Atomic Force Microscopy - Laser Scanning Confocal Microscopy imaging protocol for copepod fecal pellets

Within the marine carbon and nitrogen biogeochemical cycles, fecal pellets produced by pelagic and benthic copepods are important microbial activity hotspots as particle substrate for bacteria colonization and as sources of particulate and dissolved organic C and N. We developed a protocol combining Atomic Force Microscopy (AFM) and Laser Scanning Confocal Microscopy (LSCM) to study the peritrophic membrane structure and associated bacteria of fecal pellets produced by a benthic copepod, Paramphiascella fulvofasciata. AFM imaging revealed a fibrillar network structure of the peritrophic membrane, 0.7-5.9 nm thick similar to marine polysaccharides and a-chitin. Bacterial cell volume range was 0.006-0.117 µm3 in liquid. LSCM imaging showed a 3D-heterogeneous microenvironment. This protocol would allow high-resolution interrogation of structural changes and bacterial dynamics within the copepod fecal pellets and other heterogeneous particles such marine snow under environmental conditions

Assembling large, complex environmental metagenomes

The large volumes of sequencing data required to sample complex environments deeply pose new challenges to sequence analysis approaches. De novo metagenomic assembly effectively reduces the total amount of data to be analyzed but requires significant computational resources. We apply two pre-assembly filtering approaches, digital normalization and partitioning, to make large metagenome assemblies more comput\ ationaly tractable. Using a human gut mock community dataset, we demonstrate that these methods result in assemblies nearly identical to assemblies from unprocessed data. We then assemble two large soil metagenomes from matched Iowa corn and native prairie soils. The predicted functional content and phylogenetic origin of the assembled contigs indicate significant taxonomic differences despite similar function. The assembly strategies presented are generic and can be extended to any metagenome; full source code is freely available under a BSD license.Comment: Includes supporting informatio

Naturwall: active timber wall for renovation of existent buildings

The renovation of old buildings is actually the challenge to cope with increased effort in order to reduce climate global change, channeling more investment and awareness in this sense, defining more experimentations and find innovative solutions. The difficulty of carrying out an intervention on the existing buildings necessarily arise from the lack of information on the existing structure and the lack of coordinated processes between the multidisciplinary skills involved, as well as a difficulty to optimize the process that would make it even more competitive on the renovation work instead on the new construction. Naturwall is an innovative energy saving system for existent buildings by using wood in multifunctional components able to mitigate the environmental effort in building management. The project meant to introduce an industrialized design method in the renovation of existing build environment that highlights opportunities gave by "off site" production and parametric design approach, without neglecting the aesthetical values and the possibility to change the architectural image of residential and non residential constructions. The project aims to create a representative model of solution that will be promoted in Italy and widespread in other similar contex

Heat Treated NiP–SiC Composite Coatings: Elaboration and Tribocorrosion Behaviour in NaCl Solution

Tribocorrosion behaviour of heat-treated NiP and NiP–SiC composite coatings was investigated in a 0.6 M NaCl solution. The tribocorrosion tests were performed in a linear sliding tribometer with an electrochemical cell interface. It was analyzed the influence of SiC particles dispersion in the NiP matrix on current density developed, on coefficient of friction and on wear volume loss. The results showed that NiP–SiC composite coatings had a lower wear volume loss compared to NiP coatings. However, the incorporation of SiC particles into the metallic matrix affects the current density developed by the system during the tribocorrosion test. It was verified that not only the volume of co-deposited particles (SiC vol.%) but also the number of SiC particles per coating area unit (and consequently the SiC particles size) have made influence on the tribocorrosion behaviour of NiP–SiC composite coatings

New perspectives in cancer biology from a study of canonical and non-canonical functions of base excision repair proteins with a focus on early steps

Alterations of DNA repair enzymes and consequential triggering of aberrant DNA damage response (DDR) pathways are thought to play a pivotal role in genomic instabilities associated with cancer development, and are further thought to be important predictive biomarkers for therapy using the synthetic lethality paradigm. However, novel unpredicted perspectives are emerging from the identification of several non-canonical roles of DNA repair enzymes, particularly in gene expression regulation, by different molecular mechanisms, such as (i) non-coding RNA regulation of tumour suppressors, (ii) epigenetic and transcriptional regulation of genes involved in genotoxic responses and (iii) paracrine effects of secreted DNA repair enzymes triggering the cell senescence phenotype. The base excision repair (BER) pathway, canonically involved in the repair of non-distorting DNA lesions generated by oxidative stress, ionising radiation, alkylation damage and spontaneous or enzymatic deamination of nucleotide bases, represents a paradigm for the multifaceted roles of complex DDR in human cells. This review will focus on what is known about the canonical and non-canonical functions of BER enzymes related to cancer development, highlighting novel opportunities to understand the biology of cancer and representing future perspectives for designing new anticancer strategies. We will specifically focus on APE1 as an example of a pleiotropic and multifunctional BER protein

Hybrid stars with sequential phase transitions: the emergence of the g2_2 mode

Neutron stars are the densest objects in the Universe, with $M \sim 1.4 M_{\odot}$ and $R \sim 12$ km, and the equation of state associated to their internal composition is still unknown. The extreme conditions to which matter is subjected inside neutron stars could lead to a phase transition in their inner cores, giving rise to a hybrid compact object. The observation of $2M_{\odot}$ binary pulsars (PSR~J1614-2230, PSR~J0343$+$0432 and PSR~J0740$+$6620) strongly constraints theoretical models of the equation of state. Moreover, the detection of gravitational waves emitted during the binary neutron star merger, GW170817, and its electromagnetic counterpart, GRB170817A, impose additional constraints on the tidal deformability. In this work, we investigate hybrid stars with sequential phase transitions hadron-quark-quark in their cores. We assume that both phase transitions are sharp and analyse the rapid and slow phase conversion scenarios. For the outer core, we use modern hadronic equations of state. For the inner core we employ the constant speed of sound parametrization for quark matter. We analyze more than 3000 hybrid equations of state, taking into account the recent observational constraints from neutron stars. The effects of hadron-quark-quark phase transitions on the normal oscillation modes $f$ and $g$, are studied under the Cowling relativistic approximation. Our results show that, in the slow conversion regime, a second quark-quark phase transition gives rise to a new $g_2$~mode. We discuss the observational implications of our results associated to the gravitational waves detection and the possibility of detecting hints of sequential phase transitions and the associated $g_2$~mode.Comment: 24 pages, 9 figure

Rede de estações meteorológicas automáticas sem fio.

bitstream/item/133214/1/ID-43372-2014-TCC-MiniColetorDados.pdfTCC (Bacharelado em Ciência da Computação) - Universidade de Passo Fundo, Passo Fundo

Integrating sol-gel and carbon dots chemistry for the fabrication of fluorescent hybrid organic-inorganic films

Highly fluorescent blue and green-emitting carbon dots have been designed to be integrated into sol-gel processing of hybrid organic-inorganic materials through surface modification with an organosilane, 3-(aminopropyl)triethoxysilane (APTES). The carbon dots have been synthesised using citric acid and urea as precursors; the intense fluorescence exhibited by the nanoparticles, among the highest reported in the scientific literature, has been stabilised against quenching by APTES. When the modification is carried out in an aqueous solution, it leads to the formation of silica around the C-dots and an increase of luminescence, but also to the formation of large clusters which do not allow the deposition of optically transparent films. On the contrary, when the C-dots are modified in ethanol, the APTES improves the stability in the precursor sol even if any passivating thin silica shell does not form. Hybrid films containing APTES-functionalized C-dots are transparent with no traces of C-dots aggregation and show an intense luminescence in the blue and green range

Enzymatically active apurinic/apyrimidinic endodeoxyribonuclease 1 is released by mammalian cells through exosomes

The apurinic/apyrimidinic endodeoxyribonuclease 1 (APE1), the main AP-endonuclease of the DNA base excision repair pathway, is a key molecule of interest to researchers due to its unsuspected roles in different nonrepair activities, such as: I) adaptive cell response to genotoxic stress, ii) regulation of gene expression, and iii) processing of microRNAs, which make it an excellent drug target for cancer treatment. We and others recently demonstrated that APE1 can be secreted in the extracellular environment and that serum APE1 may represent a novel prognostic biomarker in hepatocellular and non-smallcell lung cancers. However, the mechanism by which APE1 is released extracellularly was not described before. Here, using three different approaches for exosomes isolation: Commercial kit, nickel-based isolation, and ultracentrifugation methods and various mammalian cell lines, we elucidated the mechanisms responsible for APE1 secretion. We demonstrated that APE1 p37 and p33 forms are actively secreted through extracellular vesicles (EVs), including exosomes from different mammalian cell lines. We then observed that APE1 p33 form is generated by proteasomal-mediated degradation and is enzymatically active in EVs. Finally, we revealed that the p33 form of APE1 accumulates in EVs upon genotoxic treatment by cisplatin and doxorubicin, compounds commonly found in chemotherapy pharmacological treatments. Taken together, these findings provide for the first time evidence that a functional Base Excision Repair protein is delivered through exosomes in response to genotoxic stresses, shedding new light into the complex noncanonical biological functions of APE1 and opening new intriguing perspectives on its role in cancer biology