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1.
Environ Microbiol ; 26(2): e16572, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38195068

RESUMO

Factors regulating the diversity and composition of soil microbial communities include soil properties, land cover and climate. How these factors interact at large scale remains poorly investigated. Here, we used an extensive dataset including 715 locations from 24 European countries to investigate the interactive effects of climatic region, land cover and pH on soil bacteria and fungi. We found that differences in microbial diversity and community composition between land cover types depended on the climatic region. In Atlantic, Boreal and Continental regions, microbial richness was higher in croplands and grasslands than woodlands while richness in Mediterranean areas did not vary significantly among land cover types. These differences were further related to soil pH, as a driver of bacterial and fungal richness in most climatic regions, but the interaction of pH with land cover depended on the region. Microbial community composition differed the most between croplands and woodlands in all regions, mainly due to differences in pH. In the Mediterranean region, bacterial communities in woodlands and grasslands were the most similar, whereas in other regions, grassland and cropland-associated bacteria showed more similarity. Overall, we showed that key factors interact in shaping soil microbial communities in a climate-dependent way at large scale.


Assuntos
Microbiologia do Solo , Solo , Solo/química , Bactérias/genética , Florestas , Concentração de Íons de Hidrogênio , Pradaria
2.
Glob Chang Biol ; 30(1): e16992, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37902125

RESUMO

The EU Soil Strategy 2030 aims to increase soil organic carbon (SOC) in agricultural land to enhance soil health and support biodiversity as well as to offset greenhouse gas emissions through soil carbon sequestration. Therefore, the quantification of current SOC stocks and the spatial identification of the main drivers of SOC changes is paramount in the preparation of agricultural policies aimed at enhancing the resilience of agricultural systems in the EU. In this context, changes of SOC stocks (Δ SOCs) for the EU + UK between 2009 and 2018 were estimated by fitting a quantile generalized additive model (qGAM) on data obtained from the revisited points of the Land Use/Land Cover Area Frame Survey (LUCAS) performed in 2009, 2015 and 2018. The analysis of the partial effects derived from the fitted qGAM model shows that land use and land use change observed in the 2009, 2015 and 2018 LUCAS campaigns (i.e. continuous grassland [GGG] or cropland [CCC], conversion grassland to cropland (GGC or GCC) and vice versa [CGG or CCG]) was one of the main drivers of SOC changes. The CCC was the factor that contributed to the lowest negative change on Δ SOC with an estimated partial effect of -0.04 ± 0.01 g C kg-1 year-1 , while the GGG the highest positive change with an estimated partial effect of 0.49 ± 0.02 g C kg-1 year-1 . This confirms the C sequestration potential of converting cropland to grassland. However, it is important to consider that local soil and environmental conditions may either diminish or enhance the grassland's positive effect on soil C storage. In the EU + UK, the estimated current (2018) topsoil (0-20 cm) SOC stock in agricultural land below 1000 m a.s.l was 9.3 Gt, with a Δ SOC of -0.75% in the period 2009-2018. The highest estimated SOC losses were concentrated in central-northern countries, while marginal losses were observed in the southeast.


Assuntos
Carbono , Solo , Pradaria , Agricultura , Sequestro de Carbono , Produtos Agrícolas
3.
Glob Chang Biol ; 29(19): 5706-5719, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37449367

RESUMO

Soil eukaryotes play a crucial role in maintaining ecosystem functions and services, yet the factors driving their diversity and distribution remain poorly understood. While many studies focus on some eukaryotic groups (mostly fungi), they are limited in their spatial scale. Here, we analyzed an unprecedented amount of observational data of soil eukaryomes at continental scale (787 sites across Europe) to gain further insights into the impact of a wide range of environmental conditions (climatic and edaphic) on their community composition and structure. We found that the diversity of fungi, protists, rotifers, tardigrades, nematodes, arthropods, and annelids was predominantly shaped by ecosystem type (annual and permanent croplands, managed and unmanaged grasslands, coniferous and broadleaved woodlands), and higher diversity of fungi, protists, nematodes, arthropods, and annelids was observed in croplands than in less intensively managed systems, such as coniferous and broadleaved woodlands. Also in croplands, we found more specialized eukaryotes, while the composition between croplands was more homogeneous compared to the composition of other ecosystems. The observed high proportion of overlapping taxa between ecosystems also indicates that DNA has accumulated from previous land uses, hence mimicking the land transformations occurring in Europe in the last decades. This strong ecosystem-type influence was linked to soil properties, and particularly, soil pH was driving the richness of fungi, rotifers, and annelids, while plant-available phosphorus drove the richness of protists, tardigrades, and nematodes. Furthermore, the soil organic carbon to total nitrogen ratio crucially explained the richness of fungi, protists, nematodes, and arthropods, possibly linked to decades of agricultural inputs. Our results highlighted the importance of long-term environmental variables rather than variables measured at the time of the sampling in shaping soil eukaryotic communities, which reinforces the need to include those variables in addition to ecosystem type in future monitoring programs and conservation efforts.


Assuntos
Artrópodes , Ecossistema , Animais , Solo/química , Eucariotos , Carbono , Biodiversidade , Europa (Continente) , Fungos , Microbiologia do Solo
4.
Chembiochem ; 23(12): e202200115, 2022 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-35420232

RESUMO

Protein therapeutics offer exquisite selectivity in targeting cellular processes and behaviors, but are rarely used against non-cell surface targets due to their poor cellular uptake. While cell-penetrating peptides can be used to deliver recombinant proteins to the cytosol, it is generally difficult to selectively deliver active proteins to target cells. Here, we report a recombinantly produced, intracellular protein delivery and targeting platform that uses a photocaged intein to regulate the spatio-temporal activation of protein activity in selected cells upon irradiation with light. The platform was successfully demonstrated for two cytotoxic proteins to selectively kill cancer cells after photoactivation of intein splicing. This platform can generically be applied to any protein whose activity can be disrupted by a fused intein, allowing it to underpin a wide variety of future protein therapeutics.


Assuntos
Antineoplásicos , Peptídeos Penetradores de Células , Inteínas , Processamento de Proteína , Proteínas Recombinantes
5.
Conserv Biol ; 36(5): e13930, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35510330

RESUMO

Soil biodiversity and related ecosystem functions are neglected in most biodiversity assessments and nature conservation actions. We examined how society, and particularly policy makers, have addressed these factors worldwide with a focus on Europe and explored the role of soils in nature conservation in Germany as an example. We reviewed past and current global and European policies, compared soil ecosystem functioning in- and outside protected areas, and examined the role of soils in nature conservation management via text analyses. Protection and conservation of soil biodiversity and soil ecosystem functioning have been insufficient. Soil-related policies are unenforceable and lack soil biodiversity conservation goals, focusing instead on other environmental objectives. We found no evidence of positive effects of current nature conservation measures in multiple soil ecosystem functions in Europe. In German conservation management, soils are considered only from a limited perspective (e.g., as physicochemical part of the environment and as habitat for aboveground organisms). By exploring policy, evidence, and management as it relates to soil ecosystems, we suggest an integrative perspective to move nature conservation toward targeting soil ecosystems directly (e.g., by setting baselines, monitoring soil threats, and establishing a soil indicator system).


La biodiversidad del suelo y las funciones ambientales relacionadas se dejan de lado en la mayoría de las evaluaciones de la biodiversidad y de las acciones de conservación de la naturaleza. Analizamos cómo la sociedad, y particularmente los formuladores de políticas, han abordado estos factores a nivel mundial con un enfoque en Europa y exploramos como ejemplo el papel de los suelos en la conservación de la naturaleza en Alemania. Revisamos las políticas mundiales y europeas en el pasado y en la actualidad, comparamos el funcionamiento ambiental del suelo dentro y fuera de las áreas protegidas y examinamos el papel de los suelos en la gestión de la conservación por medio del análisis de textos. La protección y la conservación de la biodiversidad y el funcionamiento ambiental del suelo han sido insuficientes. Las políticas relacionadas con el suelo son inaplicables y carecen de objetivos de conservación para su biodiversidad, pues se enfocan más bien en otros objetivos ambientales. No descubrimos evidencias de los efectos positivos de las medidas actuales de conservación en múltiples funciones ambientales del suelo en Europa. En la gestión alemana de la conservación, los suelos sólo se consideran desde una perspectiva limitada (p. ej.: como una parte físico química del ambiente y como hábitat para los organismos que habitan por encima de él). Mediante la exploración de la política, evidencias y gestión conforme se relaciona con los ecosistemas del suelo, sugerimos una perspectiva integrada para dirigir a la conservación hacia el enfoque directo sobre los ecosistemas del suelo (p. ej.: al establecer líneas base, monitorear las amenazas para el suelo y establecer un sistema indicador del suelo).


Assuntos
Biodiversidade , Conservação dos Recursos Naturais , Solo , Ecossistema , Europa (Continente)
6.
Int J Mol Sci ; 22(22)2021 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-34830153

RESUMO

A series of 3-(6-substituted phenyl-[1,2,4]-triazolo[3,4-b]-[1,3,4]-thiadiazol-3-yl)-1H-indoles (5a-l) were designed, synthesized and evaluated for anti-apoptotic Bcl-2-inhibitory activity. Synthesis of the target compounds was readily accomplished through a reaction of acyl hydrazide (1) with carbon disulfide in the presence of alcoholic potassium hydroxide to afford the corresponding intermediate potassium thiocarbamate salt (2), which underwent cyclization reaction in the presence of excess hydrazine hydrate to the corresponding triazole thiol (3). Further cyclisation reaction with substituted benzoyl chloride derivatives in the presence of phosphorous oxychloride afforded the final 6-phenyl-indol-3-yl [1,2,4]-triazolo[3,4-b]-[1,3,4]-thiadiazole compounds (5a-l). The novel series showed selective sub-micromolar IC50 growth-inhibitory activity against Bcl-2-expressing human cancer cell lines. The most potent 6-(2,4-dimethoxyphenyl) substituted analogue (5k) showed selective IC50 values of 0.31-0.7 µM against Bcl-2-expressing cell lines without inhibiting the Bcl-2-negative cell line (Jurkat). ELISA binding affinity assay (interruption of Bcl-2-Bim interaction) showed potent binding affinity for (5k) with an IC50 value of 0.32 µM. Moreover, it fulfils drug likeness criteria as a promising drug candidate.


Assuntos
Antineoplásicos/química , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Tiadiazóis/química , Triazóis/química , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Desenho de Fármacos , Células HeLa , Humanos , Concentração Inibidora 50 , Células Jurkat , Modelos Químicos , Estrutura Molecular , Terapia de Alvo Molecular/métodos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Neoplasias/patologia , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Relação Estrutura-Atividade
7.
Glob Chang Biol ; 26(4): 2584-2598, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31923343

RESUMO

Cover crops (CC) promote the accumulation of soil organic carbon (SOC), which provides multiple benefits to agro-ecosystems. However, additional nitrogen (N) inputs into the soil could offset the CO2 mitigation potential due to increasing N2 O emissions. Integrated management approaches use organic and synthetic fertilizers to maximize yields while minimizing impacts by crop sequencing adapted to local conditions. The goal of this work was to test whether integrated management, centered on CC adoption, has the potential to maximize SOC stocks without increasing the soil greenhouse gas (GHG) net flux and other agro-environmental impacts such as nitrate leaching. To this purpose, we ran the DayCent bio-geochemistry model on 8,554 soil sampling locations across the European Union. We found that soil N2 O emissions could be limited with simple crop sequencing rules, such as switching from leguminous to grass CC when the GHG flux was positive (source). Additional reductions of synthetic fertilizers applications are possible through better accounting for N available in green manures and from mineralization of soil reservoirs while maintaining cash crop yields. Therefore, our results suggest that a CC integrated management approach can maximize the agro-environmental performance of cropping systems while reducing environmental trade-offs.

8.
Mol Ther ; 27(11): 1950-1962, 2019 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-31427168

RESUMO

Lipid nanoparticles have great potential for delivering nucleic-acid-based therapeutics, but low efficiency limits their broad clinical translation. Differences in transfection capacity between in vitro models used for nanoparticle pre-clinical testing are poorly understood. To address this, using a clinically relevant lipid nanoparticle (LNP) delivering mRNA, we highlight specific endosomal characteristics in in vitro tumor models that impact protein expression. A 30-cell line LNP-mRNA transfection screen identified three cell lines having low, medium, and high transfection that correlated with protein expression when they were analyzed in tumor models. Endocytic profiling of these cell lines identified major differences in endolysosomal morphology, localization, endocytic uptake, trafficking, recycling, and endolysosomal pH, identified using a novel pH probe. High-transfecting cells showed rapid LNP uptake and trafficking through an organized endocytic pathway to lysosomes or rapid exocytosis. Low-transfecting cells demonstrated slower endosomal LNP trafficking to lysosomes and defective endocytic organization and acidification. Our data establish that efficient LNP-mRNA transfection relies on an early and narrow endosomal escape window prior to lysosomal sequestration and/or exocytosis. Endocytic profiling should form an important pre-clinical evaluation step for nucleic acid delivery systems to inform model selection and guide delivery-system design for improved clinical translation.


Assuntos
Expressão Gênica , Lipídeos/química , Nanopartículas , RNA Mensageiro/genética , Transfecção , Linhagem Celular Tumoral , Endocitose , Endossomos/metabolismo , Citometria de Fluxo , Imunofluorescência , Genes Reporter , Humanos , Concentração de Íons de Hidrogênio , Lisossomos/metabolismo , Nanopartículas/química , RNA Mensageiro/administração & dosagem , Transfecção/métodos
9.
Int J Mol Sci ; 21(23)2020 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-33256166

RESUMO

A series of 2-(1H-indol-3-yl)-5-substituted-1,3,4-oxadiazoles, 4a-m, were designed, synthesized and tested in vitro as potential pro-apoptotic Bcl-2 inhibitory anticancer agents based on our previously reported hit compounds. Synthesis of the target 1,3,4-oxadiazoles was readily accomplished through a cyclization reaction of indole carboxylic acid hydrazide 2 with substituted carboxylic acid derivatives 3a-m in the presence of phosphorus oxychloride. New compounds 4a-m showed a range of IC50 values concentrated in the low micromolar range selectively in Bcl-2 positive human cancer cell lines. The most potent candidate 4-trifluoromethyl substituted analogue 4j showed selective IC50 values of 0.52-0.88 µM against Bcl-2 expressing cell lines with no inhibitory effects in the Bcl-2 negative cell line. Moreover, 4j showed binding that was two-fold more potent than the positive control gossypol in the Bcl-2 ELISA binding affinity assay. Molecular modeling studies helped to further rationalize anti-apoptotic Bcl-2 binding and identified compound 4j as a candidate with drug-like properties for further investigation as a selective Bcl-2 inhibitory anticancer agent.


Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Desenho de Fármacos , Oxidiazóis/síntese química , Oxidiazóis/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Antineoplásicos/química , Sítios de Ligação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Modelos Moleculares , Oxidiazóis/química , Ligação Proteica/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
10.
EMBO Rep ; 18(10): 1727-1739, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28835545

RESUMO

Autophagy (macroautophagy) is a highly conserved eukaryotic degradation pathway in which cytosolic components and organelles are sequestered by specialized autophagic membranes and degraded through the lysosomal system. The autophagic pathway maintains basal cellular homeostasis and helps cells adapt during stress; thus, defects in autophagy can cause detrimental effects. It is therefore crucial that autophagy is properly regulated. In this study, we show that the cysteine protease Atg4B, a key enzyme in autophagy that cleaves LC3, is an interactor of the small GTPase Rab7b. Indeed, Atg4B interacts and co-localizes with Rab7b on vesicles. Depletion of Rab7b increases autophagic flux as indicated by the increased size of autophagic structures as well as the magnitude of macroautophagic sequestration and degradation. Importantly, we demonstrate that Rab7b regulates LC3 processing by modulating Atg4B activity. Taken together, our findings reveal Rab7b as a novel negative regulator of autophagy through its interaction with Atg4B.


Assuntos
Proteínas Relacionadas à Autofagia/metabolismo , Autofagia , Cisteína Endopeptidases/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Proteínas Relacionadas à Autofagia/genética , Cisteína Endopeptidases/genética , Regulação da Expressão Gênica , Humanos , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas rab de Ligação ao GTP/deficiência , Proteínas rab de Ligação ao GTP/genética , proteínas de unión al GTP Rab7
11.
Geoderma ; 355: 113912, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31798185

RESUMO

This paper presents the second part of the mapping of topsoil properties based on the Land Use and Cover Area frame Survey (LUCAS). The first part described the physical properties (Ballabio et al., 2016) while this second part includes the following chemical properties: pH, Cation Exchange Capacity (CEC), calcium carbonates (CaCO3), C:N ratio, nitrogen (N), phosphorus (P) and potassium (K). The LUCAS survey collected harmonised data on changes in land cover and the state of land use for the European Union (EU). Among the 270,000 land use and cover observations selected for field visit, approximately 20,000 soil samples were collected in 24 EU Member States in 2009 together with more than 2000 samples from Bulgaria and Romania in 2012. The chemical properties maps for the European Union were produced using Gaussian process regression (GPR) models. GPR was selected for its capacity to assess model uncertainty and the possibility of adding prior knowledge in the form of covariance functions to the model. The derived maps will establish baselines that will help monitor soil quality and provide guidance to agro-environmental research and policy developments in the European Union.

12.
Molecules ; 24(7)2019 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-30986908

RESUMO

The Bcl-2 protein has been studied as an anticancer drug target in recent years, due to its gatekeeper role in resisting programmed cancer cell death (apoptosis), and the design of BH3 domain mimetics has led to the clinical approval of Venetoclax (ABT-199) for the treatment of chronic lymphocytic leukaemia. In this work we extend our previous studies on the discovery of indole-based heterocycles as Bcl-2 inhibitors, to the identification of quinolin-4-yl based oxadiazole and triazole analogues. Target compounds were readily synthesized via a common aryl-substituted quinolin-4-carbonyl-N-arylhydrazine-1-carbothioamide (5a-b) intermediate, through simple variation of the basic cyclisation conditions. Some of the quinoline-based oxadiazole analogues (e.g. compound 6i) were found to exhibit sub-micromolar anti-proliferative activity in Bcl-2-expressing cancer cell lines, and sub-micromolar IC50 activity within a Bcl2-Bim peptide ELISA assay. The Bcl-2 targeted anticancer activity of 6i was further rationalised via computational molecular modelling, offering possibilities to extend this work into the design of further potent and selective Bcl-2 inhibitory heteroaromatics with therapeutic potential.


Assuntos
Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Leucemia Linfocítica Crônica de Células B/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Quinolinas/química , Apoptose/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Ensaio de Imunoadsorção Enzimática , Compostos Heterocíclicos/química , Compostos Heterocíclicos/uso terapêutico , Humanos , Hidrocarbonetos Aromáticos/química , Hidrocarbonetos Aromáticos/uso terapêutico , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Oxidiazóis/uso terapêutico , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Triazóis/uso terapêutico
13.
Bioconjug Chem ; 29(4): 1030-1046, 2018 04 18.
Artigo em Inglês | MEDLINE | ID: mdl-29481068

RESUMO

Ligand-mediated targeting and internalization of plasma membrane receptors is central to cellular function. These types of receptors have accordingly been investigated as targets to facilitate entry of diagnostic and therapeutic constructs into cells. However, there remains a need to characterize how receptor targeting agents on nanoparticles interact at surface receptors and whether it is possible to control these interactions via exogenous stimuli. Here, we describe the switchable display of the iron-transporting protein, transferrin (Tf), at the surface of thermoresponsive polymer-coated gold nanoparticles and show that internalization of the coated nanoparticles into target cells changes across temperature ranges over which transferrin is expected to be sterically "hidden" by an extended polymer chain and then "revealed" by polymer chain collapse. The switching process is dependent on the numbers of transferrin molecules and thermoresponsive polymer chains attached and whether the assay temperature is above or below the transition temperatures of the responsive polymers at the nanoparticle surfaces. Significantly, however, the control of internalization is critically reliant on overall nanoparticle colloidal stability while the thermoresponsive component of the surface undergoes conformational change. The data show that the cell entry function of complex and large biomolecule ligands can be modulated by polymer-induced accessibility change but that a simple "hide and reveal" mechanism for ligand display following polymer chain collapse is insufficient to account for nanoparticle uptake and subsequent intracellular trafficking.


Assuntos
Endocitose/efeitos dos fármacos , Substâncias Macromoleculares/química , Nanopartículas Metálicas/química , Polímeros/química , Polímeros/farmacologia , Sítios de Ligação , Entropia , Ouro/química , Células HeLa , Humanos , Interações Hidrofóbicas e Hidrofílicas , Ligantes , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Proteínas/química , Espectrofotometria Ultravioleta , Temperatura , Transferrina/química
14.
Bioorg Med Chem Lett ; 27(4): 1037-1040, 2017 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-28087272

RESUMO

A series of 5-(1H-indol-3-yl)-N-aryl-1,3,4-oxadiazol-2-amines 8a-j has been designed, synthesized and tested in vitro as potential pro-apoptotic Bcl-2-inhibitory anticancer agents based on our previous lead compound 8a. Synthesis of the target compounds was readily accomplished through a cyclisation reaction between indole-3-carboxylic acid hydrazide (5) and substituted isothiocyanates 6a-j, followed by oxidative cyclodesulfurization of the corresponding thiosemicarbazide 7a-j using 1,3-dibromo-5,5-dimethylhydantoin. Active compounds of the series 8a-j were found to have sub-micromolar IC50 values selectively in Bcl-2 expressing human cancer cell lines; notably the 2-nitrophenyl analogue 8a was found to exhibit potent activity, and compounds 8a and 8e possessed comparable Bcl-2 binding affinity (ELISA assay) to the established natural product-based Bcl-2 inhibitor, gossypol. Molecular modeling studies helped to further rationalise anti-apoptotic Bcl-2 binding, and identified compounds 8a and 8e as candidates for further development as Bcl-2 inhibitory anticancer agents.


Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Oxidiazóis/síntese química , Oxidiazóis/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Humanos
15.
Glob Chang Biol ; 22(5): 1976-84, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26679897

RESUMO

The idea of offsetting anthropogenic CO2 emissions by increasing global soil organic carbon (SOC), as recently proposed by French authorities ahead of COP21 in the 'four per mil' initiative, is notable. However, a high uncertainty still exits on land C balance components. In particular, the role of erosion in the global C cycle is not totally disentangled, leading to disagreement whether this process induces lands to be a source or sink of CO2. To investigate this issue, we coupled soil erosion into a biogeochemistry model, running at 1 km(2) resolution across the agricultural soils of the European Union (EU). Based on data-driven assumptions, the simulation took into account also soil deposition within grid cells and the potential C export to riverine systems, in a way to be conservative in a mass balance. We estimated that 143 of 187 Mha have C erosion rates <0.05 Mg C ha(-1) yr(-1), although some hot-spot areas showed eroded SOC >0.45 Mg C ha(-1) yr(-1). In comparison with a baseline without erosion, the model suggested an erosion-induced sink of atmospheric C consistent with previous empirical-based studies. Integrating all C fluxes for the EU agricultural soils, we estimated a net C loss or gain of -2.28 and +0.79 Tg yr(-1) of CO2 eq, respectively, depending on the value for the short-term enhancement of soil C mineralization due to soil disruption and displacement/transport with erosion. We concluded that erosion fluxes were in the same order of current carbon gains from improved management. Even if erosion could potentially induce a sink for atmospheric CO2, strong agricultural policies are needed to prevent or reduce soil erosion, in order to maintain soil health and productivity.


Assuntos
Dióxido de Carbono/análise , Carbono/análise , Solo/química , Agricultura , Ciclo do Carbono , Monitoramento Ambiental , Europa (Continente) , Modelos Teóricos
16.
Mol Ther ; 23(12): 1888-98, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26412588

RESUMO

A major unmet clinical need is a universal method for subcellular targeting of bioactive molecules to lysosomes. Delivery to this organelle enables either degradation of oncogenic receptors that are overexpressed in cancers, or release of prodrugs from antibody-drug conjugates. Here, we describe a general method that uses receptor crosslinking to trigger endocytosis and subsequently redirect trafficking of receptor:cargo complexes from their expected route, to lysosomes. By incubation of plasma membrane receptors with biotinylated cargo and subsequent addition of streptavidin to crosslink receptor:cargo-biotin complexes, we achieved rapid and selective lysosomal targeting of transferrin, an anti-MHC class I antibody, and the clinically approved anti-Her2 antibody trastuzumab. These three protein ligands each target a receptor with a distinct cellular function and intracellular trafficking profile. Importantly, we confirmed that crosslinking of trastuzumab increased lysosomal degradation of its cognate oncogenic receptor Her2 in breast cancer cell lines SKBR3 and BT474. These data suggest that crosslinking could be exploited for a wide range of target receptors, for navigating therapeutics through the endolysosomal pathway, for significant therapeutic benefit.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Marcação de Genes/métodos , Lisossomos/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Endocitose/efeitos dos fármacos , Feminino , Células HeLa , Humanos , Imunoconjugados/farmacologia , Ligantes , Pró-Fármacos , Transporte Proteico , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Trastuzumab/farmacologia
17.
Mol Pharm ; 12(6): 1970-9, 2015 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-25879919

RESUMO

Glycol chitosan nanogels have been widely used in gene, drug, and contrast agent delivery in an effort to improve disease diagnosis and treatment. Herein, we evaluate the internalization mechanisms and intracellular fate of previously described glycol chitosan nanogels decorated with folate to target the folate receptor. Uptake of the folate-decorated nanogel was impaired by free folate, suggesting competitive inhibition and shared internalization mechanisms via the folate receptor. Nanogel uptake was shown to occur mainly through flotillin-1 and Cdc42-dependent endocytosis. This was determined by inhibition of uptake reduction observed upon siRNA depletion of these two proteins and the pathways that they regulate. The data also suggest the involvement of the actin cytoskeleton in nanogel uptake via macropinocytosis. After 7 h of incubation with HeLa cells, approximately half of the nanogel population was localized in endolysosomal compartments, whereas the remaining 50% of the material was in undefined regions of the cytoplasm. Glycol chitosan nanogels may thus have potential as drug delivery vectors for targeting different intracellular compartments.


Assuntos
Quitosana/química , Ácido Fólico/química , Polietilenoglicóis/química , Polietilenoimina/química , RNA Interferente Pequeno/genética , Western Blotting , Eletroforese em Gel de Poliacrilamida , Endocitose/fisiologia , Citometria de Fluxo , Células HeLa , Humanos , Nanogéis
18.
Pharm Res ; 32(5): 1546-56, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25777610

RESUMO

PURPOSE: To investigate the suitability of three antimicrobial peptides (AMPs) as cell-penetrating antimicrobial peptides. METHODS: Cellular uptake of three AMPs (PK-12-KKP, SA-3 and TPk) and a cell-penetrating peptide (penetratin), all 5(6)-carboxytetramethylrhodamine-labeled, were tested in HeLa WT cells and analyzed by flow cytometry and confocal microscopy. Furthermore, the effects of the peptides on eukaryotic cell viability as well as their antimicrobial effect were tested. In addition, the disrupting ability of the peptides in the presence of bilayer membranes of different composition were analyzed. RESULTS: AMP uptake relative to penetratin was ~13% (PK-12-KKP), ~66% (SA-3) and ~50% (TPk). All four peptides displayed a punctate uptake pattern in HeLa WT cells with co-localization to lysosomes and no indication that clathrin-mediated endocytosis was the predominant uptake mechanism. TPk showed the highest antibacterial activity. SA-3 exhibited selective disruption of liposomes mimicking Gram-positive and Gram-negative membranes. CONCLUSION: PK-12-KKP is an unlikely candidate for targeting intracellular bacteria, as the eukaryotic cell-penetrating ability is poor. SA-3, affected the cellular viability to an unacceptable degree. TPk showed acceptable uptake efficiency, high antimicrobial activity and relatively low toxicity, and it is the best potential lead peptide for further development.


Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Bactérias/efeitos dos fármacos , Proteínas de Transporte/farmacologia , Peptídeos Penetradores de Células/farmacologia , Sequência de Aminoácidos , Antibacterianos/química , Antibacterianos/farmacocinética , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacocinética , Infecções Bacterianas/tratamento farmacológico , Proteínas de Transporte/química , Proteínas de Transporte/farmacocinética , Peptídeos Penetradores de Células/química , Peptídeos Penetradores de Células/farmacocinética , Endocitose , Células HeLa , Humanos , Dados de Sequência Molecular
19.
Glob Chang Biol ; 20(1): 313-26, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23765562

RESUMO

Proposed European policy in the agricultural sector will place higher emphasis on soil organic carbon (SOC), both as an indicator of soil quality and as a means to offset CO2 emissions through soil carbon (C) sequestration. Despite detailed national SOC data sets in several European Union (EU) Member States, a consistent C stock estimation at EU scale remains problematic. Data are often not directly comparable, different methods have been used to obtain values (e.g. sampling, laboratory analysis) and access may be restricted. Therefore, any evolution of EU policies on C accounting and sequestration may be constrained by a lack of an accurate SOC estimation and the availability of tools to carry out scenario analysis, especially for agricultural soils. In this context, a comprehensive model platform was established at a pan-European scale (EU + Serbia, Bosnia and Herzegovina, Croatia, Montenegro, Albania, Former Yugoslav Republic of Macedonia and Norway) using the agro-ecosystem SOC model CENTURY. Almost 164 000 combinations of soil-climate-land use were computed, including the main arable crops, orchards and pasture. The model was implemented with the main management practices (e.g. irrigation, mineral and organic fertilization, tillage) derived from official statistics. The model results were tested against inventories from the European Environment and Observation Network (EIONET) and approximately 20 000 soil samples from the 2009 LUCAS survey, a monitoring project aiming at producing the first coherent, comprehensive and harmonized top-soil data set of the EU based on harmonized sampling and analytical methods. The CENTURY model estimation of the current 0-30 cm SOC stock of agricultural soils was 17.63 Gt; the model uncertainty estimation was below 36% in half of the NUTS2 regions considered. The model predicted an overall increase of this pool according to different climate-emission scenarios up to 2100, with C loss in the south and east of the area (involving 30% of the whole simulated agricultural land) compensated by a gain in central and northern regions. Generally, higher soil respiration was offset by higher C input as a consequence of increased CO2 atmospheric concentration and favourable crop growing conditions, especially in northern Europe. Considering the importance of SOC in future EU policies, this platform of simulation appears to be a very promising tool to orient future policymaking decisions.


Assuntos
Carbono/análise , Modelos Teóricos , Solo/química , Agricultura , Sequestro de Carbono , Europa (Continente)
20.
Glob Chang Biol ; 20(11): 3557-67, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24789378

RESUMO

Bottom-up estimates from long-term field experiments and modelling are the most commonly used approaches to estimate the carbon (C) sequestration potential of the agricultural sector. However, when data are required at European level, important margins of uncertainty still exist due to the representativeness of local data at large scale or different assumptions and information utilized for running models. In this context, a pan-European (EU + Serbia, Bosnia and Herzegovina, Montenegro, Albania, Former Yugoslav Republic of Macedonia and Norway) simulation platform with high spatial resolution and harmonized data sets was developed to provide consistent scenarios in support of possible carbon sequestration policies. Using the CENTURY agroecosystem model, six alternative management practices (AMP) scenarios were assessed as alternatives to the business as usual situation (BAU). These consisted of the conversion of arable land to grassland (and vice versa), straw incorporation, reduced tillage, straw incorporation combined with reduced tillage, ley cropping system and cover crops. The conversion into grassland showed the highest soil organic carbon (SOC) sequestration rates, ranging between 0.4 and 0.8 t C ha(-1)  yr(-1) , while the opposite extreme scenario (100% of grassland conversion into arable) gave cumulated losses of up to 2 Gt of C by 2100. Among the other practices, ley cropping systems and cover crops gave better performances than straw incorporation and reduced tillage. The allocation of 12 to 28% of the European arable land to different AMP combinations resulted in a potential SOC sequestration of 101-336 Mt CO2 eq. by 2020 and 549-2141 Mt CO2 eq. by 2100. Modelled carbon sequestration rates compared with values from an ad hoc meta-analysis confirmed the robustness of these estimates.


Assuntos
Agricultura/métodos , Sequestro de Carbono , Conservação dos Recursos Naturais , Solo/química , Europa (Continente) , Modelos Teóricos
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