Metabolomics of the interaction between a consortium of entomopathogenic fungi and their target insect: Mechanisms of attack and survival.

One of the most concerning pests that attack strawberries in Brazil is Duponchelia fovealis (Zeller), a non-native moth with no registered control methods to date. Our group recently observed that a fungal consortium formed by two strains of Beauveria bassiana (Balsamo) increased the mortality of D. fovealis more than inoculation with each strain on its own. However, the molecular interaction between the fungal consortium and the caterpillars is unknown. Thus, in this work, we sought to pioneer the evaluation of the molecular interaction between a fungal consortium of B. bassiana and D. fovealis caterpillars. We aimed to understand the biocontrol process involved in this interaction and the defense system of the caterpillar. Seven days after D. fovealis were inoculated with the consortium, the dead and surviving caterpillars were analyzed using GC-MS and LC-MS. Some of the metabolites identified in dead caterpillars have primarily antioxidant action. Other metabolites may have insecticidal potential, such as diltiazem-like and tamsulosin-like compounds, as well as 2,5-dimethoxymandelic acid. In surviving caterpillars, the main mechanisms are pro-inflammatory from 2-Palmitoylglycerol metabolite and the antifungal action of the metabolite Aegle marmelos Alkaloid-C. The metabolites identified in dead caterpillars may explain the increased mortality caused by the consortium due to its antioxidant mechanism, which can suppress the caterpillars' immune system, and insecticide action. In surviving caterpillars, the main resistance mechanisms may involve the stimulus to the immunity and antifungal action.

Fungal consortium of two Beauveria bassiana strains increases their virulence, growth, and resistance to stress: A metabolomic approach.

The use of two or more microorganisms in a microbial consortium has been increasingly applied in the biological control of diseases and pests. Beauveria bassiana is one of the most widely studied fungal species in biological control, yet little is known about its role in fungal consortiums. In a previous study, our group found that a consortium formed by two strains of B. bassiana had significantly greater biocontrol potential against the polyphagous caterpillars Duponchelia fovealis (Lepidoptera: Crambidae) than either strain on its own. In this study, we use GC-MS and LC-MS/MS to evaluate and discuss the metabolomics of the consortium. A total of 21 consortium biomarkers were identified, corresponding to 14 detected by LC-MS/MS and seven by GC-MS. Antioxidant and anti-inflammatory mechanisms are the main properties of the metabolites produced by the consortium. These metabolites can depress the insect's immune system, increasing its vulnerability and, hence, the fungal virulence of the consortium. In light of these results, we propose an action model of insect mortality due to the metabolites secreted by the consortium. The model includes the inhibition of defense mechanisms such as pro-inflammatory interleukin secretion, cell migration, cell aggregation, Dif, Dorsal and Relish gene transcription, and JAK/STAT and JNK signaling pathways. It also promotes the cleaning of oxidative molecules, like ROS, NOS, and H2O2, and the induction of virulence factors.

Modulation of Photoaging-Induced Cutaneous Elastin: Evaluation of Gene and Protein Expression of Markers Related to Elastogenesis Under Different Photoexposure Conditions.

INTRODUCTION: Photoaging is the process by which ultraviolet rays gradually induce clinical and histological changes in the skin through the production and organization of biological molecules, such as elastin, which is critical to skin strength and elasticity. After exposure to radiation, elastin may undergo alternative mRNA splicing, resulting in modified proteins that contribute to the formation of aging characteristics, such as solar elastosis. The present work aimed to study two different forms of elastin under these conditions: normal elastin and elastin that had been altered in exon 26A. METHODS: These different forms of elastin were characterized for gene expression by quantitative real-time polymerase chain reaction (qPCR) and for protein expression by immunohistochemistry of ex vivo skins (from photoexposed and non-photoexposed areas) and in vitro reconstituted skin. In addition, up- and downstream molecules in the elastin signaling cascade were evaluated. RESULTS: As a result, a significant increase in the gene expression of elastin 26A was observed in both ex vivo photoexposed skin tissues and the in vitro photoexposed reconstituted skins. Additionally, significant increases in the gene expression levels of matrix metalloproteinase-12 (MMP12) and lysyl oxidase (LOX) were observed in the ex vivo skin model. The evaluation of protein expression levels of some photoaging markers on the reconstituted skin revealed increased tropoelastin and fibrillin-1 expression after photoexposure. CONCLUSION: This work contributes to a better understanding of the biological mechanisms involved in photoaging, making it possible to obtain new strategies for the development of dermocosmetic active ingredients to prevent and treat skin aging.

Triple nanoemulsion potentiates the effects of topical treatments with microencapsulated retinol and modulates biological processes related to skin aging / Tripla nanoemulsao potencializa os efeitos do tratamento topico com retinol microencapsulado e modula processos biologicos relacionados ao envelhecimento da pele

BACKGROUND: The sum of environmental and genetic factors affects the appearance and function of the skin as it ages. The identification of molecular changes that take place during skin aging provides biomarkers and possible targets for therapeutic intervention. Retinoic acid in different formulations has emerged as an alternative to prevent and repair age-related skin damage. OBJECTIVES: To understand the effects of different retinoid formulations on the expression of genes associated with biological processes that undergo changes during skin aging. METHODS: Ex-vivo skin samples were treated topically with different retinoid formulations. The modulation of biological processes associated with skin aging was measured by Reverse Transcription quantitative PCR (RT-qPCR). RESULTS: A formulation containing microencapsulated retinol and a blend of active ingredients prepared as a triple nanoemulsion provided the best results for the modulation of biological, process-related genes that are usually affected during skin aging. CONCLUSION: This association proved to be therapeutically more effective than tretinoin or microencapsulated retinol used singly. .

FUNDAMENTOS: A soma de fatores genéticos e ambientais afeta a aparência e a funcionalidade da pele ao longo do envelhecimento. O conhecimento a respeito das mudanças moleculares durante o envelhecimento fornece biomarcadores e possíveis alvos para intervenções terapêuticas. O ácido retinoico em diferentes formulações surgiu como uma alternativa para prevenir e reparar os danos da pele associados ao envelhecimento. OBJETIVOS: Avaliar comparativamente os efeitos de diferentes formulações contendo retinoides na expressão de genes associados a processos biológicos que são alterados com o envelhecimento da pele. MÉTODOS: Peles ex vivo foram topicamente tratadas com diferentes retinoides, micro e nanoencapsulados. A modulação dos processos biológicos associados ao envelhecimento da pele foi medida por PCR quantitativa, precedida de transcrição reversa (RT-qPCR). RESULTADOS: A formulação contendo uma mistura de princípios ativos incorporados em uma tripla nanoemulsão e retinol microencapsulado apresentou os melhores resultados de modulação de genes relacionados a processos biológicos que são normalmente alterados durante o envelhecimento da pele. CONCLUSÃO: Essa associação demonstrou uma maior eficácia terapêutica quando comparada ao uso isolado de tretinoína ou retinol microencapsulado. .

Triple nanoemulsion potentiates the effects of topical treatments with microencapsulated retinol and modulates biological processes related to skin aging.

BACKGROUND: The sum of environmental and genetic factors affects the appearance and function of the skin as it ages. The identification of molecular changes that take place during skin aging provides biomarkers and possible targets for therapeutic intervention. Retinoic acid in different formulations has emerged as an alternative to prevent and repair age-related skin damage. OBJECTIVES: To understand the effects of different retinoid formulations on the expression of genes associated with biological processes that undergo changes during skin aging. METHODS: Ex-vivo skin samples were treated topically with different retinoid formulations. The modulation of biological processes associated with skin aging was measured by Reverse Transcription quantitative PCR (RT-qPCR). RESULTS: A formulation containing microencapsulated retinol and a blend of active ingredients prepared as a triple nanoemulsion provided the best results for the modulation of biological, process-related genes that are usually affected during skin aging. CONCLUSION: This association proved to be therapeutically more effective than tretinoin or microencapsulated retinol used singly.

Culturable endophytic filamentous fungi from leaves of transgenic imidazolinone-tolerant sugarcane and its non-transgenic isolines.

The diversity of endophytic filamentous fungi from leaves of transgenic imidazolinone-tolerant sugarcane plants and its isoline was evaluated by cultivation followed by amplified rDNA restriction analysis (ARDRA) of randomly selected strains. Transgenic and non-transgenic cultivars and their crop management (herbicide application or manual weed control) were used to assess the possible non-target effects of genetically modified sugarcane on the fungal endophytic community. A total of 14 ARDRA haplotypes were identified in the endophytic community of sugarcane. Internal transcribed spacer (ITS) sequencing revealed a rich community represented by 12 different families from the Ascomycota phylum. Some isolates had a high sequence similarity with genera that are common endophytes in tropical climates, such as Cladosporium, Epicoccum, Fusarium, Guignardia, Pestalotiopsis and Xylaria. Analysis of molecular variance indicated that fluctuations in fungal population were related to both transgenic plants and herbicide application. While herbicide applications quickly induced transient changes in the fungal community, transgenic plants induced slower changes that were maintained over time. These results represent the first draft on composition of endophytic filamentous fungi associated with sugarcane plants. They are an important step in understanding the possible effects of transgenic plants and their crop management on the fungal endophytic community.

Contaminação fúngica em chás de camomila, erva-doce e erva-mate / Fungi contamination in the chamomile, anis and mate teas

A avaliação da qualidade microbiológica de produtos armazenados, utilizados como plantas medicinais, é fundamental para garantir a segurança alimentar, em função do potencial micotoxigênico apresentado por algumas espécies de fungos. No presente trabalho foram realizados o isolamento, a quantificação e a identificação de fungos potencialmente produtores de micotoxinas em amostras de chás de camomila (Chamomilla recutita L.), erva-doce (Pimpinella anisum L.) e erva-mate (Ilex paraguariensis St.-Hil.), adquiridas em estabelecimentos comerciais da cidade de Curitiba, PR, Brasil. As amostras foram analisadas nas formas de infusão fria, de infusão tradicional e de cocção. Não houve diferenças significativas nos valores de UFC/g nas amostras de camomila, erva-mate e erva-doce analisadas na forma de infusão. Foram observadas diferenças significativas em amostras de camomila e erva-doce em forma de infusão fria em relação às demais formas de processamento de amostras, contudo essas não foram evidentes nas amostras de erva-mate. Os principais gêneros de fungos isolados das amostras analisadas foram Aspergillus sp (35,9%); Penicillium sp (9,4%); Fusarium sp (0,21%); Rhizopus sp (11,5%), Ulocladium sp (18,4%) e Mycelia sterilia (6,84%). Entre os fungos potencialmente toxigênicos, o Aspergillus sp foi o mais frequente nos três tipos de chás, seguido de Penicillium sp e Fusarium sp. Considerando que os fungos com potencial micotoxigênico persistem mesmo após a infusão ou cocção, recomenda-se que estratégias sejam desenvolvidas para garantir a qualidade e a segurança alimentar dos produtos consumidos pela população.

Identification and colonization of endophytic fungi from soybean (Glycine max (L) Merril) under different environmental conditions

A total of 297 endophytic fungi were isolated from 1728 leaf and stem fragments collected about twenty and forty days after germination from soybean (Glycine max (L.) Merril) plants grown in the field and a greenhouse. The fungi belonged to eight groups, six dematiaceous genera (Alternaria, Cladosporium, Chaetomium, Curvularia, Drechslera and Scopulariopsis) and the non-dematiaceous genera Acremonium, Aspergillus, Colletotrichum, Fusarium, Paecilomyces and Penicillium along with some Mycelia sterilia.. There were qualitative and quantitative differences in the type and number of isolates obtained from greenhouse and field-grown plants, with more isolates being obtained from the latter. No difference was found in the number of fungi isolated from leaves and stems irrespective of where the plants was grown. For was field-grown plants, the number of isolates decreased as the plants aged and more fungi were found in tissues near the soil, while for greenhouse-grown plants the number of isolates increased as the plants aged but in this case no more fungi were isolated from those tissues nearer the soil. These results could have biotechnological relevance for the biological control of pests or plant growth promotion.

A partir de 1728 fragmentos de hastes e folhas de soja (Glycine max (L.) Merril). provenientes de plantas do campo e de casa de vegetação, coletadas cerca de 20 e 40 dias após a germinação das sementes, 297 fungos endofíticos foram isolados. Os gêneros encontrados foram: Alternaria, Cladosporium, Curvularia, Chaetomium, Scopulariopsis, Drechslera (todos dematiáceos) além de Colletotrichum, Fusarium, Acremonium, Aspergillus, Penicillium, Paecilomyces e Mycelia sterilia. Foram detectadas diferenças qualitativas e quantitativas entre os isolados, em relação a micobiota de hospedeiros provenientes do campo e de casa de vegetação com maior frequência de fungos isolados de plantas no campo em comparação com as de casa de vegetação. Houve diminuição da frequência de fungos com a idade das plantas crescidas no campo, ocorrendo o inverso com as da casa de vegetação, mas em ambos os casos, não houve diferença no número de fungos isolados de hastes e de folhas. Houve em plantas crescidas no campo uma maior incidência nas regiões próximas ao solo o que não ocorreu nas plantas provenientes da casa de vegetação. Os resultados podem ser de utilidade do ponto de vista biotecnológico pelo uso de endófitos introduzidos deliberadamente em plantas visando o controle biológico de pestes ou promoção de crescimento vegetal.