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We report the successful fabrication of a pharmaceutical cellular bank (PCB) containing magnetotactic bacteria (MTB), which belong to the Magnetospirillum gryphiswaldense MSR1 species. To produce such PCB, we amplified MTB in a minimal growth medium essentially devoid of other heavy metals than iron and of CMR (Carcinogenic, mutagenic and reprotoxic) products. The PCB enabled to acclimate MTB to such minimal growth conditions and then to produce highly pure magnetosomes composed of more than 99.9% of iron. The qualification of the bank as a PCB relies first on a preserved identity of the MTB compared with the original strain, second on genetic bacterial stability observed over 100 generations or under cryo-preservation for 16 months, third on a high level of purity highlighted by an absence of contaminating microorganisms in the PCB. Furthermore, the PCB was prepared under high-cell load conditions (9.108 cells/mL), allowing large-scale bacterial amplification and magnetosome production. In the future, the PCB could therefore be considered for commercial as well as research orientated applications in nanomedicine. We describe for the first-time conditions for setting-up an effective pharmaceutical cellular bank preserving over time the ability of certain specific cells, i.e. Magnetospirillum gryphiswaldense MSR1 MTB, to produce nano-minerals, i.e. magnetosomes, within a pharmaceutical setting.
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Magnetosomas , Magnetospirillum , Magnetospirillum/genética , Hierro , Preparaciones Farmacéuticas , Proteínas Bacterianas/genéticaRESUMEN
We report the fabrication of highly pure magnetosomes that are synthesized by magnetotactic bacteria (MTB) using pharmaceutically compatible growth media, i.e., without compounds of animal origin (yeast extracts), carcinogenic, mutagenic, or toxic for reproduction (CMR) products, and other heavy metals than iron. To enable magnetosome medical applications, these growth media are reduced and amended compared with media commonly used to grow these bacteria. Furthermore, magnetosomes are made non-pyrogenic by being extracted from these micro-organisms and heated above 400 °C to remove and denature bacterial organic material and produce inorganic magnetosome minerals. To be stabilized, these minerals are further coated with citric acid to yield M-CA, leading to fully reconstructed chains of magnetosomes. The heating properties and anti-tumor activity of highly pure M-CA are then studied by bringing M-CA into contact with PC3-Luc tumor cells and by exposing such assembly to an alternating magnetic field (AMF) of 42 mT and 195 kHz during 30 min. While in the absence of AMF, M-CA are observed to be non-cytotoxic, they result in a 35% decrease in cell viability following AMF application. The treatment efficacy can be associated with a specific absorption rate (SAR) value of M-CA, which is relatively high in cellular environment, i.e., SARcell = 253 ± 11 W/gFe, while being lower than the M-CA SAR value measured in water, i.e., SARwater = 1025 ± 194 W/gFe, highlighting that a reduction in the Brownian contribution to the SAR value in cellular environment does not prevent efficient tumor cell destruction with these nanoparticles. KEY POINTS : ⢠Highly pure magnetosomes were produced in pharmaceutically compatible growth media ⢠Non-pyrogenic and stable magnetosomes were prepared for human injection ⢠Magnetosomes efficiently destroyed prostate tumor cells in magnetic hyperthermia.
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Hipertermia Inducida , Magnetosomas , Magnetospirillum , Neoplasias de la Próstata , Masculino , Animales , Humanos , Línea Celular Tumoral , Neoplasias de la Próstata/terapia , BacteriasRESUMEN
Results of the 2018 commissioning and experimental campaigns of the new High Power Laser Facility on the Energy-dispersive X-ray Absorption Spectroscopy (ED-XAS) beamline ID24 at the ESRF are presented. The front-end of the future laser, delivering 15â J in 10â ns, was interfaced to the beamline. Laser-driven dynamic compression experiments were performed on iron oxides, iron alloys and bismuth probed by online time-resolved XAS.
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BACKGROUND: An important but rarely addressed question in nano-therapy is to know whether bio-degraded nanoparticles with reduced sizes and weakened heating power are able to maintain sufficient anti-tumor activity to fully eradicate a tumor, hence preventing tumor re-growth. To answer it, we studied magnetosomes, which are nanoparticles synthesized by magnetotactic bacteria with sufficiently large sizes (~ 30 nm on average) to enable a follow-up of nanoparticle sizes/heating power variations under two different altering conditions that do not prevent anti-tumor activity, i.e. in vitro cellular internalization and in vivo intra-tumor stay for more than 30 days. RESULTS: When magnetosomes are internalized in U87-Luc cells by being incubated with these cells during 24 h in vitro, the dominant magnetosome sizes within the magnetosome size distribution (DMS) and specific absorption rate (SAR) strongly decrease from DMS ~ 40 nm and SAR ~ 1234 W/gFe before internalization to DMS ~ 11 nm and SAR ~ 57 W/gFe after internalization, a behavior that does not prevent internalized magnetosomes to efficiently destroy U87-Luc cell, i.e. the percentage of U87-Luc living cells incubated with magnetosomes decreases by 25% between before and after alternating magnetic field (AMF) application. When 2 µl of a suspension containing 40 µg of magnetosomes are administered to intracranial U87-Luc tumors of 2 mm3 and exposed (or not) to 15 magnetic sessions (MS), each one consisting in 30 min application of an AMF of 27 mT and 198 kHz, DMS and SAR decrease between before and after the 15 MS from ~ 40 nm and ~ 4 W/gFe down to ~ 29 nm and ~ 0 W/gFe. Although the magnetosome heating power is weakened in vivo, i.e. no measurable tumor temperature increase is observed after the sixth MS, anti-tumor activity remains persistent up to the 15th MS, resulting in full tumor disappearance among 50% of treated mice. CONCLUSION: Here, we report sustained magnetosome anti-tumor activity under conditions of significant magnetosome size reduction and complete loss of magnetosome heating power.
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Antineoplásicos/química , Neoplasias Encefálicas/tratamiento farmacológico , Nanopartículas de Magnetita/química , Magnetosomas/química , Magnetospirillum/química , Animales , Línea Celular Tumoral , Supervivencia Celular , Femenino , Calefacción , Humanos , Hipertermia Inducida , Campos Magnéticos , Ratones , Ratones Desnudos , Tamaño de la Partícula , Nanomedicina Teranóstica/métodos , Distribución TisularRESUMEN
Investigation of the iron phase diagram under high pressure and temperature is crucial for the determination of the composition of the cores of rocky planets and for better understanding the generation of planetary magnetic fields. Here we present X-ray diffraction results from laser-driven shock-compressed single-crystal and polycrystalline iron, indicating the presence of solid hexagonal close-packed iron up to pressure of at least 170 GPa along the principal Hugoniot, corresponding to a temperature of 4,150 K. This is confirmed by the agreement between the pressure obtained from the measurement of the iron volume in the sample and the inferred shock strength from velocimetry deductions. Results presented in this study are of the first importance regarding pure Fe phase diagram probed under dynamic compression and can be applied to study conditions that are relevant to Earth and super-Earth cores.
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There are longstanding and ongoing controversies about the abiotic or biological origin of nanocrystals of magnetite. On Earth, magnetotactic bacteria perform biomineralization of intracellular magnetite nanoparticles under a controlled pathway. These bacteria are ubiquitous in modern natural environments. However, their identification in ancient geological material remains challenging. Together with physical and mineralogical properties, the chemical composition of magnetite was proposed as a promising tracer for bacterial magnetofossil identification, but this had never been explored quantitatively and systematically for many trace elements. Here, we determine the incorporation of 34 trace elements in magnetite in both cases of abiotic aqueous precipitation and of production by the magnetotactic bacterium Magnetospirillum magneticum strain AMB-1. We show that, in biomagnetite, most elements are at least 100 times less concentrated than in abiotic magnetite and we provide a quantitative pattern of this depletion. Furthermore, we propose a previously unidentified method based on strontium and calcium incorporation to identify magnetite produced by magnetotactic bacteria in the geological record.
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Biomarcadores/análisis , Nanopartículas de Magnetita/análisis , Magnetospirillum/química , Magnetospirillum/crecimiento & desarrollo , Oligoelementos/análisis , Análisis de Varianza , Biomarcadores/metabolismo , Técnicas de Cultivo de Célula , Fermentación , Óxido Ferrosoférrico/síntesis química , Magnetospirillum/metabolismo , Microscopía Electrónica de Transmisión , Oligoelementos/metabolismoRESUMEN
BACKGROUND: Biologics magnetics nanoparticles, magnetosomes, attract attention because of their magnetic characteristics and potential applications. The aim of the present study was to develop and characterize novel magnetosomes, which were extracted from magnetotactic bacteria, purified to produce apyrogen magnetosome minerals, and then coated with Chitosan, Neridronate, or Polyethyleneimine. It yielded stable magnetosomes designated as M-Chi, M-Neri, and M-PEI, respectively. Nanoparticle biocompatibility was evaluated on mouse fibroblast cells (3T3), mouse glioblastoma cells (GL-261) and rat glioblastoma cells (RG-2). We also tested these nanoparticles for magnetic hyperthermia treatment of tumor in vitro on two tumor cell lines GL-261 and RG-2 under the application of an alternating magnetic field. Heating, efficacy and internalization properties were then evaluated. RESULTS: Nanoparticles coated with chitosan, polyethyleneimine and neridronate are apyrogen, biocompatible and stable in aqueous suspension. The presence of a thin coating in M-Chi and M-PEI favors an arrangement in chains of the magnetosomes, similar to that observed in magnetosomes directly extracted from magnetotactic bacteria, while the thick matrix embedding M-Neri leads to structures with an average thickness of 3.5 µm2 per magnetosome mineral. In the presence of GL-261 cells and upon the application of an alternating magnetic field, M-PEI and M-Chi lead to the highest specific absorption rates of 120-125 W/gFe. Furthermore, while M-Chi lead to rather low rates of cellular internalization, M-PEI strongly associate to cells, a property modulated by the application of an alternating magnetic field. CONCLUSIONS: Coating of purified magnetosome minerals can therefore be chosen to control the interactions of nanoparticles with cells, organization of the minerals, as well as heating and cytotoxicity properties, which are important parameters to be considered in the design of a magnetic hyperthermia treatment of tumor.
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Materiales Biocompatibles Revestidos/química , Materiales Biocompatibles Revestidos/uso terapéutico , Glioma/terapia , Magnetosomas/química , Nanopartículas/química , Nanopartículas/uso terapéutico , Células 3T3 , Animales , Línea Celular Tumoral , Quitosano/química , Quitosano/uso terapéutico , Difosfonatos/química , Difosfonatos/uso terapéutico , Hipertermia Inducida , Campos Magnéticos , Magnetospirillum/química , Ratones , Polietileneimina/química , Polietileneimina/uso terapéutico , RatasRESUMEN
The leaching behavior of Pb and Zn from lead blast furnace (LBF) and imperial smelting furnace (ISF) slags sampled in the North of France was studied as a function of pHs and under two atmospheres (open air and nitrogen). The leaching of major elements from the slags was monitored as a function of pH (4, 5.5, 7, 8.5 and 10) under both atmospheres for different slag-water interaction times (1 day and 9 days). The leaching results were coupled with a geochemical model; Visual MINTEQ version 3.0, and a detailed morphological and mineralogical analysis was performed on the leached slags by scanning and transmission electron microscopy (SEM and TEM). Significant amounts of Ca, Fe and Zn were released under acidic conditions (pH 4) with a decrease towards the neutral to alkaline conditions (pH 7 and 10) for both LBF and ISF slags. On the other hand, Fe leachability was limited at neutral to alkaline pH for both slags. The concentrations of all elements increased gradually after 216 h compared to initial 24 h of leaching period. The presence of oxygen under open-air atmosphere not only enhanced oxidative weathering but also encouraged formation of secondary oxide and carbonate phases. Formation of carbonates and clay minerals was suggested by Visual MINTEQ which was further confirmed by SEM & TEM. The hydration and partial dissolution of hardystonite, as well as the destabilization of amorphous glassy matrix mainly contributed to the release of major elements, whereas the spinel related oxides were resistant against pH changes and atmospheres within the time frame concerned for both LBF and ISF slags. The total amount of Pb leached out at pH 7 under both atmospheres suggested that both LBF and ISF slags are prone to weathering even at neutral environmental conditions.
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Incineración/métodos , Residuos Industriales/análisis , Plomo/química , Material Particulado/química , Eliminación de Residuos/métodos , Contaminantes del Suelo/química , Contaminantes Químicos del Agua/química , Zinc/química , Arsénico/análisis , Carbono/análisis , Fraccionamiento Químico , Precipitación Química , Ceniza del Carbón/análisis , Monitoreo del Ambiente , Francia , Concentración de Iones de Hidrógeno , Hierro/análisis , Plomo/análisis , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Material Particulado/análisis , Contaminantes del Suelo/análisis , Acero/química , Contaminantes Químicos del Agua/análisis , Zinc/análisisRESUMEN
We report a density-functional theory (DFT)-based study of the interface of bulk water with a prototypical oxide surface, MgO(001), and focus our study on the often-overlooked surface electric field. In particular, we observe that the bare MgO(001) surface, although charge-neutral and defectless, has an intense electric field on the Å scale. The MgO(001) surface covered with 1 water monolayer (1 ML) is investigated via a supercell accounting for the experimentally-observed (2 × 3) reconstruction, stable at ambient temperature, and in which two out of six water molecules are dissociated. This 1 ML-hydrated surface is also found to have a high, albeit short-ranged, normal component of the field. Finally, the oxide/water interface is studied via room-temperature ab initio molecular dynamics (AIMD) using 34 H2O molecules between two MgO(001) surfaces. To our best knowledge this is the first AIMD study of the MgO(001)/liquid water interface in which all atoms are treated using DFT and including several layers above the first adsorbed layer. We observe that the surface electric field, averaged over the AIMD trajectories, is still very strong on the fully-wet surface, peaking at about 3 V Å(-1). Even in the presence of bulk-like water, the structure of the first layer in contact with the surface remains similar to the (2 × 3)-reconstructed ice ad-layer on MgO(001). Moreover, we observe proton exchange within the first layer, and between the first and second layers - indeed, the O-O distances close to the surface are found to be distributed towards shorter distances, a property which has been shown to directly promote proton transfer.
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Electricidad , Óxido de Magnesio/química , Simulación de Dinámica Molecular , Teoría Cuántica , Agua/química , Conformación Molecular , Protones , Propiedades de Superficie , TemperaturaRESUMEN
Batch experiments were conducted in water at 150 °C and PCO2 = 280 bar on a Mg-rich tholeiitic basalt (9.3 wt % MgO and 12.2 wt % CaO) composed of olivine, Ti-magnetite, plagioclase, and clinopyroxene. After 45 days of reaction, 56 wt % of the initial MgO had reacted with CO2 to form Fe-bearing magnesite, (Mg0.8Fe0.2)CO3, along with minor calcium carbonates. The substantial decrease in olivine content upon carbonation supports the idea that ferroan magnesite formation mainly follows from olivine dissolution. In contrast, in experiments performed under similar run durations and P/T conditions with a San Carlos olivine separate (47.8 wt % MgO) of similar grain size, only 5 wt % of the initial MgO content reacted to form Fe-bearing magnesite. The overall carbonation kinetics of the basalt was enhanced by a factor of ca. 40. This could be explained by differences in the chemical and textural properties of the secondary silica layer that covers reacted olivine grains in both types of sample. Consequently, laboratory data obtained on olivine separates might yield a conservative estimate of the true carbonation potential of olivine-bearing basaltic rocks.
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Dióxido de Carbono/química , Carbonatos/química , Compuestos de Hierro/química , Compuestos de Magnesio/química , Minerales/química , Silicatos/química , Aluminio/química , Silicatos de Aluminio/química , Arcilla , Microscopía Electrónica de Rastreo , Porosidad , Silicio/química , Soluciones , Termodinámica , Difracción de Rayos XRESUMEN
The global geochemical carbon cycle involves exchanges between the Earth's interior and the surface. Carbon is recycled into the mantle via subduction mainly as carbonates and is released to the atmosphere via volcanism mostly as CO(2). The stability of carbonates versus decarbonation and melting is therefore of great interest for understanding the global carbon cycle. For all these reasons, the thermodynamic properties and phase diagrams of these minerals are needed up to core mantle boundary conditions. However, the nature of C-bearing minerals at these conditions remains unclear. Here we show the existence of a new Mg-Fe carbon-bearing compound at depths greater than 1,800 km. Its structure, based on three-membered rings of corner-sharing (CO(4))(4-) tetrahedra, is in close agreement with predictions by first principles quantum calculations [Oganov AR, et al. (2008) Novel high-pressure structures of MgCO(3), CaCO(3) and CO(2) and their role in Earth's lower mantle. Earth Planet Sci Lett 273:38-47]. This high-pressure polymorph of carbonates concentrates a large amount of Fe((III)) as a result of intracrystalline reaction between Fe((II)) and (CO(3))(2-) groups schematically written as 4FeO + CO(2) â 2Fe(2)O(3) + C. This results in an assemblage of the new high-pressure phase, magnetite and nanodiamonds.
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Carbono/química , Carbonatos/química , Planeta Tierra , Minerales/química , Hierro/química , Magnesio/química , Estructura Molecular , Presión , Temperatura , Termodinámica , Difracción de Rayos XRESUMEN
Humanity's strive to understand why and how life appeared on planet Earth dates back to prehistoric times. At the beginning of the 19th century, empirical biology started to tackle this question yielding both Charles Darwin's Theory of Evolution and the paradigm that the crucial trigger putting life on its tracks was the appearance of organic molecules. In parallel to these developments in the biological sciences, physics and physical chemistry saw the fundamental laws of thermodynamics being unraveled. Towards the end of the 19th century and during the first half of the 20th century, the tensions between thermodynamics and the "organic-molecules-paradigm" became increasingly difficult to ignore, culminating in Erwin Schrödinger's 1944 formulation of a thermodynamics-compliant vision of life and, consequently, the prerequisites for its appearance. We will first review the major milestones over the last 200 years in the biological and the physical sciences, relevant to making sense of life and its origins and then discuss the more recent reappraisal of the relative importance of metal ions vs. organic molecules in performing the essential processes of a living cell. Based on this reassessment and the modern understanding of biological free energy conversion (aka bioenergetics), we consider that scenarios wherein life emerges from an abiotic chemiosmotic process are both thermodynamics-compliant and the most parsimonious proposed so far.
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We review the most recent and significant results published in the field of magnetotactic bacteria (MTB), in particular data relating to the use of bacterial magnetosomes in magnetic hyperthermia for the treatment of tumours. We review different methods for cultivating MTB and preparing suspensions of bacterial magnetosomes. As well as the production of magnetosomes, we also review key data on the toxicity of the magnetosomes as well as their heating and anti-tumour efficiencies. The toxicity and efficiency of magnetosomes needs to be understood and the risk-benefit ratio with which to evaluate their use in the magnetic hyperthermia treatment of tumours needs to be measured.
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Hipertermia Inducida/métodos , Magnetosomas , Neoplasias/terapia , Animales , Humanos , Fenómenos MagnéticosRESUMEN
Mn(II)-oxidizing organisms promote the biomineralization of manganese oxides with specific textures, under ambient conditions. Controlling the phases formed and their texture on a larger scale may offer environmentally relevant routes to manganese oxide synthesis, with potential technological applications, for example, for energy storage. In the present study, we sought to use biofilms to promote the formation of electroactive minerals and to control the texture of these biominerals down to the electrode scale (i.e., cm scale). We used the bacterium Pseudomonas putida strain MnB1 which can produce manganese oxide in a biofilm. We characterized the biofilm-mineral assembly using a combination of electron microscopy, synchrotron-based X-ray absorption spectroscopy, X-ray diffraction, thermogravimetric analysis and electron paramagnetic resonance spectroscopy. Under optimized conditions of biofilm growth on the surface of current collectors, mineralogical characterizations revealed the formation of several minerals including a slightly crystalline MnOx birnessite. Electrochemical measurements in a half-cell against Li(0) revealed the electrochemical signature of the Mn4+/Mn3+ redox couple indicating the electroactivity of the biomineralized biofilm without any post-synthesis chemical, physical or thermal treatment. These results provide a better understanding of the properties of biomineralized biofilms and their possible use in designing new routes for one-pot electrode synthesis.
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Thermococcales, a major order of hyperthermophilic archaea inhabiting iron- and sulfur-rich anaerobic parts of hydrothermal deep-sea vents, are known to induce the formation of iron phosphates, greigite (Fe3S4) and abundant quantities of pyrite (FeS2), including pyrite spherules. In the present study, we report the characterization of the sulfide and phosphate minerals produced in the presence of Thermococcales using X-ray diffraction, synchrotron-based X ray absorption spectroscopy and scanning and transmission electron microscopies. Mixed valence Fe(II)-Fe(III) phosphates are interpreted as resulting from the activity of Thermococcales controlling phosphorus-iron-sulfur dynamics. The pyrite spherules (absent in abiotic control) consist of an assemblage of ultra-small nanocrystals of a few ten nanometers in size, showing coherently diffracting domain sizes of few nanometers. The production of these spherules occurs via a sulfur redox swing from S0 to S-2 and then to S-1, involving a comproportionation of (-II) and (0) oxidation states of sulfur, as supported by S-XANES data. Importantly, these pyrite spherules sequester biogenic organic compounds in small but detectable quantities, possibly making them good biosignatures to be searched for in extreme environments.
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The introduction of various iron-chelating agents to the Magnetospirillum magneticum strain AMB-1 bacterial growth medium stimulated the growth of M. magneticum strain AMB-1 magnetotactic bacteria and enhanced the production of magnetosomes. After 7 days of growth, the number of bacteria and the production of magnetosomes were increased in the presence of iron-chelating agents by factors of up to â¼2 and â¼6, respectively. The presence of iron-chelating agents also produced an increase in magnetosome size and chain length and yielded improved magnetosome heating properties. The specific absorption rate of suspensions of magnetosome chains isolated from M. magneticum strain AMB-1 magnetotactic bacteria, measured under the application of an alternating magnetic field of average field strength â¼20 mT and frequency 198 kHz, increased from â¼222 W/g(Fe) in the absence of iron-chelating agent up to â¼444 W/g(Fe) in the presence of 4 µM rhodamine B and to â¼723 W/g(Fe) in the presence of 4 µM EDTA. These observations were made at an iron concentration of 20 µM and iron-chelating agent concentrations below 40 µM.
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Quelantes del Hierro/metabolismo , Magnetosomas/metabolismo , Magnetospirillum/crecimiento & desarrollo , Magnetospirillum/metabolismo , Medios de Cultivo/química , Ácido Edético/metabolismo , Rodaminas/metabolismoRESUMEN
Life can be found even in extreme environments, e.g., near black smokers on the ocean floor at temperatures up to ca. 120 °C and hydrostatic pressures of 40 MPa. To maintain vital reactions under these hostile conditions, extremophiles are interacting with the surrounding geochemical system. In this context, the stabilities of the essential energy-storing adenosine triphosphate (ATP) and adenosine diphosphate (ADP) are of fundamental importance because reactions involving adenosine phosphates constrain the conditions at which carbon-based life can exist and might be also crucial information for the search of extra-terrestrial life. Adenosine phosphates react by non-enzymatic hydrolysis, which is kinetically enhanced at high temperatures. If these abiotic hydrolysis processes are too rapid, they will most likely prevent metabolisms from relying on ATP. Here, we report on an approach used in experimental geochemistry, i.e., in situ Raman spectroscopic analyses of a fluid phase at high temperature using a hydrothermal diamond anvil cell. This combination allowed the investigation of the hydrolysis of Adenosine Tri-Phosphate (ATP) in aqueous solution at pH 3 and 7 and at temperatures of 80, 100 and 120 °C, extending the so far measured temperature range substantially. We observed Arrhenian behaviour over this temperature interval. The rate constants at 120 °C were 4.34 × 10-3 s-1 at pH 3 and 2.91 × 10-3 s-1 at pH 7. This corresponds to ATP half-lives of a few minutes. These high decomposition rates of ATP suggest that organisms must have developed a mechanism to counteract this fast reaction at high temperatures to maintain the vital processes.
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Adenosina Trifosfato , Fosfatos , Adenosina , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Carbono , Diamante , Hidrólisis , Cinética , Fosfatos/metabolismo , TemperaturaRESUMEN
Defining chemical properties of intracellular organelles is necessary to determine their function(s) as well as understand and mimic the reactions they host. However, the small size of bacterial and archaeal microorganisms often prevents defining local intracellular chemical conditions in a similar way to what has been established for eukaryotic organelles. This work proposes to use magnetite (Fe3O4) nanocrystals contained in magnetosome organelles of magnetotactic bacteria as reporters of elemental composition, pH, and redox potential of a hypothetical environment at the site of formation of intracellular magnetite. This methodology requires combining recent single-cell mass spectrometry measurements together with elemental composition of magnetite in trace and minor elements. It enables a quantitative characterization of chemical disequilibria of 30 chemical elements between the intracellular and external media of magnetotactic bacteria, revealing strong transfers of elements with active influx or efflux processes that translate into elemental accumulation (Mo, Se, and Sn) or depletion (Sr and Bi) in the bacterial internal medium of up to seven orders of magnitude relative to the extracellular medium. Using this concept, we show that chemical conditions in magnetosomes are compatible with a pH of 7.5-9.5 and a redox potential of -0.25 to -0.6 V.
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Magnetosomas , Magnetospirillum , Bacterias , Óxido Ferrosoférrico/química , Bacterias Gramnegativas , Magnetosomas/químicaRESUMEN
Halophilic microorganisms have long been known to survive within the brine inclusions of salt crystals, as evidenced by the change in color for salt crystals containing pigmented halophiles. However, the molecular mechanisms allowing this survival has remained an open question for decades. While protocols for the surface sterilization of halite (NaCl) have enabled isolation of cells and DNA from within halite brine inclusions, "-omics" based approaches have faced two main technical challenges: (1) removal of all contaminating organic biomolecules (including proteins) from halite surfaces, and (2) performing selective biomolecule extractions directly from cells contained within halite brine inclusions with sufficient speed to avoid modifications in gene expression during extraction. In this study, we tested different methods to resolve these two technical challenges. Following this method development, we then applied the optimized methods to perform the first examination of the early acclimation of a model haloarchaeon (Halobacterium salinarum NRC-1) to halite brine inclusions. Examinations of the proteome of Halobacterium cells two months post-evaporation revealed a high degree of similarity with stationary phase liquid cultures, but with a sharp down-regulation of ribosomal proteins. While proteins for central metabolism were part of the shared proteome between liquid cultures and halite brine inclusions, proteins involved in cell mobility (archaellum, gas vesicles) were either absent or less abundant in halite samples. Proteins unique to cells within brine inclusions included transporters, suggesting modified interactions between cells and the surrounding brine inclusion microenvironment. The methods and hypotheses presented here enable future studies of the survival of halophiles in both culture model and natural halite systems.