Nanocellulose-Bovine Serum Albumin Interactions in an Aqueous Medium: Investigations Using In Situ Nanocolloidal Probe Microscopy and Reactive Molecular Dynamics Simulations.

As a versatile nanomaterial derived from renewable sources, nanocellulose has attracted considerable attention for its potential applications in various sectors, especially those focused on water treatment and remediation. Here, we have combined atomic force microscopy (AFM) and reactive molecular dynamics (RMD) simulations to characterize the interactions between cellulose nanofibers modified with carboxylate or phosphate groups and the protein foulant model bovine serum albumin (BSA) at pH 3.92, which is close to the isoelectric point of BSA. Colloidal probes were prepared by modification of the AFM probes with the nanofibers, and the nanofiber coating on the AFM tip was for the first time confirmed through fluorescence labeling and confocal optical sectioning. We have found that the wet-state normalized adhesion force is approximately 17.87 ± 8.58 pN/nm for the carboxylated cellulose nanofibers (TOCNF) and about 11.70 ± 2.97 pN/nm for the phosphorylated ones (PCNF) at the studied pH. Moreover, the adsorbed protein partially unfolded at the cellulose interface due to the secondary structure's loss of intramolecular hydrogen bonds. We demonstrate that nanocellulose colloidal probes can be used as a sensitive tool to reveal interactions with BSA at nano and molecular scales and under in situ conditions. RMD simulations helped to gain a molecular- and atomistic-level understanding of the differences between these findings. In the case of PCNF, partially solvated metal ions, preferentially bound to the phosphates, reduced the direct protein-cellulose connections. This understanding can lead to significant advancements in the development of cellulose-based antifouling surfaces and provide crucial insights for expanding the pH range of use and suggesting appropriate recalibrations.

Engineered Multilayer Microcapsules Based on Polysaccharides Nanomaterials.

The preparation of microcapsules composed by natural materials have received great attention, as they represent promising systems for the fabrication of micro-containers for controlled loading and release of active compounds, and for other applications. Using polysaccharides as the main materials is receiving increasing interest, as they constitute the main components of the plant cell wall, which represent an ideal platform to mimic for creating biocompatible systems with specific responsive properties. Several researchers have recently described methods for the preparation of microcapsules with various sizes and properties using cell wall polysaccharide nanomaterials. Researchers have focused mostly in using cellulose nanomaterials as structural components in a bio-mimetic approach, as cellulose constitutes the main structural component of the plant cell wall. In this review, we describe the microcapsules systems presented in the literature, focusing on the works where polysaccharide nanomaterials were used as the main structural components. We present the methods and the principles behind the preparation of these systems, and the interactions involved in stabilizing the structures. We show the specific and stimuli-responsive properties of the reported microcapsules, and we describe how these characteristics can be exploited for specific applications.

Anisotropic Diffusion and Phase Behavior of Cellulose Nanocrystal Suspensions.

In this paper, we use dynamic light scattering in polarized and depolarized modes to determine the translational and rotational diffusion coefficients of concentrated rodlike cellulose nanocrystals in aqueous suspension. Within the range of studied concentrations (1-5 wt %), the suspension starts a phase transition from an isotropic to an anisotropic state as shown by polarized light microscopy and viscosity measurements. Small-angle neutron scattering measurements also confirmed the start of cellulose nanocrystal alignment and a decreasing distance between the cellulose nanocrystals with increasing concentration. As expected, rotational and translational diffusion coefficients generally decreased with increasing concentration. However, the translational parallel diffusion coefficient was found to show a local maximum at the onset of the isotropic-to-nematic phase transition. This is attributed to the increased available space for rods to move along their longitudinal axis upon alignment. This increased parallel diffusion coefficient thus confirms the general idea that rodlike particles gain translational entropy upon alignment while paying the price for losing rotational degrees of freedom. Once the concentration increases further, diffusion becomes more hindered even in the aligned regions due to a reduction in the rod separation distance. This leads once again to a decrease in translational diffusion coefficients. Furthermore, the relaxation rate for fast mode translational diffusion (parallel to the long particle axis) exhibited two regimes of relaxation behavior at concentrations where significant alignment of the rods is measured. We attribute this unusual dispersive behavior to two length scales: one linked to the particle length (at large wavevector q) and the other to a twist fluctuation correlation length (at low wavevector q) along the cellulose nanocrystal rods that is of a larger length when compared to the actual length of rods and could be linked to the size of aligned domains.

Thermodynamic Study of Ion-Driven Aggregation of Cellulose Nanocrystals.

The thermodynamics of interactions between cations of the second group of the periodic table and differently negatively charged cellulose nanocrystals was investigated using isothermal titration calorimetry (ITC). The interaction of cations with the negatively charged CNCs was found to be endothermic and driven by an increase in entropy upon adsorption of the ions, due to an increase in degrees of freedom gained by the surface bound water upon ion adsorption. The effect was pH-dependent, showing an increase in enthalpy for cellulose suspensions at near-neutral pH (6.5) when compared to acidic pH (2). Sulfated cellulose nanoparticles were found to readily interact with divalent ions at both pH levels. The adsorption on carboxylate nanocrystals was found to be pH dependent, showing that the carboxylic group needs to be in the deprotonated form to interact with divalent ions. For the combined system (sulfate and carboxylate present at the same time), at neutral pH, the adsorption enthalpy was higher than the value obtained from cellulose nanocrystals containing a single functional group, while the association constant was higher due to an increased favorable entropic contribution. The higher entropic contribution indicates a more restricted surface-bound water layer when multiple functionalities are present. The stoichiometric number n was nearly constant for all systems, showing that the adsorption depends almost completely on the ion valency and on the amount of ionic groups on the CNC surface, independent of the type of functional group on the CNC surface as long as it is deprotonated. In addition, we showed that the reduction in Gibbs free energy drives the ionotropic gelation of nanocellulose suspensions, and we show that ITC is able to detect gel formation at the same time as determining the critical association concentration.

Toward Improved Understanding of the Interactions between Poorly Soluble Drugs and Cellulose Nanofibers.

Cellulose nanofibers (CNFs) have interesting physicochemical and colloidal properties that have been recently exploited in novel drug-delivery systems for tailored release of poorly soluble drugs. The morphology and release kinetics of such drug-delivery systems heavily relied on the drug-CNF interactions; however, in-depth understanding of the interactions was lacking. Herein, the interactions between a poorly soluble model drug molecule, furosemide, and cationic cellulose nanofibers with two different degrees of substitution are studied by sorption experiments, Fourier transform infrared spectroscopy, and molecular dynamics (MD) simulation. Both MD simulations and experimental results confirmed the spontaneous sorption of drug onto CNF. Simulations further showed that adsorption occurred by the flat aryl ring of furosemide. The spontaneous sorption was commensurate with large entropy gains as a result of release of surface-bound water. Association between furosemide molecules furthermore enabled surface precipitation as indicated by both simulations and experiments. Finally, sorption was also found not to be driven by charge neutralization, between positive CNF surface charges and the furosemide negative charge, so that surface area is the single most important parameter determining the amount of sorbed drug. An optimized CNF-furosemide drug-delivery vehicle thus needs to have a maximized specific surface area irrespective of the surface charge with which it is achieved. The findings also provide important insights into the design principles of CNF-based filters suitable for removal of poorly soluble drugs from wastewater.

Effect of Gelation on the Colloidal Deposition of Cellulose Nanocrystal Films.

One of the most important aspects in controlling colloidal deposition is manipulating the homogeneity of the deposit by avoiding the coffee-ring effect caused by capillary flow inside the droplet during drying. After our previous work where we achieved homogeneous deposition of cellulose nanocrystals (CNCs) from a colloidal suspension by reinforcing Marangoni flow over the internal capillary flow (Gençer et al. Langmuir 2017, 33 (1), 228-234), we now set out to reduce the importance of capillary flow inside a drying droplet by inducing gelation. In this paper, we discuss the effect of gelation on the deposition pattern and on the self-assembly of CNCs during droplet drying. CNC films were obtained by drop casting CNC suspensions containing NaCl and CaCl2 salts. A mixed methodology using rheological and depolarized dynamic light scattering was applied to understand the colloidal behavior of the CNCs. In addition, analysis of the mixture's surface tension, viscosity, and yield stress of the suspensions were used to gain deeper insights into the deposition process. Finally, the understanding of the gelation behavior in the drying droplet was used to exert control over the deposit where the coffee-ring deposit can be converted to a dome-shaped deposit.

Thermodynamics of the interactions of positively charged cellulose nanocrystals with molecules bearing different amounts of carboxylate anions.

Negatively charged ions readily interact with the surface of positively charged pyridinium-grafted cellulose nanocrystals. In this work we investigated the thermodynamics of these interactions using isothermal titration calorimetry. We investigated the effect of the ionic charge, using carboxylate salts with different valence (1-4), and compared it with sodium sulfate as another delocalized ionic charge. Experiments performed using cellulose nanocrystals with three different degrees of substitution of pyridinium grafts showed that the number of ions adsorbed onto the surface of cellulose nanocrystals is directly linked to the number of grafted cationic functionalities. The adsorption of anions onto pyridinium-grafted nanocrystals was further found to be endothermic and driven by an increase in entropy upon adsorption of the anions due to the release of surface bound water. The association constant increased with the net charge of the anions from a low value for monovalent sodium acetate. Both entropy and enthalpy increased linearly with the net charge of the anions, demonstrating that a stronger depolarization occurred when anions of higher net charge were adsorbed. The values of the stoichiometric number determined were lower than the values relative to processes involving ions in solution, suggesting a more complex mode of interaction, involving hydrogen-bonding and bridge formation. However, the number of adsorbed ions was directly correlated with the amount of cationic surface grafts. This makes it possible to control the amount of surface interactions directly with the degree of substitution on the nanocrystal surfaces. Comparison with zeta potential measurements showed that zeta-potential measurements can be used as a direct method to determine the stoichiometry of binding between the positive surface grafts and the carboxylate salts.

Thermodynamic Study of the Interaction of Bovine Serum Albumin and Amino Acids with Cellulose Nanocrystals.

The interaction of bovine serum albumin (BSA) with sulfated, carboxylated, and pyridinium-grafted cellulose nanocrystals (CNCs) was studied as a function of the degree of substitution by determining the adsorption isotherm and by directly measuring the thermodynamics of interaction. The adsorption of BSA onto positively charged pyridinium-grafted cellulose nanocrystals followed Langmuirian adsorption with the maximum amount of adsorbed protein increasing linearly with increasing degree of substitution. The binding mechanism between the positively charged pyridinum-grafted cellulose nanocrystals and BSA was found to be endothermic and based on charge neutralization. A positive entropy of adsorption associated with an increase of the degree of disorder upon addition of BSA compensated for the unfavorable endothermic enthalpy and enabled formation of pyridinium-g-CNC-BSA complexes. The endothermic enthalpy of adsorption was further found to decrease as a function of increasing degree of substitution. Negatively charged cellulose nanocrystals bearing sulfate and/or carboxylic functionalities were found to not interact significantly with the BSA protein. To investigate in more detail the role of single amino acids in the adsorption of proteins onto cellulose nanocrystals, we also studied the interaction of different types of amino acids with CNCs, i.e., charged (lysine, aspartic acid), aromatic (tryptophan, tyrosine), and polar (serine) amino acids. We found that none of the single amino acids bound with CNCs irrespective of surface charge and that therefore the binding of proteins with CNCs appears to require larger amino acid sequences that induce a greater entropic contribution to stabilize binding. Single amino acids are thus not adsorbed onto cellulose nanocrystals.

Thermodynamics of interactions between cellulose nanocrystals and monovalent counterions.

Alkali and quaternary ammonium cations interact with negatively charged cellulose nanocrystals (CNCs) bearing sulfated or carboxylated functional groups. As these are some of the most commonly occurring cations CNC encounter in applications, the thermodynamic parameters of these CNC-counterion interactions were evaluated with isothermal titration calorimetry (ITC). Whereas the adsorption of monovalent counterions onto CNCs was thermodynamically favourable at all evaluated conditions as indicated by a negative Gibbs free energy, the enthalpic and entropic contributions to the CNC-ion interactions were found to be strongly dependent on the hydration characteristics of the counterion and could be correlated with the potential barrier to water exchange of the respective ions. The adsorption of chaotropic cations onto the surface was exothermic, while the interactions with kosmotropic cations were endothermic and completely entropy-driven. The interactions of CNCs with more bulky quaternary ammonium counterions were more complex, and the mechanism of interaction shifted from electrostatic interactions with surface charged groups of CNCs towards adsorption of alkyl chains onto the CNC hydrophobic planes when the alkyl chain length increased.

Ultra-Low Loading of Gold on Nickel Foam for Nitrogen Electrochemistry.

Ammonia (NH3) is widely used in various fields, and it is also considered a promising carbon free energy carrier, due to its high hydrogen content. The nitrogen reduction reaction (NRR), which converts nitrogen into ammonia by using protons from water as the hydrogen source, is receiving a lot of attention, since effective process optimization would make it possible to overcome the Haber-Bosch method. In this study, we used a solution-based approach to obtain functionalized porous Ni foam substrates with a small amount of gold (<0.1 mg cm-1). We investigated several deposition conditions and obtained different morphologies. The electrochemical performance of various catalysts on the hydrogen evolution reaction (HER) and NRR has been characterized. The ammonia production yield was determined by chronoamperometry experiments at several potentials, and the results showed a maximum ammonia yield rate of 20 µg h-1 mgcat-1 and a Faradaic efficiency of 5.22%. This study demonstrates the potential of gold-based catalysts for sustainable ammonia production and highlights the importance of optimizing deposition conditions to improve the selectivity toward HER.

Role of the Membrane Transport Mechanism in Electrochemical Nitrogen Reduction Experiments.

The electrochemical synthesis of ammonia through the nitrogen reduction reaction (NRR) is receiving much attention, since it is considered a promising alternative to the Haber-Bosch process. In NRR experiments, a Nafion membrane is generally adopted as a separator. However, its use is controversial since ammonia can be trapped in the membrane, to some extent, or even pass through it. We systematically investigate the interaction of a Nafion membrane with ammonia and with an electrolyte and compare it with Zirfon as a possible alternative separator. We show that Nafion containing ammonia can easily release it when immersed in a 0.1 M Na2SO4 ammonia-free electrolyte, due to the cation exchange mechanism (Na+-NH4+). Since Na2SO4 is a commonly adopted electrolyte for NRR experiments, this may cause serious measurement errors and non-reproducible results. The same experiments performed using the polysulfone Zirfon separator clearly show that it is immune to interactions with ammonia, because of its different ion conduction mechanism. The findings provide a deeper understanding of the choice of membrane and electrolyte to be adopted for NRR tests, and may allow one to obtain more accurate and reliable results.

Application of Sebum Lipidomics to Biomarkers Discovery in Neurodegenerative Diseases.

Lipidomics is strategic in the discovery of biomarkers of neurodegenerative diseases (NDDs). The skin surface lipidome bears the potential to provide biomarker candidates in the detection of pathological processes occurring in distal organs. We investigated the sebum composition to search diagnostic and, possibly, prognostic, biomarkers of Alzheimer's disease (AD) and Parkinson's disease (PD). The observational study included 64 subjects: 20 characterized as "probable AD with documented decline", 20 as "clinically established PD", and 24 healthy subjects (HS) of comparable age. The analysis of sebum by GCMS and TLC retrieved the amounts (µg) of 41 free fatty acids (FFAs), 7 fatty alcohols (FOHs), vitamin E, cholesterol, squalene, and total triglycerides (TGs) and wax esters (WEs). Distributions of sebum lipids in NDDs and healthy conditions were investigated with multivariate ANOVA-simultaneous component analysis (ASCA). The deranged sebum composition associated with the PD group showed incretion of most composing lipids compared to HS, whereas only two lipid species (vitamin E and FOH14:0) were discriminant of AD samples and presented lower levels than HS sebum. Thus, sebum lipid biosynthetic pathways are differently affected in PD and AD. The characteristic sebum bio-signatures detected support the value of sebum lipidomics in the biomarkers search in NDDs.

Real-World Experience with Benralizumab in Patients with Severe Eosinophilic Asthma: A Case Series.

PURPOSE: Severe eosinophilic asthma (SEA) is characterized by high eosinophilia, severe symptoms, important comorbidities, frequent exacerbations, and poor asthma control. Benralizumab, targeting the interleukin-5 receptor alpha, proved effective in inducing rapid eosinophil depletion and amelioration of symptoms and lung function; it also allowed to reduce exacerbations and the use of oral corticosteroids (OCS). The present case series, spanning different subtypes of SEA, aimed at expanding the real-world experience with benralizumab in Italy. PATIENTS AND METHODS: We collected data from SEA patients treated with benralizumab, at baseline and during treatment. We focused on the effects of benralizumab in the following conditions and endpoints: i) overlap between high-IgE and high-eosinophilic asthma; ii) presence of nasal polyposis as comorbidity; iii) corticosteroid-sparing effect; iv) patient perception. RESULTS: Ten SEA patients (females: N=7; age range: 19-70 years) referred to 8 Italian Centers and treated with benralizumab were included, presenting with several comorbidities such as non-allergic disease (8/10), atopy (3/10), high IgE (5/10) and nasal polyposis (6/10). Overall, benralizumab yielded optimal disease control in all patients, particularly in terms of rapid clinical and functional improvement, decreased systemic steroid need (OCS therapy was completely discontinued in 7 cases) and amelioration of patient quality of life, except for 1 case, in whom other conditions not related to benralizumab therapy interfered with the patient perception. CONCLUSION: Our findings further support the efficacy and safety of benralizumab observed in randomized clinical trials, providing even better results for lung function improvement.

Ni foam electrode solution impregnated with Ni-Fe X (OH) Y catalysts for efficient oxygen evolution reaction in alkaline electrolyzers.

Oxygen evolution reaction (OER) is a demanding step within the water splitting process for its requirement of a high overpotential. Thus, to overcome this unfavourable kinetics, an efficient catalyst is required to expedite the process. In this context, we report on Ni foam functionalised with low cost iron (Fe) and iron hydroxide (Fe(OH) X ), wet chemically synthesized as OER catalysts. The prepared catalyst based on iron hydroxide precipitate shows a promising performance, exhibiting an overpotential of 270 mV (at a current density of 10 mA cm-2 in 1 M KOH solution), an efficient Tafel slope of ∼50 mV dec-1 and stable chronopotentiometry. The promising performance of the anode was further reproduced in the overall water splitting reaction with a two electrode cell. The overall reaction requires a lower potential of 1.508 V to afford 10 mA cm-2, corresponding to 81.5% electrical to fuel efficiency.

Feasibility studies on si-based biosensors.

The aim of this paper is to summarize the efforts carried out so far in the fabrication of Si-based biosensors by a team of researchers in Catania, Italy. This work was born as a collaboration between the Catania section of the Microelectronic and Microsystem Institute (IMM) of the CNR, the Surfaces and Interfaces laboratory (SUPERLAB) of the Consorzio Catania Ricerche and two departments at the University of Catania: the Biomedical Science and the Biological Chemistry and Molecular Biology Departments. The first goal of our study was the definition and optimization of an immobilization protocol capable of bonding the biological sensing element on a Si-based surface via covalent chemical bonds. We chose SiO(2) as the anchoring surface due to its biocompatibility and extensive presence in microelectronic devices. The immobilization protocol was tested and optimized, introducing a new step, oxide activation, using techniques compatible with microelectronic processing. The importance of the added step is described by the experimental results. We also tested different biological molecule concentrations in the immobilization solutions and the effects on the immobilized layer. Finally a MOS-like structure was designed and fabricated to test an electrical transduction mechanism. The results obtained so far and the possible evolution of the research field are described in this review paper.

Cellulose-Cyclodextrin Co-Polymer for the Removal of Cyanotoxins on Water Sources.

With increasing global water temperatures and nutrient runoff in recent decades, the blooming season of algae lasts longer, resulting in toxin concentrations that exceed safe limits for human consumption and for recreational use. From the different toxins, microcystin-LR has been reported as the main cyanotoxin related to liver cancer, and consequently its abundance in water is constantly monitored. In this work, we report a methodology for decorating cellulose nanofibrils with ß-cyclodextrin or with poly(ß-cyclodextrin) which were tested for the recovery of microcystin from synthetic water. The adsorption was followed by Quartz Crystal Microbalance with Dissipation monitoring (QCM-D), allowing for real-time monitoring of the adsorption behavior. A maximum recovery of 196 mg/g was obtained with the modified by cyclodextrin. Characterization of the modified substrate was confirmed with Fourier Transform Infrared Spectroscopy (FT-IR), X-ray Photoelectron Spectroscopy (XPS), Thermogravimetric Analysis (TGA), and Atomic Force Microscopy (AFM).

Isothermal titration calorimetry to study the influence of citrus pectin degree and pattern of methylesterification on Zn2+ interaction.

This work explored the influence of the citrus pectin degree and pattern of methylesterification on its interaction with Zn2+ using isothermal titration calorimetry (ITC). Pectin samples with a comparable degree of methylesterification (DM) but distinct distribution patterns of non-methylesterified carboxylic groups (absolute degree of blockiness, DBabs) were produced through enzymatic (blockwise pattern) or alkaline (random pattern) demethylesterification. The pectin-Zn2+ interaction was found to be endothermic, in which a positive entropy change compensated for the unfavorable endothermic enthalpy change, driving the interaction between pectin and Zn2+. Decreasing pectin DM or increasing DBabs promoted Zn2+ binding, with the estimated binding capacity (mol Zn2+/mol GalA) and binding constant (mM-1) being mainly determined by pectin DBabs, rather than DM. ITC was found to be a useful technique to study the pectin-cation interaction, however, low DM pectin samples are sensitive to gelling and therefore more difficult to study.

Crucial aspects for the use of silicon photomultiplier devices in continuous wave functional near-infrared spectroscopy.

In this work, we investigate some major issues for the use of silicon photomultiplier (SiPM) devices in continuous wave functional near-infrared spectroscopy (CW fNIRS). We analyzed the after-pulsing effect, proposing the physical mechanism causing it, and determining its relevance for CW fNIRS. We studied the SiPM transients occurring as the SiPM device goes from the dark (LED in off state) to the illumination (LED in on state) conditions, and vice-versa. Finally, we studied the SiPM SNR in standard CW fNIRS operation.

Synaptic plasticity modulates the spontaneous recovery of locomotion after spinal cord hemisection.

Several evidences have demonstrated that adult mammals could achieve a wide range of spontaneous sensory-motor recovery after spinal cord injury by means of various forms of neuroplasticity. In this study we evaluated the possibility that after low-thoracic spinal cord hemisection in the adult rat, significant hindlimb locomotor recovery could occur, and that this recovery may be driven, at least in part, by mechanisms of synaptic plasticity. In order to address these issues, we measured the expression levels of synapsin-I and brain-derived neurotrophic factor by Western blotting, at various time points after hemisection and correlated them with the motor performance on a grid walk test. Regression analysis showed that the expression of synapsin-I was strongly correlated with the spontaneous recovery of hindlimb locomotion (R=0.78). Conversely, neither the expression levels of synapsin-I nor the locomotor recovery were associated with the expression of brain-derived neurotrophic factor. Overall results indicate that after spinal cord hemisection, substantial recovery of hindlimb locomotion could occur spontaneously, and that synaptic plasticity within spinal circuitries below the level of the lesion, could be an important mechanism involved in these processes.

Characterization of a fiber-less, multichannel optical probe for continuous wave functional near-infrared spectroscopy based on silicon photomultipliers detectors: in-vivo assessment of primary sensorimotor response.

We report development, testing, and in vivo characterization of a multichannel optical probe for continuous wave (CW) functional near-infrared spectroscopy (fNIRS) that relies on silicon photomultipliers (SiPMs) detectors. SiPMs are cheap, low voltage, and robust semiconductor light detectors with performances analogous to photomultiplier tubes (PMTs). In contrast with PMTs, SiPMs allow direct contact with the head and transfer of the analog signals through thin cables greatly increasing the system flexibility avoiding optical fibers. The coupling of SiPMs and light-emitting diodes (LEDs) made the optical probe lightweight and robust against motion artifacts. After characterization of SiPM performances, which was proven to provide a noise equivalent power below 3 fW, the apparatus was compared through an in vivo experiment to a commercial system relying on laser diodes, PMTs, and optical fibers for light probing and detection. The optical probes were located over the primary sensorimotor cortex and the similarities between the hemodynamic responses to the contralateral motor task were assessed. When compared to other state-of-the-art wearable fNIRS systems, where photodiode detectors are employed, the single photon sensitivity and dynamic range of SiPMs can fully exploit the long and variable interoptode distances needed for correct estimation of brain hemodynamics using CW-fNIRS.