Analysis of the activation of acetylcholinesterase by carbon nanoparticles using a monolithic immobilized enzyme microreactor: role of the water molecules in the active site gorge

A biochromatographic system was used to study the direct effect of carbon nanoparticles (CNPs) on the acetylcholinesterase (AChE) activity. The AChE enzyme was covalently immobilized on a monolithic CIM-disk via its NH₂ residues. Our results showed an increase in the AChE activity in presence of CNPs. The catalytic constant (k_cat) was increased while the Michaelis constant (K_m) was slightly decreased. This indicated an increase in the enzyme efficiency with increase of the substrate affinity to the active site. The thermodynamic data of the activation mechanism of the enzyme, i.e. ΔH* and ΔS*, showed no change in the substrate interaction mechanism with the anionic binding site. The increase of the enthalpy (ΔH*) and the entropy (ΔS*) with decrease in the free energy of activation (Ea) was related to structural conformation change in the active site gorge. This affected the stability of water molecules in the active site gorge and facilitated water displacement by substrate for entering to the active site of the enzyme.     

Way out of Africa: Early Pleistocene paleoenvironments inhabited by Homo erectus in Sangiran, Java

A sequence of paleosols in the Solo Basin, Central Java, Indonesia, documents the local and regional environments present when Homo erectus spread through Southeast Asia during the early Pleistocene. The earliest human immigrants encountered a low-relief lake-margin landscape dominated by moist grasslands with open woodlands in the driest landscape positions. By 1.5 Ma, large streams filled the lake and the landscape became more riverine in nature, with riparian forests, savanna, and open woodland. Paleosol morphology and carbon isotope values of soil organic matter and pedogenic carbonates indicate a long-term shift toward regional drying or increased duration of the annual dry season through the early Pleistocene. This suggests that an annual dry season associated with monsoon conditions was an important aspect of the paleoclimate in which early humans spread from Africa to Southeast Asia.     

Cd污染及其与大气CO2浓度升高、N添加复合作用对大叶相思生长的影响

为了解Cd污染胁迫下树木对CO_2浓度升高、N添加及其复合作用的响应,应用开顶箱,探讨Cd及其与CO_2、N的复合作用对大叶相思(Acacia auriculiformis)基径、树高和生物量的影响。结果表明,Cd添加抑制大叶相思基径、树高和生物量的增长,并且具有时间滞后性;大气CO_2浓度升高、N添加及CO_2+N均有缓解Cd对植物生长抑制作用的趋势,其中, N添加更能促进大叶相思基径的生长,树高生长则对CO_2升高更为敏感;在Cd污染土壤中,N添加的缓解作用最显著。因此,氮肥管理是重金属污染土地修复初期促进植物修复的重要策略。     

Les sépultures italiennes du Paléolithique supérieur. Reconstitutions du régime alimentaire

The identification of dietary habits is increasingly seen as a fundamental aspect for studying the ancient human populations. Accordingly, several projects aiming to identify Paleolithic individuals’ dietary patterns were developed to analyze the organic component of bone tissue and identify isotopic markers to reconstruct the food typology. Bone fragments from six individuals were selected for carbon and nitrogen stable isotopes analysis. The interpretation of human isotopic data was framed through a dataset of twenty-one Italian Paleolithic individuals. The isotopic data generated for the Paleolithic individuals agree with the information already provided by the archaeological record concerning the Italian hunter and gatherer communities. Their subsistence economy was essentially grounded upon the exploitation of high protein foods, either from terrestrial fauna resources or inland lacustrine or riverine species.     

IMPROV-ED trial: eHealth programme for faster recovery and reduced healthcare utilisation after CABG

Netherlands Heart Journal - After coronary artery bypass grafting (CABG), healthcare utilisation is high and is partly unplanned. eHealth applications have been proposed to reduce healthcare...     

Design of mutualistic microbial consortia for stable conversion of carbon monoxide to value-added chemicals

Carbon monoxide (CO) is a promising carbon source for producing value-added biochemicals via microbial fermentation. However, its microbial conversion has been challenging because of difficulties in genetic engineering of CO-utilizing microorganisms and, more importantly, maintaining CO consumption which is negatively affected by the toxicity of CO and accumulated byproducts. To overcome these issues, we devised mutualistic microbial consortia, co-culturing Eubacterium limosum and genetically engineered Escherichia coli for the production of 3-hydroxypropionic acid (3-HP) and itaconic acid (ITA). During the co-culture, E. limosum assimilated CO and produced acetate, a toxic by-product, while E. coli utilized acetate as a sole carbon source. We found that this mutualistic interaction dramatically stabilized and improved CO consumption of E. limosum compared to monoculture. Consequently, the improved CO consumption allowed successful production of 3-HP and ITA from CO. This study is the first demonstration of value-added biochemical production from CO using a microbial consortium. Moreover, it suggests that synthetic mutualistic microbial consortium can serve as a powerful platform for the valorization of CO.     

Carbon nanotubes promote Cr(VI) reduction by alginate-immobilized Shewanella oneidensis MR-1

Bioreduction of hexavalent chromium (Cr(VI)) into trivalent one (Cr(III)) based on microbial immobilization techniques has been recognized as a promising way to remove Cr contaminants from wastewater. However, such a bioreduction process is inefficient due to limited electron transfer through the immobilization matrix. In this study, a modified immobilization process was proposed by impregnating carbon nanotubes (CNTs) into Ca-alginate beads, which were then used to immobilize Shewanella oneidensis MR-1 for enhanced Cr(VI) reduction. Compared with the free cells and the beads without CNTs, the AL/CNT/cell beads showed up to 4 times higher reduction rates, mainly attributed to an enhanced electron transfer by the CNTs. In addition, the dose of CNTs greatly improved the stability of beads, suggesting a high feasibility of the AL/CNT/cell beads for repeated use. The optimized CNT concentration, temperature and pH for Cr(VI) reduction by the AL/CNT/cell beads were 0.5%, 30 °C and 6.0–7.0, respectively.     

Unimodal size scaling of phytoplankton growth and the size dependence of nutrient uptake and use

Phytoplankton size structure is key for the ecology and biogeochemistry of pelagic ecosystems, but the relationship between cell size and maximum growth rate (μ_max) is not yet well understood. We used cultures of 22 species of marine phytoplankton from five phyla, ranging from 0.1 to 10⁶ μm³ in cell volume (V_cell), to determine experimentally the size dependence of growth, metabolic rate, elemental stoichiometry and nutrient uptake. We show that both μ_max and carbon‐specific photosynthesis peak at intermediate cell sizes. Maximum nitrogen uptake rate (V_maxN) scales isometrically with V_cell, whereas nitrogen minimum quota scales as V_cell^0.84. Large cells thus possess high ability to take up nitrogen, relative to their requirements, and large storage capacity, but their growth is limited by the conversion of nutrients into biomass. Small species show similar volume‐specific V_maxN compared to their larger counterparts, but have higher nitrogen requirements. We suggest that the unimodal size scaling of phytoplankton growth arises from taxon‐independent, size‐related constraints in nutrient uptake, requirement and assimilation.     

Fabrication of Carbon Nanotube High-Frequency Nanoelectronic Biosensor for Sensing in High Ionic Strength Solutions

The unique electronic properties and high surface-to-volume ratios of single-walled carbon nanotubes (SWNT) and semiconductor nanowires (NW) ^1-4 make them good candidates for high sensitivity biosensors. When a charged molecule binds to such a sensor surface, it alters the carrier density⁵ in the sensor, resulting in changes in its DC conductance. However, in an ionic solution a charged surface also attracts counter-ions from the solution, forming an electrical double layer (EDL). This EDL effectively screens off the charge, and in physiologically relevant conditions ~100 millimolar (mM), the characteristic charge screening length (Debye length) is less than a nanometer (nm). Thus, in high ionic strength solutions, charge based (DC) detection is fundamentally impeded^6-8.We overcome charge screening effects by detecting molecular dipoles rather than charges at high frequency, by operating carbon nanotube field effect transistors as high frequency mixers^9-11. At high frequencies, the AC drive force can no longer overcome the solution drag and the ions in solution do not have sufficient time to form the EDL. Further, frequency mixing technique allows us to operate at frequencies high enough to overcome ionic screening, and yet detect the sensing signals at lower frequencies^11-12. Also, the high transconductance of SWNT transistors provides an internal gain for the sensing signal, which obviates the need for external signal amplifier.Here, we describe the protocol to (a) fabricate SWNT transistors, (b) functionalize biomolecules to the nanotube¹³, (c) design and stamp a poly-dimethylsiloxane (PDMS) micro-fluidic chamber¹⁴ onto the device, and (d) carry out high frequency sensing in different ionic strength solutions¹¹.     

Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry

Carbon-based nanomaterials, like carbon nanotubes (CNTs), belong to this type of nanoparticles which are very difficult to discriminate from carbon-rich cell structures and de facto there is still no quantitative method to assess their distribution at cell and tissue levels. What we propose here is an innovative method allowing the detection and quantification of CNTs in cells using a multispectral imaging flow cytometer (ImageStream, Amnis). This newly developed device integrates both a high-throughput of cells and high resolution imaging, providing thus images for each cell directly in flow and therefore statistically relevant image analysis. Each cell image is acquired on bright-field (BF), dark-field (DF), and fluorescent channels, giving access respectively to the level and the distribution of light absorption, light scattered and fluorescence for each cell. The analysis consists then in a pixel-by-pixel comparison of each image, of the 7,000-10,000 cells acquired for each condition of the experiment. Localization and quantification of CNTs is made possible thanks to some particular intrinsic properties of CNTs: strong light absorbance and scattering; indeed CNTs appear as strongly absorbed dark spots on BF and bright spots on DF with a precise colocalization.This methodology could have a considerable impact on studies about interactions between nanomaterials and cells given that this protocol is applicable for a large range of nanomaterials, insofar as they are capable of absorbing (and/or scattering) strongly enough the light.