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Anti-inflammatory agents are widely used for the treatment of inflammatory diseases. Nevertheless, the associated side effects of the available drugs make it necessary to search for new anti-inflammatory drugs. Here, we investigated the anti-inflammatory activity of solidagenone. Initially, we observed that a single dose of 30, 60, or 90 mg/kg of solidagenone did not result in mortality or elicit any discernible signs of toxicity in mice. At the same doses, solidagenone promoted a significant reduction in the migration of neutrophils in an acute peritonitis model and decreased mortality in a lipopolysaccharide-induced endotoxic shock model. Interestingly, treatment with solidagenone conferred a protective effect against leukopenia and thrombocytopenia, hematological disorders commonly observed in sepsis conditions. In addition, treatment with all the doses of solidagenone promoted a significant reduction in nitric oxide, TNF-α, and IL-1ß levels relative to the LPS-stimulated vehicle-treated cultures. Furthermore, gene expression and in silico analyses also supported the modulation of the NF-κB pathway by solidagenone. Finally, in silico pharmacokinetics predictions indicated a favorable drugability profile for solidagenone. Taken together, the findings of the present investigation show that solidagenone exhibits significant anti-inflammatory properties in acute experimental models, potentially through the modulation of the NF-κB signaling pathway.
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The traditional way of dealing with plant diseases has been the use of chemical products, but these harm the environment and are incompatible with the global effort for sustainable development. The use of Bacillus and related species in the biological control of plant diseases is a trend in green agriculture. Many studies report the positive effect of these bacteria, but a synthesis is still necessary. So, the objective of this work is to perform a meta-analysis of Bacillus biocontrol potential and identify factors that drive its efficacy. Data were compiled from articles published in journals listed in two of the main scientific databases between 2000 and 2021. Among 6159 articles retrieved, 399 research papers met the inclusion criteria for a systematic review. Overall, Bacilli biocontrol agents reduced disease by 60% compared to control groups. Furthermore, experimental tests with higher concentrations show a strong protective effect, unlike low and single concentration essays. Biocontrol efficacy also increased when used as a protective inoculation rather than therapeutic inoculation. Inoculation directly in the fruit has a greater effect than soil drenching. The size of the effect of Bacillus-based commercial products is lower than the newly tested strains. The findings presented in this study confirm the power of Bacillus-based bioinoculants and provide valuable guidance for practitioners, researchers, and policymakers seeking effective and sustainable solutions in plant disease management.
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Bacillus , Agentes de Controle Biológico , Doenças das Plantas , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia , Controle Biológico de Vetores/métodos , Agricultura/métodos , Microbiologia do Solo , Frutas/microbiologiaRESUMO
With global climate changes and the increased demand for food due to expected world population growth, genetic improvement programs have aimed at producing crops with increased yield and tolerance to environmental stresses, such as drought, salinity, and pathogens. On the other hand, genetic improvement programs via biotechnology require candidate genes that confer traits of interest to be incorporated into improved crops. In this regard, genes encoding transcription factors (TFs) can be promising since they are proteins that transcriptionally regulate the expression of target genes related to the most diverse roles in the plant, including defense against stresses. Among TFs, bZIP (basic leucine zipper) proteins regulate many developmental and physiological processes in the plant, such as seed formation, fruit ripening, nutrient assimilation, and defense response to abiotic and biotic stresses. In this review, we aim to highlight the main advances in the potential use of bZIP TFs in the genetic improvement of crops. We address this potential mainly regarding crop tolerance to stresses and other agricultural traits, such as increased yield and fruit features.
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Fatores de Transcrição de Zíper de Leucina Básica , Proteínas de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Produtos Agrícolas/genética , Produtos Agrícolas/metabolismo , Biotecnologia , Estresse Fisiológico/genética , Regulação da Expressão Gênica de PlantasRESUMO
INTRODUCTION: The Cognitive Distortions Questionnaire (CD-Quest) is an instrument that identifies logical errors or cognitive distortions and is used in trial-based cognitive therapy (TBCT). However, it had previously only been available for adults. OBJECTIVES: To develop and validate a version of the CD-Quest for teens (CD-Quest-T) aged 11 to 17 years and test its psychometric properties. METHOD: A total of 299 schoolchildren participated in the investigation. After content validity was assessed, the language was adapted for the target age group, and the length of the instrument was reduced to eight items (from the initial 15). Five cognitive therapists analyzed the content and structure of the items. Finally, to investigate the construct validity of the CD-Quest-T, the instrument was divided into a full scale and two subscales, which measure the frequency of the distortions and the intensity attributed to them, respectively. RESULTS: The overall internal consistency of the scale was α = 0.77, whereas subscale indices were α = 0.75 for the frequency scale and α = 0.73 for the intensity scale. Results from exploratory factor analysis and concurrent validity analysis indicated that the CD-Quest-T items have good psychometric properties and generate scores reliably. CONCLUSION: The psychometric properties of the CD-Quest-T demonstrate its adequacy for measurement of cognitive distortions in adolescents.
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Cognição , Idioma , Adolescente , Criança , Humanos , Psicometria , Reprodutibilidade dos Testes , Inquéritos e QuestionáriosRESUMO
Abstract Introduction The Cognitive Distortions Questionnaire (CD-Quest) is an instrument that identifies logical errors or cognitive distortions and is used in trial-based cognitive therapy (TBCT). However, it had previously only been available for adults. Objectives To develop and validate a version of the CD-Quest for teens (CD-Quest-T) aged 11 to 17 years and test its psychometric properties. Method A total of 299 schoolchildren participated in the investigation. After content validity was assessed, the language was adapted for the target age group, and the length of the instrument was reduced to eight items (from the initial 15). Five cognitive therapists analyzed the content and structure of the items. Finally, to investigate the construct validity of the CD-Quest-T, the instrument was divided into a full scale and two subscales, which measure the frequency of the distortions and the intensity attributed to them, respectively. Results The overall internal consistency of the scale was α = 0.77, whereas subscale indices were α = 0.75 for the frequency scale and α = 0.73 for the intensity scale. Results from exploratory factor analysis and concurrent validity analysis indicated that the CD-Quest-T items have good psychometric properties and generate scores reliably. Conclusion The psychometric properties of the CD-Quest-T demonstrate its adequacy for measurement of cognitive distortions in adolescents.
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The main environmental problem in urban areas, especially in Brazil, is the discharge of untreated sewage. The in vivo Drosophila melanogaster Somatic Mutation and Recombination Test (SMART) was used to assess the genotoxicity of surface waters from three different sites in the Tocantins River, Brazil. The in silico approach was used to search for known and predicted interactions between environmental chemicals found in our samples and Drosophila and human proteins. The genotoxicity tests were performed in standard (ST) and high bioactivation (HB) crosses with samples collected at two periods, the rainy and dry seasons. Mutant spot frequencies found in treatments with unprocessed water from the test sites were compared with the frequencies observed in negative controls. The collection points were represented as sites A, B and C along Tocantins River. Sites A and B are located in Porto Nacional City, whereas site C is located in Palmas City. Considering the rainy season collection, positive responses in the ST cross were observed for sites A and C (89.47% and 85% of recombination, respectively) and in the HB cross for sites A, B and C (88.24%, 84.21% and 82.35% of recombination, respectively). The positive results in the dry season were restricted to sites A and B (88.89% and 85.71% of recombination, respectively) in the HB cross. In accordance with in vivo and in silico results, we hypothesize that ribosomal proteins (RPs) in fruit fly and humans are depleted in cells exposed to heavy metal causing DNA damage and chromosome instability, increasing homologous recombination.
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Drosophila melanogaster , Rios , Animais , Brasil , Cidades , Dano ao DNA , Drosophila , Drosophila melanogaster/genética , Humanos , Água/metabolismo , Asas de Animais/metabolismoRESUMO
El síndrome de interrupción del tallo hipofisario es una anomalía congénita de probable origen genético causante de hipopituitarismo, que radiológicamente se caracteriza por hipoplasia de la adenohipófisis, neurohipófisis ectópica e interrupción del tallo hipofisario. Suele presentarse con baja estatura y deficiencia de hormonas adenohipofisarias. El retraso en el diagnóstico se relaciona con alta morbimortalidad. Se reporta el caso de una mujer de 24 años que consulta por amenorrea primaria. Presentaba talla baja y ausencia de caracteres sexuales secundarios, con genitales infantiles. Los exámenes paraclínicos mostraron compromiso de dos ejes hipofisarios. Una resonancia magnética nuclear mostró hallazgos consistentes con síndrome de interrupción de tallo hipofisario.
Pituitary stalk disruption syndrome is a congenital abnormality of probable genetic origin that causes hypopituitarism, which is radiologically characterized by adenohypophysis hypoplasia, ectopic neurohypophysis, and pituitary stalk disruption. It usuallypresents with short stature and adenohypophyseal hormone deficiency. The delay in diagnosis is related to high morbidity and mortality. The case of a 24-year-old woman who consulted for primary amenorrhea is reported. She had short stature and no secondary sexual characteristics, with infantile genitalia. Paraclinical examinations showed involvement of two pituitary axes. An MRI scan showed findings consistent with pituitary stalk disruption syndrome.
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Feminino , Adulto , Amenorreia , Hipopituitarismo , Anormalidades CongênitasRESUMO
The current increase in salinity can intensify the disparity between potential and actual crop yields, thus affecting economies and food security. One of the mitigating alternatives is plant breeding via biotechnology, where advances achieved so far are significant. Considering certain aspects when developing studies related to plant breeding can determine the success and accuracy of experimental design. Besides this strategy, halophytes with intrinsic and efficient abilities against salinity can be used as models for improving the response of crops to salinity stress. As crops are mostly glycophytes, it is crucial to point out the molecular differences between these two groups of plants, which may be the key to guiding and optimizing the transformation of glycophytes with halophytic tolerance genes. Therefore, this can broaden perspectives in the trajectory of research towards the cultivation, commercialization, and consumption of salt-tolerant crops on a large scale.
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Tolerância ao Sal , Plantas Tolerantes a Sal , Produtos Agrícolas/genética , Melhoramento Vegetal , Salinidade , Plantas Tolerantes a Sal/genéticaRESUMO
Tuberculosis is the leading cause of death amongst adults with human immunodeficiency virus (HIV) infection. The lifetime risk of tuberculosis disease for a person with latent infection is estimated at 5-10% with most cases occurring within five years of initial infection. The World Health Organization recommends isoniazid preventive therapy (IPT) for latent tuberculosis treatment, amongst other strategies. The aim was to assess tuberculosis incidence, survival (free of tuberculosis) and associated factors in HIV-positive patients. IPT was offered to participants with a positive (≥5mm) tuberculin skin test. Participants were followed from February 2003-December 2016. Kaplan-Meier was used for survival analysis. Variables with p-value ≤ 0.2 in the univariate analysis entered into the multivariate Cox-Model, keeping those with p-value ≤ 0.05. The 95% confidence interval of incidence of tuberculosis was estimated using Poisson distribution. One hundred nineteen patients completed the IPT and were followed for a median duration of 110.7 months (IQR 93.1-121.0). The probability of developing tuberculosis (10 years post-IPT) was 5.4%. Tuberculosis incidence was 0.58/100 patient/years (CI 95% 0.213-1.264). IPT over 6 months provided long-term protection against tuberculosis. AIDS-defining illness was the only statistically significant variable (HR=5.67) in the multivariate model.
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Humanos , Análise de Sobrevida , HIV , Tuberculose Latente , IsoniazidaRESUMO
This paper aims to report a case in which central giant cell granuloma (CGCG) mimicked a periapical lesion of endodontic origin. An 18-year-old female patient was referred for diagnosis and treatment of extensive radiolucent periapical lesion involving 31, 32, 33, 34 and 35 teeth. Clinically, the patient presented slight facial asymmetry and healthy teeth on the affected side with positive response to thermal vitality tests. Thus, an incisional biopsy was performed, which presented a histopathological picture characteristic of a CGCG. The endodontic treatment of the involved teeth was followed by surgical curettage of the lesion. After two years of follow-up, the patient was asymptomatic, with marked improvement in mandibular symmetry and adequate healing of the lesion. Therefore, the diagnosis of radiolucent periapical lesions must include lesions of endodontic and non-endodontic origin for better treatment planning and execution.
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Granuloma de Células Gigantes , Adolescente , Diagnóstico Diferencial , Feminino , Granuloma de Células Gigantes/diagnóstico por imagem , Granuloma de Células Gigantes/cirurgia , HumanosRESUMO
MAIN CONCLUSION: A new Piper nigrum cysteine proteinase inhibitor, PnCPI, belonging to group I of phytocystatins, with inhibitory activity against papain and growth of Fusarium solani f. sp. piperis, was isolated and characterized. Previous studies (de Souza et al. 2011) have identified a partial cDNA sequence of putative cysteine proteinase inhibitor differentially expressed in roots of black pepper (P. nigrum L.) infected by F. solani f. sp. piperis. Here, we aimed to isolate the full-length cDNA and genomic sequences of the P. nigrum cysteine proteinase inhibitor gene, named PnCPI. Sequence analyses showed that the PnCPI gene encodes a deduced protein of 108 amino acid residues with a predicted molecular mass of 12.3 kDa and isoelectric point of 6.51. Besides the LARFAV-like sequence, common to all phytocystatins, PnCPI contains three conserved motifs of the superfamily cystatin: a glycine residue at the N-terminal region, the QxVxG reactive site more centrally positioned, and one tryptophan in the C-terminal region. PnCPI, belonging to group I of phytocystatins, showed high identity with cystatins isolated from several plant species. Sequence analyses also revealed no putative signal peptide at the N-terminal of PnCPI, as well as no introns within the genomic sequence corresponding to the PnCPI coding region. Molecular modeling showed the ability of PnCPI to interact with papain, while its inhibitory activity against this protease was confirmed after heterologous expression in Escherichia coli. The effects of heat treatments on the inhibitory activity of recombinant PnCPI, rPnCPI, were evaluated. In addition, rPnCPI exhibited in vitro activity against F. solani f. sp. piperis, revealing a new cystatin with the potential antifungal application. The identification of PnCPI as a functional cystatin able to inhibit the in vitro growth of F. solani f. sp. piperis indicates other factors contributing to in vivo susceptibility of black pepper to root rot disease.
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Antifúngicos/farmacologia , Cistatinas/farmacologia , Inibidores de Cisteína Proteinase/farmacologia , Fusarium/efeitos dos fármacos , Papaína/antagonistas & inibidores , Piper nigrum/genética , Doenças das Plantas/prevenção & controle , Antifúngicos/isolamento & purificação , Clonagem Molecular , Inibidores de Cisteína Proteinase/isolamento & purificação , DNA Complementar/genética , Fusarium/enzimologia , Piper nigrum/química , Doenças das Plantas/microbiologiaRESUMO
Bud necrosis (BN) is a common disorder that affects Vitis vinifera L. and reduces its potential yield. To minimize the losses caused by BN, the double pruning management was applied in Brazilian Southeast vineyards. In this management strategy plants are pruned at the winter to promote a vegetative cycle and then, at summer, to promote the reproductive cycle at optimal environmental conditions. To investigate the relationship of BN and the double pruning management RNA-seq libraries were sequenced from healthy and necrotic tissues at four different stages of the year. The comparison of differentially expressed genes in necrotic and non-necrotic tissues showed an enhanced expression of genes related to cell death possibly induced by endophytic microorganisms in the necrotic tissues. The de novo assembly, characterization and quantification of transcripts within the RNA-seq libraries showed that genes from the endophytic fungus Alternaria alternata, responsible for the production of toxic compounds were highly expressed under BN. Here we propose a model in which unfavorable conditions and reduced carbohydrate levels in buds can promote the switch from a biotrophic lifestyle to a necrotrophic lifestyle in the endophytic fungi, which seems to be involved in the development of BN.
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Alternaria/fisiologia , Endófitos/fisiologia , Perfilação da Expressão Gênica , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Vitis/genética , Vitis/microbiologia , Necrose/genética , Brotos de Planta/genética , Brotos de Planta/microbiologia , ReproduçãoRESUMO
Macaw palm is a highly oil-producing plant, which presents high contents of free fatty acids, being a promising feedstock for biofuel production. The current chemical routes are costly and complex, involving highly harsh industrial conditions. Enzymatic processing is a potential alternative; however, it is hampered by the scarce knowledge on biocatalysts adapted to this acidic feedstock. This work describes a novel lipase isolated from the thermophilic fungus Rasamsonia emersonii (ReLip), which tolerates extreme conditions such as the presence of methanol, high temperatures, and acidic medium. Among the tested feedstocks, the enzyme showed the highest preference for macaw palm oil, producing a hydrolyzate with a final free fatty acid content of 92%. Crystallographic studies revealed a closed conformation of the helical amphipathic lid that typically undergoes conformational changes in a mechanism of interfacial activation. Such conformation of the lid is stabilized by a salt bridge, not observed in other structurally characterized homologs, which is likely involved in the tolerance to organic solvents. Moreover, the lack of conservation of the aromatic cluster IxxWxxxxxF in the lid of ReLip with the natural mutation of the phenylalanine by an alanine might be correlated with the preference of short acyl chains, although preserving catalytic activity on insoluble substrates. In addition, the presence of five acidic amino acids in the lid of ReLip, a rare property reported in other lipases, may have contributed to its ability to tolerate and be effective in acidic environments. Therefore, our work describes a new fungal biocatalyst capable of efficiently hydrolyzing macaw oil, an attractive feedstock for the production of "drop-in" biofuels, with high desirable feature for industrial conditions such as thermal and methanol tolerance, and optimum acidic pH. Moreover, the crystallographic structure was elucidated, providing a structural basis for the enzyme substrate preference and tolerance to organic solvents.
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BACKGROUND: bZIP proteins participate in the regulation of gene expression, playing crucial roles in various biological processes in plants, including response to environmental changes. Luminosity is an environmental factor of extreme importance for plant metabolism, acting as a regulator of its growth and development. Despite advances in the identification of bZIP proteins in several plant species, studies on these transcription factors in cassava are lacking. Cassava (Manihot esculenta Crantz) is one of the most important food crops in tropical and subtropical regions, mainly in developing countries, where its storage root is a major source of calories for low-income people. OBJECTIVES: Our main aim was the isolation of a cDNA sequence encoding a bZIP protein from cassava (MebZIP) as well as the in silico characterization of its nucleotide and deduced amino acid sequences. In addition, we evaluated the expression pattern of the MebZIP gene in response to light, and its possible relationship with regulation of the chalcone synthase (MeCHS) gene. METHODS: RT-PCR and 3' and 5' RACE assays were used to isolate the full-length cDNA sequence of MebZIP. Bioinformatics tools were used to characterize the nucleotide and amino acid sequences of MebZIP. Semiquantitative RT-PCR assays were used to evaluate the expression levels of MebZIP and MeCHS genes. RESULTS: We isolated the full-length cDNA sequence of MebZIP with a 1320-bp ORF encoding a deduced protein with a predicted molecular weight and isoelectric point of 47 kDa and 5.85, respectively. Comparative analyses with GenBank sequences showed high identity of MebZIP with bZIP CPRF-2 of Hevea brasiliensis (XP_021650934) and Petroselinum crispum (Q99090.2). Besides the basic region and leucine zipper domains, MebZIP contains putative conserved domains (D1- D4), found in parsley CPRF-2 and bZIP proteins closely related to this protein. Since CPRF proteins are known for their function in regulation of the CHS gene by light, we evaluated the expression levels of the MebZIP gene and the possible target gene to be regulated by MebZIP (the MeCHS gene) in cassava under light conditions. Semi-quantitative RT-PCR assays revealed that MebZIP transcription increased in response to white light, with maximum expression levels at 6 h of light exposure. On the other hand, the expression levels of the MeCHS gene were statistically constant in all samples, indicating that they were not influenced by the experimental conditions used here. CONCLUSION: The putative MebZIP protein identified in this work contains the conserved domains (bZIP, D1-D4) that indicate its functionality, thus allowing it to be considered a new member of the bZIP transcription factor CPRF-2 family. The expression levels of the MebZIP gene increased during white light exposure, indicating a potential function in light-response in cassava.
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Luz , Manihot , Proteínas de Plantas , Fatores de Transcrição , Transcrição Gênica , Manihot/genética , Manihot/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
We provide here a current overview of marmoset (Callithrix) evolution, hybridization, species biology, basic/biomedical research, and conservation initiatives. Composed of 2 subgroups, the aurita group (C aurita and C flaviceps) and the jacchus group (C geoffroyi, C jacchus, C kuhlii, and C penicillata), this relatively young primate radiation is endemic to the Brazilian Cerrado, Caatinga, and Atlantic Forest biomes. Significant impacts on Callithrix within these biomes resulting from anthropogenic activity include (1) population declines, particularly for the aurita group; (2) widespread geographic displacement, biological invasions, and range expansions of C jacchus and C penicillata; (3) anthropogenic hybridization; and (4) epizootic Yellow Fever and Zika viral outbreaks. A number of Brazilian legal and conservation initiatives are now in place to protect the threatened aurita group and increase research about them. Due to their small size and rapid life history, marmosets are prized biomedical models. As a result, there are increasingly sophisticated genomic Callithrix resources available and burgeoning marmoset functional, immuno-, and epigenomic research. In both the laboratory and the wild, marmosets have given us insight into cognition, social group dynamics, human disease, and pregnancy. Callithrix jacchus and C penicillata are emerging neotropical primate models for arbovirus disease, including Dengue and Zika. Wild marmoset populations are helping us understand sylvatic transmission and human spillover of Zika and Yellow Fever viruses. All of these factors are positioning marmosets as preeminent models to facilitate understanding of facets of evolution, hybridization, conservation, human disease, and emerging infectious diseases.
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Febre Amarela , Infecção por Zika virus , Zika virus , Animais , Brasil , Callithrix/genética , Genômica , Hibridização GenéticaRESUMO
Tobacco farming has been proving to induce poor health outcomes in agricultural workers, genomic instability being the triggering one. This study evaluated influence of PON1 (paraoxonase 1), SOD2 (superoxide dismutase), OGG1 (8-oxoguanine glycosylase), XRCC1 (X-ray repair cross-complementing protein 1), and XRCC4 (X-ray repair cross-complementing protein 4) genes polymorphisms on DNA damage in 121 subjects occupationally exposed to pesticides mixtures and nicotine at tobacco fields and 121 non-exposed individuals. Inorganic elements (Cl, P, S and Zn) and cotinine levels were found increased in farmers, confirming exposure. Results show higher frequencies of buccal micronucleus (MN), nuclear buds (NBUD), binucleated cells (BN) and damage index (comet assay), reduced telomere length (TL), and increased parameters of oxidative stress in farmers compared to non-exposed individuals. PON1 Gln/Gln genotype was associated with increased MN frequency. SOD2 Val/Val showed association with increased frequency of MN and NBUD and decreased antioxidant activity. The XRCC1 Arg/Arg showed protective effect for MN, BN and TL, which was also positively influenced by OGG1 -/Cys. MN was decreased in XRCC4 -/Ile farmers. These genotypes also showed a risk for antioxidant activity. Our study proposes that PON1 and SOD2 variants play a role in xenobiotic-metabolizing system in farmers, while base excision repair (BER) pathway could be the repair mechanism involved in genomic instability suffered by tobacco farmers.
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Arildialquilfosfatase/genética , Dano ao DNA , DNA Glicosilases/genética , Proteínas de Ligação a DNA/genética , Praguicidas/toxicidade , Superóxido Dismutase/genética , Proteína 1 Complementadora Cruzada de Reparo de Raio-X/genética , Adulto , Ensaio Cometa , Fazendeiros , Feminino , Genótipo , Humanos , Masculino , Testes para Micronúcleos , Pessoa de Meia-Idade , Exposição Ocupacional/efeitos adversos , Polimorfismo Genético , NicotianaRESUMO
Human schistosomiasis is a neglected tropical disease of great importance in public health. A large number of people are infected with schistosomiasis, making vaccine development and effective diagnosis important control strategies. A rational epitope prediction workflow using Schistosoma mansoni hypothetical proteins was previously presented by our group, and an improvement to that approach is presented here. Briefly, immunodominant epitopes from parasite membrane proteins were predicted by reverse vaccinology strategy with additional in silico analysis. Furthermore, epitope recognition was evaluated using sera of individuals infected with S. mansoni. The epitope that stood out in both in silico and in vitro assays was used to compose a rational chimeric molecule to improve immune response activation. Out of 2185 transmembrane proteins, four epitopes with high binding affinities for human and mouse MHCII molecules were selected through computational screening. These epitopes were synthesized to evaluate their ability to induce TCD4+ lymphocyte proliferation in mice. Sm204830e and Sm043300e induced significant TCD4+ proliferation. Both epitopes were submitted to enzyme-linked immunosorbent assay to evaluate their recognition by IgG antibodies from the sera of infected individuals, and epitope Sm043300 was significantly recognized in most sera samples. Epitope Sm043300 also showed good affinity for human MHCII molecules in molecular docking, and its sequence is curiously highly conserved in four S. mansoni proteins, all of which are described as G-protein-coupled receptors. In addition, we have demonstrated the feasibility of incorporating this epitope, which showed low similarity to human sequences, into a chimeric molecule. The stability of the molecule was evaluated by molecular modeling aimed at future molecule production for use in diagnosis and vaccination trials.
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Antígenos de Helmintos/imunologia , Epitopos Imunodominantes/imunologia , Schistosoma mansoni/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Anti-Helmínticos/sangue , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/genética , Linfócitos T CD4-Positivos/imunologia , Técnicas de Química Combinatória , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Feminino , Cadeias HLA-DRB1/imunologia , Proteínas de Helminto/química , Proteínas de Helminto/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Humanos , Epitopos Imunodominantes/genética , Epitopos Imunodominantes/metabolismo , Ativação Linfocitária , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Moleculares , Simulação de Acoplamento Molecular , Conformação Proteica , Receptores Acoplados a Proteínas G/química , Receptores Acoplados a Proteínas G/imunologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Schistosoma haematobium/imunologia , Schistosoma mansoni/genética , Esquistossomose mansoni/sangue , Esquistossomose mansoni/imunologia , Alinhamento de Sequência , Vacinas de Subunidades Antigênicas/imunologia , Vacinas Sintéticas/imunologiaRESUMO
Epidemiological studies indicate that living in proximity to coal mines is correlated with numerous diseases including cancer, and that exposure to PM10 and PM2.5 components could be associated with this phenomenon. However, the understanding of the mechanisms by which PM exerts its adverse effects is still incomplete and comes mainly from studies in occupationally exposed populations. The aims of this study were to: (1) evaluate DNA damage in lymphocytes assessing the cytokinesis-block micronucleus cytome assay (CBMN-cyt) parameters; (2) identify aneugenic or clastogenic effects in lymphocytes of exposed populations using CREST immunostaining for micronuclei; (3) evaluate multi-elemental composition of atmospheric particulate matter; and (4) verify relation between the DNA damage and PM2.5 and PM10 levels around the mining area. Analysis revealed a significant increase in micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells of individuals with residential proximity to open-pit coal mines compared to residents from non-mining areas. Correlation analysis demonstrated a highly significant association between PM2.5 levels, MNBN frequencies and CREST+ micronuclei induction in exposed residents. These results suggest that PM2.5 fraction generated in coal mining activities may induce whole chromosome loss (aneuploidy) preferentially, although there are also chromosome breaks. Analysis of the chemical composition of PM2.5 by PIXE demonstrated that Si, S, K and Cr concentrations varied significantly between coal mining and reference areas. Enrichment factor values (EF) showed that S, Cr and Cu were highly enriched in the coal mining areas. Compared to reference area, mining regions had also higher concentrations of extractable organic matter (EOM) related to nonpolar and polar compounds. Our results demonstrate that PM2.5 fraction represents the most important health risk for residents living near open-pit mines, underscoring the need for incorporation of ambient air standards based on PM2.5 measures in coal mining areas.