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Every plant organ contains tens of different cell types, each with a specialized function. These functions are intrinsically associated with specific metabolic flux distributions that permit the synthesis of the ATP, reducing equivalents and biosynthetic precursors demanded by the cell. Investigating such cell-type-specific metabolism is complicated by the mosaic of different cells within each tissue combined with the relative scarcity of certain types. However, techniques for the isolation of specific cells, their analysis in situ by microscopy, or modeling of their function in silico have permitted insight into cell-type-specific metabolism. In this review we present some of the methods used in the analysis of cell-type-specific metabolism before describing what we know about metabolism in several cell types that have been studied in depth; (i) leaf source and sink cells; (ii) glandular trichomes that are capable of rapid synthesis of specialized metabolites; (iii) guard cells that must accumulate large quantities of the osmolytes needed for stomatal opening; (iv) cells of seeds involved in storage of reserves; and (v) the mesophyll and bundle sheath cells of C4 plants that participate in a CO2 concentrating cycle. Metabolism is discussed in terms of its principal features, connection to cell function and what factors affect the flux distribution. Demand for precursors and energy, availability of substrates and suppression of deleterious processes are identified as key factors in shaping cell-type-specific metabolism.
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Fotossíntese , Folhas de Planta , Folhas de Planta/metabolismoRESUMO
The simplification and fragmentation of agricultural landscapes generate effects on insects at multiple spatial scales. As each functional group perceives and uses the habitat differently, the response of pest insects and their associated natural enemies to environmental changes varies. Therefore, landscape structure may have consequences on gene flow among pest populations in space. This study aimed to evaluate the effects of local and landscape factors, at multiple scales, on the local infestation, gene flow and broad dispersion dynamics of the pest insect Bemisia tabaci (Genn.) Middle East-Asia Minor 1 (MEAM-1, former biotype B) (Hemiptera: Aleyrodidae) and its associated natural enemies in a tropical agroecosystem. We evaluated the abundance of B. tabaci populations and their natural enemy community in 20 tomato farms in Brazil and the gene flow between farms from 2019 to 2021. Landscapes dominated by agriculture resulted in larger B. tabaci populations and higher gene flow, especially in conventional farms. A higher density of native vegetation patches disfavored pest populations, regardless of the management system. The results revealed that whitefly responds to intermediate spatial scales and that landscape factors interact with management systems to modulate whitefly populations on focal farms. Conversely, whitefly natural enemies benefited from higher amounts of natural vegetation at small spatial scales, while the connectivity between natural habitat patches was beneficial for natural enemies regardless of the distance from the focal farm. The resulting dispersion model predicts that the movement of whiteflies between farms increases as the amount of natural vegetation decreases. Our findings demonstrate that landscape features, notably landscape configuration, can mediate infestation episodes, as they affect pest insects and natural enemies in opposite ways. We also showed that landscape features interact with farm traits, which highlights the need for management strategies at multiple spatial scales. In conclusion, we demonstrated the importance of the conservation of natural areas as a key strategy for area-wide ecological pest management and the relevance of organic farming to benefit natural enemy communities in tropical agroecosystems.
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Agricultura , Fluxo Gênico , Fazendas , Brasil , MovimentoRESUMO
aste collectors are exposed to a wide variety of bacteria, endotoxins, fungi, allergens, particulate matter, irritating inhalants, and vehicle exhaust, making them more prone to development of chronic diseases. Although several studies described the impact of occupational exposure on the overall health of waste collectors, few investigations were conducted regarding cellular and molecular changes that may occur due to exposure. The aim of this study was to assess biomarkers of oxidative stress such as levels of reactive oxygen species (ROS), lipoperoxidation, total antioxidant capacity (TAC), apoptosis, butyrylcholinesterase (BChE) activity and mitochondrial function (MitoTrackerTM Green FM and MitoTrackerTM Red) using the peripheral blood from individuals occupationally exposed to recyclable solid waste in Southern Brazil. The study included 30 waste collectors and 30 control individuals, who did not perform activities with recognized exposure to biological and chemical substances. Waste collectors were found to exhibit in peripheral blood leukocytes (PBL) higher rates of apoptosis, increased production of ROS, and reduced mitochondrial membrane potential (MMP), associated with decreased total antioxidant capacity (TAC) and elevated activity of BChE in plasma. Therefore, evidence indicates that cytotoxicity, oxidative stress, and inflammatory responses may be involved in the multiplicity of adverse health outcomes related to contaminant exposure in waste collectors. It is thus necessary to implement and/or improve occupational health programs aimed at workers as well as mandatory inspections for the use of personal protective equipment.
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Antineoplásicos , Antioxidantes , Humanos , Resíduos Sólidos , Butirilcolinesterase , Espécies Reativas de OxigênioRESUMO
There has been a proliferation of studies that have examined the impacts of public participation in Citizen Science (CS) that respond to environmental challenges and the recovery of ecosystems, endangered species or other important natural assets. However, comparatively few studies have explored how tourists may play a critical role in the generation of CS data and thus it has been posited that many potential opportunities remain unrealised. By systematically analysing studies that have utilised tourist-generated data in response to environmental challenges or issues, this paper seeks to establish an appraisal of what has so far been established in extant literature and to identify future possibilities for the inclusion of tourists in CS. Via our literature search, a total of 45 peer-reviewed studies were identified via the PRISMA search protocol. Our findings reveal numerous positive outcomes were reported that highlight the significant, yet largely untapped, potential of tourist integration in CS, with studies also offering a range of recommendations on how tourists could be included more effectively to expand scientific knowledge. Notwithstanding, several limitations were observed, and it is critical that future CS projects that utilise tourists for data collection purposes are acutely aware of the challenges they may encounter.
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Ciência do Cidadão , Humanos , Animais , Ecossistema , Participação da Comunidade , Coleta de Dados , Espécies em Perigo de ExtinçãoRESUMO
AIMS: We searched for recurrent pathological features and molecular alterations in a retrospective series of 72 low-grade epilepsy-associated neuroepithelial tumours (LEATs) with a prominent oligodendroglioma-like component, in order to classify them according to the 2021 World Health Organization (WHO) classification of central nervous system (CNS) tumours. METHODS: Centralised pathological examination was performed as well as targeted molecular analysis of v-Raf murine sarcoma viral oncogene homologue B (BRAF) and fibroblast growth factor receptor 1 (FGFR1) by multiplexed digital polymerase chain reaction (mdPCR). DNA methylation profiling was performed in cases with sufficient DNA. In cases with no genetic alteration by mdPCR and sufficient material, RNA sequencing was done. RESULTS: We first reclassified our cohort into three groups: ganglioglioma (GG, n = 14), dysembryoplastic neuroepithelial tumours (DNTs, n = 19) and glioneuronal tumours/paediatric-type low-grade glioma (LGG) not otherwise specified (GNT/PLGG NOS, n = 39). mdPCR found an alteration in 38/72 cases. Subsequent RNA sequencing revealed a fusion transcript involving BRAF, FGFR1/2/3 or neurotrophic tyrosine kinase receptor type 2 [NTRK2] in 9/25 cases. DNA methylation profiling found 12/46 cases with a calibrated score ≥0.9. Unsupervised hierarchical clustering revealed two clusters: Cluster 1 was enriched with cases classified as DNT at histology, belonging to the LGG-DNT methylation class (MC), with haematopoietic progenitor cell antigen (CD34) negativity and FGRF1 alterations; Cluster 2 was enriched with cases classified at histology as GG, belonging to the LGG-GG MC MC, with BRAF V600E mutation and CD34 positivity. The tumours reclassified as GNT/PLGG NOS were equally distributed across both clusters. Interestingly, all polymorphous low-grade neuroepithelial tumour of the young belonged to Cluster 2, whereas diffuse LGG mitogen-activated protein kinase (MAPK) pathway-altered were equally distributed among the two clusters. This led us to build an algorithm to classify LEATs with a prominent oligodendroglioma-like component. CONCLUSIONS: Integrated histomolecular diagnosis of LEATs with a prominent oligodendroglioma-like component remains challenging. Because these tumours can be split into two major clusters of biological significance, the clinicopathological relevance of the four types recognised by the WHO CNS5 within this spectrum of tumours is questionable.
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Neoplasias Encefálicas/patologia , Encéfalo/patologia , Epilepsia/patologia , Neoplasias Neuroepiteliomatosas/patologia , Oligodendroglia/patologia , Adolescente , Adulto , Neoplasias Encefálicas/complicações , Neoplasias Encefálicas/genética , Criança , Pré-Escolar , Metilação de DNA , Epilepsia/etiologia , Epilepsia/genética , Feminino , Humanos , Lactente , Masculino , Neoplasias Neuroepiteliomatosas/complicações , Neoplasias Neuroepiteliomatosas/genética , Estudos Retrospectivos , Adulto JovemRESUMO
We wished to determine if Mycoplasma bovis infection can negatively impact milk quality and production in Holstein dairy cows. For this Research Communication, milk samples (271) from Holstein cows from 3 herds were screened for M. bovis by real-time PCR. Positive (n = 21) and negative animals (n = 21) were matched by herd, age, lactations and days in milk (DIM). Pairs were evaluated in 7 stages of lactation: D1-50, D51-100, D101-150, D151-200, D201-250, D251-300, and D ≥ 301. A mixed model was used to assess the effect of groups (M. bovis+ × M.bovis-), time (lactation) and groups × time interaction. Cows positive for M. bovis had lower average milk production per day and high somatic cells count (SCC).
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Doenças dos Bovinos , Infecções por Mycoplasma , Mycoplasma bovis , Feminino , Bovinos , Animais , Leite , Lactação , Infecções por Mycoplasma/veterináriaRESUMO
This research communication presents an automatic method for the counting of somatic cells in buffalo milk, which includes the application of a fuzzy clustering method and image processing techniques (somatic cell count with fuzzy clustering and image processing|, SCCFCI). Somatic cell count (SCC) in milk is the main biomarker for assessing milk quality and it is traditionally performed by exhaustive methods consisting of the visual observation of cells in milk smears through a microscope, which generates uncertainties associated with human interpretation. Unlike other similar works, the proposed method applies the Fuzzy C-Means (FCM) method as a preprocessing step in order to separate the images (objects) of the cells into clusters according to the color intensity. This contributes signficantly to the performance of the subsequent processing steps (thresholding, segmentation and recognition/identification). Two methods of thresholding were evaluated and the Watershed Transform was used for the identification and separation of nearby cells. A detailed statistical analysis of the results showed that the SCCFCI method is able to provide results which are consistent with those obtained by conventional counting. This method therefore represents a viable alternative for quality control in buffalo milk production.
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Búfalos , Contagem de Células/veterinária , Processamento de Imagem Assistida por Computador/métodos , Mastite/veterinária , Leite/citologia , Animais , Contagem de Células/métodos , Feminino , Mastite/patologia , Microscopia , FotografaçãoRESUMO
Diffuse gliomas with MYB or MYBL1 alterations are rare tumours mostly affecting children or young adults with long-term epilepsy. This category of glioma includes two morphological subtypes. The angiocentric subtype is characterized by an angiocentric pattern of growth and a frequent MYB:QKI fusion. The isomorphic subtype corresponds to a highly differentiated astrocytic glioma with low cellularity, low proliferation and no specific microscopic features. The diagnosis is based on the imaging, demonstrating a supratentorial tumor, associated with the confirmation of a MYB or MYBL1 rearrangement. Here, we report the case of a 7-year-old child who presented a right frontal brain lesion corresponding to an isomorphic diffuse glioma with MYBL1 alteration.
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Astrocitoma , Neoplasias Encefálicas , Glioma , Neoplasias Supratentoriais , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/genética , Criança , Glioma/diagnóstico , Glioma/genética , Humanos , Proteínas Proto-Oncogênicas , Transativadores , Adulto JovemRESUMO
The use of blends to produce hydrogels allows modulating their characteristics as mechanical properties and microstructure. This work aimed to study the properties of pectin and starch hydrogel blends. Pectin gel was homogeneous and porous, while pectin/starch blends containing 50% or more pectin exhibited denser and closer network, indicating that starch reduced the porosity of pectin network. Such characteristic was associated with higher gel hardness, cohesiveness, firmness, and water holding capacity. The influence of total biopolymer concentration and type of process (extrusion and atomization) on particle formation were also evaluated indicating that among the tested formulations, pectin 1% and starch 1% blend was the only sample able to form particles under extrusion and atomization. The addition of 5% (w/v) microparticles to the grape nectar presented no influence on rheological parameters, maintaining the pseudoplastic behavior. Both the starch addition and the amount of polymers used impacted the micro and macrostructure of pectin gels.
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The Earth is getting brighter at night, as artificial light at night (ALAN) continues to increase and extend its reach. Despite recent recognition of the damaging impacts of ALAN on terrestrial ecosystems, research on ALAN in marine systems is comparatively lacking. To further our understanding of the impacts of ALAN on marine organisms, this study examines how the reproductive fitness of the common clownfish Amphiprion ocellaris is influenced by the presence of ALAN. We assessed how exposure to low levels of ALAN affects (i) frequency of spawning, (ii) egg fertilization success, and (iii) hatching success of A. ocellaris under control (12 : 12 day-night) and treatment (12 : 12 day-ALAN) light regimes. While we found exposure to ALAN had no impact on the frequency of spawning or fertilization success, ALAN had dramatic effects on hatching. Amphiprion ocellaris eggs incubated in the presence of ALAN simply did not hatch, resulting in zero survivorship of offspring. These findings suggest ALAN can significantly reduce reproductive fitness in a benthic-spawning reef fish. Further research in this field is necessary to fully understand the extent of this impact on population and community dynamics in the wild.
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Luz , Perciformes , Animais , Ecossistema , PeixesRESUMO
Flavylium cations serve as models for the chemical and photochemical reactivity of anthocyanins, the natural plant pigment responsible for many of the red, blue and purple colors of fruits and flowers. Likewise, pyranoflavylium cations serve as models of the fundamental chromophoric moiety of pyranoanthocyanins, molecules that can form from reactions of grape anthocyanins in red wines during their maturation. In the present work, hybrid pigments are prepared by the adsorption of a series of five synthetic flavylium cations (FL) and five synthetic pyranoflavylium cations (PFL) on sepiolite clay (SEP). The FL are smaller in size than the PFL, but both can in principle fit into the tunnels and/or external grooves (with dimensions of 3.7 × 10.6 Å) of SEP. Measurements of the fluorescence quantum yields of the adsorbed dyes indicate that they are at least as fluorescent as in acidic acetonitrile solution, and in a few cases substantially more fluorescent. The observation of biexponential fluorescence decays is consistent with emission from dye molecules adsorbed at two distinct sites, presumably tunnels and grooves. These hybrid materials also have improved properties in terms of stability of the color in contact with pH 10 aqueous solution and resistance to thermal degradation of the dye. SEP thus appears to be a promising substrate for the development of highly fluorescent flavylium or pyranoflavylium cation-derived hybrid pigments with improved color and thermal stability.
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OBJECTIVE: To propose parameters to qualify the filling of the Patient Classification System proposed by Perroca. METHOD: Validation study by consensus held in 2014, in a university hospital. The sample included 10 specialist nurses, four teachers and a computer science professional, all knowledgeable in the Classification System. The information was collected and analyzed through meetings registration, where strategies were discussed to enable the systematic application of the instrument in the institution. For the construction, it was considered a consensus of 100% among the experts. RESULTS: The computerized parameter guide was elaborated with a description of what to evaluate in each item of the Perroca's instrument, standardizing the filling by the nurses. CONCLUSIONS: The parameter guide made it possible to minimize the subjectivity of the evaluators when applying the classification instrument, provided guiding elements to fill it in and qualified the nursing care benchmarking.
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Consenso , Avaliação em Enfermagem , Cuidados de Enfermagem/organização & administração , Hospitais Universitários , Humanos , Monitorização Fisiológica/métodosRESUMO
Magnetotactic bacteria (MTB) are a heterogeneous group of ubiquitous aquatic microorganisms capable of biomineralizing nano-sized, membrane-bound, magnetic iron-rich mineral particles called magnetosomes. MTB are found in chemically-stratified aquatic sediments and/or water columns with a wide range of salinities, moderate to high temperatures, and pH varying from neutral to strongly alkaline. MTB from very cold environments have not been investigated to any great degree and here we characterize MTB from the low temperature Antarctic maritime region. Sediment samples were collected at nine sampling sites within Admiralty Bay, King George Island (62°23'S 58°27'W) from 2009 to 2013. Samples from five sites contained MTB and those from two of these sites contained large number of magnetotactic cocci that were studied using electron microscopy and molecular techniques. The magnetotactic cocci contained magnetosomes either arranged as two or four chains or as a disorganized cluster. The crystalline habit and composition of all magnetosomes analyzed with high-resolution transmission electron microscopy and energy dispersive X-ray microanalysis were consistent with elongated prismatic crystals of magnetite (Fe3 O4 ). The retrieved 16S rRNA gene sequences from magnetically-enriched magnetotactic cocci clustered into three distinct groups affiliated with the Alphaproteobacteria class of the Proteobacteria. Novel sequences of each phylogenetic cluster were confirmed using fluorescent in situ hybridization. Metagenomic data analysis of magnetically-enriched magnetotactic cocci revealed the presence of mam genes and MTB-specific hypothetical protein coding genes. Sequence homology and phylogenetic analysis indicated that predicted proteins are related to those of cultivated alphaproteobacterial MTB. The consistent and continuous low temperature of the sediment where the magnetotactic cocci are present (always below 1°C) suggests that these MTB from maritime Antarctica are psychrophiles. Moreover, similar morphotypes and 16S gene sequences were retrieved from samples collected from different sites from maritime Antarctica for several years suggesting that these new strains of MTB are indigenous members of Antarctic microbiota.
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Alphaproteobacteria/isolamento & purificação , Sedimentos Geológicos/microbiologia , Água do Mar/microbiologia , Alphaproteobacteria/classificação , Alphaproteobacteria/genética , Alphaproteobacteria/crescimento & desenvolvimento , Regiões Antárticas , Meios de Cultura/química , Meios de Cultura/metabolismo , DNA Bacteriano/genética , Sedimentos Geológicos/química , Hibridização in Situ Fluorescente , Magnetossomos , Microscopia Eletrônica de Transmissão , Filogenia , RNA Ribossômico 16S/genética , Salinidade , Água do Mar/químicaRESUMO
University teaching remains an area of concern, and perhaps the most difficult discipline for both teaching and learning is evolution. The concepts that underpin evolution, although complex, have been shown to be fairly straightforward, yet students arrive at and leave university with serious misconceptions, misunderstandings related to language, and often a reluctance to learn the subject because of cultural or societal pressures. Because of the unifying power of the theory, however, it is necessary not only for biology students to have a thorough understanding of evolution, but also for them to learn it in their first year so that this knowledge can then be taken into further years of study. Rather than teaching evolution at the end of a degree program, embedding it as a semester-long first-year course will ensure that a far larger number of students are made aware of misconceptions that they have brought with them from high school. Teaching through traditional passive lectures makes learning difficult conceptual material more difficult, and needs to be replaced with more interactive lectures coupled with inquiry-based practicals and small group-learning sessions to increase student engagement and interest in the subject. A new approach in pedagogy, curriculum design, and academic staff professional development is essential, especially at this time, when enrollments across science courses in many countries around the world are in decline.
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Genética/educação , Ensino , Animais , Evolução Biológica , Biologia/educação , Humanos , UniversidadesRESUMO
Brain banks manage and store fully clinically and pathologically characterised brains. The diversity of techniques used in research projects increases. These biological resource centres are made to adapt brain tissue processing. Furthermore, the development of more sensitive techniques to analyse nucleic acids and proteins offers new fields of exploration when combined with laser capture microdissection in order to decipher the physiopathology of diseases at the cell level. In this study, our goal was to evaluate procedures and set a workflow compatible with the constraints of brain banks, from brain sampling to laser capture microdissection and pre-analytical quality assessment. We compared various methods of freezing brain tissue, focused on morphological quality preservation of brain microscopical structures and on the quality of nucleic acid or protein yields. Staining protocols combined with strategies to lower neurones autofluorescence were adapted for the same purpose. Finally, we found that laser capture microdissection is possible in the setting of brain banks. However, the entire process has to be envisioned from the autopsy to the analysis. The impact on protein or nucleic acid quality is a limitation that restricts the amount of samples available for this purpose.
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Encéfalo/patologia , Microdissecção , Neurônios/patologia , Bancos de Tecidos , Fluxo de Trabalho , Adulto , Idoso , Idoso de 80 Anos ou mais , Encefalopatias/patologia , Europa (Continente) , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neurônios/metabolismo , Mudanças Depois da Morte , Proteínas/genética , Proteínas/metabolismo , Manejo de Espécimes , Coloração e RotulagemRESUMO
Magnetotactic bacteria (MTB) produce intracellular organelles called magnetosomes which are magnetic nanoparticles composed of magnetite (Fe3O4) or greigite (Fe3S4) enveloped by a lipid bilayer. The synthesis of a magnetosome is through a genetically controlled process in which the bacterium has control over the composition, direction of crystal growth, and the size and shape of the mineral crystal. As a result of this control, magnetosomes have narrow and uniform size ranges, relatively specific magnetic and crystalline properties, and an enveloping biological membrane. These features are not observed in magnetic particles produced abiotically and thus magnetosomes are of great interest in biotechnology. Most currently described MTB have been isolated from saline or brackish environments and the availability of their genomes has contributed to a better understanding and culturing of these fastidious microorganisms. Moreover, genome sequences have allowed researchers to study genes related to magnetosome production for the synthesis of magnetic particles for use in future commercial and medical applications. Here, we review the current information on the biology of MTB and apply, for the first time, a genome mining strategy on these microorganisms to search for secondary metabolite synthesis genes. More specifically, we discovered that the genome of the cultured MTB Magnetovibrio blakemorei, among other MTB, contains several metabolic pathways for the synthesis of secondary metabolites and other compounds, thereby raising the possibility of the co-production of new bioactive molecules along with magnetosomes by this species.
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Magnetossomos/química , Rhodospirillaceae/química , Produtos Biológicos/isolamento & purificação , Fenômenos Magnéticos , Rhodospirillaceae/genéticaRESUMO
The biomineralization of magnetosomes in Magnetospirillum gryphiswaldense and other magnetotactic bacteria occurs only under suboxic conditions. However, the mechanism of oxygen regulation and redox control of biosynthesis of the mixed-valence iron oxide magnetite [FeII(FeIII)2O4] is still unclear. Here, we set out to investigate the role of aerobic respiration in both energy metabolism and magnetite biomineralization of M. gryphiswaldense. Although three operons encoding putative terminal cbb3-type, aa3-type, and bd-type oxidases were identified in the genome assembly of M. gryphiswaldense, genetic and biochemical analyses revealed that only cbb3 and bd are required for oxygen respiration, whereas aa3 had no physiological significance under the tested conditions. While the loss of bd had no effects on growth and magnetosome synthesis, inactivation of cbb3 caused pleiotropic effects under microaerobic conditions in the presence of nitrate. In addition to their incapability of simultaneous nitrate and oxygen reduction, cbb3-deficient cells had complex magnetosome phenotypes and aberrant morphologies, probably by disturbing the redox balance required for proper growth and magnetite biomineralization. Altogether, besides being the primary terminal oxidase for aerobic respiration, cbb3 oxidase may serve as an oxygen sensor and have a further role in poising proper redox conditions required for magnetite biomineralization.
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Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Óxido Ferroso-Férrico/metabolismo , Magnetospirillum/enzimologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Regulação Bacteriana da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Genoma Bacteriano , Magnetospirillum/genética , Magnetospirillum/metabolismo , OxirreduçãoAssuntos
Carcinoma Hepatocelular/diagnóstico por imagem , Neoplasias Hepáticas/diagnóstico por imagem , Pancreatite/diagnóstico , Ascite/diagnóstico , Ascite/etiologia , Carcinoma Hepatocelular/complicações , Humanos , Neoplasias Hepáticas/complicações , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Magnetotactic bacteria are capable of synthesizing magnetosomes only under oxygen-limited conditions. However, the mechanism of the aerobic repression on magnetite biomineralization has remained unknown. In Escherichia coli and other bacteria, Fnr (fumarate and nitrate reduction regulator) proteins are known to be involved in controlling the switch between microaerobic and aerobic metabolism. Here, we report on an Fnr-like protein (MgFnr) and its role in growth metabolism and magnetite biomineralization in the alphaproteobacterium Magnetospirillum gryphiswaldense. RESULTS: Deletion of Mgfnr not only resulted in decreased N2 production due to reduced N2O reductase activity, but also impaired magnetite biomineralization under microaerobic conditions in the presence of nitrate. Overexpression of MgFnr in the WT also caused the synthesis of smaller magnetite particles under anaerobic and microaerobic conditions in the presence of nitrate. These data suggest that proper expression of MgFnr is required for WT-like magnetosome synthesis, which is regulated by oxygen. Analyses of transcriptional gusA reporter fusions revealed that besides showing similar properties to Fnr proteins reported in other bacteria, MgFnr is involved in the repression of the expression of denitrification genes nor and nosZ under aerobic conditions, possibly owing to several unique amino acid residues specific to MTB-Fnr. CONCLUSIONS: We have identified and thoroughly characterized the first regulatory protein mediating denitrification growth and magnetite biomineralization in response to different oxygen conditions in a magnetotactic bacterium. Our findings reveal that the global oxygen regulator MgFnr is a genuine O2 sensor. It is involved in controlling expression of denitrification genes and thereby plays an indirect role in maintaining proper redox conditions required for magnetite biomineralization.
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Desnitrificação , Óxido Ferroso-Férrico/metabolismo , Regulação Bacteriana da Expressão Gênica , Magnetospirillum/genética , Magnetospirillum/metabolismo , Fatores de Transcrição/metabolismo , Aerobiose , AnaerobioseRESUMO
Most magnetotactic bacteria (MB) produce stable, single-domain magnetite nanocrystals with species-specific size, shape and chain arrangement. In addition, most crystals are elongated along the [111] direction, which is the easy axis of magnetization in magnetite, chemically pure and structurally perfect. These special characteristics allow magnetite crystal chains from MB to be recognized in environmental samples including old sedimentary rocks. Ferromagnetic resonance (FMR) has been proposed as a powerful and practical tool for screening large numbers of samples possibly containing magnetofossils. Indeed, several studies were recently published on FMR of cultured MB, mainly Magnetospirillum gryphiswaldense. In this work, we examined both uncultured magnetotactic cocci and the cultured MB M. gryphiswaldense using transmission electron microscopy (TEM) and FMR from 10 K to room temperature (RT). The TEM data supported the FMR spectral characteristics of our samples. The FMR spectra of both bacteria showed the intrinsic characteristics of magnetite produced by MB, such as extended absorption at the low field region of the spectra and a Verwey transition around 100 K. As previously observed, the spectra of M. gryphiswaldense isolated crystals were more symmetrical than the spectra obtained from whole cells, reflecting the loss of chain arrangement due to the small size and symmetrical shape of the crystals. However, the FMR spectra of magnetic crystals isolated from magnetotactic cocci were very similar to the FMR spectra of whole cells, because the chain arrangement was maintained due to the large size and prismatic shape of the crystals. Our data support the use of FMR spectra to detect magnetotactic bacteria and magnetofossils in samples of present and past environments. Furthermore, the spectra suggest the use of the temperature transition of spectral peak-to-peak intensity to obtain the Verwey temperature for these systems.