Linking biofilm spatial structure to real-time microscopic oxygen decay imaging

This is an Accepted Manuscript of an article published by Taylor & Francis Group in Biofouling on 2018, available online at: http://www.tandfonline.com/10.1080/08927014.2017.1423474Two non-destructive techniques, confocal laser scanning microscopy (CLSM) and planar optode (VisiSens imaging), were combined to relate the fine-scale spatial structure of biofilm components to real-time images of oxygen decay in aquatic biofilms. Both techniques were applied to biofilms grown for seven days at contrasting light and temperature (10/20°C) conditions. The geo-statistical analyses of CLSM images indicated that biofilm structures consisted of small (~100 µm) and middle sized (~101 µm) irregular aggregates. Cyanobacteria and EPS (extracellular polymeric substances) showed larger aggregate sizes in dark grown biofilms while, for algae, aggregates were larger in light-20°C conditions. Light-20°C biofilms were most dense while 10°C biofilms showed a sparser structure and lower respiration rates. There was a positive relationship between the number of pixels occupied and the oxygen decay rate. The combination of optodes and CLMS, taking advantage of geo-statistics, is a promising way to relate biofilm architecture and metabolism at the micrometric scale.Peer ReviewedPostprint (author's final draft

Lexical learning and dysgraphia in a group of adults with developmental dyslexia

We investigated the ability to learn new words in a group of 22 adults with developmental dyslexia/dysgraphia and the relationship between their learning and spelling problems. We identified a deficit that affected the ability to learn both spoken and written new words (lexical learning deficit). There were no comparable problems in learning other kinds of representations (lexical/semantic and visual) and the deficit could not be explained in terms of more traditional phonological deficits associated with dyslexia (phonological awareness, phonological STM). Written new word learning accounted for further variance in the severity of the dysgraphia after phonological abilities had been partialled out. We suggest that lexical learning may be an independent ability needed to create lexical/formal representations from a series of independent units. Theoretical and clinical implications are discussed. © 2005 Psychology Press Ltd

A mechanistic model (BCC-PSSICO) to predict changes in the hydraulic properties for bio-amended variably saturated soils

This is the peer reviewed version of the following article: [Carles Brangarí, A., X. Sanchez-Vila, A. Freixa, A. M. Romaní, S. Rubol, and D. Fernàndez-Garcia (2017), A mechanistic model (BCC-PSSICO) to predict changes in the hydraulic properties for bio-amended variably saturated soils, Water Resour. Res., 53, 93–109, doi:10.1002/2015WR018517], which has been published in final form at http://onlinelibrary.wiley.com/doi/10.1002/2015WR018517/abstract. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.The accumulation of biofilms in porous media is likely to influence the overall hydraulic properties and, consequently, a sound understanding of the process is required for the proper design and management of many technological applications. In order to bring some light into this phenomenon we present a mechanistic model to study the variably saturated hydraulic properties of bio-amended soils. Special emphasis is laid on the distribution of phases at pore-scale and the mechanisms to retain and let water flow through, providing valuable insights into phenomena behind bioclogging. Our approach consists in modeling the porous media as an ensemble of capillary tubes, obtained from the biofilm-free water retention curve. This methodology is extended by the incorporation of a biofilm composed of bacterial cells and extracellular polymeric substances (EPS). Moreover, such a microbial consortium displays a channeled geometry that shrinks/swells with suction. Analytical equations for the volumetric water content and the relative permeability can then be derived by assuming that biomass reshapes the pore space following specific geometrical patterns. The model is discussed by using data from laboratory studies and other approaches already existing in the literature. It can reproduce (i) displacements of the retention curve toward higher saturations and (ii) permeability reductions of distinct orders of magnitude. Our findings also illustrate how even very small amounts of biofilm may lead to significant changes in the hydraulic properties. We, therefore, state the importance of accounting for the hydraulic characteristics of biofilms and for a complex/more realistic geometry of colonies at the pore-scale.Peer ReviewedPostprint (author's final draft

A bilayer coarse-fine infiltration system minimizes bioclogging: the relevance of depth-dynamics

Bioclogging is a main concern in infiltration systems as it may significantly shorten the service life of these low-technology water treatment methods. In porous media, biofilms grow to clog partially or totally the pore network. Dynamics of biofilm accumulation (e.g., by attachment, detachment, advective transport in depth) and their impact on both surface and deep bioclogging are still not yet fully understood. To address this concern, a 104 day-long outdoor infiltration experiment in sand tanks was performed, using secondary treated wastewater and two grain size distributions (GSDs): a monolayer system filled with fine sand, and a bilayer one composed by a layer of coarse sand placed on top of a layer of fine sand. Biofilm dynamics as a function of GSD and depth were studied through cross-correlations and multivariate statistical analyses using different parameters from biofilm biomass and activity indices, plus hydraulic parameters measured at different depths. Bioclogging (both surface and deep) was found more significant in the monolayer fine system than in the bilayer coarse-fine one, possibly due to an early low-cohesive biofilm formation in the former, driven by lower porosity and lower fluxes; under such conditions biomass is favorably detached from the top layer, transported and accumulated in depth, so that new biomass might colonize the surface. On the other hand, in the bilayer system, fluxes are highest, and the biofilm is still in a growing phase, with low biofilm detachment capability from the top sand layer and high microbial activity in depth, resulting in low bioclogging. Overall, the bilayer coarse-fine system allows infiltrating higher volume of water per unit of surface area than the monolayer fine one, minimizing surface and deep bioclogging, and thus increasing the longevity and efficiency of infiltration systems.Peer ReviewedPostprint (author's final draft

Bilayer infiltration system combines benefits from both coarse and fine sands promoting nutrient accumulation in sediments and increasing removal rates

This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in Environmental science and technology, copyright © American Chemical Society after peer review. To access the final edited and published work see https://pubs.acs.org/doi/abs/10.1021/acs.est.8b00771.Infiltration systems are treatment technologies based on water percolation through porous media where biogeochemical processes take place. Grain size distribution (GSD) acts as a driver of these processes and their rates and influences nutrient accumulation in sediments. Coarse sands inhibit anaerobic reactions such as denitrification and could constrain nutrient accumulation in sediments due to smaller specific surface area. Alternatively, fine sands provide higher nutrient accumulation but need a larger area available to treat the same volume of water; furthermore, they are more susceptible to bioclogging. Combining both sand sizes in a bilayer system would allow infiltrating a greater volume of water and the occurrence of aerobic/anaerobic processes. We studied the performance of a bilayer coarse-fine system compared to a monolayer fine one - by triplicate - in an outdoor infiltration experiment to close the C-N-P cycles simultaneously in terms of mass balances. Our results confirm that the bilayer coarse-fine GSD promotes nutrient removal by physical adsorption and biological assimilation in sediments, and further it enhances biogeochemical process rates (2-fold higher than the monolayer system). Overall, the bilayer coarse-fine system allows treating a larger volume of water per surface unit achieving similar removal efficiencies as the fine system.Peer ReviewedPostprint (author's final draft

Interaction between physical heterogeneity and microbial processes in subsurface sediments: a laboratory-scale column experiment

This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in Environmental science and technology, copyright © American Chemical Society after peer review. To access the final edited and published work see [insert ACS Articles on Request author-directed link to Published Work, see http://pubs.acs.org/doi/abs/10.1021/acs.est.6b06506Physical heterogeneity determines interstitial fluxes in porous media. Nutrients and organic matter distribution in depth influence physicochemical and microbial processes occurring in subsurface. Columns 50 cm long were filled with sterile silica sand following five different setups combining fine and coarse sands or a mixture of both mimicking potential water treatment barriers. Water was supplied continuously to all columns during 33 days. Hydraulic conductivity, nutrients and organic matter, biofilm biomass, and activity were analyzed in order to study the effect of spatial grain size heterogeneity on physicochemical and microbial processes and their mutual interaction. Coarse sediments showed higher biomass and activity in deeper areas compared to the others; however, they resulted in incomplete denitrification, large proportion of dead bacteria in depth, and low functional diversity. Treatments with fine sediment in the upper 20 cm of the columns showed high phosphorus retention. However, low hydraulic conductivity values reported in these sediments seemed to constraint biofilm activity and biomass. On the other hand, sudden transition from coarse-to-fine grain sizes promoted a hot-spot of organic matter degradation and biomass growth at the interface. Our results reinforce the idea that grain-size disposition in subsurface sandy sediments drives the interstitial fluxes, influencing microbial processes.Peer ReviewedPostprint (author's final draft

L’adenocarcinoma dell’appendice ileo-ciecale: presentazione di un caso clinico e revisione della letteratura

L’adenocarcinoma dell’appendice ileo-ciecale è una neoplasia di rara osservazione rappresentando meno dello 0,5% di tutti i tumori dell’apparato gastrointestinale. Nella maggior parte dei casi viene diagnosticato all’esame istologico definitivo di un’appendice asportata per flogosi, talora invece rappresenta un reperto del tutto inatteso, documentato da biopsie estemporanee, in corso di intervento chirurgico eseguito per sospetta appendicite acuta o altra patologia non appendicolare. La storia naturale di tale neoplasia è fortemente condizionata dalle peculiari caratteristiche anatomiche del viscere che ne favoriscono la precoce diffusione e una notevole tendenza alla perforazione. Si associa frequentemente ad altre neoplasie primitive, sincrone o metacrone, a localizzazione colo-rettale o extraintestinale. Il trattamento chirurgico oncologicamente corretto è l’emicolectomia destra che può essere eseguita come prima procedura, nei casi in cui la neoplasia venga diagnosticata pre- o intraoperatoriamente, o come seconda procedura, due-tre settimane dopo l’appendicectomia, qualora soltanto l’esame istologico dell’appendice asportata riveli la presenza dell’adenocarcinoma. L’emicolectomia destra è il trattamento chirurgico più idoneo in tutti gli istotipi (colico, mucinoso, adenocarcinoide), in presenza di perforazione ed anche nelle neoplasie allo stadio A di Dukes. Durante l’atto operatorio è necessario effettuare un’accurata esplorazione della cavità addominale per la ricerca di neoplasie sincrone, mentre dopo l’intervento i pazienti dovranno essere sottoposti ad un follow-up regolare e prolungato nel tempo onde diagnosticare precocemente eventuali neoplasie metacrone. Riportiamo il caso di una donna di 78 anni con adenocarcinoma dell’appendice scoperto casualmente in corso di intervento chirurgico eseguito per un quadro di occlusione intestinale da sospetta neoplasia del cieco

Immune Checkpoints and Innovative Therapies in Glioblastoma

Targeting the Immune Checkpoint molecules (ICs; CTLA-4, PD-1, PD-L1/2, and others) which provide inhibitory signals to T cells, dramatically improves survival in hard-to-treat tumors. The establishment of an immunosuppressive environment prevents endogenous immune response in glioblastoma; therefore, manipulating the host immune system seems a reasonable strategy also for this tumor. In glioma patients the accumulation of CD4+/CD8+ T cells and Treg expressing high levels of CTLA-4 and PD-1, or the high expression of PD-L1 in glioma cells correlates with WHO high grade and short survival. Few clinical studies with IC inhibitors (ICis) were completed so far. Notably, the first large-scale randomized trial (NCT 02017717) that compared PD-1 blockade and anti-VEGF, did not show an OS increase in the patients treated with anti-PD-1. Several factors could have contributed to the failure of this trial and must be considered to design further clinical studies. In particular the possibility of targeting at the same time different ICs was pre-clinically tested in an animal model were inhibitors against IDO, CTLA-4 and PD-L1 were combined and showed persistent and significant antitumor effects in glioma-bearing mice. It is reasonable to hypothesize that the immunological characterization of the tumor in terms of type and level of expressed IC molecules on the tumor and TIL may be useful to design the optimal ICi combination for a given subset of tumor to overcome the immunosuppressive milieu of glioblastoma and to efficiently target a tumor with such high cellular complexity

Multitarget-Directed Gallium(III) Tris(acyl-pyrazolonate) Complexes Induce Ferroptosis in Cancer Cells via Dysregulation of Cell Redox Homeostasis and Inhibition of the Mevalonate Pathway

A series of Ga(Q(n))(3) coordination compounds have been synthesized, where HQ(n) is 1-phenyl-3-methyl-4-RC(=O)-pyrazolo-5-one. The complexes have been characterized through analytical data, NMR and IR spectroscopy, ESI mass spectrometry, elemental analysis, X-ray crystallography, and density functional theory (DFT) studies. Cytotoxic activity against a panel of human cancer cell lines was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, with interesting results in terms of both cell line selectivity and toxicity values compared with cisplatin. The mechanism of action was explored by spectrophotometric, fluorometric, chromatographic, immunometric, and cytofluorimetric assays, SPR biosensor binding studies, and cell-based experiments. Cell treatment with gallium(III) complexes promoted several cell death triggering signals (accumulation of p27, PCNA, PARP fragments, activation of the caspase cascade, and inhibition of the mevalonate pathway) and induced changes in cell redox homeostasis (decreased levels of GSH/GPX4 and NADP(H), increased reactive oxygen species (ROS) and 4-hydroxynonenal (HNE), mitochondrial damage, and increased activity of CPR and CcO), identifying ferroptosis as the mechanism responsible for cancer cell death

GEN-O-MA project: an Italian network studying clinical course and pathogenic pathways of moyamoya disease—study protocol and preliminary results

Background: GENetics of mOyaMoyA (GEN-O-MA) project is a multicenter observational study implemented in Italy aimed at creating a network of centers involved in moyamoya angiopathy (MA) care and research and at collecting a large series and bio-repository of MA patients, finally aimed at describing the disease phenotype and clinical course as well as at identifying biological or cellular markers for disease progression. The present paper resumes the most important study methodological issues and preliminary results. Methods: Nineteen centers are participating to the study. Patients with both bilateral and unilateral radiologically defined MA are included in the study. For each patient, detailed demographic and clinical as well as neuroimaging data are being collected. When available, biological samples (blood, DNA, CSF, middle cerebral artery samples) are being also collected for biological and cellular studies. Results: Ninety-eight patients (age of onset mean ± SD 35.5 ± 19.6 years; 68.4% females) have been collected so far. 65.3% of patients presented ischemic (50%) and haemorrhagic (15.3%) stroke. A higher female predominance concomitantly with a similar age of onset and clinical features to what was reported in previous studies on Western patients has been confirmed. Conclusion: An accurate and detailed clinical and neuroimaging classification represents the best strategy to provide the characterization of the disease phenotype and clinical course. The collection of a large number of biological samples will permit the identification of biological markers and genetic factors associated with the disease susceptibility in Italy