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1.
Proc Natl Acad Sci U S A ; 119(1)2022 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-34983846

RESUMO

Many soil-, water-, and plant-associated bacterial species from the orders Xanthomonadales, Burkholderales, and Neisseriales carry a type IV secretion system (T4SS) specialized in translocating effector proteins into other gram-negative species, leading to target cell death. These effectors, known as X-Tfes, carry a carboxyl-terminal domain of ∼120 residues, termed XVIPCD, characterized by several conserved motifs and a glutamine-rich tail. Previous studies showed that the XVIPCD is required for interaction with the T4SS coupling protein VirD4 and for T4SS-dependent translocation. However, the structural basis of the XVIPCD-VirD4 interaction is unknown. Here, we show that the XVIPCD interacts with the central all-alpha domain of VirD4 (VirD4AAD). We used solution NMR spectroscopy to solve the structure of the XVIPCD of X-TfeXAC2609 from Xanthomonas citri and to map its interaction surface with VirD4AAD Isothermal titration calorimetry and in vivo Xanthomonas citri versus Escherichia coli competition assays using wild-type and mutant X-TfeXAC2609 and X-TfeXAC3634 indicate that XVIPCDs can be divided into two regions with distinct functions: the well-folded N-terminal region contains specific conserved motifs that are responsible for interactions with VirD4AAD, while both N- and carboxyl-terminal regions are required for effective X-Tfe translocation into the target cell. The conformational stability of the N-terminal region is reduced at and below pH 7.0, a property that may facilitate X-Tfe unfolding and translocation through the more acidic environment of the periplasm.


Assuntos
Antibacterianos/química , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/química , Escherichia coli/química , Sistemas de Secreção Tipo IV/antagonistas & inibidores , Sistemas de Secreção Tipo IV/química , Xanthomonas/química , Proteínas de Bactérias/genética , Escherichia coli/genética , Modelos Moleculares , Mutação , Ressonância Magnética Nuclear Biomolecular , Domínios Proteicos , Relação Estrutura-Atividade , Sistemas de Secreção Tipo IV/genética , Xanthomonas/genética
2.
J Basic Microbiol ; 64(2): e2300444, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38051942

RESUMO

The aim of this study was to investigate the production, stability and applicability of colorants produced by filamentous fungi isolated from soil samples from the Amazon. Initially, the isolates were evaluated in a screening for the production of colorants. The influences of cultivation and nutritional conditions on the production of colorants by fungal isolates were investigated. The colorants produced by selected fungal isolates were chemically characterized using the Liquid Chromatography-Mass Spectrometry technique. The antimicrobial and cytotoxic activities, stability evaluation and applicability of the colorants were investigated. As results, we observed that the isolates Penicillium sclerotiorum P3SO224, Clonostachys rosea P2SO329 and Penicillium gravinicasei P3SO332 stood out since they produced the most intense colorants. Compounds produced by Penicillium sclerotiorum P3SO224 and Clonostachys rosea P2SO329 were identified as sclerotiorin and penicillic acid. The colorant fraction (EtOAc) produced by these species has antimicrobial activity, stability at temperature and at different pHs, stability when exposure to light and UV, and when exposed to different concentrations of salts, as well as being nontoxic and having the ability to dye fabrics and be used as a pigment in creams and soap. Considering the results found in this study, it was concluded that fungi from the soil in the Amazon have the potential to produce colorants with applications in the textile and pharmaceutical industries.


Assuntos
Anti-Infecciosos , Hypocreales , Penicillium , Pigmentos Biológicos/química , Fungos/química , Solo
3.
Int J Mol Sci ; 25(3)2024 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-38338793

RESUMO

Hypothyroidism compromises the testicular redox status and is associated with reduced sperm quality and infertility in men. In this regard, studies have demonstrated the antioxidant potential of kisspeptin in reproductive and metabolic diseases. In this study, we evaluate the effects of kisspeptin-10 (Kp10) on the testicular redox, as well as mediators of the unfolded protein response (UPR) in adult rats with hypothyroidism. Adult male Wistar rats were randomly separated into the Control (n = 15), Hypo (n = 13) and Hypo + Kp10 (n = 14) groups, and hypothyroidism was induced with 6-propyl-2-thiouracil (PTU) for three months. In the last month, half of the hypothyroid animals received Kp10. Testis samples were collected for enzymatic, immunohistochemical and/or gene evaluation of mediators of oxidative stress (TBARs, lipid hydroperoxides (LOOH), ROS, peroxynitrite, SOD, CAT and GPX), endoplasmic reticulum stress (GRP78, ATF6, PERK, CHOP, HO-1 and sXBP1) and antiapoptocytes (BCL-2). Hypothyroidism increased apoptosis index, TBARS and LOOH concentrations, and reduced testicular gene expression of Sod1, Sod2 and Gpx1, as well as the expression of Grp78, Atf6, Ho1 and Chop. Treatment with Kp10, in turn, reduced testicular apoptosis and the production of peroxynitrite, while increased SOD1 and GPX ½ expression, and enzymatic activity of CAT, but did not affect the lower expression of UPR mediators caused by hypothyroidism. This study demonstrated that hypothyroidism causes oxidative stress and dysregulated the UPR pathway in rat testes and that, although Kp10 does not influence the low expression of UPR mediators, it improves the testicular redox status, configuring it as an important antioxidant factor in situations of thyroid dysfunction.


Assuntos
Antioxidantes , Hipotireoidismo , Humanos , Ratos , Masculino , Animais , Antioxidantes/metabolismo , Testículo/metabolismo , Kisspeptinas/metabolismo , Ratos Wistar , Superóxido Dismutase-1/genética , Chaperona BiP do Retículo Endoplasmático , Ácido Peroxinitroso/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Sêmen/metabolismo , Oxirredução , Hipotireoidismo/tratamento farmacológico , Hipotireoidismo/metabolismo , Estresse Oxidativo , Resposta a Proteínas não Dobradas
4.
Environ Geochem Health ; 46(5): 154, 2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38592573

RESUMO

Contamination of aquatic ecosystems by potentially toxic elements (PTEs) is a concerning environmental issue, given their persistence, toxicity potential, and ability to accumulate in living organisms. Several studies have been conducted to assess the contamination of aquatic ecosystems by PTEs, using pollution and ecological risk indices that rely on the concentration of these elements in aquatic sediments. However, many of these studies use global reference values for calculating the indices, which can lead to misleading interpretations due to substantial variations in PTEs concentrations influenced by the geological characteristics of each region. Therefore, the use of regional reference values is more appropriate when available. This study aimed to investigate variations in the results of five indices, employing global, regional, and quality reference values, based on sediment samples collected from rivers in the Ipanema National Forest, a protected area in Brazil exposed to various anthropogenic pressures. The results revealed that elements such as Al, Fe, and Mn exceeded the limits allowed by legislation in water samples, while As and Cr surpassed the limits in sediment samples. Comparative analysis highlighted significant discrepancies in the results of the indices when global reference values were used compared to regional and quality reference values, especially for As and Ba. Thus, this study underscores the importance of establishing specific regional values for an accurate assessment of sediment quality and the risks associated with contamination by PTEs in different regions worldwide.


Assuntos
Conservação dos Recursos Naturais , Ecossistema , Brasil , Valores de Referência , Poluição Ambiental
5.
PLoS Pathog ; 17(11): e1010067, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34784389

RESUMO

Human ascariasis is the most prevalent but neglected tropical disease in the world, affecting approximately 450 million people. The initial phase of Ascaris infection is marked by larval migration from the host's organs, causing mechanical injuries followed by an intense local inflammatory response, which is characterized mainly by neutrophil and eosinophil infiltration, especially in the lungs. During the pulmonary phase, the lesions induced by larval migration and excessive immune responses contribute to tissue remodeling marked by fibrosis and lung dysfunction. In this study, we investigated the relationship between SIgA levels and eosinophils. We found that TLR2 and TLR4 signaling induces eosinophils and promotes SIgA production during Ascaris suum infection. Therefore, control of parasite burden during the pulmonary phase of ascariasis involves eosinophil influx and subsequent promotion of SIgA levels. In addition, we also demonstrate that eosinophils also participate in the process of tissue remodeling after lung injury caused by larval migration, contributing to pulmonary fibrosis and dysfunction in re-infected mice. In conclusion, we postulate that eosinophils play a central role in mediating host innate and humoral immune responses by controlling parasite burden, tissue inflammation, and remodeling during Ascaris suum infection. Furthermore, we suggest that the use of probiotics can induce eosinophilia and SIgA production and contribute to controlling parasite burden and morbidity of helminthic diseases with pulmonary cycles.


Assuntos
Ascaríase/imunologia , Ascaris suum/imunologia , Eosinófilos/fisiologia , Imunoglobulina A Secretora/metabolismo , Pneumonia/prevenção & controle , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Ascaríase/metabolismo , Ascaríase/parasitologia , Feminino , Imunoglobulina A Secretora/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Pneumonia/imunologia , Pneumonia/parasitologia , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética
6.
Nitric Oxide ; 134-135: 61-71, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-37059259

RESUMO

Production of nitric oxide (NO) by LPS-activated macrophages is due to a complex cellular signaling initiated by TLR4 that leads to the transcription of IFN-ß, which activates IRF-1 and STAT-1, as well as to the activation of NF-κB, required for iNOS transcription. High concentrations of LPS can also be uptaken by scavenger receptors (SRs), which, in concert with TLR4, leads to inflammatory responses. The mechanisms by which TLR4 and SRs interact, and the pathways activated by this interaction in macrophages are not elucidated. Therefore, our main goal was to evaluate the role of SRs, particularly SR-A, in LPS-stimulated macrophages for NO production. We first showed that, surprisingly, LPS can induce the expression of iNOS and the production of NO in TLR4-/- mice, provided exogenous IFN-ß is supplied. These results indicate that LPS stimulate receptors other than TLR4. The inhibition of SR-A using DSS or neutralizing antibody to SR-AI showed that SR-A is essential for the expression of iNOS and NO production in stimulation of TLR4 by LPS. The restoration of the ability to express iNOS and produce NO by addition of rIFN-ß to inhibited SR-A cells indicated that the role of SR-AI in LPS-induced NO production is to provide IFN-ß, probably by mediating the internalization of LPS/TLR4, and the differential inhibition by DSS and neutralizing antibody to SR-AI suggested that other SRs are also involved. Our results reinforce that TLR4 and SR-A act in concert in LPS activation and demonstrated that, for the production of NO, it does mainly by synthesizing IRF-3 and also by activating the TRIF/IRF-3 pathway for IFN-ß production, essential for LPS-mediated transcription of iNOS. Consequently STAT-1 is activated, and IRF-1 is expressed, which together with NF-κB from TLR4/MyD88/TIRAP, induce iNOS synthesis and NO production. SUMMARY SENTENCE: TLR4 and SRs act in concert activating IRF-3 to transcribe IFN-ß and activate STAT-1 to produce NO by LPS-activated macrophages.


Assuntos
NF-kappa B , Óxido Nítrico , Camundongos , Animais , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Receptor 4 Toll-Like/metabolismo , Lipopolissacarídeos , Macrófagos/metabolismo , Receptores Depuradores/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo
7.
J Clin Densitom ; 26(1): 97-103, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36543669

RESUMO

BACKGROUND: Despite the well-established role of muscle in maintaining bone mass and physical performance, there are still few studies that assess the relationship between muscle groups and specific bone sites and its contribution to physical function in older people. The aim of the study is to verify the association between hip and knee's muscle strength and bone mineral density of the femoral neck (BMD-FN) and investigated which muscle strength that are related with BMD-FN is associated with physical function in older women. METHODOLOGY: This is a cross-sectional study of 94 women over the age of 60 years, physically independent. Muscle strength of the hip and knee was evaluated using an isokinetic dynamometer (Biodex, USA) to obtain peak torque (PT). The BMD-FN was obtained using dual-energy x-ray absorption densitometry (DXA). Physical function was evaluated by the Timed Up and Go (TUG), gait speed (GS), five times stand to sit (5TSS) and single-leg stance (SS). To verify the association between muscle strength (independent variable) and the BMD-FN (dependent variable) and the relationship between muscle strength (independent variable) and clinical tests (dependent variable), the multiple linear regression was carried out, adjusted for age, time of menopause, height and level of physical activity. All of the analyses were carried out by the SPSS 17.0 software, with a level of p < 0.05 significance. RESULTS: We found an association between the hip abductors PT and the BMD-FN and between hip abductors PT and all clinical tests. CONCLUSION: We observe that hip abductor muscle strength contributes to BMD-FN and to performance in several clinical tests in older women. These findings reinforce the importance of this musculature not only for improving balance and gait, but also it can be a strategy to maintain/improve bone mass in the femoral neck in this population.


Assuntos
Densidade Óssea , Músculo Esquelético , Humanos , Feminino , Idoso , Pessoa de Meia-Idade , Densidade Óssea/fisiologia , Estudos Transversais , Músculo Esquelético/diagnóstico por imagem , Músculo Esquelético/fisiologia , Absorciometria de Fóton , Colo do Fêmur/diagnóstico por imagem , Colo do Fêmur/fisiologia
8.
Curr Microbiol ; 80(9): 289, 2023 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-37462778

RESUMO

Trichoderma is an important fungal genus, known mainly for its potential for the biological control of phytopathogens. Accurate identification of these fungi is essential for research and applications involving them, to be addressed correctly. The objectives of this study were to isolate, identify, and report the species richness of Trichoderma species that occur in the soil of different regions of Pernambuco, Brazil. DNA sequences of portions of the translation elongation factor 1-α (TEF1) gene region were generated for 56 isolates of Trichoderma, obtained from the Zona da Mata, Agreste, and Sertão regions of Pernambuco. According to the phylogenetic analysis based on these sequences, these fungi belong to two Sections-Trichoderma (35 isolates) and Pachybasidium (21 isolates). These fungi have been resolved in nine species, including Trichoderma afroharzianum, Trichoderma asperelloides, Trichoderma asperellum, Trichoderma koningiopsis, and five possible new species to be confirmed in further studies. This study shows that the soils of Pernambuco host a diversity of Trichoderma species and consequently of biological resources with potential for application in agriculture.


Assuntos
Trichoderma , Filogenia , Análise de Sequência de DNA , Solo , Brasil
9.
Clin Oral Investig ; 27(3): 943-953, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36629964

RESUMO

OBJECTIVES: This systematic review investigated the prevalence of tooth wear between patients with and without Down syndrome. METHODS: Six databases (Embase, LILACS, Livivo, PubMed, Scopus and Web of Science) and grey literature (Google Scholar, OpenGrey and ProQuest) were searched until March 7, 2022. Observational studies were included to assess the differences in tooth wear prevalence and/or severity in Down syndrome and non-syndromic controls. Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) recommendations were followed. Three reviewers independently screened studies, extracted data, assessed the methodological quality (Joanna Briggs Institute) and graded the certainty of evidence through the Grading of Recommendations, Assessment, Development, Evaluation (GRADE) approach. Results were summarized through meta-analyses using a random-effects model. The protocol was registered at PROSPERO (CRD42021266997). RESULTS: Of the 1382 records identified, six cross-sectional studies were included. Individuals with Down syndrome had a higher prevalence and were more likely to have tooth wear than individuals without Down syndrome (44% × 15%; OR = 4.43; 95% CI 3.17-6.18; p < 0.00001; I2 = 8%). Also, the severity of tooth wear was higher in the Down syndrome group (n = 275) compared with the controls (n = 294). The certainty of evidence analysis was very low. CONCLUSIONS: Based on very low certainty of the evidence, patients with Down syndrome had a higher prevalence and likelihood and severity of tooth wear when compared to those without Down syndrome. CLINICAL RELEVANCE: Screening early tooth wear lesions should be carefully done in the Down syndrome population since its occurrence is remarkable compared to the general population.


Assuntos
Síndrome de Down , Atrito Dentário , Desgaste dos Dentes , Humanos , Prevalência , Estudos Transversais
10.
Int J Paediatr Dent ; 33(6): 577-584, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37017581

RESUMO

BACKGROUND: Parents believe that teething is associated with signs and symptoms, which may induce them to give medications that could harm their children. Some children may require alleviation of symptoms and overall attention. AIM: To assess parents' beliefs in and attitudes toward teething. DESIGN: Through electronic databases and gray literature, this systematic review identified cross-sectional studies reporting parents' beliefs in, knowledge about, and attitudes toward the signs and symptoms of primary tooth eruption in children aged between 0 and 36 months. Three reviewers independently selected the studies, collected the information, assessed methodological quality, and checked for accuracy with disagreements solved by a fourth reviewer. The Agency of Research and Quality in Health questionnaire for cross-sectional studies was used for quality assessment. Descriptive analysis with median and interquartile ranges was adopted. RESULTS: Twenty-nine studies comprising 10 524 participants from all geographic regions were included. The methodological quality of the studies was moderate. Most parents have beliefs in signs and symptoms during dentition, the most reported symptom being the desire to bite. Oral rehydration was the most exposed attitude in the studies included. Only a small proportion of parents reported no attitude. CONCLUSIONS: The majority of parents believed in at least one sign or symptom associated with teething, and only few of them would do nothing or just wait for the signs or symptoms to pass, with no difference among countries (Protocol doi: 10.17605/OSF.IO/S2KZ3).


Assuntos
Conhecimentos, Atitudes e Prática em Saúde , Erupção Dentária , Criança , Humanos , Recém-Nascido , Lactente , Pré-Escolar , Estudos Transversais , Pais , Inquéritos e Questionários
11.
Infect Immun ; 90(2): e0059521, 2022 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-34807734

RESUMO

Ascariasis is a neglected tropical disease that is widespread in the world and has important socioeconomic impacts. The presence of various stages of worm development in the pulmonary and intestinal mucosae induces a humoral and cellular immune response. However, although there is much evidence of the protective role of mucosal immunity against various pathogens, including helminths, there is still a gap in the knowledge about the immune response and the mechanisms of action that are involved in protection against diseases, especially in the initial phase of ascariasis. Thus, the aim of this study was to evaluate the kinetic aspects of the immune parasitological parameters in intestinal and pulmonary mucosae in male mice with early ascariasis. Therefore, two mouse strains that showed different susceptibilities to ascariasis (BALB/c and C57BL/6J) when experimentally infected with 2,500 infective eggs of Ascaris suum from time point 0 were examined: the immune parasitological parameters were evaluated each 2 days after infection over a period of 12 days. The results were suggestive of a synergetic action of intestinal and pulmonary secretory IgA (S-IgA) contributing to protection against early ascariasis by reducing the amount of migrating larvae as well as the influx of leukocytes in the lung and the consequent impairment of pulmonary capacity.


Assuntos
Ascaríase , Ascaris suum , Parasitos , Pneumonia , Doenças dos Suínos , Animais , Ascaris suum/genética , Patrimônio Genético , Imunoglobulina A Secretora , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Suínos
12.
Emerg Infect Dis ; 28(3): 548-555, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35081022

RESUMO

To assess whether high-dose coronavirus disease (COVID-19) convalescent plasma (CCP) transfusion may benefit patients with severe COVID-19, we conducted a multicenter randomized trial in Brazil. Patients with severe COVID-19 who were within 10 days of initial symptom onset were eligible. Patients in the CCP group received 3 daily doses of CCP (600 mL/d) in addition to standard treatment; control patients received standard treatment only. Primary outcomes were death rates at days 30 and 60 of study randomization. Secondary outcomes were ventilator-free days and hospital-free days. We enrolled 107 patients: 36 CCP and 71 control. At day 30, death rates were 22% for CCP and 25% for the control group; at day 60, rates were 31% for CCP and 35% for control. Needs for invasive mechanical ventilation and durations of hospital stay were similar between groups. We conclude that high-dose CCP transfused within 10 days of symptom onset provided no benefit for patients with severe COVID-19.


Assuntos
COVID-19 , COVID-19/terapia , Humanos , Imunização Passiva/efeitos adversos , Plasma , SARS-CoV-2 , Resultado do Tratamento , Soroterapia para COVID-19
13.
Parasite Immunol ; 44(1-2): e12899, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34861047

RESUMO

Interactions between parasites during co-infections are often complex and can impact immunization and treatment programmes, as well as disease outcomes and morbidity. However, little is known about these interactions and the mechanisms involved. In this study, a coproparasitological survey was carried out in school-age children living in endemic areas of parasitic infection in the state of Sergipe, Northeastern Brazil. Anti-helminth-specific and total secretory immunoglobulin-A (SIgA) levels were measured in stool and saliva samples and were compared in children presenting monoparasitism, polyparasitism (helminths and/or intestinal protozoa) and no infections. The survey showed that protozoa were more prevalent than helminths, and that there was a high frequency of polyparasitism in the studied population, mainly from combinations of protozoan species. Although less frequent, combinations between species of protozoa and helminths were also observed. The levels of salivary SIgA in these co-infected individuals were lower than the average observed in infections with helminths alone. Although the children participating in this survey were asymptomatic, and it was, therefore, not possible to evaluate the impact of salivary SIgA reduction on the diseases, and the study highlights the need for further investigations of co-infections by intestinal parasites and the effects on immune response induced by the interactions between different parasites.


Assuntos
Anti-Helmínticos , Helmintíase , Enteropatias Parasitárias , Animais , Brasil/epidemiologia , Criança , Fezes/parasitologia , Helmintíase/epidemiologia , Helmintíase/parasitologia , Humanos , Imunoglobulina A Secretora , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/parasitologia , Prevalência , Solo/parasitologia
14.
Transfus Med ; 32(3): 248-251, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34085363

RESUMO

OBJECTIVES: Evaluate the impact of ABO histo-blood group type on COVID-19 severity. BACKGROUND: ABO histo-blood type has been associated with different outcomes in infectious diseases. It has also shown a higher proportion of type A patients with SARS-CoV-2. In this observational study, extracted from an ongoing clinical trial on the efficacy of convalescent plasma transfused in COVID-19 patients, we describe the impact of ABO blood type on the risk of developing severe COVID-19. MATERIALS AND METHODS: Seventy-two consecutive patients (37 type A, 23 type O, 11 type B, 1 type AB) with severe (respiratory failure) COVID-19 were included. Control group was composed of 160 individuals randomly selected from the same populational basis. RESULTS: Blood group A was overrepresented (51.39%) in the patient group in relation to the control group (30%), whereas blood group O was less represented (31.94%) in patient than in control group (48%). Odds ratio (A vs. O) was 2.581 (1.381-4.817), CI 95%; p = 0.004. Also, blood group A patients appeared to have more severe disease, given by the scores of the Sequential Organ Failure Assessment and Simplified Acute Physiologic Score 3 (p = 0.036 and p = 0.058, respectively). CONCLUSION: Histo-blood type A is associated with a higher risk of developing severe COVID-19 in relation to blood type O.


Assuntos
COVID-19 , Sistema ABO de Grupos Sanguíneos , COVID-19/terapia , Humanos , Imunização Passiva , Fatores de Risco , SARS-CoV-2 , Soroterapia para COVID-19
15.
Nucleic Acids Res ; 48(5): 2312-2331, 2020 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-32020195

RESUMO

Eukaryotic protein synthesis generally initiates at a start codon defined by an AUG and its surrounding Kozak sequence context, but the quantitative importance of this context in different species is unclear. We tested this concept in two pathogenic Cryptococcus yeast species by genome-wide mapping of translation and of mRNA 5' and 3' ends. We observed thousands of AUG-initiated upstream open reading frames (uORFs) that are a major contributor to translation repression. uORF use depends on the Kozak sequence context of its start codon, and uORFs with strong contexts promote nonsense-mediated mRNA decay. Transcript leaders in Cryptococcus and other fungi are substantially longer and more AUG-dense than in Saccharomyces. Numerous Cryptococcus mRNAs encode predicted dual-localized proteins, including many aminoacyl-tRNA synthetases, in which a leaky AUG start codon is followed by a strong Kozak context in-frame AUG, separated by mitochondrial-targeting sequence. Analysis of other fungal species shows that such dual-localization is also predicted to be common in the ascomycete mould, Neurospora crassa. Kozak-controlled regulation is correlated with insertions in translational initiation factors in fidelity-determining regions that contact the initiator tRNA. Thus, start codon context is a signal that quantitatively programs both the expression and the structures of proteins in diverse fungi.


Assuntos
Códon de Iniciação/química , Cryptococcus/genética , Regulação Fúngica da Expressão Gênica , Genoma Fúngico , Iniciação Traducional da Cadeia Peptídica , Aminoacil-tRNA Sintetases/genética , Aminoacil-tRNA Sintetases/metabolismo , Candida albicans/genética , Candida albicans/metabolismo , Mapeamento Cromossômico , Códon de Iniciação/metabolismo , Cryptococcus/metabolismo , Neurospora crassa/genética , Neurospora crassa/metabolismo , Fases de Leitura Aberta , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismo , Especificidade da Espécie
16.
Proc Natl Acad Sci U S A ; 116(48): 24056-24065, 2019 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-31712417

RESUMO

Viruses have transformed our understanding of mammalian RNA processing, including facilitating the discovery of the methyl-7-guanosine (m7G) cap on the 5' end of RNAs. The m7G cap is required for RNAs to bind the eukaryotic translation initiation factor eIF4E and associate with the translation machinery across plant and animal kingdoms. The potyvirus-derived viral genome-linked protein (VPg) is covalently bound to the 5' end of viral genomic RNA (gRNA) and associates with host eIF4E for successful infection. Divergent models to explain these observations proposed either an unknown mode of eIF4E engagement or a competition of VPg for the m7G cap-binding site. To dissect these possibilities, we resolved the structure of VPg, revealing a previously unknown 3-dimensional (3D) fold, and characterized the VPg-eIF4E complex using NMR and biophysical techniques. VPg directly bound the cap-binding site of eIF4E and competed for m7G cap analog binding. In human cells, VPg inhibited eIF4E-dependent RNA export, translation, and oncogenic transformation. Moreover, VPg formed trimeric complexes with eIF4E-eIF4G, eIF4E bound VPg-luciferase RNA conjugates, and these VPg-RNA conjugates were templates for translation. Informatic analyses revealed structural similarities between VPg and the human kinesin EG5. Consistently, EG5 directly bound eIF4E in a similar manner to VPg, demonstrating that this form of engagement is relevant beyond potyviruses. In all, we revealed an unprecedented modality for control and engagement of eIF4E and show that VPg-RNA conjugates functionally engage eIF4E. As such, potyvirus VPg provides a unique model system to interrogate eIF4E.


Assuntos
Fator de Iniciação 4E em Eucariotos/química , Potyvirus/genética , Biossíntese de Proteínas/fisiologia , RNA/química , Ribonucleoproteínas/química , Proteínas Virais/química , Sítios de Ligação , Ligação Competitiva , Linhagem Celular , Fator de Iniciação 4E em Eucariotos/metabolismo , Humanos , Modelos Moleculares , Ressonância Magnética Nuclear Biomolecular , Dobramento de Proteína , Capuzes de RNA/química , Processamento Pós-Transcricional do RNA , Ribonucleoproteínas/metabolismo , Proteínas Virais/metabolismo , Proteínas Virais/fisiologia
17.
Sensors (Basel) ; 22(3)2022 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-35161583

RESUMO

The impact of diet and digestive disorders in flatus composition remains largely unexplored. This is partially due to the lack of standardized sampling collection methods, and the easy atmospheric contamination. This paper describes a method to quantitatively determine the major gases in flatus and their application in a nutritional intervention. We describe how to direct sample flatus into Tedlar bags, and simultaneous analysis by gas chromatography-thermal conductivity detection (GC-TCD). Results are analyzed by univariate hypothesis testing and by multilevel principal component analysis. The reported methodology allows simultaneous determination of the five major gases with root mean measurement errors of 0.8% for oxygen (O2), 0.9% for nitrogen (N2), 0.14% for carbon dioxide (CO2), 0.11% for methane (CH4), and 0.26% for hydrogen (H2). The atmospheric contamination was limited to 0.86 (95% CI: [0.7-1.0])% for oxygen and 3.4 (95% CI: [1.4-5.3])% for nitrogen. As an illustration, the method has been successfully applied to measure the response to a nutritional intervention in a reduced crossover study in healthy subjects.


Assuntos
Flatulência , Metano , Dióxido de Carbono , Cromatografia Gasosa , Estudos Cross-Over , Dieta , Humanos , Condutividade Térmica
18.
J Oral Rehabil ; 49(7): 701-711, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35340028

RESUMO

BACKGROUND: Non-human primates are the closest animal models to humans regarding genetics, physiology and behaviour. Marmoset monkeys in particular are one of the most versatile species for biomedical research. OBJECTIVE: To assess the craniofacial growth and development of the masticatory function in the common marmoset (Callithrix jacchus), from birth to the fourth month of life through minimally invasive cineradiographic imaging. METHODS: Ten individuals were followed-up from 0 to 4 months of age regarding craniofacial growth and masticatory function assessed by cineradiography. For the experimental procedure, we used a microfocal X-ray source apparatus and a beryllium fast-response image-intensifier. RESULTS: The duration of the masticatory cycles was stable across age groups. Chewing a very soft Castella cake or the slightly harder Marshmallow did not change the masticatory cycle in the time domain. On the other hand, linear and angular measurements of the jaw-opening movement showed a tendency for bigger movements at the latter stages of craniofacial growth. Qualitative analysis showed that marmosets had a small preference for Castella over Marshmallow, that they most often bit off pieces of food to chew with their posterior teeth, that they manipulated the food with their hands, and that they chewed the food continuously. CONCLUSION: We observed critical developmental events during the first 3 months of life in marmosets. Cineradiographic imaging in marmosets may provide valuable information on craniofacial form and function for basic and preclinical research models.


Assuntos
Callithrix , Cinerradiografia , Animais , Callithrix/fisiologia , Alimentos , Humanos , Mastigação/fisiologia
19.
Eur J Dent Educ ; 26(1): 131-137, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33735486

RESUMO

INTRODUCTION: The use of teaching resources and digital objects has gradually been incorporated into dental schools. This study aimed to evaluate the digital anatomy table in teaching-learning process of the temporomandibular joint (TMJ) anatomy and the student's perception regarding this resource. MATERIALS AND METHODS: A cross-sectional study was conducted on 41 undergraduate dental students. Knowledge assessment tests were applied at different time intervals: before and after the TMJ theoretical class, after the practical class on prosected specimens and after the use of digital anatomy table. The medians of the scores obtained in the three groups (theoretical class, practical class and digital table) were descriptively analysed and submitted to Wilcoxon, Kruskal-Wallis and Student-Newman-Keuls statistical tests. The perception survey was conducted at the end of the study. RESULTS: When the teaching strategies were compared amongst each other, the medians of the scores with the digital anatomy table were significantly higher than after the theoretical and practical classes. At the end of the research, there were no differences amongst the three groups (theoretical class, practical class and digital table). Regarding the perception, the majority of the students reported that the digital anatomy table helped them to understand the content of the theoretical class. CONCLUSION: It was concluded that learning with the use of the digital anatomy table did not increase the knowledge of dental students with respect to the TMJ anatomy. The students' perceptions of the digital anatomy table were positive and that it could be used as an additional resource in the teaching-learning process.


Assuntos
Anatomia , Educação de Graduação em Medicina , Estudantes de Medicina , Anatomia/educação , Estudos Transversais , Currículo , Educação em Odontologia , Avaliação Educacional , Humanos , Aprendizagem , Ensino , Articulação Temporomandibular
20.
Environ Monit Assess ; 194(10): 758, 2022 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-36085206

RESUMO

Trace and potentially toxic elements represent one class of food contaminants that has stimulated research. In markets, two main methods of growing vegetables are generally available: conventional and organic. Conventional farming has been the target of some concerns about the use of agrochemicals, especially the excessive use of pesticides, whereas organic agriculture minimizes the use of agrochemicals. As the main route for potentially toxic elements' absorption by humans is by food intake, it is important to evaluate if the method of cultivation influences their concentrations. This study evaluated the levels of potentially toxic elements and nutrients on four leafy vegetables: curly lettuce, collard greens, escarole, and rocket, cultivated by conventional and organic farming. We found that Al, Ba, Fe, and Sr levels were higher in conventional samples, whereas K, Pb, and Zn were higher in organic. Amongst the elements analysed, values of Fe, Al, and K were around 0.2, 0.3, and 70 g kg-1, respectively, except in collard greens, in which the values were lower. On the other hand, Ba, Sr, and Mn presented higher concentration in collard greens compared to the other vegetables in conventional cultivation (~ 35, 80, and 120 mg kg-1, respectively). The principal component analysis result shows that the samples were grouped according to the type of vegetable, regardless of the type of cultivation. Despite this, the evaluation of the cultivation by different types of farming is important in order to choose the healthiest option.


Assuntos
Monitoramento Ambiental , Verduras , Agroquímicos , Humanos , Lactuca , Folhas de Planta
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