RESUMO
This study aimed to develop microemulsions (MEs) containing α-bisabolol for the topical treatment of cutaneous leishmaniasis (CL). Initially, pseudoternary phase diagrams were developed using α-bisabolol as the oil phase, Eumulgin® CO 40 as the surfactant, Polymol® HE as the co-surfactant, and distilled water as the aqueous phase. Two transparent liquid systems (TLS) containing 5% of α-bisabolol were selected and characterized (F5E25 and F5EP25). Next, skin permeation and retention assays were performed using Franz cells. The interaction of the formulation with the stratum corneum (SC) was evaluated using the FTIR technique. The cytotoxicity was evaluated in murine peritoneal macrophages. Finally, the antileishmanial activity of microemulsions was determined in promastigotes and amastigotes of L. amazonensis (strain MHOM/BR/77/LTB 0016). As a result, the selected formulations showed isotropy, nanometric size (below 25 nm), Newtonian behavior and pH ranging from 6.5 to 6.9. The MEs achieved a 2.5-fold increase in the flux and skin-permeated amount of α-bisabolol. ATR-FTIR results showed that microemulsions promoted fluidization and extraction of lipids and proteins of the stratum corneum, increasing the diffusion coefficient and partition coefficient of the drug in the skin. Additionally, F5E25 and F5EP25 showed higher activity against promastigotes (IC50 13.27 and 18.29, respectively) compared to unencapsulated α-bisabolol (IC50 53.8). Furthermore, F5E25 and F5EP25 also showed antileishmanial activity against intracellular amastigotes of L. amazonensis, with IC50 50 times lower than free α-bisabolol and high selectivity index (up to 15). Therefore, the systems obtained are favorable to topical administration, with significant antileishmanial activity against L. amazonensis promastigotes and amastigotes, being a promising system for future in vivo trials.
Assuntos
Emulsões , Macrófagos Peritoneais , Sesquiterpenos Monocíclicos , Sesquiterpenos , Pele , Animais , Sesquiterpenos Monocíclicos/farmacologia , Sesquiterpenos Monocíclicos/química , Emulsões/química , Camundongos , Sesquiterpenos/farmacologia , Sesquiterpenos/química , Pele/parasitologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/parasitologia , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/parasitologia , Espectroscopia de Infravermelho com Transformada de Fourier , Absorção Cutânea/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Feminino , Leishmania/efeitos dos fármacos , Tensoativos/farmacologia , Tensoativos/química , Antiprotozoários/farmacologia , Antiprotozoários/químicaRESUMO
The diversity of Geminiviridae and Alphasatellitidae species in tomatoes was assessed via high-throughput sequencing of 154 symptomatic foliar samples collected from 2002 to 2017 across seven Brazilian biomes. The first pool (BP1) comprised 73 samples from the North (13), Northeast (36), and South (24) regions. Sixteen begomoviruses and one Topilevirus were detected in BP1. Four begomovirus-like contigs were identified as putative novel species (NS). NS#1 was reported in the semi-arid (Northeast) region and NS#2 and NS#4 in mild subtropical climates (South region), whereas NS#3 was detected in the warm and humid (North) region. The second pool (BP2) comprised 81 samples from Southeast (39) and Central-West (42) regions. Fourteen viruses and subviral agents were detected in BP2, including two topileviruses, a putative novel begomovirus (NS#5), and two alphasatellites occurring in continental highland areas. The five putative novel begomoviruses displayed strict endemic distributions. Conversely, tomato mottle leaf curl virus (a monopartite species) displayed the most widespread distribution occurring across the seven sampled biomes. The overall diversity and frequency of mixed infections were higher in susceptible (16 viruses + alphasatellites) in comparison to tolerant (carrying the Ty-1 or Ty-3 introgressions) samples, which displayed 9 viruses. This complex panorama reinforces the notion that the tomato-associated Geminiviridae diversity is yet underestimated in Neotropical regions.
Assuntos
Geminiviridae , Metagenômica , Filogenia , Doenças das Plantas , Solanum lycopersicum , Solanum lycopersicum/virologia , Brasil , Doenças das Plantas/virologia , Geminiviridae/genética , Geminiviridae/classificação , Geminiviridae/isolamento & purificação , Animais , Variação Genética , Genoma Viral , Begomovirus/genética , Begomovirus/classificação , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
Two novel tomato-infecting begomoviruses were discovered via high-throughput sequencing in Brazil. Both viruses were also Sanger-sequenced and displayed DNA-A components phylogenetically related to New World bipartite begomoviruses. The names tomato golden net virus (ToGNV) and tomato yellow net virus (ToYNV) were proposed. The majority of the New World begomoviruses has bipartite genomes. However, extensive analyses revealed that ToGNV and ToYNV have monopartite genomes, because no cognate DNA-B components were detected. Hence, they may comprise a unique group of monopartite New World begomoviruses, which have enormous biological, molecular, and plant breeding interest.
Assuntos
Begomovirus , Solanum lycopersicum , Begomovirus/genética , Melhoramento Vegetal , Brasil , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
Eggplant (Solanum melongena L.) is an economically important vegetable crop in Brazil, especially in family-based farming. Eggplant hybrids 'Ciça' and 'Napoli' (≈ 400 plants) were detected exhibiting virus-like symptoms (5-20% incidence) in field surveys (2015-2018) in Brasília-DF (Figure 1). Symptoms included chlorosis, mosaic and apical leaf deformation. Six symptomatic leaf samples were collected from fruit-bearing plants (around 100 days after planting) aiming at verifying the potential orthotospovirus infection. Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was carried out with polyclonal antibodies (produced at Centro Nacional de Pesquisa de Hortaliças - CNPH) against the N gene coat protein of the three major orthotospoviruses: tomato spotted wilt orthotospovirus (TSWV), groundnut ringspot orthotospovirus (GRSV) and, tomato chlorotic spot orthotospovirus (TCSV). Strong serological reactions were observed only against GRSV antibodies in the extracts from symptomatic samples, but not in the controls. To confirm the causal agent of those symptoms, total RNA was extracted from infected leaf samples via the standard Trizol® (Sigma) protocol and subsequently used in a two-step reverse transcriptase polymerase chain reaction (RT-PCR) approach. Synthesis of the cDNA was carried out with the J13 primer (5'-CCC GGA TCC AGA GCA AT-3') (Cortez et al., 2001) followed by PCR assays with the primer pair BR60 (5'-AGA GCA ATC GTG TCA-3`) and BR65 (5`-ATC AAG CCT TCT GAA AGT CAT-3') (Eiras et al., 2002). This primer set amplifies a fragment of 453 bp including the 3' untranslated region at the 3' terminus of the S RNA and the protein N-coding gene of at least five species: TSWV, GRSV, TCSV, chrysanthemum stem necrosis orthotospovirus (CSNV) and zucchini lethal chlorosis orthotospovirus (ZLCV). In addition, GRSV-specific primers (LNA Reis, unpublished) were used for amplification of all three segments: L segment: LF/LR (5'-AAC AGG ATT CAG CAA TAT GG-3'/ 5'-AAT TCC TTG AAG ACA ATT GTG T -3'); M segment: MF/MR (5'-TTT GTC CAA CCA TAC CAG ACC C- 3' / 5'-GGC TTC AAT AAA GGC TTG GG-3') and, S segment: SF/SR (5'-TTC AAA CTC AGT TGT ACT CTG A-3'/5'-TTA CTT TCG ATC TGG TTG AA- 3'). Amplicons with 509 bp (MT043204), 289 bp (MT043205), and 901 bp (MT043203) were obtained for L, M and S segments of the eggplant isolate DF-687. PCR amplicons corresponding to a segment of the N-coding gene (396 bp) of a second eggplant isolate (BJL02; MK176337) were obtained with the primer pair BR60/BR65 and subjected to Sanger dideoxy sequencing at CNPH. Alignments of nucleotide sequences of both isolates revealed identity levels varying around 99% to the corresponding genomic regions of a large set of GRSV isolates from GenBank database. PCR assays using total RNA as template yielded 494 bp amplicons solely with GRSV-specific primers (Webster et al., 2011), but no products were obtained with TSWV-specific primers (Adkins and Rosskopf, 2002), confirming the former as the sole causal agent of the field symptoms. Leaves of eggplant cv. 'Ciça' and indicator hosts, including Nicotiana rustica, Capsicum chinense 'PI 159236' (with the Tsw gene), and S. lycopersicum cv. Santa Clara were rub inoculated with extracts prepared from eggplant samples naturally infected with GRSV. Mosaic, necrotic ringspots and systemic leaf deformation symptoms were observed around ten days after inoculation on newly emerged leaves of all inoculated plants. GRSV infection was confirmed by DAS-ELISA and RT-PCR ten days after inoculation. Eggplant was erroneously listed as a host of GRSV in Brazil (Kitajima, 2020). Hence, this is the first report of eggplant infection by this virus in South America. No significant yield losses have been observed in eggplant due to GRSV infection since the overall symptoms are often mild. However, this natural host of GRSV might impact disease management strategies since eggplant is quite often cultivated under family-based farming conditions as a companion crop of highly susceptible tomato, sweet-pepper, and lettuce cultivars. References: Adkins, S., and Rosskopf, E. N. 2002. Plant Dis. 86: 1310. Cortez, I., et al. 2001. Arch. Virol. 146:265. Eiras, M. et al., 2002. Fitopatol. Bras. 27:285. Kitajima, E.W. 2020. Biota Neotrop. 20: e2019932. Webster, C. G., et al. 2011. Virology 413: 216.
RESUMO
Conventional treatments for leishmaniasis have caused serious adverse effects, poor tolerance, development of resistant strains. Natural products have been investigated as potential therapeutic alternatives. The cashew nut shell liquid (CNSL) is a natural source of phenolic compounds with several biological activities, where cardanol (CN) is considered one of the most important and promising compounds. This study aimed to evaluate antileishmanial, cytotoxic and immunomodulatory activities of CNSL and CN. Both showed antileishmanial potential, with IC50 for CNSL and CN against Leishmania infantum: 148.12 and 56.74 µg/mL; against Leishmania braziliensis: 85.71 and 64.28 µg/mL; against Leishmania major: 153.56 and 122.31 µg/mL, respectively. The mean cytotoxic concentrations (CC50) of CNSL and CN were 37.51 and 31.44 µg/mL, respectively. CNSL and CN significantly reduced the percentage of infected macrophages, with a selectivity index (SI) >20 for CN. CNSL and cardanol caused an increase in phagocytic capacity and lysosomal volume. Survival rates of Zophobas morio larvae at doses of 3; 30 and 300 mg/kg were: 85%, 75% and 60% in contact with CNSL and 85%, 60% and 40% in contact with CN, respectively. There was a significant difference between the survival curves of larvae when treated with CN, demonstrating a significant acute toxicity for this substance. Additional investigations are needed to evaluate these substances in the in vivo experimental infection model.
Assuntos
Anacardium , Antineoplásicos , Nozes , Fenóis/toxicidadeRESUMO
Isopropyl gallate (IPG) is a polyphenol obtained from alterations in the gallic acid molecule via acid catalysis with previously reported leishmanicidal and trypanocidal activities. The present study aims to evaluate in silico binding activity towards some targets for antileishmanial chemotherapy against Leishmania major species, and ADMET parameters for IPG, as well as in vitro antileishmanial and cytotoxic effects. Molecular docking was performed using AutoDockVina and BIOVIA Discovery Studio software, whereas in silico analysis used SwissADME, PreADMET and admetSAR software. In vitro antileishmanial activity on promastigotes and amastigotes of Leishmania major, cytotoxicity and macrophages activation were assessed. IPG exhibited affinity for pteridine reductase (PTR1; -8.2 kcal/mol) and oligopeptidase B (OPB; -8.0 kcal/mol) enzymes. ADMET assays demonstrated good lipophilicity, oral bioavailability, and skin permeability, as well as non-mutagenic, non-carcinogenic properties and low risk of cardiac toxicity for IPG. Moreover, IPG inhibited the in vitro growth of promastigotes (IC50 = 90.813 µM), presented significant activity against amastigotes (IC50 = 13.45 µM), promoted low cytotoxicity in macrophages (CC50 = 1260 µM), and increased phagocytic capacity. These results suggest IPG is more selectively toxic to the parasite than to mammalian cells. IPG demonstrated acceptable in silico pharmacokinetics parameters, and reduced infection and infectivity in parasitized macrophages, possibly involving macrophage activation pathways and inhibition of leishmania enzymes.
RESUMO
BACKGROUND: Leishmaniasis is a parasitosis with a wide incidence in developing countries. The drugs which are indicated for the treatment of this infection usually are able to promote high toxicity. PURPOSE: A combination of limonene and carvacrol, monoterpenes present in plants with antiparasitic activity may constitute an alternative for the treatment of these diseases. METHODS: In this study, the antileishmania activity against Leishmania major, cytotoxicity tests, assessment of synergism, parasite membrane damage tests as well as molecular docking and immunomodulatory activity of limonene-carvacrol (Lim-Car) combination were evaluated. RESULTS: The Lim-Car combination (5:0; 1:1; 1:4; 2:3; 3:2; 4:1 and 0:5) showed potential antileishmania activity, with mean inhibitory concentration (IC50) ranging from 5.8 to 19.0 µg.mL-1. They demonstrated mean cytotoxic concentration (CC50) ranging from 94.1 to 176.0 µg.mL-1, and did not show significant hemolytic effect. In the investigation of synergistic interaction, the 4:1 Lim-Car combination showed better fractional inhibitory concentration (FIC) index as well as better activity on amastigotes and IS. The samples caused considerable damage to the parasite membrane this monoterpene activity seems to be more related to Trypanothione Reductase (TryR) enzyme interaction, demonstrated in the molecular docking assay. In addition, the 4:1 Lim-Car combination stimulated macrophage activation, and showed at was the best association, with reduction of infection and infectivity of parasitized macrophages. CONCLUSION: The 4:1 Lim-Car combination appears to be a promising candidate as a monotherapeutic antileishmania agent.
Assuntos
Antiprotozoários/toxicidade , Cimenos/toxicidade , Fatores Imunológicos/toxicidade , Leishmania major/efeitos dos fármacos , Limoneno/toxicidade , Animais , Sobrevivência Celular/efeitos dos fármacos , DNA Polimerase Dirigida por DNA/metabolismo , Combinação de Medicamentos , Sinergismo Farmacológico , Eritrócitos/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Lisossomos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/parasitologia , Simulação de Acoplamento Molecular , NADH NADPH Oxirredutases/metabolismo , Proteínas de Protozoários/metabolismo , OvinosRESUMO
A pandemia de Covid-19 impactou a vida dos indivíduos, causando adoecimento, sofrimento psíquico e morte de milhares de pessoas. Propõe-se apresentar, neste artigo, estratégias metodológicas de pesquisa-intervenção com objetivo de rastrear casos de transtorno mental comum (TMC) em trabalhadores(as) no estado da Bahia no contexto da pandemia. Foi realizada revisão bibliográfica para delineamento da proposta de vigilância em saúde a partir dos descritores "transtornos mentais", "Covid-19", "pandemia por Coronavírus e transtorno mental" e "Covid-19", com busca na base Medline e no site Google Acadêmico. Delimitou-se ações em duas dimensões específicas: (1) rastreamento dos casos de TMC a partir dos sistemas de informação e-SUS VE e Sivep-Gripe; articulação com entidades representativas de trabalhadores(as) e rastreamento de tentativas de suicídios e óbitos por causas externas a partir de dados secundários; (2) investigação da relação do transtorno mental com o trabalho e sua consequente notificação em sistema de informação. O rastreamento dos TMC em trabalhadores(as) permitirá conhecer a magnitude do evento na população. O delineamento de projeto piloto como uma estratégia de ação possibilita a identificação de facilitadores e barreiras no processo de implantação, tendo em vista a adequação e/ou ampliação da intervenção para outras regiões do estado, no âmbito da Rede Estadual de Saúde do Trabalhador. A articulação interinstitucional ensino-serviço no desenvolvimento do projeto piloto amplia a capacidade de compreensão e delimitação de respostas da vigilância em saúde à problemática, subsidiando ações individuais e coletivas, de assistência e prevenção dos transtornos mentais na população de trabalhadores(as).
The Covid-19 pandemic affected the peoples' life, causing illness, psychological distress and death. This article presents methodological research-intervention strategies to survey cases of common mental disorder (CMD) in workers of the state of Bahia relating it to the pandemic. A Bibliographic review was conducted to outline the health surveillance proposal based on the descriptors: "transtornos mentais", "Covid-19", "pandemia por Coronavírus e transtorno mental" and Covid-19, with search on the Medline database and Google Scholar website. Actions were delimited in specific dimensions: (1) searching for CMD cases from the e-SUS VE and Sivep-Gripe information systems; articulation with representative bodies of workers and search for suicide attempts and deaths from external causes based on secondary data; (2) investigating of the relationship between mental disorder and work and its consequent notification in an information system. The survey of CMD in workers will allow knowing the magnitude of the event in the population. The design of a pilot project as an action strategy allows the identification of facilitators and barriers in the implementation process considering the adapting and/or expanding needs of the intervention to other regions within the State Workers' Health Network. The interinstitutional teaching-service articulation expands the capacity for understanding and delimiting health surveillance responses, supporting individual and collective actions, assistance and prevention of mental disorders in the worker population.
La pandemia del Covid-19 afectó a la vida de las personas causando enfermedades, angustia psicológica y muerte. En este artículo se propone presentar estrategias metodológicas de investigación-intervención con el objetivo de rastrear casos de trastorno mental común (TMC) en trabajadores del estado de Bahía en el contexto de la pandemia. Se realizó una revisión bibliográfica para perfilar la propuesta de vigilancia de la salud a partir de los descriptores: "trastornos mentales"; "Covid-19"; "Pandemia de coronavirus y trastorno mental" y "Covid-19", con búsqueda en la base de datos Medline y el sitio web de Google Scholar. Las acciones se delimitaron en dos dimensiones específicas: (1) seguimiento de casos de TMC en los sistemas de información e-SUS VE y Sivep-Gripe; articulación con órganos representativos de trabajadores y seguimiento de intentos de suicidio y muertes por causas externas con base en datos secundarios; (2) investigación de la relación entre trastorno mental y trabajo y su consiguiente notificación en un sistema de información. El rastreo de los trastornos mentales en trabajadores permitirá conocer su magnitud en la población. El diseño de un proyecto piloto como estrategia de acción posibilita la identificación de facilitadores y barreras en el proceso de implementación con miras a adaptar y/o expandir la intervención a otras regiones del estado, en la Red Estadual de Salud de los Trabajadores. La articulación interinstitucional docencia-servicio en el desarrollo del proyecto piloto amplía la capacidad para comprender y delimitar las respuestas de vigilancia en salud a la problemática, subsidiando acciones individuales y colectivas, de atención y prevención de trastornos mentales en la población de trabajadores.
Assuntos
Humanos , Masculino , Feminino , Saúde Mental , Vigilância em Saúde Pública , COVID-19/psicologia , Transtornos Mentais/epidemiologia , Projetos de Pesquisa , Brasil/epidemiologia , Programas de Rastreamento , Saúde Ocupacional , Transtornos Mentais/diagnósticoRESUMO
Apesar de incipientes, estudos têm contribuído com orientações a respeito de procedimentos para o afastamento de trabalhadores expostos direta ou indiretamente ao SARS-CoV-2, com sintomas de Síndrome Gripal ou de Síndrome Respiratória Aguda Grave ou que testaram positivo para Covid-19. Ainda há lacunas sobre as condutas necessárias para avaliação do retorno ao trabalho que extrapolem a abordagem clínica. Este estudo visa orientar a tomada de decisão, o planejamento e a adoção de medidas eficazes de proteção à saúde dos trabalhadores que retornam ao ambiente de trabalho após adoecimento por Covid-19. Realizou-se revisão da produção técnico-científica sobre Covid-19, afastamento e retorno ao trabalho e sistematização em documento orientador sobre o tema. Propõe-se elementos essenciais para a avaliação de retorno ao trabalho após adoecimento por Covid-19 mediante desenvolvimento de ações sincrônicas nas dimensões indivíduo, ambiente de trabalho e atores sociais. Tais ações consideram desde a real situação de saúde do(a) trabalhador(a) até as condições do ambiente de trabalho para o qual o(a) trabalhador(a) irá retornar, mediadas pelos atores envolvidos. A decisão sobre o tempo de afastamento do trabalho pautada apenas em critérios clínicos tende a ser restritiva e insuficiente frente à complexidade do processo de adoecimento por Covid-19, devendo-se incorporar aspectos psicossociais e adequações nos locais de trabalho.
Although incipient, studies have contributed with guidance regarding procedures for the removal of workers exposed directly or indirectly to SARS-CoV-2; with symptoms of Flu Syndrome or Severe Acute Respiratory Syndrome; and/or tested positive for Covid-19. There is still a gap on the necessary conducts for returning to work that go beyond the clinical approach. This study aims at guiding decision-making, planning and adoption of effective measures to protect the health of workers that return to the work environment after falling ill by Covid-19. A review of the technical-scientific production on Covid-19, removal and return to work was conducted, and systematized in a guiding document on the subject. Essential elements are proposed for the assessment of return to work after illness by Covid-19 by the development of synchronous actions in the individual, work environment and social actor dimensions. Such actions consider the real health situation of the worker and the conditions of the work environment to which the worker will return, mediated by the actors involved. The decision on the time away from work based only on clinical criteria tends to be restrictive and insufficient considering the complexity of the illness process by Covid-19, and psychosocial aspects and adaptations in the workplace should be incorporated.
Aunque incipientes, los estudios han contribuido con la orientación sobre los procedimientos del alejamiento de trabajadores expuestos directa o indirectamente al SARS-CoV-2; con síntomas de síndrome de gripe o síndrome respiratorio agudo severo, o que dieron positivo para Covid-19. Aún existen lagunas en las conductas necesarias para la reinserción al trabajo que van más allá del abordaje clínico. Este estudio tiene como objetivo orientar la toma de decisiones, la planificación y la adopción de medidas efectivas de prevención a la salud de los trabajadores que regresan al entorno laboral después de enfermarse por Covid-19. Se realizó una revisión de la producción técnico-científica sobre Covid-19, alejamiento y vuelta al trabajo y una sistematización en un documento guía sobre el tema. Se proponen elementos esenciales para evaluar la reinserción al trabajo después de la enfermedad por Covid-19 mediante el desarrollo de acciones sincrónicas en las dimensiones individual, de ambiente laboral y de actores sociales. Tales acciones contemplan desde la situación real de salud del trabajador hasta las condiciones del entorno laboral al que retornará el trabajador, mediadas por los actores involucrados. La decisión sobre el tiempo de baja laboral basada únicamente en criterios clínicos tiende a ser restrictiva e insuficiente ante la complejidad del proceso de enfermedad por Covid-19, debiendo incorporarse aspectos psicosociales y adaptaciones en el ámbito laboral.
Assuntos
Condições de Trabalho , Saúde Ocupacional , Infecções por Coronavirus , Síndrome Respiratória Aguda Grave , Influenza Humana , Pandemias , Retorno ao TrabalhoRESUMO
Megacoelomyces (type species: Megacoelomyces sanchezii), an ascomycete asexual morph infecting Myrcia fenzliana (Myrtaceae) from the Brazilian Cerrado, is described as a new genus in the Phaeosphaeriaceae (Pleosporales, Dothideomycetes, Ascomycota), based on multilocus phylogeny (three nuclear ribosomal DNA and two protein-coding genes) in addition to morphological (light and scanning electron microscopy) and ecological data.