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1.
Mar Drugs ; 21(5)2023 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-37233466

RESUMO

Cassiopea andromeda (Forsskål, 1775), commonly found across the Indo-Pacific Ocean, the Red Sea, and now also in the warmest areas of the Mediterranean Sea, is a scyphozoan jellyfish that hosts autotrophic dinoflagellate symbionts (family Symbiodiniaceae). Besides supplying photosynthates to their host, these microalgae are known to produce bioactive compounds as long-chain unsaturated fatty acids, polyphenols, and pigments, including carotenoids, with antioxidant properties and other beneficial biological activities. By the present study, a fractionation method was applied on the hydroalcoholic extract from two main body parts (oral arms and umbrella) of the jellyfish holobiont to obtain an improved biochemical characterization of the obtained fractions from the two body parts. The composition of each fraction (i.e., proteins, phenols, fatty acids, and pigments) as well as the associated antioxidant activity were analyzed. The oral arms proved richer in zooxanthellae and pigments than the umbrella. The applied fractionation method was effective in separating pigments and fatty acids into a lipophilic fraction from proteins and pigment-protein complexes. Therefore, the C. andromeda-dinoflagellate holobiont might be considered as a promising natural source of multiple bioactive compounds produced through mixotrophic metabolism, which are of interest for a wide range of biotechnological applications.


Assuntos
Cnidários , Cifozoários , Animais , Cifozoários/química , Antioxidantes/farmacologia , Antioxidantes/química , Proteínas , Ácidos Graxos
2.
Foods ; 11(24)2022 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-36553715

RESUMO

This study describes the set-up and optimization of a fermentation strategy applied to a composite raw material containing jellyfish biomass as the principal ingredient. New fermented food was developed by combining fresh jellyfish Rhizostoma pulmo and the sequential solid-state submerged liquid fermentation method used in Asian countries for processing a high-salt-containing raw material. Aspergillus oryzae was used to drive the first fermentation, conducted in solid-state conditions, of a jellyfish-based product, here named Jelly paste. The second fermentation was performed by inoculating the Jelly paste with different selected bacteria and yeasts, leading to a final product named fermented Jellyfish paste. For the first time, a set of safety parameters necessary for monitoring and describing a jellyfish-based fermented food was established. The new fermented products obtained by the use of Debaryomyces hansenii BC T3-23 yeast strain and the Bacillus amyloliquefaciens MS3 bacterial strain revealed desirable nutritional traits in terms of protein, lipids and total phenolic content, as well as valuable total antioxidant activity. The obtained final products also showed a complex enzyme profile rich in amylase, protease and lipase activities, thus making them characterized by unique composite sensory odor descriptors (umami, smoked, dried fruit, spices).

3.
Foods ; 11(17)2022 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-36076882

RESUMO

Edible jellyfish are a traditional Southeast Asian food, usually prepared as a rehydrated product using a salt and alum mixture, whereas they are uncommon in Western Countries and considered as a novel food in Europe. Here, a recently developed, new approach for jellyfish processing and stabilization with calcium salt brining was upgraded by modifying the pre-treatment step of freshly caught jellyfish and successfully applied to several edible species. Treated jellyfish obtained by the application of the optimized version of this method respected both quality and safety parameters set by EU law, including no pathogenic microorganisms, absence or negligible levels of histamine and of total volatile basic nitrogen, no heavy metals; and the total bacterial, yeast, and mold counts were either negligible or undetectable. Jellyfish treated by the presented method exhibited unique protein content, amino acid and fatty acid profiles, antioxidant activity, and texture. The optimized method, initially set up on Rhiszostoma pulmo, was also successfully applied to other edible jellyfish species (such as Cotylorhiza tuberculata, Phyllorhiza punctata, and Rhopilema nomadica) present in the Mediterranean Sea. This study discloses an innovative process for the preparation of jellyfish-based food products for potential future distribution in Europe.

4.
Front Nutr ; 8: 718798, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34497822

RESUMO

Jellyfish, marketed and consumed as food in The Far East, are traditionally processed using salt and alum mixtures. In recent years, the interest of Western consumers in jellyfish (JF) as a food source is increasing. In Europe [European Union (EU)], JF-derived food products are regulated by a novel food law, but methods for JF treatment and processing have not been developed yet. In this study, a protocol for the stabilization and processing of JF into semi-finished food products without the use of alum is proposed for the first time. Safety and quality parameters, together with a series of technological and nutritional traits, were used to monitor the proposed process and for the characterization of the JF-derived products. Calcium lactate (E327), calcium citrate (E333), and calcium acetate (E263), which are food thickening/stabilizing agents allowed by EU regulations, were used in order to control the presence of possible microbial pathogens and spoilage species. The use of calcium lactate and citrate led to an increase in texture values (~1.7-1.8-fold higher than in starting raw materials) and in several nutritional traits such as antioxidant activity, and protein and fatty acid content. In particular, the combination of JF treatments with calcium salts and phenolic compounds resulted in an antioxidant activity increase of up to 8-fold, protein concentration increase of up to 2.6-fold, fatty acid composition maintenance, and a ω6/ω3 ratio lower than 1. For the first time, the application of phenolic compounds to improve JF technological and nutritional features was verified. This study proposes a new procedure for JF treatment and stabilization useful for future potential food applications in Western countries.

5.
Mar Drugs ; 19(9)2021 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-34564160

RESUMO

Increasing frequency of native jellyfish proliferations and massive appearance of non-indigenous jellyfish species recently concur to impact Mediterranean coastal ecosystems and human activities at sea. Nonetheless, jellyfish biomass may represent an exploitable novel resource to coastal communities, with reference to its potential use in the pharmaceutical, nutritional, and nutraceutical Blue Growth sectors. The zooxanthellate jellyfish Cassiopea andromeda, Forsskål, 1775 (Cnidaria, Rhizostomeae) entered the Levant Sea through the Suez Canal and spread towards the Western Mediterranean to reach Malta, Tunisia, and recently also the Italian coasts. Here we report on the biochemical characterization and antioxidant activity of C. andromeda specimens with a discussion on their relative biological activities. The biochemical characterization of the aqueous (PBS) and hydroalcoholic (80% ethanol) soluble components of C. andromeda were performed for whole jellyfish, as well as separately for umbrella and oral arms. The insoluble components were hydrolyzed by sequential enzymatic digestion with pepsin and collagenase. The composition and antioxidant activity of the insoluble and enzymatically digestible fractions were not affected by the pre-extraction types, resulting into collagen- and non-collagen-derived peptides with antioxidant activity. Both soluble compounds and hydrolyzed fractions were characterized for the content of proteins, phenolic compounds, and lipids. The presence of compounds coming from the endosymbiont zooxanthellae was also detected. The notable yield and the considerable antioxidant activity detected make this species worthy of further study for its potential biotechnological sustainable exploitation.


Assuntos
Suplementos Nutricionais , Cifozoários , Animais , Antioxidantes , Organismos Aquáticos , Ecossistema , Mar Mediterrâneo
6.
Plants (Basel) ; 9(8)2020 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-32731416

RESUMO

The production of crocin, an uncommon and valuable apocarotenoid with strong biological activity, was obtained in a cell suspension culture of saffron (Crocus sativus L.) established from style-derived calli to obtain an in-vitro system for metabolite production. Salycilic acid (SA) was used at different concentrations to elicit metabolite production, and its effect was analyzed after a 4 days of treatment. HPLC-DAD analysis was used for total crocin quantification while the Folin-Ciocâlteu method was applied for phenolic compounds (PC) content. Interestingly, despite cell growth inhibition, a considerable exudation was observed when the highest SA concentration was applied, leading to a 7-fold enhanced production of crocin and a 4-fold increase of phenolics compared to mock cells. The maximum antioxidant activity of cell extracts was evidenced after SA 0.1 mM elicitation. Water-soluble extracts of saffron cells at concentrations of 1, 0.5, and 0.1 µg mL-1 showed significant inhibitory effects on MDA-MB-231 cancer cell viability. The heterologous vacuolar markers RFP-SYP51, GFPgl133Chi, and AleuRFP, were transiently expressed in protoplasts derived from the saffron cell suspensions, revealing that SA application caused a rapid stress effect, leading to cell death. Cell suspension elicitation with SA on the 7th day of the cell growth cycle and 24 h harvest time was optimized to exploit these cells for the highest increase of metabolite production in saffron cells.

7.
Cells ; 8(6)2019 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-31212874

RESUMO

Microglia cells are the primary immune population of the central nervous system with a role in the regulation of several physiological and pathological conditions. Upon appropriate stimulation, microglia cells can be polarized in a pro-inflammatory M1-like or anti-inflammatory M2-like status. Biological processes and pathways engaged in microglia polarization are starting to be elucidated. To help clarify this, we used a liquid chromatography-mass spectrometry (LC-MS/MS) label free approach to characterize the proteomic profile of human microglia cell line (CHME-5) stimulated with gamma-interferon (IFN-γ) and interleukin-4 (IL-4) to induce a M1 or M2 phenotype, respectively. Outside the classical M1/M2 polarization markers, the M1 status appears to center around the activation of a classical inflammatory response and through the activation of multiple signaling pathways. M2 polarization resulted in a different pattern of protein modulation related to RNA and cellular metabolic processes. Together, our findings provide information regarding the protein changes specific to M1 and M2 activation states, and potentially link the polarization of microglia cells to the acquisition of a specific proteomic profile.


Assuntos
Interferon gama/farmacologia , Interleucina-4/farmacologia , Microglia/metabolismo , Proteínas/metabolismo , Linhagem Celular , Humanos , Ativação de Macrófagos/efeitos dos fármacos , Redes e Vias Metabólicas/efeitos dos fármacos , Microglia/efeitos dos fármacos , Fenótipo , Proteômica , Transdução de Sinais/efeitos dos fármacos , Transcrição Gênica/efeitos dos fármacos
8.
Mar Drugs ; 17(2)2019 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-30813405

RESUMO

The jellyfish Rhizostoma pulmo, Macrì 1778 (Cnidaria, Rhizostomae) undergoes recurrent outbreaks in the Mediterranean coastal waters, with large biomass populations representing a nuisance or damage for marine and maritime activities. A preliminary overview of the antioxidant activity (AA) of R. pulmo proteinaceous compounds is provided here based on the extraction and characterization of both soluble and insoluble membrane-fractioned proteins, the latter digested by sequential enzymatic hydrolyses with pepsin and collagenases. All jellyfish proteins showed significant AA, with low molecular weight (MW) proteins correlated with greater antioxidant activity. In particular, collagenase-hydrolysed collagen resulted in peptides with MW lower than 3 kDa, ranging 3⁻10 kDa or 10⁻30 kDa, with AA inversely proportional to MW. No cytotoxic effect was detected on cultured human keratinocytes (HEKa) in a range of protein concentration 0.05⁻20 µg/mL for all tested protein fractions except for soluble proteins higher than 30 kDa, likely containing the jellyfish venom compounds. Furthermore, hydrolyzed jellyfish collagen peptides showed a significantly higher AA and provided a greater protective effect against oxidative stress in HEKa than the hydrolyzed collagen peptides from vertebrates. Due to a high reproductive potential, jellyfish may represent a potential socioeconomic opportunity as a source of natural bioactive compounds, with far-reaching beneficial implications. Eventually, improvements in processing technology will promote the use of untapped marine biomasses in nutraceutical, cosmeceutical, and pharmaceutical fields, turning marine management problems into a more positive perspective.


Assuntos
Antioxidantes/farmacologia , Peptídeos/análise , Peptídeos/farmacologia , Cifozoários/química , Animais , Antioxidantes/análise , Antioxidantes/química , Antioxidantes/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/metabolismo , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Peptídeos/isolamento & purificação
9.
FEBS J ; 286(4): 688-709, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30657636

RESUMO

Mitochondrial dysfunction is a key feature of cancer and is frequently associated with increased aggressiveness and metastatic potential. Recent evidence has brought to light a metabolic rewiring that takes place during the epithelial-to-mesenchymal transition (EMT), a process that drives the invasive capability of malignant tumors, and highlights a mechanistic link between mitochondrial dysfunction and EMT that has been only partially investigated. In this study, we characterized mitochondrial function and bioenergetic status of cultured human breast cancer cell lines, including luminal-like and basal-like subtypes. Through a combination of biochemical and functional studies, we demonstrated that basal-like cell lines exhibit impaired, but not completely inactive, mitochondrial function, and rely on a consequent metabolic switch to glycolysis to support their ATP demand. These altered metabolic activities are linked to modifications of key electron transport chain proteins and a significant increase in levels of reactive oxygen species compared to luminal cells. Furthermore, we observed that the stable knockdown of EMT markers caused functional changes in mitochondria that result in acquisition of a hybrid glycolysis/OXPHOS phenotype in cancer cells as a means to sustain their metabolic demand.


Assuntos
Neoplasias da Mama/patologia , Carcinoma Basocelular/patologia , Reprogramação Celular , Metabolismo Energético , Transição Epitelial-Mesenquimal , Mitocôndrias/patologia , Fosforilação Oxidativa , Trifosfato de Adenosina/metabolismo , Neoplasias da Mama/classificação , Neoplasias da Mama/metabolismo , Carcinoma Basocelular/metabolismo , Feminino , Glicólise , Humanos , Mitocôndrias/metabolismo , Oxirredução , Consumo de Oxigênio , Fenótipo , Espécies Reativas de Oxigênio/metabolismo , Células Tumorais Cultivadas
10.
Physiol Plant ; 165(2): 198-208, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30051613

RESUMO

Multiple stresses are becoming common challenges in modern agriculture due to environmental changes. A large set of phytochemicals collectively known as oxylipins play a key role in responses to several stresses. Understanding the fine-tuned plant responses to multiple and simultaneous stresses could open new perspectives for developing more tolerant varieties. We carried out the molecular and biochemical profiling of genes, proteins and active compounds involved in oxylipin metabolism in response to single/combined salt and wounding stresses on Medicago truncatula. Two new members belonging to the CYP74 gene family were identified. Gene expression profiling of each of the six CYP74 members indicated a tissue- and time-specific expression pattern for each member in response to single/combined salt and wounding stresses. Notably, hormonal profiling pointed to an attenuated systemic response upon combined salt and leaf wounding stresses. Combined, these results confirm the important role of jasmonates in legume adaptation to abiotic stresses and point to the existence of a complex molecular cross-talk among signals generated by multiple stresses.


Assuntos
Medicago truncatula/fisiologia , Oxilipinas/metabolismo , Cloreto de Sódio/farmacologia , Estresse Fisiológico , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genoma de Planta , Medicago truncatula/efeitos dos fármacos , Medicago truncatula/genética , Família Multigênica , Filogenia , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética
11.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1864(3): 344-357, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30578966

RESUMO

Several studies have identified a specific metabolic program that is associated with the process of epithelial-mesenchymal transition (EMT). Whereas much is known about the association between glucose metabolism and EMT, the contribution of lipid metabolism is not still completely understood. Here, we studied epithelial and mesenchymal breast cancer cells by proteomic and lipidomic approaches and identified significant differences that characterised these models concerning specific metabolic enzymes and metabolites including fatty acids and phospholipids. Higher levels of monounsaturated fatty acids together with increased expression of enzymes of de novo fatty acid synthesis is the distinct signature of epithelial with respect to mesenchymal cells that, on the contrary, show reduced lipogenesis, higher polyunsaturated fatty acids level and increased expression of genes involved in the triacylglycerol (TAG) synthesis and lipid droplets formation. In the mesenchymal model, the diacylglycerol acyltransferase (DGAT)-1 appears to be the major enzyme involved in TAG synthesis and inhibition of DGAT1, but not DGAT2, drastically reduces the incorporation of labeled palmitate into TAG. Moreover, knockdown of ß-catenin demonstrated that this metabolic phenotype in under the control of a network of transcriptional factors and that ß-catenin has a specific role in the regulation of lipid metabolism in mesenchymal cells.


Assuntos
Neoplasias da Mama/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Metabolismo dos Lipídeos/fisiologia , Linhagem Celular Tumoral , Ácidos Graxos/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Metabolismo dos Lipídeos/genética , Lipogênese , Metaboloma , Fosfolipídeos , Proteômica , Transcriptoma/genética , Transcriptoma/fisiologia , Triglicerídeos/metabolismo , beta Catenina/metabolismo , beta Catenina/fisiologia
12.
Oxid Med Cell Longev ; 2018: 7512159, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30524660

RESUMO

Hepatocellular carcinoma (HCC) accounts for over 80% of liver cancer cases and is highly malignant, recurrent, drug-resistant, and often diagnosed in the advanced stage. It is clear that early diagnosis and a better understanding of molecular mechanisms contributing to HCC progression is clinically urgent. Metabolic alterations clearly characterize HCC tumors. Numerous clinical parameters currently used to assess liver functions reflect changes in both enzyme activity and metabolites. Indeed, differences in glucose and acetate utilization are used as a valid clinical tool for stratifying patients with HCC. Moreover, increased serum lactate can distinguish HCC from normal subjects, and serum lactate dehydrogenase is used as a prognostic indicator for HCC patients under therapy. Currently, the emerging field of metabolomics that allows metabolite analysis in biological fluids is a powerful method for discovering new biomarkers. Several metabolic targets have been identified by metabolomics approaches, and these could be used as biomarkers in HCC. Moreover, the integration of different omics approaches could provide useful information on the metabolic pathways at the systems level. In this review, we provided an overview of the metabolic characteristics of HCC considering also the reciprocal influences between the metabolism of cancer cells and their microenvironment. Moreover, we also highlighted the interaction between hepatic metabolite production and their serum revelations through metabolomics researches.


Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/terapia , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/terapia , Redes e Vias Metabólicas , Animais , Carcinoma Hepatocelular/metabolismo , Humanos , Neoplasias Hepáticas/metabolismo , Metabolômica , Prognóstico
13.
Biotechnol J ; 13(11): e1700699, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29663725

RESUMO

Several techniques can be used to monitor cell dynamism after a perturbation. Among these, Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D) offers the great advantage to study the mechanical properties of cells in real-time and with a great sensitivity. Here, we used QCM-D to investigate the effects of two cytoskeleton-targeting agents, cytochalasin D (CytoD) and Y27632, on human MCF-7 cells. Cell adhesion on the sensor surface, crucial for in-flow experiments, was obtained by covalent adsorption of a fibronectin (FN) film, an extracellular matrix (ECM) protein. Direct analysis of MCF-7 cells on FN-coated sensor, shows a specific cellular response that was revealed and quantified by QCM-D after drugs exposure. Notably, upon treatment with Y27632, we observed a two-regime dissipation behavior that we associated with specific modifications of actin filaments and signaling proteins providing a link between biophysical and molecular mechanisms. Overall, this approach opens new opportunities for studying cellular response to mechanical cues in different biological conditions.


Assuntos
Técnicas Citológicas/métodos , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/fisiologia , Técnicas de Microbalança de Cristal de Quartzo/métodos , Amidas/farmacologia , Citocalasina D/farmacologia , Fibronectinas , Humanos , Células MCF-7 , Piridinas/farmacologia
14.
J Cell Physiol ; 233(8): 6207-6223, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29327509

RESUMO

Peripheral nerve regeneration is regulated through the coordinated spatio-temporal activation of multiple cellular pathways. In this work, an integrated proteomics and bioinformatics approach was employed to identify differentially expressed proteins at the injury-site of rat sciatic nerve at 20 days after damage. By a label-free liquid chromatography mass-spectrometry (LC-MS/MS) approach, we identified 201 differentially proteins that were assigned to specific canonical and disease and function pathways. These include proteins involved in cytoskeleton signaling and remodeling, acute phase response, and cellular metabolism. Metabolic proteins were significantly modulated after nerve injury to support a specific metabolic demand. In particular, we identified a group of proteins involved in lipid uptake and lipid storage metabolism. Immunofluorescent staining for acyl-CoA diacylglycerol acyltransferase 1 (DGAT1) and DAGT2 expression provided evidence for the expression and localization of these two isoforms in Schwann cells at the injury site in the sciatic nerve. This further supports a specific local regulation of lipid metabolism in peripheral nerve after damage.


Assuntos
Regeneração Nervosa/fisiologia , Doenças do Sistema Nervoso/metabolismo , Nervo Isquiático/metabolismo , Animais , Cromatografia Líquida/métodos , Biologia Computacional/métodos , Diacilglicerol O-Aciltransferase/metabolismo , Feminino , Metabolismo dos Lipídeos/fisiologia , Proteômica/métodos , Ratos , Ratos Sprague-Dawley , Células de Schwann/metabolismo , Transdução de Sinais/fisiologia , Espectrometria de Massas em Tandem/métodos
15.
Plant Physiol ; 176(2): 1531-1546, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29203558

RESUMO

Lipid droplets (LDs) are ubiquitous organelles in plant cells, but their physiological roles are largely unknown. To gain insight into the function of LDs in plants, we have characterized the Arabidopsis homologs of SEIPIN proteins, which are crucial factors for LD biogenesis in yeast and animals. SEIPIN1 is expressed almost exclusively in embryos, while SEIPIN2 and SEIPIN3 have broader expression profiles with maximal levels in embryos and pollen, where LDs accumulate most abundantly. Genetic analysis demonstrates that all three SEIPINs contribute to proper LD biogenesis in embryos, whereas in pollen, only SEIPIN2 and SEIPIN3 play a significant role. The double seipin2 seipin3 and triple seipin mutants accumulate extremely enlarged LDs in seeds and pollen, which hinders their subsequent mobilization during germination. Interestingly, electron microscopy analysis reveals the presence of nuclear LDs attached to type I nucleoplasmic reticulum in triple seipin mutant embryos, supporting that SEIPINs are essential for maintaining the correct polarity of LD budding at the nuclear envelope, restricting it to the outer membrane. In pollen, the perturbations in LD biogenesis and turnover are coupled to reduced germination in vitro and with lower fertilization efficiency in vivo. In seeds, germination per se is not affected in seipin2 seipin3 and triple seipin mutants, but there is a striking increase in seed dormancy levels. Our findings reveal the relevance of SEIPIN-dependent LD biogenesis in pollen transmission and in adjusting the timing of seed germination, two key adaptive traits of great importance in agriculture.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Gotículas Lipídicas/metabolismo , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Cloroplastos/metabolismo , Subunidades gama da Proteína de Ligação ao GTP/genética , Subunidades gama da Proteína de Ligação ao GTP/metabolismo , Germinação , Pólen/genética , Pólen/fisiologia , Sementes/genética , Sementes/fisiologia
16.
Front Physiol ; 8: 544, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28798698

RESUMO

ß-catenin plays an important role as regulatory hub in several cellular processes including cell adhesion, metabolism, and epithelial mesenchymal transition. This is mainly achieved by its dual role as structural component of cadherin-based adherens junctions, and as a key nuclear effector of the Wnt pathway. For this dual role, different classes of proteins are differentially regulated via ß-catenin dependent mechanisms. Here, we applied a liquid chromatography-mass spectrometry (LC-MS/MS) approach to identify proteins modulated after ß-catenin knockdown in the breast cancer cell line MCF-7. We used a label free analysis to compare trypsin-digested proteins from CTR (shCTR) and ß-catenin knockout cells (shßcat). This led to the identification of 98 differentially expressed proteins, 53 of them were up-regulated and 45 down-regulated. Loss of ß-catenin induced morphological changes and a significant modulation of the expression levels of proteins associated with primary metabolic processes. In detail, proteins involved in carbohydrate metabolism and tricarboxylic acid cycle were found to be down-regulated, whereas proteins associated to lipid metabolism were found up-regulated in shßcat compared to shCTR. A loss of mitochondrial mass and membrane potential was also assessed by fluorescent probes in shßcat cells with respect to the controls. These data are consistent with the reduced expression of transcriptional factors regulating mitochondrial biogenesis detected in shßcat cells. ß-catenin driven metabolic reprogramming resulted also in a significant modulation of lipogenic enzyme expression and activity. Compared to controls, ß-catenin knockout cells showed increased incorporation of [1-14C]acetate and decreased utilization of [U-14C]glucose for fatty acid synthesis. Our data highlight a role of ß-catenin in the regulation of metabolism and energy homeostasis in breast cancer cells.

17.
Mol Biosyst ; 13(6): 1131-1141, 2017 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-28429008

RESUMO

Resveratrol, a naturally occurring phytoalexin, has long been known to play an important regulatory role in key functions in cell physiology. This multifunctional role of resveratrol is explained by its ability to interact with several targets of various cell pathways. In the recent past, synthetic chemical modifications have been made in an attempt to enhance the biological effects of resveratrol, including its anti-cancer properties. In this study, we investigated the molecular mechanisms of action of novel trans-restricted analogues of resveratrol in which the C-C double bond of the natural derivative has been replaced by diaryl-substituted imidazole analogues. In ovarian cancer models, the results of in vitro screening revealed that the resveratrol analogues exhibited enhanced anti-proliferative properties compared with resveratrol. We found that the resveratrol analogues also significantly inhibited Akt and MAPK signalling and reduced the migration of IL-6 and EGF-treated cells. Finally, in ascite-derived cancer cells, we demonstrated that the resveratrol analogues reduced the expression of epithelial mesenchymal transition (EMT) markers. Collectively, these findings indicate the enhanced anti-cancer properties of the resveratrol analogues.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Ovarianas/metabolismo , Estilbenos/farmacologia , Antineoplásicos Fitogênicos/química , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Feminino , Humanos , Interleucina-6/farmacologia , Resveratrol , Transdução de Sinais/efeitos dos fármacos , Estilbenos/química
18.
Springerplus ; 4: 49, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25674504

RESUMO

The plant phenol trans-resveratrol, which is mainly found in grape, displays a wide range of biological effects. A cell suspension culture was developed from calli of grape leaves of Vitis vinifera cv. Negramaro in order to study the bioproduction of resveratrol. The effects of a number of secondary plant metabolism elicitors, namely chitosan, methyl jasmonate, jasmonic acid, coronatine, and 12-oxo-phytodienoic acid, were tested on this cell suspension culture. The identification and quantification of stilbenes was achieved with high performance liquid chromatography, with both spectrophotometric and mass spectrometric detection. Of the tested elicitors, methyl jasmonate was the most effective in inducing the biosynthesis of approximately 4 mg g(-1) dry weight (about 60 mg L(-1)) of resveratrol. Conversely, 12-oxo-phytodienoic acid, jasmonic acid, and coronatine were able to trigger the synthesis of approximately 20 mg g(-1) dry weight (200-210 mg L(-1)) of viniferins. Taken together, our results show for the first time different modulatory effects of closely-related jasmonates on stilbene biosynthesis.

19.
Int J Mol Sci ; 16(1): 2174-86, 2015 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-25608652

RESUMO

In this work we explored the possibility of using genetically modified Arabidopsis thaliana plants as a rapid and low-cost screening tool for evaluating human anticancer drugs action and efficacy. Here, four different inhibitors with a validated anticancer effect in humans and distinct mechanism of action were screened in the plant model for their ability to interfere with the cytoskeletal and endomembrane networks. We used plants expressing a green fluorescent protein (GFP) tagged microtubule-protein (TUA6-GFP), and three soluble GFPs differently sorted to reside in the endoplasmic reticulum (GFPKDEL) or to accumulate in the vacuole through a COPII dependent (AleuGFP) or independent (GFPChi) mechanism. Our results demonstrated that drugs tested alone or in combination differentially influenced the monitored cellular processes including cytoskeletal organization and endomembrane trafficking. In conclusion, we demonstrated that A. thaliana plants are sensitive to the action of human chemotherapeutics and can be used for preliminary screening of drugs efficacy. The cost-effective subcellular imaging in plant cell may contribute to better clarify drugs subcellular targets and their anticancer effects.


Assuntos
Antineoplásicos/farmacologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Custos e Análise de Custo , Avaliação Pré-Clínica de Medicamentos/economia , Avaliação Pré-Clínica de Medicamentos/métodos , Células Vegetais/efeitos dos fármacos , Arabidopsis/citologia , Proteínas de Fluorescência Verde/metabolismo , Humanos , Células Vegetais/metabolismo , Epiderme Vegetal/citologia , Plantas Geneticamente Modificadas
20.
Front Microbiol ; 5: 570, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25389422

RESUMO

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is constantly increasing. In the Greek style, table olives are obtained by spontaneous fermentations, without any chemical debittering treatment. Evolution of sugars, organic acids, alcohols, mono, and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A protocol was developed and applied aimed at the technological characterization of lactic acid bacteria (LAB) and yeast strains as possible candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars. The study of the main physic-chemical parameters and volatile compounds during fermentation helped to determine chemical descriptors that may be suitable for monitoring olive fermentation. In both the analyzed table olive cultivars, aldehydes proved to be closely related to the first stage of fermentation (30 days), while higher alcohols (2-methyl-1-propanol; 3-methyl-1-butanol), styrene, and o-cymene were associated with the middle stage of fermentation (90 days) and acetate esters with the final step of olive fermentation (180 days).

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