Defensin-like peptides from Capsicum chinense induce increased ROS, loss of mitochondrial functionality, and reduced growth of the fungus Colletotrichum scovillei.

In the present study, we identified and characterized two defensin-like peptides in an antifungal fraction obtained from Capsicum chinense pepper fruits and inhibited the growth of Colletotrichum scovillei, which causes anthracnose. AMPs were extracted from the pericarp of C. chinense peppers and subjected to ion exchange, molecular exclusion, and reversed-phase in a high-performance liquid chromatography system. We investigated the endogenous increase in reactive oxygen species (ROS), the loss of mitochondrial functioning, and the ultrastructure of hyphae. The peptides obtained from the G3 fraction through molecular exclusion chromatography were subsequently fractionated using reverse-phase chromatography, resulting in the isolation of fractions F1, F2, F3, F4, and F5. The F1-Fraction suppressed C. scovillei growth by 90, 70.4, and 44% at 100, 50, and 25 µg mL-1, respectively. At 24 h, the IC50 and minimum inhibitory concentration were 21.5 µg mL-1 and 200 µg mL-1, respectively. We found an increase in ROS, which may have resulted in an oxidative burst, loss of mitochondrial functioning, and cytoplasm retraction, as well as an increase in autophagic vacuoles. MS/MS analysis of the F1-Fraction indicated the presence of two defensin-like proteins, and we were able to identify the expression of three defensin sequences in our C. chinense fruit extract. The F1-Fraction was also found to inhibit the activity of insect α-amylases. In summary, the F1-Fraction of C. chinense exhibits antifungal activity against a major pepper pathogen that causes anthracnose. These defensin-like compounds are promising prospects for further research into antifungal and insecticide biotechnology applications. © 2024 Society of Chemical Industry.

Inhibition of Serine Protease, α-Amylase and Growth of Phytopathogenic Fungi by Antimicrobial Peptides from Capsicum chinense Fruits.

Plant fungal diseases cause major problems for the global economy. Antimicrobial peptides have aroused great interest in the control of phytopathogens, as they are natural molecules and have a broad spectrum of inhibitory activity. Herein, we have tried to identify and characterize antimicrobial peptides present in fruits of Capsicum chinense and to evaluate their enzymatic and antifungal activities. The retained fraction obtained in the anion exchange chromatography with strong antifungal activity was subjected to molecular exclusion chromatography and obtained four fractions named G1, G2, G3, and G4. The 6.0-kDa protein band of G2 showed similarity with protease inhibitors type II, and it was able to inhibit 100% of trypsin and α-amylase activities. The protein band with approximately 6.5 kDa of G3 showed similarity with sequences of protease inhibitors from genus Capsicum and showed growth inhibition of 48% for Colletotrichum lindemuthianum, 49% for Fusarium lateritium, and 51% for F. solani and F. oxysporum. Additionally, G3 causes morphological changes, membrane permeabilization, and ROS increase in F. oxysporum cells. The 9-kDa protein band of G4 fraction was similar to a nsLTP type 1, and a protein band of 6.5 kDa was similar to a nsLTP type 2. The G4 fraction was able to inhibit 100% of the activities of glycosidases tested and showed growth inhibition of 35 and 50% of F. oxysporum and C. lindemuthianum, respectively. C. chinense fruits have peptides with antifungal activity and enzyme inhibition with biotechnological potential.

Potent Anti-Candida Fraction Isolated from Capsicum chinense Fruits Contains an Antimicrobial Peptide That is Similar to Plant Defensin and is Able to Inhibit the Activity of Different α-Amylase Enzymes.

Antimicrobial peptides (AMPs) are molecules present in several life forms, possess broad-spectrum of inhibitory activity against pathogenic microorganisms, and are a promising alternative to combat the multidrug resistant pathogens. The aim of this work was to identify and characterize AMPs from Capsicum chinense fruits and to evaluate their inhibitory activities against yeasts of the genus Candida and α-amylases. Initially, after protein extraction from fruits, the extract was submitted to anion exchange chromatography resulting two fractions. Fraction D1 was further fractionated by molecular exclusion chromatography, and three fractions were obtained. These fractions showed low molecular mass peptides, and in fraction F3, only two protein bands of approximately 6.5 kDa were observed. Through mass spectrometry, we identified that the lowest molecular mass protein band of fraction F3 showed similarity with AMPs from plant defensin family. We named this peptide CcDef3 (Capsicum chinense defensin 3). The antifungal activity of these fractions was analyzed against yeasts of the genus Candida. At 200 µg/mL, fraction F1 inhibited the growth of C. tropicalis by 26%, fraction F2 inhibited 35% of the growth of C. buinensis, and fraction F3 inhibited all tested yeasts, exhibiting greater inhibition activity on the growth of the yeast C. albicans (86%) followed by C. buinensis (69%) and C. tropicalis (21%). Fractions F1 and F2 promoted membrane permeabilization of all tested yeasts and increased the endogenous induction of reactive oxygen species (ROS) in C. buinensis and C. tropicalis, respectively. We also observed that fraction F3 at a concentration of 50 µg/mL inhibited the α-amylase activities of Tenebrio molitor larvae by 96% and human salivary by 100%. Thus, our results show that fraction F3, which contains CcDef3, is a very promising protein fraction because it has antifungal potential and is able to inhibit the activity of different α-amylase enzymes.

Boas práticas de transparência, informatização e comunicação social no licenciamento ambiental brasileiro: uma análise comparada dos websites dos órgãos licenciadores estaduais / Good practices of transparency, electronic procedures, and social communication in Brazil's environmental impact assessment and licensing system: a comparative analysis of the websites of state-level agencies

RESUMO: O objetivo deste artigo foi analisar as boas práticas de transparência, informatização e comunicação social evidentes nos websites dos órgãos públicos estaduais e do distrito federal responsáveis pelo controle da avaliação de impacto e do licenciamento ambiental. A metodologia baseou-se em análises de conteúdo dos websites dos 27 entes federados, com base em uma lista de verificação de 28 boas práticas. Foram identificados níveis diversificados de atendimento a cada uma das boas práticas analisadas. O estudo também mostrou que os websites dos órgãos licenciadores estaduais variam muito em termos de atendimento às práticas analisadas, sugerindo a existência de diferenças significativas na capacidade institucional dos órgãos. Diversas boas práticas, sobretudo as relacionadas aos controles informatizados de processos e de participação social, têm um claro potencial de replicação em diferentes jurisdições. O artigo conclui com uma discussão das implicações da pesquisa e com sugestões de novos estudos.

ABSTRACT: This article analyzed the good practices of transparency, electronic procedures and social communication in the websites of State-level Public Agencies in charge of Environmental Impact Assessment (EIA) and Environmental Licensing in their jurisdictions. The methodology was based on content analyses of the websites of all 26 states and the federal district, based on a checklist of 28 good practices. It was found diverse levels of implementation of each good practice analyzed. The study also showed that the websites vary greatly in terms of good practice adoption, thus suggesting the existence of significant differences in institutional capacity across State-level agencies. Many good practices, particularly those related to electronic procedures and social participation, have a clear potential for replication across jurisdictions. The article concludes with a discussion of the research implications and suggesting future research agendas.

Perfil epidemiológico das pacientes com câncer de mama atendidas no Hospital Geral de Palmas, Tocantins, no período de 2004 a 2009 / Epidemiological profile of patients with breast cancer at the GeneralHospital of Palmas, Tocantins, in the period from 2004 to 2009

O objetivo do presente estudo é descrever o perfil epidemiológico das pacientes com diagnóstico de câncer de mama tratadas no Hospital Geral Público de Palmas, Tocantins, no período de 2004 a 2009 e contribuir para a melhoria das informações e formulações de medidas educacionais relacionadas a esse problema de saúde pública. Foi realizado estudo retrospectivo de corte transversal no serviço de Registro Hospitalar de Câncer do Hospital Geral Público de Palmas. Foram incluídas todas as pacientes diagnosticadas e tratadas com câncer de mama no período de 2004 a 2009, excluindo homens acometidos. As informações foram coletadas por meio da análise das fichas do Registro Hospitalar de Câncer disponibilizadas pelo Instituto Nacional do Câncer (INCA). As fichas se referem a 168 pacientes, das quais 70,3% encontravam-se entre 41 e 60 anos. Dessas, 57,1% são procedentes do interior do estado. Em relação ao estadiamento, os níveis II e III correspondiam a 74,4% dos casos analisados. Quanto ao grau de instrução, 11,9% possuíam ensino superior e 7,1% eram analfabetas. Os principais métodos de tratamento foram: quimioterapia (71,4%), cirurgia (47,6%) e hormonioterapia (30,4%), sendo que 32,5% das pacientes terminaram o tratamento com a doença estável e 34,5% com remissão completa do tumor. Os resultados da pesquisa mostraram que a faixa etária mais acometida correspondia às mulheres ainda em idade laboral, grande parte em estadio avançado. A maioria das pacientes do interior recebeu diagnóstico tardio. Mulheres cujo tratamento foi realizado nos estadios iniciais da doença obtiveram melhores resultados.

To describe of this study is to describe the epidemiological profile of patients diagnosed with breast cancer treated at the Palmas Public General Hospital, in Palmas, Tocantins, Brazil, during the period of 2004?2009, and to contribute to the improvement of information and formulation of educational actions related to this public health problem. A retrospective cross-sectional study was developed at the Cancer Hospital Registry service in the HGPP. All breast cancer patients diagnosed and treated in the period from 2004 to 2009, excluding affected men, were included. Data were collected by means of the analysis of the Cancer Hospital Registry records provided by the National Cancer Institute (INCA). Data refer to 168 patients, of which 70.3% were between 41?60 years of age; 57.1% of these were from the inland Tocantins. Regarding the staging, levels II and III accounted for 74.4% of the analyzed cases. Concerning the education degree, 11.9% had higher education and 7.1% were illiterate. The main treatment strategies were: chemotherapy (71.4%), surgery (47.6%), and hormone therapy (30.4%), so that 32.5% of patients have completed the treatment with stable disease and 34.5% with complete remission of the tumor. Research results show that the most affected age group correspond to working-age women, largely in advanced stage of cancer. Most of the patients from inland received a late diagnosis. Women whose treatment was performed in the early stages of the disease have demonstrated better results.