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Memory engrams are formed through experience-dependent remodeling of neural circuits, but their detailed architectures have remained unresolved. Using 3D electron microscopy, we performed nanoscale reconstructions of the hippocampal CA3-CA1 pathway following chemogenetic labeling of cellular ensembles with a remote history of correlated excitation during associative learning. Projection neurons involved in memory acquisition expanded their connectomes via multi-synaptic boutons without altering the numbers and spatial arrangements of individual axonal terminals and dendritic spines. This expansion was driven by presynaptic activity elicited by specific negative valence stimuli, regardless of the co-activation state of postsynaptic partners. The rewiring of initial ensembles representing an engram coincided with local, input-specific changes in the shapes and organelle composition of glutamatergic synapses, reflecting their weights and potential for further modifications. Our findings challenge the view that the connectivity among neuronal substrates of memory traces is governed by Hebbian mechanisms, and offer a structural basis for representational drifts.
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The patterns of synaptic connectivity and physiological properties of diverse neuron types are shaped by distinct gene sets. Our study demonstrates that, in the mouse forebrain, the transcriptional profiles of inhibitory GABAergic interneurons are regulated by Nr4a1, an orphan nuclear receptor whose expression is transiently induced by sensory experiences and is required for normal learning. Nr4a1 exerts contrasting effects on the local axonal wiring of parvalbumin- and somatostatin-positive interneurons, which innervate different subcellular domains of their postsynaptic partners. The loss of Nr4a1 activity in these interneurons results in bidirectional, cell-type-specific transcriptional switches across multiple gene families, including those involved in surface adhesion and repulsion. Our findings reveal that combinatorial synaptic organizing codes are surprisingly flexible and highlight a mechanism by which inducible transcription factors can influence neural circuit structure and function.
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Neurônios GABAérgicos , Interneurônios , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares , Animais , Interneurônios/metabolismo , Neurônios GABAérgicos/metabolismo , Neurônios GABAérgicos/fisiologia , Camundongos , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Somatostatina/metabolismo , Somatostatina/genética , Parvalbuminas/metabolismo , Camundongos Knockout , Masculino , Sinapses/metabolismoRESUMO
For decades, the expression of immediate early genes (IEGs) such as FOS has been the most widely used molecular marker representing neuronal activation. However, to date, there is no equivalent surrogate available for the decrease of neuronal activity. Here, we developed an optogenetic-based biochemical screen in which population neural activities can be controlled by light with single action potential precision, followed by unbiased phosphoproteomic profiling. We identified that the phosphorylation of pyruvate dehydrogenase (pPDH) inversely correlated with the intensity of action potential firing in primary neurons. In in vivo mouse models, monoclonal antibody-based pPDH immunostaining detected activity decreases across the brain, which were induced by a wide range of factors including general anesthesia, chemogenetic inhibition, sensory experiences, and natural behaviors. Thus, as an inverse activity marker (IAM) in vivo, pPDH can be used together with IEGs or other cell-type markers to profile and identify bi-directional neural dynamics induced by experiences or behaviors.
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Encéfalo , Neurônios , Camundongos , Animais , Fosforilação , Encéfalo/metabolismo , Neurônios/fisiologia , Oxirredutases/genética , Oxirredutases/metabolismo , Piruvatos/metabolismo , Genes PrecocesRESUMO
The study is dedicated to the consideration of lower alkyl ethers of glycerol as potential components of low-melting technical fluids (e.g., heat transfer fluids, hydraulic fluids, aircraft de-icing fluids, etc.). Four isomeric mixtures of glycerol ethers (GMME-monomethyl; GDME-dimethyl; GMEE-monoethyl; GDEE-diethyl) were synthesized from epichlorohydrin and methanol/ethanol in the presence of sodium and subjected to detailed characterization as pure compounds and as aqueous solutions (30-90 vol%). The temperature and concentration dependencies of density, viscosity, cloud point, boiling range, specific heat capacity, thermal conductivity, and rubber swelling were obtained. On the basis of the data obtained, a comparison was made between the aqueous solutions of glycerol ethers and of other common bases for low-melting liquids (glycerol, ethylene glycol, and propylene glycol). Pure glycerol ethers could potentially be used as technical fluids in a very wide temperature range-from -114 to 150 °C. It was further demonstrated that in low temperature applications (e.g., in low-temperature chiller systems) the glycerol-ether-based aqueous heat transfer fluids could provide enhanced efficiency when compared to the glycerol- or propylene-glycol-based ones due to their lower viscosities and favorable environmental properties.
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The homogeneous acetic acid synthesis-type Ru-Co-Li/N-methylpyrrolidone catalyst for CO and H2 transformations has been studied at moderately high pressures. For 1CO:2H2, low acetic acid selectivity has been observed, along with remarkable methyl acetate selectivity, the absence of aldehydes and ethyl acetate and sharp deviations from the Anderson-Schultz-Flory distribution for both alcaohols and long-chain hydrocarbons. For 1CO:1H2 and slightly elevated pressure, acetic acid selectivity slightly increased, notable ethyl acetate formation was detected, and both long-chain hydrocarbons and alcohols disappeared. Hypotheses are discussed about the direct parallel formation of all observed product groups (hydrocarbons, alcohols, esters, and acetic acid) and hydrocarbon chain growth limitations according to the formed Ru-Co cluster size in the presence of the aforementioned catalytic system.
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The annual production of plastic waste is a serious ecological problem as it causes substantial pollution of the environment. Polyethylene terephthalate, a material usually found in disposable plastic bottles, is one of the most popular material used for packaging in the world. In this paper, it is proposed to recycle polyethylene terephthalate waste bottles into benzene-toluene-xylene fraction using a heterogeneous nickel phosphide catalyst formed in situ during the polyethylene terephthalate recycling process. The catalyst obtained was characterized using powder X-ray diffraction, high-resolution transmission electron microscopy, and X-ray photoelectron spectroscopy techniques. The catalyst was shown to contain a Ni2P phase. Its activity was studied in a temperature range of 250-400 °C and a H2 pressure range of 5-9 MPa. The highest selectivity for benzene-toluene-xylene fraction was 93% at quantitative conversion.
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For decades, the expression of immediate early genes (IEGs) such as c- fos has been the most widely used molecular marker representing neuronal activation. However, to date, there is no equivalent surrogate available for the decrease of neuronal activity (i.e., inhibition). Here, we developed an optogenetic-based biochemical screen in which population neural activities can be controlled by light with single action potential precision, followed by unbiased phosphoproteomic profiling. We identified that the phosphorylation of pyruvate dehydrogenase (pPDH) inversely correlated with the intensity of action potential firing in primary neurons. In in vivo mouse models, monoclonal antibody-based pPDH immunostaining detected neuronal inhibition across the brain induced by a wide range of factors including general anesthesia, sensory experiences, and natural behaviors. Thus, as an in vivo marker for neuronal inhibition, pPDH can be used together with IEGs or other cell-type markers to profile and identify bi-directional neural dynamics induced by experiences or behaviors.
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Bifunctional catalysts are a major type of heterogeneous catalytic systems that have been widely investigated for biomass upgrading. In this work, Ru-catalysts based on sulfonated porous aromatic frameworks (PAFs) were used in the hydrodeoxygenation (HDO) of lignin-derived compounds: guaiacol, veratrole, and catechol. The relationship between the activity of metal nanoparticles and the content of acid sites in synthesized catalysts was studied. Herein, their synergy was demonstrated in the Ru-PAF-30-SO3H/5-COD catalyst. The results revealed that this catalytic system promoted partial hydrogenation of lignin-based compounds to ketones without any further transformations. The design of the Ru-PAF-30-SO3H/5-COD catalytic system opens a promising route to the selective conversion of lignin model compounds to cyclohexanone.
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The present study is dedicated to the experimental verification of a concept for the hydrogenolysis of glycerol over in situ-generated Cu dispersed particles (Cu-DP). The Cu-DP were generated by in situ reduction of a precursor salt (Cu(OAc)2, CuSO4, CuCl2) in the presence of KOH and were active in glycerol conversion under hydrogen (T = 200-220 °C, p(H2) = 1-4 MPa), where 1,2-propylene glycol (PG) and lactic acid (LA) were detected to be the main products. The influence of the reaction conditions (temperature, hydrogen pressure, reaction time, catalyst-to-feed ratio and the KOH/Cu ratio) on the yields of the products is described. It was shown that the selectivity between the PG and LA could be tuned by changing p(H2) or by the KOH amount, i.e., higher yields of LA corresponded to lower p(H2) and higher alkalinity of the reaction media. The activity of the in situ-generated Cu-DP was found to be comparable to that of an industrial Cu-Cr2O3 catalyst. The Cu-DP catalysts were characterized by XRD, XPS, HRTEM and SEM. During the reaction, the catalyst evolved by the sintering and recrystallization of the separate Cu-DP; the crystallite sizes after 1 and 15 h reaction times amounted to 35 and 49 nm, respectively.
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Glicerol , Propilenoglicol , Glicerol/química , Propilenoglicol/química , Hidrogênio/química , Temperatura , CatáliseRESUMO
Bulk MoP and WP were investigated and compared in guaiacol hydrodeoxygenation to phenol. The catalysts obtained were studied by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and temperature-programmed desorption of NH3 (NH3-TPD) analyses. MoP was shown to be more active than WP. However, WP was more selective in phenol production. Guaiacol conversion using MoP was 90-98%. The highest selectivity for phenol was 66% (340 °C). By increasing the temperature to 380 °C, phenol selectivity decreased to 31%, while selectivity for cyclohexane increased to 29%. Thus, MoP was active not only in hydrodeoxygenation but also in hydrogenation. Guaiacol conversion over WP was 53-90%. The highest selectivity for phenol was 84% (380 °C). Hydrogenation products were also detected but with low selectivity. Thus, WP was active in the partial hydrodeoxygenation of guaiacol and was more suitable for the selective production of phenol than MoP. It was shown that after a 30 h recycling test, the activity of MoP did not decrease (1st and 5th cycle conversion value was 91%), while the activity of WP reduced (1st and 5th cycle conversion values were 81 and 64%, respectively). However, the activity of both catalysts at average conversion values decreased. Selectivity for phenol remained unaltered over both catalysts. It was supposed that catalyst activity decreased due to partial destruction of the crystalline phosphide phase and the surface phosphide oxidation to phosphate.
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Neuronal activity initiates signaling cascades that culminate in diverse outcomes including structural and functional neuronal plasticity, and metabolic changes. While studies have revealed activity-dependent neuronal cell type-specific transcriptional changes, unbiased quantitative analysis of cell-specific activity-induced dynamics in newly synthesized proteins (NSPs) synthesis in vivo has been complicated by cellular heterogeneity and a relatively low abundance of NSPs within the proteome in the brain. Here we combined targeted expression of mutant MetRS (methionine tRNA synthetase) in genetically defined cortical glutamatergic neurons with tight temporal control of treatment with the noncanonical amino acid, azidonorleucine, to biotinylate NSPs within a short period after pharmacologically induced seizure in male and female mice. By purifying peptides tagged with heavy or light biotin-alkynes and using direct tandem mass spectrometry detection of biotinylated peptides, we quantified activity-induced changes in cortical glutamatergic neuron NSPs. Seizure triggered significant changes in â¼300 NSPs, 33% of which were decreased by seizure. Proteins mediating excitatory and inhibitory synaptic plasticity, including SynGAP1, Pak3, GEPH1, Copine-6, and collybistin, and DNA and chromatin remodeling proteins, including Rad21, Smarca2, and Ddb1, are differentially synthesized in response to activity. Proteins likely to play homeostatic roles in response to activity, such as regulators of proteastasis, intracellular ion control, and cytoskeleton remodeling proteins, are activity induced. Conversely, seizure decreased newly synthetized NCAM, among others, suggesting that seizure induced degradation. Overall, we identified quantitative changes in the activity-induced nascent proteome from genetically defined cortical glutamatergic neurons as a strategy to discover downstream mediators of neuronal plasticity and generate hypotheses regarding their function.SIGNIFICANCE STATEMENT Activity-induced neuronal and synaptic plasticity are mediated by changes in the protein landscape, including changes in the activity-induced newly synthesized proteins; however, identifying neuronal cell type-specific nascent proteome dynamics in the intact brain has been technically challenging. We conducted an unbiased proteomic screen from which we identified significant activity-induced changes in â¼300 newly synthesized proteins in genetically defined cortical glutamatergic neurons within 20 h after pharmacologically induced seizure. Bioinformatic analysis of the dynamic nascent proteome indicates that the newly synthesized proteins play diverse roles in excitatory and inhibitory synaptic plasticity, chromatin remodeling, homeostatic mechanisms, and proteasomal and metabolic functions, extending our understanding of the diversity of plasticity mechanisms.
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Aminoacil-tRNA Sintetases , Proteoma , Masculino , Feminino , Camundongos , Animais , Proteoma/metabolismo , Proteômica/métodos , Biotina/metabolismo , Neurônios/metabolismo , Plasticidade Neuronal/fisiologia , Aminoácidos/metabolismo , Metionina/metabolismo , Alcinos/metabolismo , Convulsões/metabolismo , Aminoacil-tRNA Sintetases/genética , Aminoacil-tRNA Sintetases/metabolismo , Moléculas de Adesão de Célula Nervosa/metabolismo , Proteínas Ativadoras de ras GTPase/metabolismoRESUMO
The recent development of solvent- and polymer-based brain-clearing techniques has advanced our ability to visualize the mammalian nervous system in three dimensions. However, it remains challenging to image the mammalian body en bloc. Here we developed HYBRiD (hydrogel-based reinforcement of three-dimensional imaging solvent-cleared organs (DISCO)), by recombining components of organic- and polymer-based clearing pipelines. We achieved high transparency and protein retention, as well as compatibility with direct fluorescent imaging and immunostaining in cleared mammalian bodies. Using parvalbumin- and somatostatin-Cre models, we demonstrated the utility of HYBRiD for whole-body imaging of genetically encoded fluorescent reporters without antibody enhancement of signals in newborn and juvenile mice. Using K18-hACE2 transgenic mice, HYBRiD enabled perfusion-free clearing and visualization of SARS-CoV-2 infection in a whole mouse chest, revealing macroscopic and microscopic features of viral pathology in the same sample. HYBRiD offers a simple and universal solution to visualize large heterogeneous body parts or entire animals for basic and translational research.
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COVID-19 , Hidrogéis , Animais , Imageamento Tridimensional/métodos , Mamíferos , Camundongos , Polímeros , SARS-CoV-2 , SolventesRESUMO
The present review compiles the advances in the dendritic catalysis within the last two decades, in particular concerning heterogeneous dendrimer-based catalysts and their and application in various processes, such as hydrogenation, oxidation, cross-coupling reactions, etc. There are considered three main approaches to the synthesis of immobilized heterogeneous dendrimer-based catalysts: (1) impregnation/adsorption on silica or carbon carriers; (2) dendrimer covalent grafting to various supports (silica, polystyrene, carbon nanotubes, porous aromatic frameworks, etc.), which may be performed in a divergent (as a gradual dendron growth on the support) or convergent way (as a grafting of whole dendrimer to the support); and (3) dendrimer cross-linking, using transition metal ions (resulting in coordination polymer networks) or bifunctional organic linkers, whose size, polarity, and rigidity define the properties of the resulted material. Additionally, magnetically separable dendritic catalysts, which can be synthesized using the three above-mentioned approaches, are also considered. Dendritic catalysts, synthesized in such ways, can be stored as powders and be easily separated from the reaction medium by filtration/centrifugation as traditional heterogeneous catalysts, maintaining efficiency as for homogeneous dendritic catalysts.
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Positron emission tomography (PET) allows biomolecular tracking but PET monitoring of brain networks has been hampered by a lack of suitable reporters. Here, we take advantage of bacterial dihydrofolate reductase, ecDHFR, and its unique antagonist, TMP, to facilitate in vivo imaging in the brain. Peripheral administration of radiofluorinated and fluorescent TMP analogs enabled PET and intravital microscopy, respectively, of neuronal ecDHFR expression in mice. This technique can be used to the visualize neuronal circuit activity elicited by chemogenetic manipulation in the mouse hippocampus. Notably, ecDHFR-PET allows mapping of neuronal projections in non-human primate brains, demonstrating the applicability of ecDHFR-based tracking technologies for network monitoring. Finally, we demonstrate the utility of TMP analogs for PET studies of turnover and self-assembly of proteins tagged with ecDHFR mutants. These results establish opportunities for a broad spectrum of previously unattainable PET analyses of mammalian brain circuits at the molecular level.
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Encéfalo/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos/química , Tetra-Hidrofolato Desidrogenase/genética , Animais , Encéfalo/citologia , Callithrix , Radioisótopos de Carbono/química , Radioisótopos de Flúor/química , Genes Reporter , Células HEK293 , Humanos , Masculino , Camundongos Endogâmicos C57BL , Imagem Molecular/métodos , Rede Nervosa/diagnóstico por imagem , Proteínas/análise , Proteínas/metabolismo , Compostos Radiofarmacêuticos/síntese química , Tetra-Hidrofolato Desidrogenase/metabolismo , Trimetoprima/análogos & derivados , Trimetoprima/químicaRESUMO
Store-operated calcium entry (SOCE) is a mechanism for calcium influx through the plasma membrane in response to release of free calcium from the endoplasmic reticulum. Two recent studies revealed how SOCE regulates the exocytosis of neurotransmitter vesicles at central synapses.
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Chemical synapses of shared cellular origins have remarkably heterogeneous structures, but how this diversity is generated is unclear. Here, we use three-dimensional (3D) electron microscopy and artificial intelligence algorithms for image processing to reconstruct functional excitatory microcircuits in the mouse hippocampus and microcircuits in which neurotransmitter signaling is permanently suppressed with genetic tools throughout the lifespan. These nanoscale analyses reveal that experience is dispensable for morphogenesis of synapses with different geometric shapes and contents of membrane organelles and that arrangement of morphologically distinct connections in local networks is stochastic. Moreover, loss of activity increases the variability in sizes of opposed pre- and postsynaptic structures without disrupting their alignments, suggesting that inherently variable weights of naive connections become progressively matched with repetitive use. These results demonstrate that mechanisms for the structural diversity of neuronal synapses are intrinsic and provide insights into how circuits essential for memory storage assemble and integrate information.
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Imageamento Tridimensional , Microscopia Eletrônica , Nanotecnologia , Sinapses/ultraestrutura , Animais , Axônios/metabolismo , Dendritos/metabolismo , Camundongos , Modelos Neurológicos , Organelas/metabolismo , Organelas/ultraestrutura , Processos EstocásticosRESUMO
Nanosized catalyst dispersions have significant potential for improving hydrocarbon production from carbon monoxide and hydrogen via Fischer-Tropsch synthesis, an essential alternative to the use of petroleum as a raw material. New dispersed cobalt catalysts and dispersed-phase cobalt-based catalysts with Pd, Al2O3, or ZrO2 additives for the Fischer-Tropsch synthesis were synthesized in the present work. A dispersed cobalt phase was prepared in a heavy paraffin medium using ex situ and in situ approaches through thermal decomposition of a nitrate precursor at various temperatures. Analyses showed that an increase in the temperature for catalytic suspension formation from 215 to 260°C enlarged the particles in the dispersed phase from 190 to 264 nm, which was probably due to increased agglomeration at elevated temperatures. The rheological properties of the obtained catalytic suspensions can be described by the Bingham equation. Furthermore, the concentration of the dispersed phase had a direct impact on the structure of the entire catalytic system. Ultrafine suspensions of palladium-promoted catalytic systems were tested for the Fischer-Tropsch synthesis. The overall yield of C5+ hydrocarbons was as high as 50 g/m3, and the productivity of the Pd-promoted catalytic systems reached 270-290 g/(kgCo · h).
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In this study approaches for chemical conversions of the renewable compounds 1,2-propanediol (1,2-PD) and 2,3-butanediol (2,3-BD) that yield the corresponding cyclic ketals and glycol ethers have been investigated experimentally. The characterization of the obtained products as potential green solvents and gasoline components is discussed. Cyclic ketals have been obtained by the direct reaction of the diols with lower aliphatic ketones (1,2-PD + acetone â 2,2,4-trimethyl-1,3-dioxolane (TMD) and 2,3-BD + butanone-2 â 2-ethyl-2,4,5-trimethyl-1,3-dioxolane (ETMD)), for which the ΔH0r, ΔS0r and ΔG0r values have been estimated experimentally. The monoethers of diols could be obtained through either hydrogenolysis of the pure ketals or from the ketone and the diol via reductive alkylation. In the both reactions, the cyclic ketals (TMD and ETMD) have been hydrogenated in nearly quantitative yields to the corresponding isopropoxypropanols (IPP) and 3-sec-butoxy-2-butanol (SBB) under mild conditions (T = 120-140 °C, p(H2) = 40 bar) with high selectivity (>93%). Four products (TMD, ETMD, IPP and SBB) have been characterized as far as their physical properties are concerned (density, melting/boiling points, viscosity, calorific value, evaporation rate, Antoine equation coefficients), as well as their solvent ones (Kamlet-Taft solvatochromic parameters, miscibility, and polymer solubilization). In the investigation of gasoline blending properties, TMD, ETMD, IPP and SBB have shown remarkable antiknock performance with blending antiknock indices of 95.2, 92.7, 99.2 and 99.7 points, respectively.
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Butileno Glicóis/química , Gasolina , Propilenoglicóis/química , Solventes/química , Acetona/química , Algoritmos , Butileno Glicóis/síntese química , Fenômenos Químicos , Técnicas de Química Sintética , Glicóis , Modelos Químicos , Propilenoglicóis/síntese química , Solventes/síntese químicaRESUMO
As- and Cl-containing impurities are highly detrimental to sulfided catalysts in hydrotreating processes. To prevent the irreversible loss of activity of the main sulfide catalysts by As and Cl contaminants, a protective double-layered guard bed catalyst is applied. Two types of mesoporous silica supports (SBA-15 and MCF) were used to obtain sorption-catalytic materials. The high specific surface area of the supports allowed for a significant increase in access to the active catalyst centers. The NiMo/SBA-15/Al2O3 and NiMg/MCF/Al2O3 sorption-catalytic materials demonstrated high activity and stability over 48 h for the simultaneous removal of As and Cl. The catalytic materials allowed for reducing the concentrations of As and Cl to less than 0.1 ppm in the diesel fraction under the following conditions: 5.0 MPa pressure, 2.0 h-1 LHSV, 300 L/L H2-to-substrate volume ratio, and 360 °C.
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Mg, Ca, and Ba catalysts supported on structured mesoporous silica oxides types MCM-41 and Al-SBA-15 were synthesized and investigated in sulfone cracking for sulfur removal from oxidized diesel fuel. Functional materials and catalysts were characterized by low-temperature nitrogen adsorption/desorption, transmission electron microscopy, and inductively coupled plasma atomic emission spectroscopy techniques. Catalytic tests were carried out in fixed-bed and batch reactors with a model compound dibenzothiophene sulfone and oxidized diesel fraction as a feed. MgO/MCM-41 and MgO/Al-MCM-41 possess high activity in sulfone cracking. The sulfur content in the diesel fraction decreases from initial 450 up to 100 ppmw. Catalysts can be regenerated for reuse in several cycles and may be potentially scaled up for industrial applications.