Cardosins improve neuronal regeneration after cell disruption: a comparative expression study

Duarte, Ana Sofia; Duarte, Emília P.; Correia, António; Pires, Euclides; Barros, Marlene

doi:10.1007/s10565-008-9058-x

Cited by 5 publications

(4 citation statements)

References 29 publications

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“…Neurons treated with cardosins exhibit normal morphology and function when compared to cultures prepared with the widely used trypsin-dissociation method (Duarte et al, 2007). Furthermore, use of cardosins resulted in an improvement of neuronal regeneration at an early stages of culture after cell dissociation (Duarte et al, 2009).…”

Section: Biotechnological and Biomedical Applicationsmentioning

confidence: 99%

Properties and applications of phytepsins from thistle flowers

et al. 2013

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Section: Biotechnological and Biomedical Applicationsmentioning

confidence: 99%

Properties and applications of phytepsins from thistle flowers

et al. 2013

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“…Gelatinolytic activity was assessed by zymography, as described previously [ 48 ], with slight modifications. Gelatin [1% ( w / v )] was used.…”

Section: Methodsmentioning

confidence: 99%

Lasiodiplodia theobromae as a Producer of Biotechnologically Relevant Enzymes

Félix

Libório

Nunes

et al. 2018

IJMS

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Phytopathogenic fungi are known to produce several types of enzymes usually involved in plant cell wall degradation and pathogenesis. The increasing of global temperature may induce fungi, such as Lasiodiplodia theobromae (L. theobromae), to alter its behavior. Nonetheless, there is only limited information regarding the effect of temperature on L. theobromae production of enzymes. The need for new, thermostable enzymes, that are biotechnologically relevant, led us to investigate the effect of temperature on the production of several extracellular enzymatic activities by different L. theobromae strains. Fungi were grown at 25 °C, 30 °C and 37 °C and the enzymatic activities were detected by plate assays, quantified by spectrophotometric methods and characterized by zymography. The thermostability (25–80 °C) of the enzymes produced was also tested. Strains CAA019, CBS339.90, LA-SOL3, LA-SV1 and LA-MA-1 produced amylases, gelatinases, caseinases, cellulases, lipases, laccases, xylanases, pectinases and pectin liases. Temperature modulated the expression of the enzymes, and this effect was more visible when fungi were grown at 37 °C than at lower temperatures. Contrary to proteolytic and endoglucanolytic activities, whose highest activities were detected when fungi were grown at 30 °C, lipolytic activity was not detected at this growth temperature. Profiles of proteases and endoglucanases of fungi grown at different temperatures were characterized by zymography. Enzymes were shown to be more thermostable when fungi were grown at 30 °C. Proteases were active up to 50 °C and endoglucanases up to 70 °C. Lipases were the least stable, with activities detected up to 45 °C. The enzymatic profiles detected for L. theobromae strains tested showed to be temperature and strain-dependent, making this species a good target for biotechnological applications.

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“…Gelatinolytic and caseinolytic activity was assessed by zymography, using 1% (w/v) gelatine or 1% (w/v) casein, as described previously ( Duarte et al, 2009 ; Esteves et al, 2014 ), with slight modifications. After electrophoresis, the gel was incubated overnight, at room temperature, in 1.5 mM Tris, pH 8.8, 1 M NaCl, 1 M CaCl 2 , 2 mM ZnCl 2 , pH 7.4, stained with Coomassie Brilliant Blue R-250 [(in 50% ethanol (v/v), 10% acetic acid (v/v)] and destained with 25% ethanol (v/v), 5% acetic acid (bgv/v).…”

Section: Methodsmentioning

confidence: 99%

Temperature Modulates the Secretome of the Phytopathogenic Fungus Lasiodiplodia theobromae

et al. 2016

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Environmental alterations modulate host–microorganism interactions. Little is known about how climate changes can trigger pathogenic features on symbiont or mutualistic microorganisms. Current climate models predict increased environmental temperatures. The exposing of phytopathogens to these changing conditions can have particularly relevant consequences for economically important species and for humans. The impact on pathogen/host interaction and the shift on their biogeographical range can induce different levels of virulence in new hosts, allowing massive losses in agricultural and health fields. Lasiodiplodia theobromae is a phytopathogenic fungus responsible for a number of diseases in various plants. It has also been described as an opportunist pathogen in humans, causing infections with different levels of severity. L. theobromae has a high capacity of adaptation to different environments, such as woody plants, moist argillaceous soils, or even humans, being able to grow and infect hosts in a wide range of temperatures (9–39°C). Nonetheless, the effect of an increase of temperature, as predicted in climate change models, on L. theobromae is unknown. Here we explore the effect of temperature on two strains of L. theobromae – an environmental strain, CAA019, and a clinical strain, CBS339.90. We show that both strains are cytotoxic to mammalian cells but while the environmental strain is cytotoxic mainly at 25°C, the clinical strain is cytotoxic mainly at 30 and 37°C. Extracellular gelatinolytic, xylanolytic, amylolytic, and cellulolytic activities at 25 and 37°C were characterized by zymography and the secretome of both strains grown at 25, 30, and 37°C were characterized by electrophoresis and by Orbitrap LC-MS/MS. More than 75% of the proteins were identified, mostly enzymes (glycosyl hydrolases and proteases). The strains showed different protein profiles, which were affected by growth temperature. Also, strain specific proteins were identified, such as a putative f5/8 type c domain protein – known for being involved in pathogenesis – by strain CAA019 and a putative tripeptidyl-peptidase 1 protein, by strain CBS339.90. We showed that temperature modulates the secretome of L. theobromae. This modulation may be associated with host-specificity requirements. We show that the study of abiotic factors, such as temperature, is crucial to understand host/pathogen interactions and its impact on disease.

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Cardosins improve neuronal regeneration after cell disruption: a comparative expression study

Cited by 5 publications

References 29 publications

Properties and applications of phytepsins from thistle flowers

Properties and applications of phytepsins from thistle flowers

Lasiodiplodia theobromae as a Producer of Biotechnologically Relevant Enzymes

Temperature Modulates the Secretome of the Phytopathogenic Fungus Lasiodiplodia theobromae

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