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1.
Int J Mol Sci ; 25(16)2024 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-39201682

RESUMO

Liver fibrosis, characterized by excessive extracellular matrix deposition, is driven by activated hepatic stellate cells (HSCs). Due to the limited availability of anti-fibrotic drugs, the research on therapeutic agents continues. Here we have investigated Moringa oleifera Lam. (MO), known for its various bioactive properties, for anti-fibrotic effects. This study has focused on 1-phenyl-2-pentanol (1-PHE), a compound derived from MO leaves, and its effects on LX-2 human hepatic stellate cell activation. TGF-ß1-stimulated LX-2 cells were treated with MO extract or 1-PHE, and the changes in liver fibrosis markers were assessed at both gene and protein levels. Proteomic analysis and molecular docking were employed to identify potential protein targets and signaling pathways affected by 1-PHE. Treatment with 1-PHE downregulated fibrosis markers, including collagen type I alpha 1 chain (COL1A1), collagen type IV alpha 1 chain (COL4A1), mothers against decapentaplegic homologs 2 and 3 (SMAD2/3), and matrix metalloproteinase-2 (MMP2), and reduced the secretion of matrix metalloproteinase-9 (MMP-9). Proteomic analysis data showed that 1-PHE modulates the Wnt/ß-catenin pathway, providing a possible mechanism for its effects. Our results suggest that 1-PHE inhibits the TGF-ß1 and Wnt/ß-catenin signaling pathways and HSC activation, indicating its potential as an anti-liver-fibrosis agent.


Assuntos
Células Estreladas do Fígado , Cirrose Hepática , Moringa oleifera , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Humanos , Moringa oleifera/química , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Linhagem Celular , Proteômica/métodos , Simulação de Acoplamento Molecular , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Antifibróticos/farmacologia , Fator de Crescimento Transformador beta1/metabolismo , Transdução de Sinais/efeitos dos fármacos
2.
Immunology ; 170(3): 359-373, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37340593

RESUMO

A significant number of babies present transiently with low protein kinase C zeta (PKCζ) levels in cord blood T cells (CBTC), associated with reduced ability to transition from a neonatal Th2 to a mature Th1 cytokine bias, leading to a higher risk of developing allergic sensitisation, compared to neonates whose T cells have 'normal' PKCζ levels. However, the importance of PKCζ signalling in regulating their differentiation from a Th2 to a Th1 cytokine phenotype propensity remains undefined. To define the role of PKCζ signalling in the regulation of CBTC differentiation from a Th2 to a Th1cytokine phenotype we have developed a neonatal T cell maturation model which enables the cells to develop to CD45RA- /CD45RO+ T cells while maintaining the Th2 immature cytokine bias, despite having normal levels of PKCζ. The immature cells were treated with phytohaemagglutinin, but in addition with phorbol 12-myristate 13-acetate (PMA), an agonist which does not activate PKCζ. This was compared to development in CBTC in which the cells were transfected to express constitutively active PKCζ. The lack of PKCζ activation by PMA was monitored by western blot for phospho-PKCζ and translocation from cell cytosol to the membrane by confocal microscopy. The findings demonstrate that PMA fails to activate PKCζ in CBTC. The data show that CBTC matured under the influence of the PKC stimulator, PMA, maintain a Th2 cytokine bias, characterised by robust IL-4 and minimal interferon gamma production (IFN-γ), and lack of expression of transcriptional factor, T-bet. This was also reflected in the production of a range of other Th2/Th1 cytokines. Interestingly, introduction of a constitutively active PKCζ mutant into CBTC promoted development towards a Th1 profile with high IFN-γ production. The findings demonstrate that PKCζ signalling is essential for the immature neonatal T cells to transition from a Th2 to a Th1 cytokine production bias.


Assuntos
Interferon gama , Células Th1 , Recém-Nascido , Humanos , Interferon gama/metabolismo , Células Th1/metabolismo , Sangue Fetal , Citocinas/metabolismo , Diferenciação Celular , Antígenos Comuns de Leucócito , Células Th2/metabolismo
3.
Planta ; 259(1): 1, 2023 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-37966555

RESUMO

MAIN CONCLUSION: SMAX/SMXL family genes were successfully identified and characterized in the chickpea and lentil and gene expression data revealed several genes associated with the modulation of plant branching and powerful targets for use in transgenesis and genome editing. Strigolactones (SL) play essential roles in plant growth, rooting, development, and branching, and are associated with plant resilience to abiotic and biotic stress conditions. Likewise, karrikins (KAR) are "plant smoke-derived molecules" that act in a hormonal signaling pathway similar to SL playing an important role in seed germination and hairy root elongation. The SMAX/SMXL family genes are part of these two signaling pathways, in addition to some of these members acting in a still little known SL- and KAR-independent signaling pathway. To date, the identification and functional characterization of the SMAX/SMXL family genes has not been performed in the chickpea and lentil. In this study, nine SMAX/SMXL genes were systematically identified and characterized in the chickpea and lentil, and their expression profiles were explored under different unstressless or different stress conditions. After a comprehensive in silico characterization of the genes, promoters, proteins, and protein-protein interaction network, the expression profile for each gene was determined using a meta-analysis from the RNAseq datasets and complemented with real-time PCR analysis. The expression profiles of the SMAX/SMXL family genes were very dynamic in different chickpea and lentil organs, with some genes assuming a tissue-specific expression pattern. In addition, these genes were significantly modulated by different stress conditions, indicating that SMAX/SMXL genes, although working in three distinct signaling pathways, can act to modulate plant resilience. Most CaSMAX/SMXL and partner genes such as CaTiE1 and CaLAP1, have a positive correlation with the plant branching level, while most LcSMAX/SMXL genes were less correlated with the plant branching level. The SMXL6, SMXL7, SMXL8, TiE1, LAP1, BES1, and BRC1 genes were highlighted as powerful targets for use in transgenesis and genome editing aiming to develop chickpea and lentil cultivars with improved architecture. Therefore, this study presented a detailed characterization of the SMAX/SMXL genes in the chickpea and lentil, and provided new insights for further studies focused on each SMAX/SMXL gene.


Assuntos
Cicer , Lens (Planta) , Lens (Planta)/genética , Cicer/genética , Biotecnologia , Edição de Genes , Desenvolvimento Vegetal
4.
Int J Mol Sci ; 24(4)2023 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-36834600

RESUMO

Vegetable and ornamental plants represent a very wide group of heterogeneous plants, both herbaceous and woody, generally without relevant salinity-tolerant mechanisms. The cultivation conditions-almost all are irrigated crops-and characteristics of the products, which must not present visual damage linked to salt stress, determine the necessity for a deep investigation of the response of these crops to salinity stress. Tolerance mechanisms are linked to the capacity of a plant to compartmentalize ions, produce compatible solutes, synthesize specific proteins and metabolites, and induce transcriptional factors. The present review critically evaluates advantages and disadvantages to study the molecular control of salt tolerance mechanisms in vegetable and ornamental plants, with the aim of distinguishing tools for the rapid and effective screening of salt tolerance levels in different plants. This information can not only help in suitable germplasm selection, which is very useful in consideration of the high biodiversity expressed by vegetable and ornamental plants, but also drive the further breeding activities.


Assuntos
Melhoramento Vegetal , Verduras , Produtos Agrícolas , Tolerância ao Sal/fisiologia , Salinidade
5.
J Sci Food Agric ; 103(11): 5472-5480, 2023 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-37046389

RESUMO

BACKGROUND: Biofortification of vegetables is an important innovation technique in the horticultural sector. Vegetables can be a vector of different minor elements that have beneficial effects on human health. Selenium (Se) is an important element for human nutrition and plays a significant role in defence mechanisms. The aim of this work was to investigate the effect of Se in the nutrient solutions on the crop biofortification ability, yield, and quality parameters of four baby leafy vegetables destined to the minimally processed industry. Experiments were performed on lamb's lettuce, lettuce, wild rocket, and spinach. These crops were cultivated in the floating systems with nutrient solution enriched with 0, 2.6, 3.9, and 5.2 µmol L-1 Se provided as sodium selenate. RESULTS: At harvest, Se concentrations, yield, nitrate concentration, sugars, and some mineral elements were measured. Data collected and analyses showed that yield, nitrate, sucrose, and reducing sugars were not affected by Se treatments, even if varied among species. Se concentrations linearly increased in leaves of different species by increasing the Se concentration in the nutrient solution. Rocket was the species with the highest accumulation ability and reached a concentration of 11 µg g-1 fresh weight Se in plants grown with 5.2 µmol L-1 Se. CONCLUSION: A floating system with Se-enriched nutrient solution is an optimal controlled growing biofortification system for leafy vegetables. The accumulation ability decreased in different species in the order wild rocket, spinach, lettuce, and lamb's lettuce, highlighting a crop-dependent behaviour and their attitude to biofortification. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Assuntos
Selênio , Verduras , Humanos , Biofortificação/métodos , Selênio/análise , Nitratos/análise , Lactuca , Nutrientes/análise , Folhas de Planta/química
6.
Clin Exp Immunol ; 207(3): 329-335, 2022 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-35553639

RESUMO

Primary immunodeficiency disorders comprise a rare group of mostly monogenic disorders caused by inborn errors of immunity. The majority can be identified by either Sanger sequencing or next generation sequencing. Some disorders result from large insertions or deletions leading to copy number variations (CNVs). Sanger sequencing may not identify these mutations. Here we present droplet digital PCR as an alternative cost-effective diagnostic method to identify CNV in these genes. The data from patients with large deletions of NFKB1, SERPING1, and SH2D1A are presented.


Assuntos
Variações do Número de Cópias de DNA , Doenças da Imunodeficiência Primária , Variações do Número de Cópias de DNA/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Reação em Cadeia da Polimerase
7.
Int J Mol Sci ; 22(9)2021 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-34063174

RESUMO

Low Protein Kinase C zeta (PKCζ) levels in cord blood T cells (CBTC) have been shown to correlate with the development of allergic sensitization in childhood. However, little is known about the mechanisms responsible. We have examined the relationship between the expression of different levels of PKCζ in CBTC and their development into mature T cell cytokine producers that relate to allergy or anti-allergy promoting cells. Maturation of naïve CBTC was initiated with anti-CD3/-CD28 antibodies and recombinant human interleukin-2 (rhIL-2). To stimulate lymphocyte proliferation and cytokine production the cells were treated with Phytohaemagglutinin (PHA) and Phorbol myristate acetate (PMA). Irrespective of the PKCζ levels expressed, immature CBTC showed no difference in lymphocyte proliferation and the production of T helper 2 (Th2) cytokine interleukin-4 (IL-4) and Th1 cytokine, interferon-gamma (IFN-γ), and influenced neither their maturation from CD45RA+ to CD45RO+ cells nor cell viability/apoptosis. However, upon maturation the low PKCζ expressing cells produced low levels of the Th1 cytokines, IFN-γ, IL-2 and tumour necrosis factor-alpha (TNF), no changes to levels of the Th2 cytokines, IL-4, IL-5 and IL-13, and an increase in the Th9 cytokine, IL-9. Other cytokines, lymphotoxin-α (LT-α), IL-10, IL-17, IL-21, IL-22 and Transforming growth factor-beta (TGF-ß) were not significantly different. The findings support the view that low CBTC PKCζ levels relate to the increased risk of developing allergic diseases.


Assuntos
Sangue Fetal/citologia , Proteína Quinase C/metabolismo , Linfócitos T/enzimologia , Células Th1/citologia , Células Th1/metabolismo , Apoptose , Diferenciação Celular , Proliferação de Células , Sobrevivência Celular , Citocinas , Humanos , Células Th2/citologia , Células Th2/metabolismo
8.
Int J Mol Sci ; 22(23)2021 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-34884454

RESUMO

Cord blood T cells (CBTC) from a proportion of newborns express low/deficient levels of some protein kinase C (PKC) isozymes, with low levels of PKCζ correlating with increased risk of developing allergy and associated decrease in interferon-gamma (IFN-γ) producing T cells. Interestingly, these lower levels of PKCζ were increased/normalized by supplementing women during pregnancy with n-3 polyunsaturated fatty acids. However, at present, we have little understanding of the transient nature of the deficiency in the neonate and how PKCζ relates to other PKC isozymes and whether their levels influence maturation into IFN-γ producing T cells. There is also no information on PKCζ isozyme levels in the T cell subpopulations, CD4+ and CD8+ cells. These issues were addressed in the present study using a classical culture model of neonatal T cell maturation, initiated with phytohaemagglutinin (PHA) and recombinant human interleukin-2 (rhIL-2). Of the isozymes evaluated, PKCζ, ß2, δ, µ, ε, θ and λ/ι were low in CBTCs. The PKC isozyme deficiencies were also found in the CD4+ and CD8+ T cell subset levels of the PKC isozymes correlated between the two subpopulations. Examination of changes in the PKC isozymes in these deficient cells following addition of maturation signals showed a significant increase in expression within the first few hours for PKCζ, ß2 and µ, and 1-2 days for PKCδ, ε, θ and λ/ι. Only CBTC PKCζ isozyme levels correlated with cytokine production, with a positive correlation with IFN-γ, interleukin (IL)-2 and tumour necrosis factor-alpha (TNF), and a negative association with IL-9 and IL-10. The findings reinforce the specificity in using CBTC PKCζ levels as a biomarker for risk of allergy development and identify a period in which this can be potentially 'corrected' after birth.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Sangue Fetal/citologia , Proteína Quinase C/genética , Feminino , Sangue Fetal/imunologia , Idade Gestacional , Humanos , Recém-Nascido , Interferon gama/metabolismo , Interleucina-10/metabolismo , Interleucina-2/metabolismo , Interleucina-9/metabolismo , Masculino , Fito-Hemaglutininas/farmacologia , Gravidez , Fator de Necrose Tumoral alfa/metabolismo
9.
J Sci Food Agric ; 101(14): 6134-6142, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34402534

RESUMO

BACKGROUND: Fresh-cut vegetables are subjected to multiple stressing agents including: (i) slicing, which induces cellular decompartmentalization; (ii) low refrigeration temperatures, responsible for chilling injury in the most sensitive products (e.g. tomatoes), and (iii) storage time because tissue senescence and aging can occur and reduce the shelf-life. In tomato slices, one of the most important issues is the membrane, which is responsible for several disorders related to the alteration of physiological processes, including ethylene biosynthesis. RESULTS: Electrolyte leakage and the content of thiobarbituric acid reactive substances in sliced tomatoes increased over time at two storage temperatures (4 °C and 15 °C) compared with intact fruit for the commercial variety (cultivar) Jama used as reference. However, in the tomato Italian landrace Canestrino, electrolyte leakage in sliced fruits increased after 120 h of storage compared to intact tomatoes, while the thiobarbituric acid reactive substance content increased rapidly over time at both storage temperatures. In the packages, higher ethylene content and carbon dioxide concentrations were detected in sliced tomatoes compared with intact fruits for both genotypes. In the most sensitive genotype for slicing (Jama), phospholipase C activity increased in tomato slices after 24 h of storage, but phospholipase D reached a higher value only at 168 h after processing at 4 °C of storage. CONCLUSIONS: The results evidence that the main damage in slices of full ripe tomatoes is more related to cutting, rather than chilling injury due to storage temperatures, with differences related to the genotype. Slicing enhanced membrane catabolism, ethylene production, and enzyme activity of phospholipases with a significant genotype effect. © 2021 Society of Chemical Industry.


Assuntos
Membrana Celular/química , Manipulação de Alimentos/métodos , Solanum lycopersicum/química , Membrana Celular/genética , Membrana Celular/metabolismo , Etilenos/metabolismo , Armazenamento de Alimentos , Frutas/química , Frutas/genética , Frutas/metabolismo , Genótipo , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Temperatura
10.
Physiol Mol Biol Plants ; 27(5): 1059-1072, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34103849

RESUMO

The yield and quality of leafy vegetables can be compromised by reduced water availability. Glutamic acid is involved in different biological processes and among them it plays an important role in chlorophyll and proline biosynthesis. The aim of this work was to evaluate the possible efficacy of glutamic acid in counteracting water stress in romaine lettuce. Lettuce plants were grown in pots filled with substrate and subjected to water deprivation. A glutamic acid solution (1.9 mM) was applied as foliar treatment, both in stressed and non-stressed plants. The effect of the treatment was evaluated at different time points during the experiment in order to evaluate changes at a molecular, physiological, biochemical and agronomic level. Yield was reduced by 35% in stressed plants, while no significant changes in quality parameters were observed, except for nitrate content, which increased under water stress. At a molecular level, the expression of genes encoding for ROS scavenging enzymes was monitored but, apparently, glutamic acid did not significantly prevent the water stress response. Slightly positive effects deriving from glutamic acid application were found for nitrate and proline contents, suggesting that a possible mode of action of glutamic acid would involve a role for these molecules. Further studies are required, also on other crop species, for confirming these results. Different concentrations and application modes should be also tested.

11.
Physiol Mol Biol Plants ; 26(4): 649-660, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32255929

RESUMO

Market is increasingly demanding vegetables with high quality and nutraceutical characteristics. It was demonstrated that leafy vegetables can get benefit from biostimulants, for the reduction of nitrate concentration and the increment of antioxidants, with potential benefit for human health. The research purpose was to investigate on the role of a novel plant-based biostimulant in affecting nitrogen and carbon metabolism in wild rocket (Diplotaxis tenuifolia L.). Foliar spray treatments were performed with extracts obtained from borage (Borago officinalis L.) leaves and flowers. To evaluate the treatments effect, in vivo determinations (chlorophyll a fluorescence and chlorophyll content) were performed. At harvest, nitrate concentration, sucrose, total sugars, chlorophyll, and carotenoids levels were measured in leaves. In order to characterize the mechanism of action also at molecular level, a set of genes encoding for some of the key enzymes implicated in nitrate and carbon metabolism was selected and their expression was measured by qRT-PCR. Interesting results concerned the increment of sucrose, coherent with a high value of Fv/Fm, in addition to a significant reduction of nitrate and ABA than control, and an enhanced NR in vivo activity. Also, genes expression was influenced by extracts, with a more pronounced effect on N related genes.

12.
Fish Shellfish Immunol ; 92: 871-880, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31299464

RESUMO

The neutrophil oxidative respiratory burst response is a key component of the innate immune system responsible for killing microbial pathogens. Since fish rely on the innate immune system for health, monitoring the respiratory burst activity may be an effective means of gauging fish health status. Here we report that the respiratory burst of Asian seabass neutrophils can be measured in whole blood by the dihydrorhodamine (DHR)-123 reduction assay and flow cytometry. Neutrophils responded to phorbol myristate acetate (PMA) in a concentration dependent manner with significant respiratory burst activity at 100-1000 nM. Other known neutrophil agonists, such as bacterial lipopolysaccharide, tumor necrosis factor, the tripeptide f-met-leu-phe and zymosan, did not induce a significant DHR reduction. Thus, the findings enable us to propose that the DHR-123 flow cytometry whole blood assay, incorporating PMA as a stimulator, would not only facilitate future studies into fish blood neutrophil research but provides a simple, rapid and reliable assay for gauging fish natural immunity status and health.


Assuntos
Bass/fisiologia , Citometria de Fluxo/veterinária , Imunidade Inata , Neutrófilos/fisiologia , Explosão Respiratória/fisiologia , Animais , Citometria de Fluxo/métodos , Oxirredução , Rodaminas/química
13.
Cancer Immunol Immunother ; 67(9): 1461-1472, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30014244

RESUMO

INTRODUCTION: In this phase I study using a 3 + 3 dose escalation design, the safety, dose-limiting toxicity (DLT), immunogenicity and efficacy of intravenous Lipovaxin-MM-a multi-component dendritic cell-targeted liposomal vaccine against metastatic melanoma-was investigated. METHODS: Twelve subjects with metastatic cutaneous melanoma were recruited in three cohorts. Patients in Cohort A (n = 3) and Cohort B (n = 3) received three doses of 0.1 and 1 mL of Lipovaxin-MM, respectively, every 4 weeks. Patients in Cohort C (n = 6) received four doses of 3 mL vaccine weekly. Immunologic assessments of peripheral blood were made at regular intervals and included leukocyte subsets, cytokine levels, and Lipovaxin-MM-specific T-cell and antibody reactivities. Tumor responses were assessed by RECIST v1.0 at screening, then 8 weekly in Cohorts A and B and 6 weekly in Cohort C. RESULTS: Of a total of 94 adverse events (AEs) reported in ten subjects, 43 AEs in six subjects were considered to be possibly or probably vaccine-related. Most (95%) vaccine-related AEs were grade 1 or 2, two (5%) grade 3 vaccine-related AEs of anemia and lethargy were recorded, and higher grade AEs and DLTs were not observed. No consistent evidence of vaccine-specific humoral or cellular immune responses was found in post-immunization blood samples. One patient had a partial response, two patients had stable disease, and the remaining patients had progressive disease. CONCLUSIONS: Lipovaxin-MM was well tolerated and without clinically significant toxicity. Immunogenicity of Lipovaxin-MM was not detected. Partial response and stable disease were observed in one and two patients, respectively.


Assuntos
Vacinas Anticâncer/administração & dosagem , Células Dendríticas/imunologia , Melanoma/terapia , Neoplasias Cutâneas/terapia , Adulto , Idoso , Vacinas Anticâncer/efeitos adversos , Vacinas Anticâncer/imunologia , Células Dendríticas/efeitos dos fármacos , Relação Dose-Resposta Imunológica , Feminino , Humanos , Lipossomos/administração & dosagem , Lipossomos/imunologia , Masculino , Melanoma/imunologia , Pessoa de Meia-Idade , Neoplasias Cutâneas/imunologia , Melanoma Maligno Cutâneo
14.
J Immunol ; 194(6): 2855-61, 2015 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-25687755

RESUMO

The complement receptor Ig (CRIg) is selectively expressed by macrophages. This receptor not only promotes the rapid phagocytosis of bacteria by macrophages but also has anti-inflammatory and immunosuppressive functions. Previous findings have suggested that protein kinase C (PKC) may be involved in the regulation of CRIg expression in human macrophages. We have now examined the role of PKCα in CRIg expression in human monocyte-derived macrophages (MDM). Macrophages nucleofected with plasmid containing short hairpin RNA against PKCα showed markedly reduced expression of PKCα, but normal PKCζ expression, by Western blotting analysis, and vice versa. PKCα-deficient MDM showed increased expression of CRIg mRNA and protein (both the long and short form), an increase in phagocytosis of complement-opsonized Candida albicans, and decreased production of TNF-α and IL-6. TNF-α caused a marked decrease in CRIg expression, and addition of anti-TNF mAb to the TNF-α-producing MDMs increased CRIg expression. PKCα-deficient macrophages also showed significantly less bacterial LPS-induced downregulation of CRIg. In contrast, cells deficient in PKCα showed decreased expression of CR type 3 (CR3) and decreased production of TNF-α and IL-6 in response to LPS. MDM developed under conditions that increased expression of CRIg over CR3 showed significantly reduced production of TNF-α in response to opsonized C. albicans. The findings indicate that PKCα promotes the downregulation of CRIg and upregulation of CR3 expression and TNF-α and IL-6 production, a mechanism that may promote inflammation.


Assuntos
Macrófagos/imunologia , Monócitos/imunologia , Proteína Quinase C-alfa/imunologia , Receptores de Complemento/imunologia , Anti-Inflamatórios/imunologia , Anti-Inflamatórios/farmacologia , Western Blotting , Candida albicans/imunologia , Adesão Celular/efeitos dos fármacos , Adesão Celular/genética , Adesão Celular/imunologia , Células Cultivadas , Dexametasona/imunologia , Dexametasona/farmacologia , Regulação para Baixo/imunologia , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/imunologia , Humanos , Interleucina-1beta/imunologia , Interleucina-1beta/metabolismo , Interleucina-6/imunologia , Interleucina-6/metabolismo , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/farmacologia , Antígeno de Macrófago 1/genética , Antígeno de Macrófago 1/imunologia , Antígeno de Macrófago 1/metabolismo , Macrófagos/citologia , Macrófagos/metabolismo , Monócitos/citologia , Monócitos/metabolismo , Proteína Quinase C-alfa/genética , Proteína Quinase C-alfa/metabolismo , Interferência de RNA , Receptores de Complemento/genética , Receptores de Complemento/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
15.
J Exp Bot ; 67(20): 5919-5931, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27591432

RESUMO

Flowers are complex systems whose vegetative and sexual structures initiate and die in a synchronous manner. The rapidity of this process varies widely in flowers, with some lasting for months while others such as Hibiscus rosa-sinensis survive for only a day. The genetic regulation underlying these differences is unclear. To identify key genes and pathways that coordinate floral organ senescence of ephemeral flowers, we identified transcripts in H. rosa-sinensis floral organs by 454 sequencing. During development, 2053 transcripts increased and 2135 decreased significantly in abundance. The senescence of the flower was associated with increased abundance of many hydrolytic genes, including aspartic and cysteine proteases, vacuolar processing enzymes, and nucleases. Pathway analysis suggested that transcripts altering significantly in abundance were enriched in functions related to cell wall-, aquaporin-, light/circadian clock-, autophagy-, and calcium-related genes. Finding enrichment in light/circadian clock-related genes fits well with the observation that hibiscus floral development is highly synchronized with light and the hypothesis that ageing/senescence of the flower is orchestrated by a molecular clock. Further study of these genes will provide novel insight into how the molecular clock is able to regulate the timing of programmed cell death in tissues.


Assuntos
Flores/crescimento & desenvolvimento , Hibiscus/crescimento & desenvolvimento , Transcriptoma/fisiologia , Envelhecimento/fisiologia , Cálcio/fisiologia , Ritmo Circadiano/fisiologia , Flores/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Genes de Plantas/fisiologia , Hibiscus/genética , Hibiscus/fisiologia , Análise de Sequência com Séries de Oligonucleotídeos , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/fisiologia
16.
Plant Mol Biol ; 87(1-2): 169-80, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25425166

RESUMO

Flower senescence is a fascinating natural process that represents the final developmental stage in the life of a flower. Plant hormones play an important role in regulating the timing of flower senescence. Ethylene is a trigger and usually accelerates the senescence rate, while cytokinins are known to delay it. The aim of this work was to study the effect of 6-benzylaminopurine (BA) on petal senescence by transcript profile comparison after 3 or 6 h using a cross-species method by hybridizing petunia samples to a 4 × 44 K Agilent tomato array. The relative content of ethylene, abscisic acid, anthocyanins, total carotenoids and total phenols that determine the physiological behaviours of the petal tissue were measured. BA treatment prolonged the flower life and increased the concentrations of phenols and anthocyanins, while total carotenoids did not increase and were lower than the control. The ethylene biosynthetic and perception gene expressions were studied immediately after treatment until 24 h and all genes were repressed, while ethylene production was strongly induced after 4 days. The microarray analyses highlighted that BA strongly affected gene regulation after 3 h, but only 14% of genes remained differentially expressed after 6 h. The most affected pathways and genes were those related to stress, such as heat shock proteins, abscisic acid (ABA) catabolism and its signalling pathway, lipid metabolism and antioxidant defence systems. A gene annotation enrichment analysis using DAVID showed that the most important gene clusters were involved in energy generation and conservation processes. In addition to the ethylene pathway, cytokinins seem to be strongly involved the regulation of the ABA response in flower tissues.


Assuntos
Citocininas/fisiologia , Flores/fisiologia , Genes de Plantas , Petunia/fisiologia , Transcriptoma , Compostos de Benzil , Carotenoides/metabolismo , Etilenos/metabolismo , Cinetina/farmacologia , Petunia/genética , Petunia/metabolismo , Fenóis/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Purinas , Reação em Cadeia da Polimerase em Tempo Real
18.
J Exp Bot ; 66(5): 1511-26, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25614658

RESUMO

Fleshy fruits develop from an unripe organ that needs to be protected from damage to a ripe organ that attracts frugivores for seed dispersal through production of volatile organic compounds (VOCs). Thus, different responses to wounding damage are predicted. The aim of this study was to discover whether wound-induced changes in the transcriptome and VOC production alter as tomato transitions from unripe to ripe. Transcript changes were analysed 3h post-wounding using microarray analysis in two commercial salad-tomato (Solanum lycopersicum L.) cultivars: Luna Rossa and AVG, chosen for their high aroma production. This was followed by quantitative PCR on Luna Rossa genes involved in VOC biosynthesis and defence responses. VOCs elicited by wounding at different ripening stages were analysed by solid phase micro extraction and gas chromatography-mass spectrometry. Approximately 4000 differentially expressed genes were identified in the cultivar AVG and 2500 in Luna Rossa. In both cultivars the majority of genes were up-regulated and the most affected pathways were metabolism of terpenes, carotenoids, and lipids. Defence-related genes were mostly up-regulated in immature stages of development, whereas expression of genes related to VOCs changed at riper stages. More than 40 VOCs were detected and profiles changed with ripening stage. Thus, both transcriptome and VOC profiles elicited by wounding depend on stage of ripening, indicating a shift from defence to attraction.


Assuntos
Frutas/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Solanum lycopersicum/crescimento & desenvolvimento , Compostos Orgânicos Voláteis/metabolismo , Frutas/química , Frutas/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Solanum lycopersicum/química , Solanum lycopersicum/metabolismo , Proteínas de Plantas/metabolismo , Compostos Orgânicos Voláteis/química
19.
Physiol Plant ; 153(2): 269-83, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24943920

RESUMO

Blueberry (Vaccinium corymbosum) is a fruit very much appreciated by consumers for its antioxidant potential and health-promoting traits. Its beneficial potential properties are mainly due to a high content of anthocyanins and their amount can change after elicitation with methyl jasmonate. The aim of this work is to evaluate the changes in expression of several genes, accumulation of phenolic compounds and alterations in antioxidant potential in two different blueberry cultivars ('Duke' and 'Blueray') in response to methyl jasmonate (0.1 mM). Results showed that 9 h after treatment, the expression of phenylalanine ammonium lyase, chalcone synthase and anthocyanidin synthase genes was stimulated more in the 'Blueray' variety. Among the phenols measured an increase was recorded also for epicatechin and anthocyanin concentrations. 'Duke' is a richer sourche of anthocyanins compared to 'Blueray', treatment with methyl jasmonate promoted in 'Blueray' an increase in pigments as well as in the antioxidant potential, especially in fully ripe berries, but treated 'Duke' berries had greater levels, which were not induced by methyl jasmonate treatment. In conclusion, methyl jasmonate was, in some cases, an effective elicitor of phenolic metabolism and gene expression in blueberry, though with different intensity between cultivars.


Assuntos
Acetatos/farmacologia , Mirtilos Azuis (Planta)/genética , Mirtilos Azuis (Planta)/metabolismo , Ciclopentanos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oxilipinas/farmacologia , Fenóis/metabolismo , Antocianinas/metabolismo , Compostos de Bifenilo/metabolismo , Mirtilos Azuis (Planta)/efeitos dos fármacos , Carboidratos/análise , Flavonóis/metabolismo , Sequestradores de Radicais Livres/farmacologia , Genes de Plantas , Picratos/metabolismo , Extratos Vegetais/farmacologia , Propanóis/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Estilbenos/metabolismo , Sacarose/metabolismo
20.
Heliyon ; 10(11): e32569, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38961974

RESUMO

Sword lily is regarded as a useful and commercially demanding cut flower crop; hence, assessing its responses to abiotic stress, particularly salt stress, is vital. Melatonin (MT) exhibits stress tolerance in crop plants and is an emerging stress relieving alternative to chemicals. Nevertheless, the possible process underlying the effects of MT under salt stress has yet to be fully elucidated in plants. Herein, the salt stress (SS) mitigation potential of MT was assessed in a commercially important cut flower, sword lily. Melatonin, expressed as MT1, MT2, MT3, and MT4, was administered at concentrations of 0.2, 0.4, 0.6, and 0.8 mM. The results revealed that SS (5 dS m-1) restricted the growth and physiological aspects of sword lily. Furthermore, malondialdehyde (MDA), hydrogen peroxide (H2O2), membrane permeability, endogenous proline, and soluble protein contents were enhanced in SS. MT application improved morphological traits, photosynthetic pigments, and corm traits. The application of MT mitigated the effects of SS stress in Gladiolus grandiflorus plants by improving growth and photosynthetic pigments. MT application under SS improved the reducing and non-reducing sugar and NPK contents of the sword lily. Furthermore, MT improved the levels of secondary metabolites, such as anthocyanins, flavonoids, and ascorbic acid, in sword lily. Moreover, MT supplementation ameliorated salt-induced oxidative stress in the gladiolus, as depicted by a decrease in stress markers (EL, MDA, and H2O2) and an increase in defense-related enzymes (POD, CAT, and SOD) with highest increase in the MT3 treatment under salinity stress. The SOD and CAT enzyme activities were 3-3.6-fold higher in the MT3 under stress than the control. In conclusion, MT applications on cut flowers can be an effective strategy to reduce salt stress and can be used to regulate salinity stress in cut flower production. MT can be used as a safe alternative to other agrochemicals to maintain the growth and flower quality of sword lilies, with beneficial effects during vase life.

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