPs are mainly induced by two specific signaling pathways, Toll and immune deficiency (IMD). Here, we characterize the expression profile of four genes from both pathways and describe the importance of AgraRelish in the immune defense of Anthonomus grandis against the entomopathogenic fungus Metarhizium anisopliae by RNA interference (RNAi). RESULTS: To characterize the pathway that is activated early during the A. grandis-M. anisopliae interaction, we assessed the expression profiles of AgraMyD88 and AgraDorsal (Toll pathway), AgraIMD and AgraRelish (IMD pathway), and several AMP genes. Interestingly, we found that IMD pathway genes are upregulated early, and Toll pathway genes are upregulated just 3 days after inoculation (DAI). Furthermore, nine AMPs were upregulated 24 h after fungus inoculation, including attacins, cecropins, coleoptericins, and defensins. AgraRelish knockdown resulted in a reduction in median lethal time (LT50 ) for M. anisopliae-treated insects of around 2 days compared to control treatments. In addition, AgraRelish remained knocked down at 3 DAI. Finally, we identified that AgraRelish knockdown increased fungal loads at 2 DAI compared to control treatments, possibly indicating a faster infection. CONCLUSIONS: Our data indicate the influence of the IMD pathway on the antifungal response in A. grandis. Combining biocontrol and RNAi could significantly improve cotton boll weevil management. Hence, AgraRelish is a potential target for the development of biotechnological tools aimed at improving the efficacy of M. anisopliae against A. grandis.
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Metarhizium , Gorgulhos , Animais , Biotecnologia , Insetos , Metarhizium/genética , Interferência de RNARESUMO
The formation of biomolecular coronas around nanoparticles as soon as they come in contact with biological media is nowadays well accepted. The self-developed biological outer surfaces can affect the targeting capability of the colloidal carriers as well as their cytotoxicity and cellular uptake behavior. In this framework, we explored the structural features and biological consequences of protein coronas around block copolymer assemblies consisting of a common pH-responsive core made by poly[2-(diisopropylamino) ethyl methacrylate] (PDPA) and hydrophilic shells of different chemical natures: zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) or highly hydrophilic poly(ethylene oxide) (PEO) and poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA). We demonstrated the presence of â¼50 nm protein coronas around the nanoparticles regardless of the chemical nature of the polymeric shells. The thickness is understood as the sum of the soft and hard layers and it is the actual interface seen by the cells. Although the soft corona composition is difficult to determine because the proteins are loosely bound to the outer surface of the assemblies, the tightly bound proteins (hard corona) could be identified and quantified. The compositional analysis of the hard corona demonstrated that human serum albumin (HSA), immunoglobulin G (IgG) and fibrinogen are the main components of the protein coronas, and serotransferrin is present particularly in the protein corona of the zwitterionic-stabilized assemblies. The protein coronas substantially reduce the cellular uptake of the colloidal particles due to their increased size and the presence of HSA which is known to reduce nanoparticle-cell adhesion. On the other hand, their existence also reduces the levels of cytotoxicity of the polymeric assemblies, highlighting that protein coronas should not be always understood as artifacts that need to be eliminated due to their positive outputs.
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Fenômenos Mecânicos , Nanopartículas/química , Coroa de Proteína/química , Adesão Celular , Humanos , Concentração de Íons de Hidrogênio , Polímeros/química , Propriedades de SuperfícieRESUMO
Atrazine is an herbicide commonly used in several countries. Due to its long half-life, associated with its use in large scales, atrazine residues remain as environmental pollutants in water bodies. Phytoremediation is often pointed out as an interesting approach to remove atrazine from the aquatic environment, but its practical application is limited by the high toxicity of this herbicide. Here, we characterize the damages triggered by atrazine in Pistia stratiotes, evaluating the role of nitric oxide (NO), a cell-signaling molecule, in increasing the tolerance to the pollutant and the phytoremediation potential of this species. Pistia stratiotes plants were exposed to four treatments: Control; Sodium nitroprusside (SNP) (0.05 mg L-1); Atrazine (ATZ) (150 µg L-1) and ATZ + SNP. The plants remained under those conditions for 24 h for biochemical and physiological analysis and 3 days for the evaluation of relative growth rate. The presence of atrazine in plant cells triggered a series of biochemical and physiological damages, such as the increase in the generation of reactive oxygen species, damages to cell membranes, photosynthesis impairment, and negative carbon balance. Despite this, the plants maintained greater growth rates than other aquatic macrophytes exposed to atrazine and showed high bioconcentration and translocation factors. The addition of SNP, a NO donor, decreased the herbicide toxicity, with an increase of over 60% in the IC50 value (Inhibitor Concentration). Indeed, the NO signaling action was able to increase the tolerance of plants to atrazine, which resulted in increments in pollutant uptake and translocation, with the maintenance of overall cell (e.g. membranes) and organs (root system) structure, and the functioning of central physiological processes (e.g. photosynthesis). These factors allowed for more quickly and efficient removal of the pollutant from the environment, reducing costs, and increasing the viability of the phytoremediation process.