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1.
FASEB J ; 38(3): e23454, 2024 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-38315457

RESUMEN

Mitochondria shape intracellular Ca2+ signaling through the concerted activity of Ca2+ uptake via mitochondrial calcium uniporters and efflux by Na+ /Ca2+ exchangers (NCLX). Here, we describe a novel relationship among NCLX, intracellular Ca2+ , and autophagic activity. Conditions that stimulate autophagy in vivo and in vitro, such as caloric restriction and nutrient deprivation, upregulate NCLX expression in hepatic tissue and cells. Conversely, knockdown of NCLX impairs basal and starvation-induced autophagy. Similarly, acute inhibition of NCLX activity by CGP 37157 affects bulk and endoplasmic reticulum autophagy (ER-phagy) without significant impacts on mitophagy. Mechanistically, CGP 37157 inhibited the formation of FIP200 puncta and downstream autophagosome biogenesis. Inhibition of NCLX caused decreased cytosolic Ca2+ levels, and intracellular Ca2+ chelation similarly suppressed autophagy. Furthermore, chelation did not exhibit an additive effect on NCLX inhibition of autophagy, demonstrating that mitochondrial Ca2+ efflux regulates autophagy through the modulation of Ca2+ signaling. Collectively, our results show that the mitochondrial Ca2+ extrusion pathway through NCLX is an important regulatory node linking nutrient restriction and autophagy regulation.


Asunto(s)
Señalización del Calcio , Calcio , Clonazepam/análogos & derivados , Tiazepinas , Señalización del Calcio/fisiología , Calcio/metabolismo , Intercambiador de Sodio-Calcio , Mitocondrias/metabolismo , Autofagia , Sodio/metabolismo
2.
J Biol Chem ; 299(3): 102904, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36642177

RESUMEN

Calcium (Ca2+) is a key regulator in diverse intracellular signaling pathways and has long been implicated in metabolic control and mitochondrial function. Mitochondria can actively take up large amounts of Ca2+, thereby acting as important intracellular Ca2+ buffers and affecting cytosolic Ca2+ transients. Excessive mitochondrial matrix Ca2+ is known to be deleterious due to opening of the mitochondrial permeability transition pore (mPTP) and consequent membrane potential dissipation, leading to mitochondrial swelling, rupture, and cell death. Moderate Ca2+ within the organelle, on the other hand, can directly or indirectly activate mitochondrial matrix enzymes, possibly impacting on ATP production. Here, we aimed to determine in a quantitative manner if extra- or intramitochondrial Ca2+ modulates oxidative phosphorylation in mouse liver mitochondria and intact hepatocyte cell lines. To do so, we monitored the effects of more modest versus supraphysiological increases in cytosolic and mitochondrial Ca2+ on oxygen consumption rates. Isolated mitochondria present increased respiratory control ratios (a measure of oxidative phosphorylation efficiency) when incubated with low (2.4 ± 0.6 µM) and medium (22.0 ± 2.4 µM) Ca2+ concentrations in the presence of complex I-linked substrates pyruvate plus malate and α-ketoglutarate, respectively, but not complex II-linked succinate. In intact cells, both low and high cytosolic Ca2+ led to decreased respiratory rates, while ideal rates were present under physiological conditions. High Ca2+ decreased mitochondrial respiration in a substrate-dependent manner, mediated by mPTP. Overall, our results uncover a Goldilocks effect of Ca2+ on liver mitochondria, with specific "just right" concentrations that activate oxidative phosphorylation.


Asunto(s)
Calcio , Mitocondrias , Fosforilación Oxidativa , Animales , Ratones , Calcio/metabolismo , Mitocondrias/metabolismo
3.
Environ Res ; 233: 116435, 2023 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-37331556

RESUMEN

In this study, samples of bromeliad Tillandsia usneoides (n = 70) were transplanted and exposed for 15 and 45 days in 35 outdoor residential areas in Brumadinho (Minas Gerais state, Brazil) after one of the most severe mining dam collapses in the world. Trace elements aluminum (Al), arsenic (As), chromium (Cr), copper (Cu), iron (Fe), mercury (Hg), manganese (Mn), nickel (Ni) and zinc (Zn) were quantified by atomic absorption spectrometry. Scanning electron microscope generated surface images of T. usneoides fragments and particulate matter (PM2.5, PM10 and PM > 10). Aluminum, Fe and Mn stood out from the other elements reflecting the regional geological background. Median concentrations in mg kg-1 increased (p < 0.05) between 15 and 45 days for Cr (0.75), Cu (1.23), Fe (474) and Mn (38.1), while Hg (0.18) was higher at 15 days. The exposed-to-control ratio revealed that As and Hg increased 18.1 and 9.4-fold, respectively, not showing a pattern associated only with the most impacted sites. The PM analysis points to a possible influence of the prevailing west wind on the increase of total particles, PM2.5 and PM10 in transplant sites located to the east. Brazilian public health dataset revealed increase in cases of some cardiovascular and respiratory diseases/symptoms in Brumadinho in the year of the dam collapse (1.38 cases per 1000 inhabitants), while Belo Horizonte capital and its metropolitan region recorded 0.97 and 0.37 cases, respectively. Although many studies have been carried out to assess the consequences of the tailings dam failure, until now atmospheric pollution had not yet been evaluated. Furthermore, based on our exploratory analysis of human health dataset, epidemiological studies are required to verify possible risk factors associated with the increase in hospital admissions in the study area.


Asunto(s)
Contaminantes Atmosféricos , Mercurio , Metales Pesados , Tillandsia , Oligoelementos , Humanos , Oligoelementos/análisis , Material Particulado/análisis , Tillandsia/química , Brasil , Monitoreo Biológico , Salud Pública , Aluminio , Contaminantes Atmosféricos/análisis , Monitoreo del Ambiente/métodos , Cromo/análisis , Mercurio/análisis , Manganeso/análisis , Metales Pesados/análisis
4.
Int J Syst Evol Microbiol ; 72(10)2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-36268793

RESUMEN

The morphology, 16S rRNA gene phylogeny and 16S-23S rRNA gene ITS secondary structures of three strains of marine Cyanobacteria, isolated from inter- and subtidal environments from north Portugal were studied, resulting in the description of Zarconia navalis gen. nov., sp. nov. (Oscillatoriales incertae sedis), Romeriopsis navalis gen. nov., sp. nov. (Leptolyngbyaceae) and Romeriopsis marina sp. nov., named under the International Code of Nomenclature for algae, fungi, and plants. No diacritical morphological characters were found for the new genera and species. The 16S rRNA gene maximum-likelihood and Bayesian phylogenies supported that the genus Zarconia is a member of the Oscillatoriales, morphologically similar to the genera Microcoleus and Phormidium, but distant from them. The genus Romeriopsis is positioned within the Leptolyngbyaceae (Synechococcales) and is closely related to Alkalinema. The secondary structures of the D1-D1', Box B, V2 and V3 helices corroborate the phylogenetic results. Furthermore, our study supports previous observations of polyphyletic Oscillatoriales families and reinforces the need for their taxonomic revision.


Asunto(s)
Cianobacterias , Ácidos Grasos , Humanos , ARN Ribosómico 16S/genética , Filogenia , ADN Bacteriano/genética , Técnicas de Tipificación Bacteriana , Análisis de Secuencia de ADN , Teorema de Bayes , Portugal , Composición de Base , Ácidos Grasos/química
5.
J Phycol ; 56(1): 208-216, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31643075

RESUMEN

The McMurdo Dry Valleys constitute the largest ice-free region of Antarctica and one of the most extreme deserts on Earth. Despite the low temperatures, dry and poor soils and katabatic winds, some microbes are able to take advantage of endolithic microenvironments, inhabiting the pore spaces of soil and constituting photosynthesis-based communities. We isolated a green microalga, Endolithella mcmurdoensis gen. et sp. nov, from an endolithic sandstone sample collected in the McMurdo Dry Valleys (Victoria Land, East Antarctica) during the K020 expedition, in January 2013. The single non-axenic isolate (E. mcmurdoensis LEGE Z-009) exhibits cup-shaped chloroplasts, electron-dense bodies, and polyphosphate granules but our analysis did not reveal any diagnostic morphological characters. On the basis of phylogenetic analysis of the 18S rRNA (SSU) gene, the isolate was found to represent a new genus within the family Chlorellaceae.


Asunto(s)
Chlorella , Extremófilos , Regiones Antárticas , Filogenia , ARN Ribosómico 18S , Análisis de Secuencia de ADN
6.
Environ Microbiol ; 21(11): 4411-4424, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31573125

RESUMEN

Marine biofouling has severe economic impacts and cyanobacteria play a significant role as early surface colonizers. Despite this fact, cyanobacterial biofilm formation studies in controlled hydrodynamic conditions are scarce. In this work, computational fluid dynamics was used to determine the shear rate field on coupons that were placed inside the wells of agitated 12-well microtiter plates. Biofilm formation by three different cyanobacterial strains was assessed at two different shear rates (4 and 40 s-1 ) which can be found in natural ecosystems and using different surfaces (glass and perspex). Biofilm formation was higher under low shear conditions, and differences obtained between surfaces were not always statistically significant. The hydrodynamic effect was more noticeable during the biofilm maturation phase rather than during initial cell adhesion and optical coherence tomography showed that different shear rates can affect biofilm architecture. This study is particularly relevant given the cosmopolitan distribution of these cyanobacterial strains and the biofouling potential of these organisms.


Asunto(s)
Biopelículas , Cianobacterias/fisiología , Hidrodinámica
7.
Mol Cell Biochem ; 462(1-2): 11-23, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31446616

RESUMEN

BRCA-1 is a nuclear protein involved in DNA repair, transcriptional regulation, and cell cycle control. Its involvement in other cellular processes has been described. Here, we aimed to investigate the role of BRCA-1 in macrophages M(LPS), M(IL-4), and tumor cell-induced differentiation. We used siRNAs to knockdown BRCA-1 in RAW 264.7 macrophages exposed to LPS, IL-4, and C6 glioma cells conditioned medium (CMC6), and evaluated macrophage differentiation markers and functional phagocytic activity as well as DNA damage and cell survival in the presence and absence of BRCA-1. LPS and CMC6, but not by IL-4, increased DNA damage in macrophages, and this effect was more pronounced in BRCA-1-depleted cells, including M(IL-4). BRCA-1 depletion impaired expression of pro-inflammatory cytokines, TNF-α and IL-6, and reduced the phagocytic activity of macrophages in response to LPS. In CMC6-induced differentiation, BRCA-1 knockdown inhibited TNF-α and IL-6 expression which was accompanied by upregulation of the anti-inflammatory markers IL-10 and TGF-ß and reduced phagocytosis. In contrast, M(IL-4) phenotype was not affected by BRCA-1 status. Molecular docking predicted that the conserved BRCA-1 domain BRCT can interact with the p65 subunit of NF-κB. Immunofluorescence assays showed that BRCA-1 and p65 co-localize in the nucleus of LPS-treated macrophages and reporter gene assay showed that depletion of BRCA-1 decreased LPS and CMC6-induced NF-κB transactivation. IL-4 had no effect upon NF-κB. Taken together, our findings suggest a role of BRCA-1 in macrophage differentiation and phagocytosis induced by LPS and tumor cells secretoma, but not IL-4, in a mechanism associated with inhibition of NF-κB.


Asunto(s)
Proteína BRCA1/metabolismo , Polaridad Celular , Inflamación/patología , Activación de Macrófagos , Macrófagos/metabolismo , Macrófagos/patología , FN-kappa B/metabolismo , Animales , Biomarcadores/metabolismo , Ciclo Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Polaridad Celular/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Medios de Cultivo Condicionados/farmacología , Daño del ADN , Inflamación/metabolismo , Lipopolisacáridos , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Metaloproteinasas de la Matriz/metabolismo , Ratones , Fagocitosis/efectos de los fármacos , Células RAW 264.7 , ARN Interferente Pequeño/metabolismo , Ratas
8.
Phytother Res ; 33(5): 1394-1403, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30868680

RESUMEN

Obesity is a metabolic disorder associated with adverse health consequences that has increased worldwide at an epidemic rate. This has encouraged many people to utilize nonprescription herbal supplements for weight loss without knowledge of their safety or efficacy. However, mounting evidence has shown that some herbal supplements used for weight loss are associated with adverse effects. Guarana seed powder is a popular nonprescription dietary herb supplement marketed for weight loss, but no study has demonstrated its efficacy or safety when administered alone. Wistar rats were fed four different diets (low-fat diet and Western diet with or without guarana supplementation) for 18 weeks. Metabolic parameters, gut microbiota changes, and toxicity were then characterized. Guarana seed powder supplementation prevented weight gain, insulin resistance, and adipokine dysregulation induced by Western diet compared with the control diet. Guarana induced brown adipose tissue expansion, mitochondrial biogenesis, uncoupling protein-1 overexpression, AMPK activation, and minor changes in gut microbiota. Molecular docking suggested a direct activation of AMPK by four guarana compounds tested here. We propose that brown adipose tissue activation is one of the action mechanisms involved in guarana supplementation-induced weight loss and that direct AMPK activation may underlie this mechanism. In summary, guarana is an attractive potential therapeutic agent to treat obesity.


Asunto(s)
Adipoquinas/metabolismo , Tejido Adiposo Pardo/efectos de los fármacos , Resistencia a la Insulina , Paullinia/química , Animales , Dieta Alta en Grasa/efectos adversos , Dieta Occidental , Suplementos Dietéticos , Humanos , Masculino , Simulación del Acoplamiento Molecular , Obesidad/metabolismo , Ratas , Ratas Wistar , Aumento de Peso , Pérdida de Peso/efectos de los fármacos
9.
Cell Physiol Biochem ; 42(6): 2507-2522, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28848092

RESUMEN

BACKGROUND/AIMS: Heat shock protein 70 (HSP70) has been recently described with extracellular actions, where it is actively released in inflammatory conditions. Acting as DAMPs (damage associated molecular pattern), extracellular HSP70 (eHSP70) interacts with membrane receptors and activates inflammatory pathways. At this context, the receptor for advanced glycation endproducts (RAGE) emerges as a possible candidate for interaction with eHSP70. RAGE is a pattern-recognition receptor and its expression is increased in several diseases related to a chronic pro-inflammatory state. One of the main consequences of RAGE ligand-binding is the ERK1/2 (extracellular signal-regulated kinases)-dependent activation of NF-kB (nuclear factor kappa B), which leads to expression of TNF-α (tumor necrosis factor alpha) and other cytokines. The purpose of this work is to elucidate if eHSP70 is able to evoke RAGE-dependent signaling using A549 human lung cancer cells, which constitutively express RAGE. METHODS: Immunoprecipitation and protein proximity assay were utilized to demonstrate the linkage between RAGE and eHSP70. To investigate RAGE relevance on cell response to eHSP70, siRNA was used to knockdown the receptor expression. Signaling pathways activation were evaluated by western blotting, gene reporter luciferase and real time quantitative PCR. RESULTS: Protein eHSP70 shown to be interacting physically with the receptor RAGE in our cell model. Treatment with eHSP70 caused ERK1/2 activation and NF-κB transactivation impaired by RAGE knockdown. Moreover, the stimulation of pro-inflammatory cytokines expression by eHSP70 was inhibited in RAGE-silenced cells. Finally, conditioned medium of eHSP70-treated A549 cells caused differential effects in monocytes cytokine expression when A549 RAGE expression is inhibited. CONCLUSIONS: Our results evidence eHSP70 as a novel RAGE agonist capable of influence the cross-talk between cancer and immune system cells.


Asunto(s)
Proteínas HSP70 de Choque Térmico/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , FN-kappa B/metabolismo , Receptor para Productos Finales de Glicación Avanzada/metabolismo , Células A549 , Citocinas/genética , Citocinas/metabolismo , Proteínas HSP70 de Choque Térmico/genética , Proteínas HSP70 de Choque Térmico/farmacología , Humanos , Immunoblotting , Inmunoprecipitación , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , FN-kappa B/genética , Fosforilación , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptor para Productos Finales de Glicación Avanzada/antagonistas & inhibidores , Receptor para Productos Finales de Glicación Avanzada/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/farmacología , Transducción de Señal/efectos de los fármacos , Activación Transcripcional , Células U937
10.
Mol Phylogenet Evol ; 111: 18-34, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-28279808

RESUMEN

Aiming at increasing the knowledge on marine cyanobacteria from temperate regions, we previously isolated and characterized 60 strains from the Portuguese foreshore and evaluate their potential to produce secondary metabolites. About 15% of the obtained 16S rRNA gene sequences showed less than 97% similarity to sequences in the databases revealing novel biodiversity. Herein, seven of these strains were extensively characterized and their classification was re-evaluated. The present study led to the proposal of five new taxa, three genera (Geminobacterium, Lusitaniella, and Calenema) and two species (Hyella patelloides and Jaaginema litorale). Geminobacterium atlanticum LEGE 07459 is a chroococcalean that shares morphological characteristics with other unicellular cyanobacterial genera but has a distinct phylogenetic position and particular ultrastructural features. The description of the Pleurocapsales Hyella patelloides LEGE 07179 includes novel molecular data for members of this genus. The filamentous isolates of Lusitaniella coriacea - LEGE 07167, 07157 and 06111 - constitute a very distinct lineage, and seem to be ubiquitous on the Portuguese coast. Jaaginema litorale LEGE 07176 has distinct characteristics compared to their marine counterparts, and our analysis indicates that this genus is polyphyletic. The Synechococcales Calenema singularis possess wider trichomes than Leptolyngbya, and its phylogenetic position reinforces the establishment of this new genus.


Asunto(s)
Cianobacterias/clasificación , Océano Atlántico , Cianobacterias/citología , Cianobacterias/genética , Cianobacterias/ultraestructura , ADN Bacteriano/genética , Genes Bacterianos , Funciones de Verosimilitud , Fijación del Nitrógeno/genética , Filogenia , Portugal , ARN Ribosómico 16S/genética , Especificidad de la Especie
11.
J Toxicol Environ Health A ; 78(7): 432-42, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25785557

RESUMEN

The production of bioactive compounds either toxic or with pharmacological applications by cyanobacteria is well established. However, picoplanktonic forms within this group of organisms have rarely been studied in this context. In this study, the toxicological potential of picocyanobacteria from a clade of marine Cyanobium strains isolated from the Portuguese coast was examined using different biological models. First, strains were identified by applying morphological and molecular approaches and cultured under lab conditions. A crude extract and three fractions reflecting a preliminary segregation of lipophilic metabolites were tested for toxicity with the marine microalga Nannochloropsis sp., the bacteria Pseudomonas sp., the brine shrimp Artemia salina, and fertilized eggs of the sea urchin Paracentrotus lividus. No significant apparent adverse effects were noted against Artemia salina. However, significant adverse effects were found in all other assays, with an inhibition of Nannochloropsis sp. and Pseudomonas sp. growth and marked reduction in Paracentrotus lividus larvae length. The results obtained indicated that Cyanobium genus may serve as a potential source of interesting bioactive compounds and emphasize the importance of also studying smaller picoplanktonic fractions of marine cyanobacteria.


Asunto(s)
Organismos Acuáticos/microbiología , Cianobacterias/aislamiento & purificación , Microalgas/microbiología , Animales , Artemia/microbiología , Bioensayo , Productos Biológicos/química , Productos Biológicos/farmacología , Clonación Molecular , Cianobacterias/clasificación , Cianobacterias/metabolismo , ADN Bacteriano/genética , Larva/microbiología , Filogenia , Análisis de Secuencia de ADN
12.
Appl Microbiol Biotechnol ; 98(19): 8073-82, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25085613

RESUMEN

Cyanobacteria blooms are since early times a cause for environmental concern because of their negative impact through the release of odors, water discoloration, and more dangerously through the release of toxic compounds (i.e. the cyanotoxins) that can affect both human and animal welfare. Surveillance of the aquatic ecosystems is therefore obligatory, and methods to achieve such require a prompt answer not only regarding the species that are producing the blooms but also the cyanotoxins that are being produced and/or released. Moreover, besides this well-known source of possible intoxication, it has been demonstrated the existence of several other potential routes of exposure, either for humans or other biota such as through food additives and in terrestrial environments (in plants, lichens, biological soil crusts) and the recognition of their harmful impact on less studied ecosystems (e.g. coral reefs). Nowadays, the most frequent approaches to detect toxic cyanobacteria and/or their toxins are the chemical-, biochemical-, and molecular-based methods. Above their particular characteristics and possible applications, they all bring to the environmental monitoring several aspects that are needed to be discussed and scrutinized. The end outcome of this review will be to provide newer insights and recommendations regarding the methods needed to apply in an environmental risk assessment program. Therefore, a current state of the knowledge concerning the three methodological approaches will be presented, while highlighting positive and negative aspects of each of those methods within the purpose of monitoring or studying cyanobacteria and their toxins in the environment.


Asunto(s)
Toxinas Bacterianas/análisis , Cianobacterias/aislamiento & purificación , Microbiología Ambiental , Monitoreo del Ambiente/métodos , Toxinas Bacterianas/metabolismo , Cianobacterias/química , Cianobacterias/genética , Cianobacterias/metabolismo
13.
J Stroke Cerebrovasc Dis ; 23(5): 1262-4, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24144594

RESUMEN

We describe a case of a 34-year-old man with a sudden development of right hemiparesis and aphasia because of infarction of the left middle cerebral artery that was submitted to intravenous (IV) recombinant tissue plasminogen activator and mechanical thrombectomy. Transesophageal echocardiogram showed a small mass on the anterior leaflet of the mitral valve. Cardiac surgery was performed, and histological examination of the removed material was consistent with cardiac papillary fibroelastoma (CPF). Experience in using IV thrombolysis for the treatment of embolic stroke because of CPF is limited. To the best of our knowledge, only 3 patients are reported in literature in whom acute ischemic stroke and associated CPF were treated with thrombolytic therapy. A discussion of the efficacy of IV thrombolysis and the possible superiority of mechanical thrombectomy is included.


Asunto(s)
Fibrinolíticos/administración & dosificación , Fibroma/complicaciones , Neoplasias Cardíacas/complicaciones , Infarto de la Arteria Cerebral Media/terapia , Embolia Intracraneal/terapia , Trombectomía , Terapia Trombolítica , Activador de Tejido Plasminógeno/administración & dosificación , Adulto , Angiografía Cerebral/métodos , Ecocardiografía Transesofágica , Fibroma/diagnóstico , Neoplasias Cardíacas/diagnóstico , Humanos , Infarto de la Arteria Cerebral Media/diagnóstico , Infarto de la Arteria Cerebral Media/etiología , Infusiones Intravenosas , Embolia Intracraneal/diagnóstico , Embolia Intracraneal/etiología , Imagen por Resonancia Magnética , Masculino , Proteínas Recombinantes/administración & dosificación , Resultado del Tratamiento
14.
Mar Drugs ; 11(12): 4902-16, 2013 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-24351973

RESUMEN

Cyanobactins are a recently recognized group of ribosomal cyclic peptides produced by cyanobacteria, which have been studied because of their interesting biological activities. Here, we have used a PCR-based approach to detect the N-terminal protease (A) gene from cyanobactin synthetase gene clusters, in a set of diverse cyanobacteria from our culture collection (Laboratory of Ecotoxicology, Genomics and Evolution (LEGE) CC). Homologues of this gene were found in Microcystis and Rivularia strains, and for the first time in Cuspidothrix, Phormidium and Sphaerospermopsis strains. Phylogenetic relationships inferred from available A-gene sequences, including those obtained in this work, revealed two new groups of phylotypes, harboring Phormidium, Sphaerospermopsis and Rivularia LEGE isolates. Thus, this study shows that, using underexplored cyanobacterial strains, it is still possible to expand the known genetic diversity of genes involved in cyanobactin biosynthesis.


Asunto(s)
Proteínas Bacterianas/genética , Cianobacterias/genética , Variación Genética/genética , Péptidos Cíclicos/genética , Serina Endopeptidasas/genética , Familia de Multigenes/genética , Filogenia
15.
Mar Drugs ; 11(4): 1316-35, 2013 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-23609580

RESUMEN

Marine cyanobacteria, notably those from tropical regions, are a rich source of bioactive secondary metabolites. Tropical marine cyanobacteria often grow to high densities in the environment, allowing direct isolation of many secondary metabolites from field-collected material. However, in temperate environments culturing is usually required to produce enough biomass for investigations of their chemical constituents. In this work, we cultured a selection of novel and diverse cyanobacteria isolated from the Portuguese coast, and tested their organic extracts in a series of ecologically-relevant bioassays. The majority of the extracts showed activity in at least one of the bioassays, all of which were run in very small scale. Phylogenetically related isolates exhibited different activity profiles, highlighting the value of microdiversity for bioprospection studies. Furthermore, LC-MS analyses of selected active extracts suggested the presence of previously unidentified secondary metabolites. Overall, the screening strategy employed here, in which previously untapped cyanobacterial diversity was combined with multiple bioassays, proved to be a successful strategy and allowed the selection of several strains for further investigations based on their bioactivity profiles.


Asunto(s)
Cianobacterias/química , Filogenia , Bioensayo , Cromatografía Liquida , Cianobacterias/metabolismo , Espectrometría de Masas , Portugal
16.
Rev Port Cardiol ; 32(2): 149-52, 2013 Feb.
Artículo en Portugués | MEDLINE | ID: mdl-23351919

RESUMEN

Among cardiovascular diseases, pericardial disease has specific characteristics. Its etiology, diagnosis and medical management are not as well understood as in coronary and valvular heart disease. In most cases, its cause is benign, although the proportion decreases with more severe clinical presentation. The authors present the case of a 35-year-old man with no relevant past medical history, who went to the emergency department with what appeared to be an idiopathic case of acute pericarditis. However, over the following five months, there was an unfavorable evolution to constrictive pericarditis, requiring pericardiectomy. The final diagnosis was only made following surgery - a rare case of a primary pericardial tumor, a mesothelioma.


Asunto(s)
Cardiopatías/complicaciones , Mesotelioma/complicaciones , Pericarditis Constrictiva/etiología , Pericardio , Adulto , Cardiopatías/diagnóstico , Humanos , Masculino , Mesotelioma/diagnóstico
17.
Microb Ecol ; 63(1): 85-95, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21947429

RESUMEN

An increasing body of evidence points out that allelopathy may be an important process shaping microbial communities in aquatic ecosystems. Cyanobacteria have well-documented allelopathic properties, mainly derived from the evaluation of the activity of allelopathic extracts or pure compounds towards monocultures of selected target microorganisms. Consequently, little is known regarding the community dynamics of microorganisms associated with allelopathic interactions. In this laboratory-based study, a Microcystis spp.-dominated microbial community from a freshwater lake was exposed, for 15 days, to exudates from the cyanobacterium Oscillatoria sp. strain LEGE 05292 in laboratory conditions. This cyanobacterium is known to produce the allelochemicals portoamides, which were among the exuded compounds. The community composition was followed (by means of polymerase chain reaction followed by denaturing gradient gel electrophoresis and microscopic analyses) and compared to that of a non-exposed situation. Following exposure, clear differences in the community structure were observed, in particular for cyanobacteria and unicellular eukaryotic taxa. Interestingly, distinct Microcystis genotypes present in the community were differentially impacted by the exposure, highlighting the fine-scale dynamics elicited by the exudates. These results support a role for cyanobacterial allelochemicals in the structuring of aquatic microbial communities.


Asunto(s)
Lagos/microbiología , Consorcios Microbianos/efectos de los fármacos , Microcystis/efectos de los fármacos , Oscillatoria/metabolismo , Feromonas/farmacología , Genes de ARNr , Genotipo , Microcystis/genética , Microcystis/crecimiento & desarrollo , Microcystis/aislamiento & purificación , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genética
18.
Sci Rep ; 12(1): 492, 2022 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-35017549

RESUMEN

Philaenus spumarius is a vector of Xylella fastidiosa, one of the most dangerous plants pathogenic bacteria worldwide. There is currently no control measure against this pathogen. Thus, the development of vector control strategies, like generalist predators, such as spiders, could be essential to limit the spread of this vector-borne pathogen. In this study, a polymerase chain reaction (PCR)-based approach was developed to principally detect DNA of P. spumarius in the spider's gut. Accordingly, 20 primer pairs, targeting the mitochondrial cytochrome oxidase I (COI) and cytochrome b (cytB) genes, were tested for specificity, sensitivity, and efficiency in detecting P. spumarius DNA. Overall, two primer sets, targeting COI gene (COI_Ph71F/COI_Ph941R) and the cytB gene (cytB_Ph85F/cytB_Ph635R), showed the highest specificity and sensitivity, being able to amplify 870 pb and 550 bp fragments, respectively, with P. spumarius DNA concentrations 100-fold lower than that of the DNA of non-target species. Among these two primer sets, the cytB_Ph85F/cytB_Ph635R was able to detect P. spumarius in the spider Xysticus acerbus, reaching 50% detection success 82 h after feeding. The feasibility of this primer set to detect predation of P. spumarius by spiders was confirmed in the field, where 20% of the collected spiders presented positive amplifications.


Asunto(s)
ADN/genética , Hemípteros/genética , Insectos Vectores/genética , Arañas/fisiología , Animales , Cartilla de ADN/genética , Conducta Alimentaria , Tracto Gastrointestinal/metabolismo , Hemípteros/metabolismo , Insectos Vectores/metabolismo , Control Biológico de Vectores , Reacción en Cadena de la Polimerasa , Conducta Predatoria
19.
Sci Rep ; 12(1): 2646, 2022 02 16.
Artículo en Inglés | MEDLINE | ID: mdl-35173202

RESUMEN

Cork oak is a tree species with ecological importance that contributes to economic and social development in the Mediterranean region. Cork oak decline is a major concern for forest sustainability and has negative impacts on cork oak growth and production. This event has been increasingly reported in the last decades and seems to be related with climate changes. Biscogniauxia mediterranea is an endophytic fungus of healthy cork oak trees that turns into a pathogen in trees weaken by environmental stress. Understanding the drivers of B. mediterranea populations diversity and differentiation is expected to allow a better control of cork oak decline and preserve forest sustainability. Endophyte isolates from different cork oak forests were identified as B. mediterranea and their genetic diversity was evaluated using phylogenetic and microsatellite-primed PCR analyses. Genetic diversity and variability of this fungus was correlated with environmental/phytosanitary conditions present in forests/trees from which isolates were collected. High genetic diversity and variability was found in B. mediterranea populations obtained from different forests, suggesting some degree of isolation by distance. Bioclimate was the most significant effect that explained the genetic variability of B. mediterranea, rather than precipitation or temperature intensities alone or disease symptoms. These findings bring new implications for the changing climate to cork oak forests sustainability, cork production and quality.


Asunto(s)
Ascomicetos/genética , Ascomicetos/patogenicidad , Cambio Climático , Bosques , Variación Genética , Filogenia , Quercus/microbiología , Ascomicetos/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos
20.
RSC Adv ; 12(41): 26846-26858, 2022 Sep 16.
Artículo en Inglés | MEDLINE | ID: mdl-36320853

RESUMEN

This work reports the application of Mn-doped Co3O4 oxides in the electrocatalytic oxygen evolution reaction (OER). The materials were characterized by structural, morphological, and electrochemical techniques. The oxides with higher Co : Mn molar ratio presented a lower electron transfer resistance, and consequently the most promising OER activities. Pure Co3O4 shows an overpotential at j = 10 mA cm-2 of 761, 490, and 240 mV, at pH 1, 7, and 14, respectively, and a high TOF of 1.01 × 10-1 s-1 at pH 14. Tafel slopes around 120 mV dec-1 at acidic pH and around 60 mV dec-1 at alkaline pH indicate different OER mechanisms. High stability for Co3O4 was achieved for up to 15 h in all pHs, and no change in the structure and morphology after the electrocatalysis was observed. The reported excellent OER activity of the Mn-Co oxides in a wide pH range is important to broaden the practical applicability in different electrolyte solutions.

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