RESUMO
Since human angiotensin-converting enzyme 2 (ACE2) serves as a primary receptor for SARS-CoV-2, characterizing ACE2 regions that allow SARS-CoV-2 to enter human cells is essential for designing peptide-based antiviral blockers and elucidating the pathogenesis of the virus. We identified and synthesized a 25-mer mimetic peptide (encompassing positions 22-46 of the ACE2 alpha-helix α1) implicated in the S1 receptor-binding domain (RBD)-ACE2 interface. The mimetic (wild-type, WT) ACE2 peptide significantly inhibited SARS-CoV-2 infection of human pulmonary Calu-3 cells in vitro. In silico protein modeling predicted that residues F28, K31, F32, F40, and Y41 of the ACE2 alpha-helix α1 are critical for the original, Delta, and Omicron strains of SARS-CoV-2 to establish the Spike RBD-ACE2 interface. Substituting these residues with alanine (A) or aspartic acid (D) abrogated the antiviral protective effect of the peptides, indicating that these positions are critical for viral entry into pulmonary cells. WT ACE2 peptide, but not the A or D mutated peptides, exhibited significant interaction with the SARS-CoV-2 S1 RBD, as shown through molecular dynamics simulations. Through identifying the critical amino acid residues of the ACE2 alpha-helix α1, which is necessary for the Spike RBD-ACE2 interface and mobilized during the in vitro viral infection of cells, we demonstrated that the WT ACE2 peptide protects susceptible K18-hACE2 mice against in vivo SARS-CoV-2 infection and is effective for the treatment of COVID-19.
Assuntos
Enzima de Conversão de Angiotensina 2 , COVID-19 , Peptídeos , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Enzima de Conversão de Angiotensina 2/metabolismo , Enzima de Conversão de Angiotensina 2/química , Humanos , Animais , SARS-CoV-2/efeitos dos fármacos , COVID-19/virologia , Camundongos , Glicoproteína da Espícula de Coronavírus/metabolismo , Glicoproteína da Espícula de Coronavírus/química , Peptídeos/farmacologia , Peptídeos/química , Peptídeos/uso terapêutico , Tratamento Farmacológico da COVID-19 , Antivirais/farmacologia , Antivirais/química , Linhagem Celular , Pneumonia/tratamento farmacológico , Pneumonia/virologia , Pneumonia/prevenção & controle , Pulmão/virologia , Pulmão/patologia , FemininoRESUMO
Spodoptera frugiperda control methods have proved to be inefficient, which justifies the search for new control measures. In this search for botanical insecticides for controlling S. frugiperda, the following were evaluated: (i) the toxicity of essential oils (EOs) from Cinnamodendron dinisii, Eugenia uniflora, and Melaleuca armillaris; (ii) the effect of EOs on life table parameters against S. frugiperda; (iii) the chemical characterization of EOs; and (iv) the in silico interaction of the chemical constituents present in the three EOs with the molecular targets of S. frugiperda. The EO from E. uniflora had the lowest LD50 (1.19 µg of EO/caterpillar). The major compounds bicyclogermacrene (18.64%) in C. dinisii and terpinolene (57.75%) in M. armillaris are highly predicted to interact with the octopamine receptor (OctpR). The compound 1,8-cineole (21.81%) in M. armillaris interacts mainly with a tolerant methoprene receptor (MET) and curzerene (41.22%) in E. uniflora, which acts on the OctpR receptor. Minor compounds, such as nerolidol in C. dinisii and ß-elemene in E. uniflora, are highly ranked for multiple targets: AChE, MET, OctpR, and 5-HT1. It was concluded that the EO from E. uniflora negatively affects several biological parameters of S. frugiperda development and is promising as an active ingredient in formulations for controlling this insect pest.
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Deleção de Sequência , Talassemia alfa , Humanos , Talassemia alfa/genética , Brasil , Masculino , alfa-Globinas/genética , Criança , Feminino , Hemoglobina H/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Mimosa caesalpiniifolia (Sansão-do-Campo) is a native species of the caatinga in northeastern Brazil that has been studied for its potential anti-inflammatory and antidepressant activity. It is popularly consumed as a medicinal plant and its pharmacological benefits are evidenced in the literature. AIM OF THE STUDY: The present work was carried out to promote the chemical profile and evaluate the pharmacological activity of the dry extract and the ethyl acetate fraction obtained from the dry leaves of Mimosa caesalpiniifolia. MATERIALS AND METHODS: The leaves were collected in the municipality of Alfenas-MG and subjected to drying, followed by division in a knife mill. The preparation of the dry extract was carried out by the extraction method using simple percolation and the fraction was obtained by liquid-liquid partition. Part of the extractive solution was concentrated in a rotary evaporator followed by a drying process using the spray technique with the addition of colloidal silicon dioxide. The dry extract (33.33%) showed a higher yield in mass when compared to the yield of the ethyl acetate fraction (19.67%). The in vivo pharmacological evaluation was conducted with a total of 82 male Wistar rats that underwent cecal ligation and perforation surgery to induce the inflammatory process. One week after surgery, these animals were treated for 7 days with the dry extract and the ethyl acetate fraction and submitted to behavioral tests (open field and forced swimming). RESULTS: The chemical results were obtained through analysis by HPLC-PDA coupled to a mass spectrometer, enabling the verification of the presence of phenolic acids, flavonoids, aglycones, and glycosides, in addition to tannins. This corroborates with data present in the literature for the genus Mimosa sp. Some compounds had their structure determined, where they were identified as catechin (m/z 288.97), cassiaocidentalin A (m/z 560.75), and procyanidin B2 [(epi)catechin-(epi)catechin; m/z 576.83)]. It was found that the animals that were submitted to the treatment did not present statistically significant results, demonstrating that the pharmacological action evaluated in the test was not highlighted in this type of experiment. The groups that underwent treatment had an aggravated locomotor activity. CONCLUSIONS: The results found with the chemical study contributed to the knowledge of the plant species studied. On the other hand, further studies are needed to provide a better understanding of the pharmacological evaluation of Mimosa caesalpiniifolia.
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Acetatos , Catequina , Mimosa , Ratos , Animais , Ratos Wistar , Mimosa/química , Extratos Vegetais/farmacologia , Extratos Vegetais/análise , Folhas de Planta/químicaRESUMO
Climate factors, pesticides, and landscape in coffee agroecosystems directly affect the populations of the coffee leaf miner and its parasitoids. This study aimed to investigate the effects of climate factors, insecticide use, and landscape on natural parasitism, parasitoid diversity, and infestation of L. coffeella in coffee plantations in the Planalto region, Bahia, Brazil. Mined leaves were collected monthly in six coffee plantations with varying edge density, vegetation cover, landscape diversity in scales of 500 to 3000 m of radius, insecticide use, and climate factors. Closterocerus coffeellae, and Proacrias coffeae (Eulophidae) predominated in the pest's natural parasitism. Our record is the first for the occurrence of Stiropius reticulatus, Neochrysocharis sp. 1, Neochrysocharis sp. 2, and Zagrammosoma sp. in Bahia. Higher temperature and larger forest cover increased the coffee leaf miner infestation. Higher rainfall values, insecticide use, and landscape diversity decreased the pest infestations. Natural parasitism and species diversity are favoured by increase in temperature, forest cover, and edge density, while increase in rainfall, insecticide use, and landscape diversity lead them to decrease.The natural parasitism and diversity of parasitoid species of the coffee leaf miner have been enhancing in the areas with greater forest cover and edge density associated with low use of insecticides. The areas composed of different lands with annual croplands surrounding the coffee plantations showed less natural parasitism and parasitoid species diversity. The ecosystem services provided by C. coffeellae and P. coffeae in coffee crops areas require conservation and these species are potential bioproducts for applied biological control programmes.
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Coffea , Inseticidas , Mariposas , Animais , Ecossistema , FlorestasRESUMO
The chemical recycling of poly(ethylene terephthalate) (PET) residues was performed via glycolysis with ethylene glycol (EG) over Mg-Fe and Mg-Al oxide catalysts derived from layered double hydroxides. Catalysts prepared using the high supersaturation method (h.s.c.) presented a higher surface area and larger particles, but this represented less PET conversion than those prepared by the low supersaturation method (l.s.c.). This difference was attributed to the smaller mass transfer limitations inside the (l.s.c.) catalysts. An artificial neural network model well fitted the PET conversion and bis(2-hydroxyethyl) terephthalate (BHET) yield. The influence of Fe in place of Al resulted in a higher PET conversion of the Mg-Fe-h.s.c. catalyst (~95.8%) than of Mg-Al-h.s.c. (~63%). Mg-Fe catalysts could be reused four to five times with final conversions of up to 97% with reaction conditions of EG: PET = 5:1 and catalyst: PET = 0.5%. These results confirm the Mg-Fe oxides as a biocompatible novel catalyst for the chemical recycling of PET residues to obtain non-toxic BHET for further polymerization, and use in food and beverage packaging.
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CRISPR is a revolutionary gene editing technology that has enabled scientists worldwide to explore the cell's genetic blueprint in an unprecedented easy way. In this chapter, we will briefly present the history behind the development of this innovative tool, how it emerged from a natural bacterial mechanism for antiviral defense, its key components (Cas9 endonuclease and single guide RNA), mode of action (DNA cleavage and repair via NHEJ or HDR), and versatility (acting on single- or double-stranded DNA or RNA) for diverse purposes beyond gene editing such as stochastic marking, digital encoding, high-fidelity SNP genotyping, programmed chromosome fission/fusion, gene mapping, nucleic acid detection, regulation of gene expression, DNA/RNA labeling or tracking, and more.
Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Sistemas CRISPR-Cas/genética , Proteína 9 Associada à CRISPR/metabolismo , RNA , Quebras de DNA de Cadeia Dupla , DNA/genéticaRESUMO
Several medicinal plants have drawn the attention of researchers by its phytochemical composition regarding their potential for treating chronic complications of diabetes mellitus. In this context, plants of the Myrtaceae family popularly used in Brazil for the treatment of diabetes mellitus, including Eugenia sonderiana, have shown beneficial effects due to the presence of phenolic compounds and saponins in their chemical constitution. Thus, the present work aimed to perform the phytochemical characterization of the hydroethanolic extract of E. sonderiana leaves using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), along with in vitro and in vivo studies of antidiabetic activity. The chemical characterization revealed the presence of phenolic compounds, flavonoids, neolignans, tannins, and saponins. In addition, the extract exhibited minimum inhibitory concentrations of alpha-amylase and alpha-glycosidase higher than the acarbose in the in vitro tests. Also, the in vivo tests revealed a slight increase in body mass in diabetic rats, as well as a significant decrease in water and feed consumption provided by the extract. Regarding serum biochemical parameters, the extract showed significant activity in decreasing the levels of glucose, hepatic enzymes, and triglycerides, in addition to maintaining HDL cholesterol levels within normal ranges, protecting the cell membranes against oxidative damage. Thus, the extract of E. sonderiana leaves was considered promising pharmaceutical ingredient in the production of a phytotherapy medication.
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Diabetes Mellitus Experimental , Eugenia , Saponinas , Ratos , Animais , Hipoglicemiantes/uso terapêutico , Extratos Vegetais/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Compostos Fitoquímicos/uso terapêutico , Fenóis/farmacologia , Folhas de Planta/química , Saponinas/uso terapêuticoRESUMO
OBJECTIVES: This study compared the relationships of social determinants with cardiometabolic risk in different socioeconomic contexts: sociopolitically unstable Venezuela (VE) and stable Czechia (CZ). DESIGN: cross-sectional analysis involving two population-based studies. SETTING: Brno, Czechia and 23 cities of Venezuela. PARTICIPANTS: 25-64 years old subjects from CZ (2013-2014, n=1579, 56% females) and VE (2014-2017, n=1652, 70% females). MAIN OUTCOME MEASURES: The composite cardiometabolic risk score (CMRS) (scaled 0-8) was calculated using eight biomarkers (body mass index, waist circumference, blood glucose, systolic and diastolic blood pressure, total and high-density lipoprotein-cholesterol, triglycerides). Social characteristics included education in both countries, income in CZ and a composite measure of social position (SP) in VE. Sex stratified ordinal regression examined the social gradient in having less favourable CMRS. RESULTS: In CZ, men and women with low education and women with low income had higher odds of higher CMRS compared with those with high education and income with OR 1.45 (95% CI 1.01 to 2.21), 2.29 (95% CI 1.62 to 3.24) and 1.69 (95% CI 1.23 to 2.35). In VE, women with low education and low SP had higher odds to have higher CMRS OR 1.47 (95% CI 1.09 to 1.97) and 1.51 (95% CI 1.16 to 1.97), while men with low education and low SP had lower odds to have higher CMRS OR 0.64 (95% CI 0.41 to 1.00) and 0.61 (95% CI 0.40 to 0.97), compared with those with high education and high SP. Independently of age, sex and socioeconomic characteristics, Venezuelans had higher odds to have higher CMRS than Czechs (OR 2.70; 95% CI 2.37 to 3.08). CONCLUSIONS: The results suggest that the associations of socioeconomic status indices and cardiometabolic risk differed between CZ and VE, likely reflecting differences in the social environment among countries. Further research is needed to confirm and quantify these differences.
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Doenças Cardiovasculares , Masculino , Humanos , Feminino , Adulto , Pessoa de Meia-Idade , Estudos Transversais , Doenças Cardiovasculares/epidemiologia , República Tcheca/epidemiologia , Venezuela/epidemiologia , Fatores de Risco , Classe Social , Índice de Massa CorporalRESUMO
Type 1 diabetes (T1D) arises from autoimmune-mediated destruction of insulin-producing ß-cells leading to impaired insulin secretion and hyperglycemia. T1D is accompanied by DNA damage, oxidative stress, and inflammation, although there is still scarce information about the oxidative stress response and DNA repair in T1D pathogenesis. We used the microarray method to assess mRNA expression profiles in peripheral blood mononuclear cells (PBMCs) of 19 T1D patients compared to 11 controls and identify mRNA targets of microRNAs that were previously reported for T1D patients. We found 277 differentially expressed genes (220 upregulated and 57 downregulated) in T1D patients compared to controls. Analysis by gene sets (GSA and GSEA) showed an upregulation of processes linked to ROS generation, oxidative stress, inflammation, cell death, ER stress, and DNA repair in T1D patients. Besides, genes related to oxidative stress responses and DNA repair (PTGS2, ATF3, FOSB, DUSP1, and TNFAIP3) were found to be targets of four microRNAs (hsa-miR-101, hsa-miR148a, hsa-miR-27b, and hsa-miR-424). The expression levels of these mRNAs and microRNAs were confirmed by qRT-PCR. Therefore, the present study on differential expression profiles indicates relevant biological functions related to oxidative stress response, DNA repair, inflammation, and apoptosis in PBMCs of T1D patients relative to controls. We also report new insights regarding microRNA-mRNA interactions, which may play important roles in the T1D pathogenesis.