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BackgroundHuman milk is a vehicle for bioactive compounds and beneficial bacteria which promote the establishment of a healthy gut microbiome of newborns, especially of preterm infants. Pasteurized donor human milk (PDHM) is the second-best option when preterm mother’s own milk is unavailable. Since pasteurization affect the microbiological quality of donor milk, PDHM was inoculated with different preterm milk samples and then incubated, in order to evaluate the effect in terms of bacterial growth, human milk microbiome and proteolytic phenomena.MethodsIn an in-vitro study PDHM was inoculated at 10% v/v using ten preterm milk samples. Microbiological, metataxonomic and peptidomic analyses, on preterm milk samples at the baseline (T0), on PDHM and on inoculated milk (IM) samples at T0, after 2 h (T1) and 4 h (T2) of incubation at 37 °C, were conducted.ResultsIM samples at T2 showed a Total Bacterial Count not significantly different (p > 0.01) compared to preterm milk samples. At T2 lactic acid bacteria level was restored in all IM. After inoculation, metataxonomic analysis in IM samples showed that Proteobacteria remained the predominant phylum while Firmicutes moved from 3% at T1 to 9.4% at T2. Peptidomic profile of IM resembled that of PDHM, incubated for the same time, in terms of number and type of peptides.ConclusionThe study demonstrated that inoculation of PDHM with mother’s own milk could restore bacterial growth and personalize human milk microbiome in PDHM. This effect could be beneficial because of the presence of maternal probiotic bacteria which make PDHM more similar to mother’s own milk.

Monoatomic nanosheets can form 2-dimensional channels with tunable chemical properties, for ion storage and filtering applications. Here, we demonstrate transport of K+, Na+, and Li+ cations and F- and Cl- anions on the centimeter scale in graphene oxide membranes (GOMs), triggered by an electric bias. Besides ion transport, the GOM channels foster also the aggregation of the selected ions in salt crystals, whose composition is not the same as that of the pristine salt present in solution, highlighting the difference between the chemical environment in the 2D channels and in bulk solutions.

Charge injection in Organic Field Effect Transistors is investigated by monitoring in real-time the electrical conduction, while in situ growing pentacene films. The source and drain electrodes are made of gold functionalized with alkanethiol Self-Assembled Monolayers (SAMs), whose number of carbon atoms ranges from 5 to 10. Measuring the current flowing into the channel allows us to establish a correlation between the charge injection and the pentacene film growth. In particular, the thickness dependence of both the charge mobility and the charge carrier accumulation can be explained in terms of the molecular order at the interface between the pentacene films and the SAMs. Specifically, the threshold voltage, contact resistance and temperature-dependent constant exhibit parabolic trends for increasing alkanethiol chain length. Conversely, both the band mobility and the charge mobility are characterized by oscillations that follow an odd–even trend with a linear decay for increasing number of the carbon atoms. These results corroborate the theoretical interpretation, previously proposed by Stoliar et al., that the odd–even effect is ascribable to the order degree and orientation of the pentacene molecules interacting with the SAMs, while the parabolic trend depends on the chain length of the alkanethiol SAMs.

The photochemical core of every photosynthetic apparatus is the reaction center, a transmembrane enzyme that converts photons into charge-separated states across the biological membrane with an almost unitary quantum yield. We present a light-driven organic transistor architecture, which converts light into electrical current by exploiting the efficiency of this biological machinery. Proper surface tailoring enables the integration of the bacterial reaction center as photoactive element in organic transistors, allowing the transduction of its photogenerated voltage into photomodulation of the output current up to two orders of magnitude. The device architecture, termed Light-driven Electrolyte-Gated Organic Transistor (LEGOT), is the prototype of a new generation of low-power hybrid bio-optoelectronic organic devices.

The controlled release of cell differentiating agents is crucial in many aspects of regenerative medicine. Here we propose the use of hybrid calcite single crystals as micro-carriers for the controlled and localized release of retinoic acid, which is entrapped within the crystalline lattice. The release of retinoic acid occurs only in the proximity of stem cells, upon dissolution of the calcite hybrid crystals that are dispersed in the fibrin scaffold. These hybrid crystals provide a sustained dosage of the entrapped agent. The environment provided by this composite scaffold enables differentiation towards neuronal cells that form a three-dimensional neuronal network.

Objectives To detect possible severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) RNA contamination of inanimate surfaces in areas at high risk of aerosol formation by patients with coronavirus disease 2019 (COVID-19). Methods Sampling was performed in the emergency unit and the sub-intensive care ward. SARS-CoV-2 RNA was extracted from swabbed surfaces and objects and subjected to real-time RT-PCR targeting RNA-dependent RNA polymerase and E genes. Virus isolation from positive samples was attempted in vitro on Vero E6 cells. Results Twenty-six samples were collected and only two were positive for low-level SARS-CoV-2 RNA, both collected on the external surface of continuous positive airway pressure helmets. All transport media were inoculated onto susceptible cells, but none induced a cytopathic effect on day 7 of culture. Conclusions Even though daily contact with inanimate surfaces and patient fomites in contaminated areas may be a medium of infection, our data obtained in real-life conditions suggest that it might be less extensive than hitherto recognized.

Low-cost, minimally invasive sensors able to provide real-time monitoring of wound infection can enable the optimization of healthcare resources in chronic wounds management. Here, a novel printed organic electrochemical transistors (OECT) biosensor for monitoring uric acid (UA), a bacterial infection biomarker in wounds, is demonstrated in artificial wound exudate. The sensor exploits the enzymatic conversion of UA to 5-hydroxyisourate, catalyzed by Uricase entrapped in a dual-ionic-layer hydrogel membrane casted onto the gate. The sensor response is based on the catalytic oxidation of the hydrogen peroxide, generated as part of the Uricase regeneration process, at the Pt modified gate. The proposed dual membrane avoids the occurrence of nonspecific faradic reactions as, for example, the direct oxidation of UA or other electroactive molecules that would introduce a potentially false negative response. The biosensor is robust and its response is reproducible both in phosphate buffer saline and in complex solutions mimicking the wound exudate. The sensor has a high sensitivity in the range encompassing the pathological levels of UA in wounds (Funding Agencies|Swedish Foundation for Strategic Research (BioCom Lab) [RIT15-0119]; Swedish Foundation for Strategic ResearchSwedish Foundation for Strategic Research; Knut and Alice Wallenberg FoundationKnut & Alice Wallenberg Foundation

It is well known that amphiphilic cationic beta-cyclodextrins (am beta CDs) form nanovesicles able to release their cargo in aqueous solution upon applying different stimuli. In addition they can be selectively positioned onto substrates by unconventional soft lithography. This makes them a powerful tool for designing environments where different cues can be externally supplied to the cells helping to achieve good control of their fate. Lithographically controlled wetting (LCW) of am beta CD nanovesicles loaded with fluorescein isothiocyanate (FITC), am beta CD/FITC, has been used here to fabricate geometrically functionalized surfaces, thus achieving multiscale control of the cell environment. The am beta CD functionalization was strongly influenced by the surface energy of the underlying substrates that, according to their hydrophobicity, orient the am beta CD in a different way, thus "offering" different portions to the cells. The structure of the pattern was characterized both over large scales exploiting the FITC fluorescence and at the nanoscale by atomic force microscopy. Cell guidance and aCD/FITC cell internalization were demonstrated in human neuroblastoma SHSY5Y cells.

The water dispersion of the conductive polymer poly(3,4-ethylenedioxythiophene):poly(styrenesulfonic acid) (PEDOT:PSS) is one of the most used material precursors in organic electronics also thanks to its industrial production. There is a growing interest for conductive polymers that could be alternative surrogates or replace PEDOT:PSS in some applications. A recent study by our group compared electrodeposited PEDOT:Nafion vs PEDOT:PSS in the use for neural recordings. Here, we introduce an easy and reproducible synthetic protocol to prepare a water dispersion of PEDOT:Nafion. The conductivity of the pristine material is on the order of 2 S cm-1 and was improved up to ≈6 S cm-1 upon treatment with ethylene glycol. Faster ion transfer was assessed by electrochemical impedance spectroscopy (EIS), and, interestingly, an improved adhesion was observed for coatings of the new PEDOT:Nafion dispersion on glass substrates, even without the addition of the silane cross-linker needed for PEDOT:PSS. As proof of concept, we demonstrate the use of this novel water dispersion of PEDOT:Nafion in three different organic electronic device architectures, namely, an organic electrochemical transistor (OECT), a memristor, and an artificial synapse.

Objective: This study aimed to assess the role of Tocilizumab therapy (TCZ) in terms of ICU admission and mortality rate of critically ill patients with severe COVID-19 pneumonia. Design: Patients with COVID-19 pneumonia were prospectively enrolled in SMAtteo COvid19 REgistry (SMACORE). A retrospective analysis of patients treated with TCZ matched using propensity score to patients treated with Standard Of Care (SOC) was conducted. Setting: The study was conducted at IRCCS Policlinico San Matteo Hospital, Pavia, Italy, from March 14, 2020 to March 27, 2020. Participants: Patients with a confirmed diagnosis of COVID-19 hospitalized in our institution at the time of TCZ availability. Interventions: TCZ was administered to 21 patients. The first administration was 8 mg/kg (up to a maximum 800 mg per dose) of Tocilizumab intravenously, repeated after 12 h if no side effects were reported after the first dose. Main Outcomes and Measures: ICU admission and 7-day mortality rate. Secondary outcomes included clinical and laboratory data. Results: There were 112 patients evaluated (82 were male and 30 were female, with a median age of 63.55 years). Using propensity scores, the 21 patients who received TCZ were matched to 21 patients who received SOC (a combination of hydroxychloroquine, azithromycin and prophylactic dose of low weight heparin). No adverse event was detected following TCZ administration. This study found that treatment with TCZ did not significantly affect ICU admission (OR 0.11; 95% CI between 0.00 and 3.38; p = 0.22) or 7-day mortality rate (OR 0.78; 95% CI between 0.06 and 9.34; p = 0.84) when compared with SOC. Analysis of laboratory measures showed significant interactions between time and treatment regarding C-Reactive Protein (CRP), alanine aminotransferase (ALT), platelets and international normalized ratio (INR) levels. Variation in lymphocytes count was observed over time, irrespective of treatment. Conclusions: TCZ administration did not reduce ICU admission or mortality rate in a cohort of 21 patients. Additional data are needed to understand the effect(s) of TCZ in treating patients diagnosed with COVID-19.

Peptide aggregation into oligomers and fibrillar architectures is a hallmark of severe neurodegenerative pathologies, diabetes mellitus or systemic amyloidoses. The polymorphism of amyloid forms and their distribution are both effectors that potentially modulate the disease, thus it is important to understand the molecular basis of protein amyloid disorders through the interaction of the different amyloid forms with neural cells and tissues. Here we explore the effect of amyloid fibrils on the human neuroblastoma (SH-SY5Y) cell line in vitro. We control the kinetic of fibrillization of insulin at low pH and higher temperature. We use a multiscale characterization via fluorescence microscopy and multimodal scanning probe microscopy to correlate the number of cells and their morphology, with the finer details of the insulin deposits. Our results show that insulin aggregates deposited on neuroblastoma cell cultures lead to a progressive modification and decreased number of cells that correlates with the degree of fibrillization. SPM unravels that the aggregates strongly interact with the cell membrane, forming a stiff encase that possibly leads to an increased cell membrane stiffness and deficit in the metabolic exchanges between the cells and their environment. The presence of fibrils does not affect the number of cells at 24h whereas drop down to 60% is observed after 48h of incubation.

open access article Climate change threats make it difficult to perform reliable and quick predictions on floods forecasting. This gives rise to the need of having advanced methods, e.g., computational intelligence tools, to improve upon the results from flooding events simulations and, in turn, design best practices for riverbed maintenance. In this context, being able to accurately estimate the roughness coefficient, also known as Manning’s n coefficient, plays an important role when computational models are employed. In this piece of research, we propose an optimal approach for the estimation of ‘n’. First, an objective function is designed for measuring the quality of ‘candidate’ Manning’s coefficients relative to specif cross-sections of a river. Second, such function is optimised to return coefficients having the highest quality as possible. Five well-known meta-heuristic algorithms are employed to achieve this goal, these being a classic Evolution Strategy, a Differential Evolution algorithm, the popular Covariance Matrix Adaptation Evolution Strategy, a classic Particle Swarm Optimisation and a Bayesian Optimisation framework. We report results on two real-world case studies based on the Italian rivers ‘Paglia’ and ‘Aniene’. A comparative analysis between the employed optimisation algorithms is performed and discussed both empirically and statistically. From the hydrodynamic point of view, the experimental results are satisfactory and produced within significantly less computational time in comparison to classic methods. This shows the suitability of the proposed approach for optimal estimation of the roughness coefficient and, in turn, for designing optimised hydrological models.

none 3 no Electric field; Density functional theory; organic field effect transistors; dimerization process Density functional theory (DFT) calculations were used to assess the effect of electric fields of varying magnitudes and directions on the molecular structure of pentacene and other acenes. The aim is to understand the response of acenes in organic field effect transistors, specifically the structure of the first monolayer(s) deposited on the gate dielectric, where the transversal electric field and the charge carrier density are largest and charge transport occurs. Pentacene cycloaddition can be enhanced by the application of electric fields oriented along the direction of the forming bonds. Dimerization is likely to occur in low-density, disordered domains, such as grain boundaries or terrace edges. Together with other factors, dimerization could affect device performance leading to an irreversible decrease of mobility due to the creation of new trap states. Matta, Micaela; Biscarini, Fabio; Zerbetto, Francesco Matta, Micaela; Biscarini, Fabio; Zerbetto, Francesco

Microelectrode arrays are used for recording and stimulation in neurosciences both in vitro and in vivo. The electrodeposition of conductive polymers, such as poly(3,4-ethylene dioxythiophene) (PEDOT), is widely adopted to improve both the in vivo recording and the charge injection limit of metallic microelectrodes. The workhorse of conductive polymers in the neurosciences is PEDOT:PSS, where PSS represents polystyrene-sulfonate. In this paper, the counterion is the fluorinated polymer Nafion, so the composite PEDOT:Nafion is deposited onto a flexible neural microelectrode array. PEDOT:Nafion coated electrodes exhibit comparable in vivo recording capability to the reference PEDOT:PSS, providing a large signal-to-noise ratio in a murine animal model. Importantly, PEDOT:Nafion exhibits a minimized polarization during electrical stimulation, thereby resulting in an improved charge injection limit equal to 4.4 mC cm-2 , almost 80% larger than the 2.5 mC cm-2 that is observed for PEDOT:PSS.

A multilayer self-assembled monolayer (MuL-SAM) composed by i) cysteamine, ii) 4-formilphenyl boronic acid and iii) dopamine has been used to functionalize polycrystalline Au. Its electrochemical characterization shows how this surface engineering makes Au reactive toward dopamine. Kelvin probe measurements have shown how MuL-SAM tunes the surface potential of the bare Au. Such measurements are correlated with the electrical performances of organic field-effect transistors (OFETs), whose source/drain electrodes have been functionalized by MuL-SAM. Such devices, operated as charge-injection organic gauges (CIOGs), have shown sensitivity to the concentration of dopamine in the range of [1, 50] nM.

Optically active (&minus, )589ethyl (S)-2-phenylbutyl thioether, (&minus, )(S)C-Et(PhBu)S (I), and its new diastereoisomeric mercury (II) chloride adduct, 1:2, (&minus, )[(S)S(S)C-Et(PhBu)S.(HgCl2)2]2, (II) were stereoselectively synthesized, the absorbance (UV) and circular dichroism (CD) spectra were measured and the crystal and molecular structure of complex (II) was determined by single-crystal X-ray diffraction. Two different Hg centres are present whose coordination environments are built by two short bonds to chloride ligands in one case, and to one chloride and one sulphur in the other one. These originate digonal units. Electroneutrality is achieved by a further chlorine, which can be considered prevalently ionic and bonded to the two Hg centres, forming square bridging systems nearly perpendicular to the digonal molecules. The coordination polyhedra can be interpreted as 2 + 4 tetragonally-compressed octahedra with the four longer contacts lying in the equatorial plane. IR spectroscopic data are consistent with the presence of one bent and one linear Cl&ndash, Hg&ndash, Cl moiety. The absolute configurations at both stereogenic centres of the formed diastereoisomeric complex (II) are (S). The (S)S absolute configuration at the stereogenic sulphur atom bonded to the mercury(II) atom in complex (II) has been related with the negative Cotton effect assigned in its circular dichroism (CD) spectrum to a charge-transfer transition at ca. 230 nm. The stereoselective oxidation of (I) and (II) with hydrogen peroxide, induced by the stereogenic carbon atom (S)C of the enantiopure sulphide, gave (&minus, )598ethyl (S)C-2-phenylbutyl(S)S-sulphoxide, (&minus, )598[(S)S(S)C-Et(PhBu)SO], (III), having 18.1% de. Oxidations carried out in the presence of a 200 molar excess of mercury(II) chloride gave (&minus, )598ethyl (S)C-2-phenylbutyl(R)S-sulphoxide, (&minus, ) 598[(R)S(S)C-Et(PhBu)SO], (IV) with 31% de, showing the cooperative influence of mercury(II) chloride on the selectivity of the oxidation reaction.

Palm oil is widely used in the food industry for its chemical/physical properties, low cost and wide availability. Its widespread use has provoked an intense debate about whether it is a potential danger to human health. In a careful review of the scientific literature, we focused on nutritional characteristics and health effects of the use of palm oil with regards to children, seeking to determine whether there is evidence that justifies fears about the health effects of palm oil. Our review showed that palm oil represents a significant source of saturated fatty acids, to which scientific evidence attributes negative health effects when used in excess, especially with regards to cardiovascular diseases. However, to date, there is no evidence about the harmful effects of palm oil on the health of children. Nevertheless, palm oil has possible ill health effects linked to its composition of fatty acids: its consumption is not correlated to risk factors for cardiovascular diseases in young people with a normal weight and cholesterol level; the elderly and patients with dyslipidaemia or previous cardiovascular events or hypertension are at a greater risk. Therefore, the matter is not palm oil itself but the fatty-acid-rich food group to which it belongs. The most important thing is to consume no more than 10% of saturated fatty acids, regardless of their origin and regardless of one’s age. Correct information based on a careful analysis of the scientific evidence, rather than a focus on a singular presumed culprit substance, should encourage better lifestyles.

The quantification of urea is of the utmost importance not only in medical diagnosis, where it serves as a potential indicator of kidney and liver disfunction, but also in food safety and environme ...

In the formation reaction of Ni(2+) with the chiral racemic ligand, (R)(R)bdtp(-)/(S)(S)bdtp(-), bdtp(-) = [SSPOCH)CH(3))CH(CH(3))O](-), cyclo- O,O'-[1,2-dimethylethylene] dithiophosphato ion, the meso-complex Ni[(R)(R)(lambda)bdtp][(S)(S)(delta)-bdtp] is stereoselectively produced. The meso-complex was compared with the enantiopure crystals of (+)(589)Ni[(R)(R)(lambda)bdtp](2) or (-)(589)Ni[(S)(S)(delta)bdtp](2), as well as racemic crystals, rac-(+/-)Ni[bdtp](2), which were prepared from the solution containing the two enantiomers in a 1:1 ratio. Dissociation constants in solutions indicate different stability of the meso and enantiopure complexes depending on the solvent, whereas a more efficient crystal packing, weak H-bonding, and nonbonding interactions contribute to stabilization of the meso-species over the racemic one. Molecular structures show that the outer five-membered ligand ring adopts the half-chair conformation C(2) with either the lambda or the delta chirality and the methyl groups are in equatorial (e) positions. Enantiopure ligands of (+)(589)Ni[(R)(R)(lambda)bdtp](2) and (-)(589)Ni[(S)(S)(delta)bdtp](2) induce chirality into the symmetric SSNiSS chromophore with slightly helical distortion. Thus, their CD spectra exhibit weak negative or positive Cotton effects at 662 nm. CD spectra in L(+)- and D(-)diethyltartrate of the meso-complex and racemic crystal, rac-(+/-)Ni[bdtp](2), exhibit different weak Cotton effects of opposite sign. Complexes dissociate in methanol; rac-(+/-)Ni[bdtp](2) in methanol undergoes a crystallization-induced second-order asymmetric transformation which finally yields crystals of the meso-Ni[(R)(R)(lambda)bdtp][(S)(S)(delta)bdtp] complex.

The amorphous aggregation of A?1-40 peptide is addressed by using micromolding in capillaries. Both the morphology and the size of the aggregates are modulated by changing the contact angle of the sub-micrometric channel walls. Upon decreasing the hydrophilicity of the channels, the aggregates change their morphology from small aligned drops to discontinuous lines, thereby keeping their amorphous structure. A?1-40 fibrils are observed at high contact angles.

We describe a potentiometric sensor based on Electrolyte-Gated Organic Field-Effect Transistor (EGOFET) for "in vitro'' detection of dopamine. The sensing element of this device resides at the Au gate-aqueous solution interface by means of a self-assembled monolayer (SAM) composed by cysteamine and 4-formylphenyl boronic acid. The covalent and selective adsorption of dopamine induces a surface dipole potential which shifts the electrode work function and modulates the double layer capacitance. As a result, our device is capable to detect dopamine up to pico-molar concentration showing higher sensitivity with respect to other approaches. For this reason the interface engineering of our EGOFET gate is a promising route for diagnostic applications. (c) 2012 Elsevier B.V. All rights reserved.

none 14 si Biorecognition is a central event in biological processes in the living systems that is also widely exploited in technological and health applications. We demonstrate that the Electrolyte Gated Organic Field Effect Transistor (EGOFET) is an ultrasensitive and specific device that allows us to quantitatively assess the thermodynamics of biomolecular recognition between a human antibody and its antigen, namely, the inflammatory cytokine TNFα at the solid/liquid interface. The EGOFET biosensor exhibits a superexponential response at TNFα concentration below 1 nM with a minimum detection level of 100 pM. The sensitivity of the device depends on the analyte concentration, reaching a maximum in the range of clinically relevant TNFα concentrations when the EGOFET is operated in the subthreshold regime. At concentrations greater than 1 nM the response scales linearly with the concentration. The sensitivity and the dynamic range are both modulated by the gate voltage. These results are explained by establishing the correlation between the sensitivity and the density of states (DOS) of the organic semiconductor. Then, the superexponential response arises from the energy-dependence of the tail of the DOS of the HOMO level. From the gate voltage-dependent response, we extract the binding constant, as well as the changes of the surface charge and the effective capacitance accompanying biorecognition at the electrode surface. Finally, we demonstrate the detection of TNFα in human-plasma derived samples as an example for point-of-care application. Berto, Marcello; Casalini, Stefano; Di Lauro, Michele; Marasso, Simone L; Cocuzza, Matteo; Perrone, Denis; Pinti, Marcello; Cossarizza, Andrea; Pirri, Candido F; Simon, Daniel T; Berggren, Magnus; Zerbetto, Francesco; Bortolotti, Carlo A; Biscarini, Fabio Berto, Marcello; Casalini, Stefano; Di Lauro, Michele; Marasso, Simone L; Cocuzza, Matteo; Perrone, Denis; Pinti, Marcello; Cossarizza, Andrea; Pirri, Candido F; Simon, Daniel T; Berggren, Magnus; Zerbetto, Francesco; Bortolotti, Carlo A; Biscarini, Fabio

Background: Patient stratification to match individual patients with the most effective drug treatment is still a major open challenge in cancer care. For instance, cytarabine is the main drug used for AML treatment but 30% of patients fail to respond to this agent. Laboratory developed tests determining ex-vivo cellular response to cytotoxic anticancer drugs have demonstrated good correlations with clinical response, sometimes surpassing the predictive power of molecular and genetic profiling. Standardizing sample processing to remove operator-dependent biases and maintaining live cells in a functional status that closely resembles in-vivo function are major challenges affecting these tests. Here we present the open microwell (OMW) platform, a microfluidic-based system that integrates the entire process of ex-vivo testing of anticancer drug efficacy and enables drug testing in the clinical setting prior to therapy administration. The concept was validated for the first time on 13 AML patients at Sant'Orsola hospital, Italy, showing the possibility to initiate the analysis readily after sample collection, thus minimizing drifts in cell function that typically start occurring within hours from sampling, and provide results in about 24 hours, with a fully-automated system. Methods: 13 refractory, relapsed or newly diagnosed AML patients were enrolled in the study. 2 ml of fresh bone marrow in EDTA and 1 ml of serum blood were collected from each patient. White blood cells (WBC) were separated from bone marrow by standard Ficoll-Paque, suspended in medium additioned with 2% autologous serum and loaded in microfluidic chips featuring 16 microchannels and 1200 microwells/channel (70 μm diameter), open at the bottom end (Fig. 1A-B). After settling down in microwells, cells were stained with CMAC cell tracker, anti-human-CD34/CD45 fluorescently-labeled antibodies and Propidium Iodide (PI) by injecting the reagents in the microchannels. Cytarabine or combination therapies (FLAI-3, FLAI-5, FLA, MEC-4) were then injected in the microchannels and cells were incubated for 24 hours in the system (Fig. 1C). Four channels were used per therapeutic condition, including reference (ref), high (ref x 10) and low (ref/10) dosages plus a non-treated channel as reaction control. Finally, a custom software was used to detect cells in images, classify AML blasts and analyze cell death (Fig. 1D). Drug efficacy was determined by evaluation of both cell depletion and apoptosis induction. Results: We first validated the OMW platform against gold standard (FacsAria flow cytometer) for the detection of tumor cells and measurement of cell viability and apoptosis. Count of blast frequency was carried out on 5 patient samples using anti-CD34/CD45 staining. Results (Fig. 2B) show a correlation between the gold standard and OMW (n=5, R2=0.99, p Conclusion: The OMW platform was able to count AML blasts and analyze viability and apoptosis showing high concordance with conventional diagnostics represented by flow cytometry. The assay requires 30 μl of bone marrow sample and simple manual pre-processing. Profiling of patient response to specific drugs or combination is provided within 24h. These features make the OMW platform suitable for bedside analysis, to evaluate drug activity on tumor cells within a heterogeneous sample and promptly guide personalized treatment in hematologic malignancies, with a first proof achieved on AML patients. LRo and AF equally contributed. Disclosures Rocchi: CellPly S.r.l.: Employment. Faenza:CellPly S.r.l.: Employment. Rambelli:CellPly S.r.l.: Employment. Guadagnuolo:CellPly S.r.l.: Employment. Pecorari:CellPly S.r.l.: Employment. Giulianelli:CellPly S.r.l.: Employment. Biscarini:CellPly S.r.l.: Employment. Martinelli:Novartis: Speakers Bureau; Roche: Consultancy, Speakers Bureau; Ariad: Consultancy, Speakers Bureau; BMS: Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; Amgen: Consultancy, Speakers Bureau; MSD: Consultancy; Celgene: Consultancy, Speakers Bureau; Genentech: Consultancy. Guerrieri:CellPly S.r.l.: Equity Ownership. Bocchi:CellPly S.r.l.: Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

Electrolyte-gated organic field-effect transistors (EGOFETs), based on ultrathin pentacene films on quartz, were operated with electrolyte solutions whose pH was systematically changed. Transistor parameters exhibit nonmonotonic variation versus pH, which cannot be accounted for by capacitive coupling through the Debye–Helmholtz layer. The data were fitted with an analytical model of the accumulated charge in the EGOFET, where Langmuir adsorption was introduced to describe the pH-dependent charge buildup at the quartz surface. The model provides an excellent fit to the threshold voltage and transfer characteristics as a function of the pH, which demonstrates that quartz acts as a second gate controlled by pH and is mostly effective from neutral to alkaline pH. The effective capacitance of the device is always greater than the capacitance of the electrolyte, thus highlighting the role of the substrate as an important active element for amplification of the transistor response.

A novel fully organic bioelectronic device is presented and validated as electronic transducer and current stimulator for brain implants. The device integrates polymeric electrodes made of poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) on paper thin foils, resulting in a high surface-to-volume ratio architecture that exhibits high sensitivity to interfacial ionic transport phenomena. The prototyping technique herein presented yields devices for the bidirectional communication with biological systems whose dimensionality can be controlled according to the desired application. Transduction of ultra-low local-field potentials and delivery of voltage pulse-trains alike those used in deep-brain stimulation are herein assessed, paving the way towards novel theranostic strategies for the treatment of Parkinson’s Disease and other severe neurodegenerative and/or traumatic pathologies of the central nervous system.

Cytokines are small proteins that play fundamental roles in inflammatory processes in the human body. In particular, interleukin (IL)-6 is a multifunctional cytokine, whose increased levels are associated with infection, cancer, and inflammation. The quantification of IL-6 is therefore of primary importance in early stages of inflammation and in chronic diseases, but standard techniques are expensive, time-consuming, and usually rely on fluorescent or radioactive labels. Organic electronic devices and, in particular, organic field-effect transistors (OFETs) have been proposed in the recent years as novel platforms for label-free protein detection, exploiting as sensing unit surface-immobilized antibodies or aptamers. Here, the authors report two electrolyte-gated OFETs biosensors for IL-6 detection, featuring monoclonal antibodies and peptide aptamers adsorbed at the gate. Both strategies yield biosensors that can work on a wide range of IL-6 concentrations and exhibit a remarkable limit of detection of 1 pM. Eventually, electrolyte gated OFETs responses have been used to extract and compare the binding thermodynamics between the sensing moiety, immobilized at the gate electrode, and IL-6. (C) 2017 American Vacuum Society. Funding Agencies|IT MIUR Bilateral Project Italy/Sweden "Poincare"; joint INFM-CNR Project; Life Science Department; "Fondazione di Vignola"

Scanning tunnelling microscopy (STM) studies have found stripe-like domains on gold nanoparticles (NPs) coated with certain binary mixtures of ligand molecules. The majority of these NPs' properties have been investigated for particles in solvents. Yet, most STM studies are for NPs in a dry state. Images of the same particles in air and liquid have not been obtained yet. In this work, a judicious choice of ligand molecules led to NPs with close-to-ideal STM imaging conditions in air and in 1-phenyloctane (PO). Large datasets under both conditions were acquired and rapidly evaluated through power spectral density (PSD) analysis. The result is a quantitative comparison of stripe-like domains in air and PO on the same NPs. PSD analysis determines a characteristic length-scale for these domains of similar to 1.0 nm in air and in PO showing persistence of striped domains in these two media. A length scale of similar to 0.7 nm for homoligand NPs was found.

Extracellular electrode recording demonstrates acid-triggered electrical activity in glioma cell populations., Glioma patients often suffer from epileptic seizures because of the tumor’s impact on the brain physiology. Using the rat glioma cell line C6 as a model system, we performed long-term live recordings of the electrical activity of glioma populations in an ultrasensitive detection method. The transducer exploits large-area electrodes that maximize double-layer capacitance, thus increasing the sensitivity. This strategy allowed us to record glioma electrical activity. We show that although glioma cells are nonelectrogenic, they display a remarkable electrical burst activity in time. The low-frequency current noise after cell adhesion is dominated by the flow of Na+ ions through voltage-gated ion channels. However, after an incubation period of many hours, the current noise markedly increased. This electric bursting phenomenon was not associated with apoptosis because the cells were viable and proliferative during the period of increased electric activity. We detected a rapid cell culture medium acidification accompanying this event. By using specific inhibitors, we showed that the electrical bursting activity was prompted by extracellular pH changes, which enhanced Na+ ion flux through the psalmotoxin 1–sensitive acid-sensing ion channels. Our model of pH-triggered bursting was unambiguously supported by deliberate, external acidification of the cell culture medium. This unexpected, acidosis-driven electrical activity is likely to directly perturb, in vivo, the functionality of the healthy neuronal network in the vicinity of the tumor bulk and may contribute to seizures in glioma patients.

The interactions of a biodegradable scaffold with cells or living tissues depend on the time-evolution of the nanoscale properties of the scaffold. We present an in situ quantitative study on the early-stage swelling and degradation of poly(lactic-co-glycolic acid) (PLGA). A novel metrology scheme based on force microscopy measurements of the patterns of PLGA nanostructures is developed to characterize the evolution of topography, volume and nanomechanical properties. The volume and nanoscale roughness show an oscillating behaviour during the first eight days of immersion; at a later stage, we observe a continuous decrease of the volume. The effective Young's modulus exhibits a monotonic decrease from an initial value of about 2.4 GPa down to 9 MPa at day 14. The oscillating behaviour of the volume before the onset of full degradation is explained by a coupled diffusion-swelling mechanism. The appearance of a second maximum in the volume evolution results from the competition between swelling and degradation., This work was founded by the European Union Seventh Framework Programme [FP7/2007–2013] under the grant agreement no. 280772, project iONE-FP7 and MINECO (Spain), project CSD2010-00024.

© 2015 American Chemical Society. Thin films of organic semiconductors have been widely studied at different length scales for improving the electrical response of devices based on them. Hitherto, a lot of knowledge has been gained about how molecular packing, morphology, grain boundaries, and defects affect the charge transport in organic thin film transistors. However, little is known about the impact of an electric field on the organic semiconductor microstructure and the consequent effect on the device performances. To fill this gap, we investigated the evolution of the structure of pentacene thin film transistors during device operation by in situ real time X-ray diffraction measurements and theoretical calculations. We observed for the first time the occurrence of a reversible structural strain taking place during the bias application mainly due to reorientation at the terrace edges of monolayer islands under the effect of electrical field. Strain exhibits the same trend of the threshold voltage hinting to the existence of a direct correlation between the phenomenon of bias stress and the structural modification., The authors acknowledge A. Shehu for his collaboration at the early stage of the project, F. Borgatti for the fruitful discussion, J. R. Plaisier, G. Zerauschek and A. Lausi for optimizing the experimental setup at the MCX-ELETTRA beamline, and V. L.R. Jacques and P. Evans for technical support at the ID01-ESRF beamline. Financial support for this research was by the National Project N−CHEM, Flagship NANOMAX, by the DGI (Spain) with Project BE-WELL CTQ2013-40480-R, Generalitat de Catalunya (2014-SGR-17), the ERC StG 2012-306826, and by the Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBERBBN).

The squid pen (gladius) from the Loligo vulgaris was used for preparation of β-chitin materials characterized by different chemical, micro- and nano-structural properties that preserved, almost completely the macrostructural and the mechanical ones. The β-chitin materials obtained by alkaline treatment showed porosity, wettability and swelling that are a function of the duration of the treatment. Microscopic, spectroscopic and synchrotron X-ray diffraction techniques showed that the chemical environment of the N-acetyl groups of the β-chitin chains changes after the thermal alkaline treatment. As a consequence, the crystalline packing of the β-chitin is modified, due to the intercalation of water molecules between β-chitin sheets. Potential applications of these β-chitin materials range from the nanotechnology to the regenerative medicine. The use of gladii, which are waste products of the fishing industry, has also important environmental implications.

Four thiolated oligoarylene molecules (i) 4-methoxy-terphenyl-4 ''-methanethiol (MTM), (ii) 4-methoxy-terphenyl-3 '',5 ''-dimethanethiol (MTD), (iii) 4-nitro-terphenyl-4 ''-methanethiol (NTM), and (iv) 4-nitro-terphenyl-3 '',5 ''-dimethanethiol (NTD) were synthesized and self-assembled as monolayers (SAMs) on polycrystalline Au electrodes of organic field-effect transistors (OFETs). SAMs were characterized by contact angle and AC/DC electrochemical measurements, whereas atomic force microscopy was used for imaging the pentacene films grown on the coated electrodes. The electrical properties of functionalized OFETs, the electrochemical SAMs features and the morphology of pentacene films were correlated to the molecular organization of the thiolated oligoarylenes on Au, as calculated by means of the density functional theory. This multi-methodological approach allows us to associate the systematic replacement of the SAM anchoring head group (viz. methanethiol and dimethanethiol) and/or terminal tail group (viz. nitro-, -NO2, and methoxy, -OCH3) with the change of the electrical features. The dimethanethiol head group endows SAMs with higher resistive features along with higher surface tensions compared with methanethiol. Furthermore, the different number of thiolated heads affects the kinetics of Au passivation as well as the pentacene morphology. On the other hand, the nitro group confers further distinctive properties, such as the positive shift of both threshold and critical voltages of OFETs with respect to the methoxy one. The latter experimental evidence arise from its electron-withdrawing capability, which has been verified by both DFT calculations and DC electrochemical measurements

A series of hydrophilic alkanethiols, HS(CH2)(n)OH with n = 3, 4, 6, 8, 9 and 11, have been self-assembled on gold electrodes of pentacene-based thin-film transistors. The multiparametric and ultra-sensitive response of these devices allows us to characterize both charge-injection and electrical stability moving from vacuum to air. The decay exponent beta of charge injection is found to be 0.7-1.1 angstrom(-1) in agreement with earlier measurements of charge tunneling done by electrochemistry and large-area molecular junctions. We find that the intermediate chain lengths (n = 4 and 6) yield an optimum response as they represent the best compromise in terms of decrease of the contact-resistance and bias stress.

This study is an investigation of the effect of biocompatible glutathione-covered gold nanoparticles (AuSG_7) with an average size of 3 nm on the amyloid fibrils of hen egg-white lysozyme. The anti-amyloid activity of AuSG_7 nanoparticles on this protein was monitored with thioflavin T assay, atomic force microscopy and transmission electron microscopy. The study found that AuSG_7 nanoparticles in vitro depolymerize the amyloid aggregates and inhibit lysozyme aggregate formation. The ability to inhibit amyloid formation and promote amyloid disassembly has concentration-dependent characteristics: the concentration of nanoparticles at which inhibition is half maximal (IC 50 ) was found to be 6.19 μg/mL, and the concentration at which depolymerization is half maximal (DC 50 ) was found to be 8.26 μg/mL.

A whole organic artificial synapse has been fabricated by patterning PEDOT:PSS electrodes on PDMS that are biased in frequency to yield a STP response. The timescale of the STP response is shown to be sensitive to the concentration of dopamine, DA, a neurotransmitter relevant for monitoring the development of Parkinson's disease and potential locoregional therapies. The sensitivity of the sensor towards DA has been validated comparing signal variation in the presence of DA and its principal interfering agent, ascorbic acid, AA. The whole organic synapse is biocompatible, soft and flexible, and is attractive for implantable devices aimed to real-time monitoring of DA concentration in bodily fluids. This may open applications in chronic neurodegenerative diseases such as Parkinson's disease. EU 7th Framework Programme [FP7/2007–2013]; Grant Agreement No. 280772; IT MIUR Bilateral Project IT/SE “Poincaré”