Atomistic Simulations of Functionalized Nano-Materials for Biosensors Applications

Nanoscale biosensors, a highly promising technique in clinical analysis, can provide sensitive yet label-free detection of biomolecules. The spatial and chemical specificity of the surface coverage, the proper immobilization of the bioreceptor as well as the underlying interfacial phenomena are crucial elements for optimizing the performance of a biosensor. Due to experimental limitations at the microscopic level, integrated cross-disciplinary approaches that combine in silico design with experimental measurements have the potential to present a powerful new paradigm that tackles the issue of developing novel biosensors. In some cases, computational studies can be seen as alternative approaches to assess the microscopic working mechanisms of biosensors. Nonetheless, the complex architecture of a biosensor, associated with the collective contribution from “substrate–receptor–analyte” conjugate in a solvent, often requires extensive atomistic simulations and systems of prohibitive size which need to be addressed. In silico studies of functionalized surfaces also require ad hoc force field parameterization, as existing force fields for biomolecules are usually unable to correctly describe the biomolecule/surface interface. Thus, the computational studies in this field are limited to date. In this review, we aim to introduce fundamental principles that govern the absorption of biomolecules onto functionalized nanomaterials and to report state-of-the-art computational strategies to rationally design nanoscale biosensors. A detailed account of available in silico strategies used to drive and/or optimize the synthesis of functionalized nanomaterials for biosensing will be presented. The insights will not only stimulate the field to rationally design functionalized nanomaterials with improved biosensing performance but also foster research on the required functionalization to improve biomolecule–surface complex formation as a whole.     

Axitinib in Ponatinib-Resistant B-Cell Acute Lymphoblastic Leukemia Harboring a T315L Mutation

Adult acute lymphoblastic leukemia (ALL) with BCR-ABL1 rearrangement (Philadelphia chromosome, Ph) is a hematological aggressive disease with a fatal outcome in more than 50% of cases. Tyrosine kinase inhibitors (TKIs) targeting the activity of BCR-ABL1 protein have improved the prognosis; however, relapses are frequent because of acquired somatic mutations in the BCR-ABL1 kinase domain causing resistance to first, second and third generation TKIs. Axitinib has shown in vitro and ex vivo activity in blocking ABL1; however, clinical trials exploring its efficacy in ALL are missing. Here, we presented a 77-year-old male with a diagnosis of Ph positive ALL resistant to ponatinib and carrying a rare threonine to leucine (T315L) mutation on BCR-ABL1 gene. The patient was treated with axitinib at 5 mg/twice daily as salvage therapy showing an immediate although transient benefit with an overall survival of 9.3 months. Further dose-finding and randomized clinical trials are required to assess the real efficacy of axitinib for adult Ph positive ALL resistant to third generation TKIs.     

Light emitting devices based on Si nanoclusters： the integration with a photonic-crystal and electroluminescence properties

We present the properties and potentialities of light emitting devices based on amorphous Si nanoclusters. Amorphousnanostructures may constitute an interesting alternative to Si nanocrystals for the monolithic integration of optical andelectrical functions in Si technology. In fact, they exhibit an intense room temperature electroluminescence (EL). The ELproperties of these devices have been studied as a function of current and of temperature. Moreover, to improve theextraction efficiency of the light, we have integrated the emitting system with a 2D photonic crystal structure opportunelyfabricated by using conventional optical lithography to reduce the total internal reflection of the emitted light. The extractionefficiency in such devices increases by a factor of 4 at a resonance wavelength.     

ATP and a mitochondrial electrochemical gradient are required for functional activity of the steroidogenic acute regulatory (StAR) protein in isolated mitochondria

The Steroidogenic Acute Regulatory (StAR) protein has been put forth as the rapidly synthesized, cycloheximide-sensitive protein that is required for the transport of cholesterol to the inner mitochondrial membrane and the P450scc enzyme and thereby acutely regulates steroidogenesis in steroidogenic tissues. In this study, several of the factors that may be required for StAR activity were examined using an in vitro system. Lysates from StAR-transfected COS-1 cells were added to mitochondria isolated from MA-10 Leydig tumor cells. Results obtained demonstrated that StAR-containing cell lysate increased steroidogenesis in isolated mitochondria, but failed to do so in the presence of m-CCCP, apyrase, or AMP-PNP, suggesting that StAR function requires ATP hydrolysis as well as an electrochemical gradient for maximal steroidogenic activity.     

Goat farm variability affects milk Fourier-transform infrared spectra used for predicting coagulation properties

Driven by the large amount of goat milk destined for cheese production, and to pioneer the goat cheese industry, the objective of this study was to assess the effect of farm in predicting goat milk-coagulation and curd-firmness traits via Fourier-transform infrared spectroscopy. Spectra from 452 Sarda goats belonging to 14 farms in central and southeast Sardinia (Italy) were collected. A Bayesian linear regression model was used, estimating all spectral wavelengths' effects simultaneously. Three traditional milk-coagulation properties [rennet coagulation time (min), time to curd firmness of 20 mm (min), and curd firmness 30 min after rennet addition (mm)] and 3 curd-firmness measures modeled over time [rennet coagulation time estimated according to curd firmness change over time (RCT_eq), instant curd-firming rate constant, and asymptotical curd firmness] were considered. A stratified cross validation (SCV) was assigned, evaluating each farm separately (validation set; VAL) and keeping the remaining farms to train (calibration set) the statistical model. Moreover, a SCV, where 20% of the goats randomly taken (10 replicates per farm) from the VAL farm entered the calibration set, was also considered (SCV₈₀). To assess model performance, coefficient of determination (R²_VAL) and the root mean squared error of validation were recorded. The R²_VAL varied between 0.14 and 0.45 (instant curd-firming rate constant and RCT_eq, respectively), albeit the standard deviation was approximating half of the mean for all the traits. Although average results of the 2 SCV procedures were similar, in SCV₈₀, the maximum R²_VAL increased at about 15% across traits, with the highest observed for time to curd firmness of 20 mm (20%) and the lowest for RCT_eq (6%). Further investigation evidenced important variability among farms, with R²_VAL for some of them being close to 0. Our work outlined the importance of considering the effect of farm when developing Fourier-transform infrared spectroscopy prediction equations for coagulation and curd-firmness traits in goats.     

Label-free probing of G-quadruplex formation by surface-enhanced Raman scattering

In this work, we establish the use of surface-enhanced Raman scattering (SERS) as a label-free analytical technique for the direct detection of G-quadruplex formation. In particular, we demonstrate that SERS analysis allows the evaluation of the relative stability of G quadruplexes that differ for the number of G tetrads and investigate several structural features of quadruplexes, such as the orientation of glycosidic bonds, the identification of distortions in the sugar-phosphate backbone, and the degree of hydrogen-bond solvation. Herein, the fluctuation of the SERS spectra, due to the specific interaction of vibrational modes with the SERS-active substrate, is quantitatively analyzed before and after quadruplex formation. The results of this study suggest a perpendicular orientation of the quadruplexes (with or without the 3'-tetra end linker) with respect to the silver colloidal surface, which opens new perspectives for the use of SERS as a label-free analytical tool for the study of the binding mode between quadruplexes and their ligands.     

Increasing Cold Tolerance of Cactus Pear Fruit by High-Temperature Conditioning and Film Wrapping

Pre-storage high-temperature conditioning (HTC, 38 °C, and 95% RH for 24 h) and individual film wrapping (IFW) with a perforated polyolefinic heat-shrinkable film were used as individual treatments or in combination to mitigate chilling injury of first crop cactus pear cv ‘Gialla’. The fruit was stored for 21 days at either 2 or 8 °C (CS) plus 1 week of simulated marketing conditions (SMC) at 20 °C. The reduction in peel disorders and decay in HTC-treated fruit stored at 2 °C was comparable to that detected in control fruit stored at 8 °C. IFW was more efficient than HTC in reducing peel disorders, almost completely inhibited weight loss, and preserved freshness in fruit stored at 8 °C as well as in those stored at 8 °C. The internal quality of the fruit (pH, total soluble solids, titratable acidity, vitamin C, polyphenols, betaxanthins, antioxidant capacity, acetaldehyde, and ethanol) was slightly affected by treatments. Combining HTC with IFW did not improve fruit tolerance to low temperature but reduced decay with respect to individual treatments. IFW with a perforated film in combination with HTC is a good means of overcoming the stringent conditions of cold quarantine treatments, maintaining fruit freshness and reducing decay in cold-stored cactus pears.     

Preparation of drug nanocrystals embedded in mannitol microcrystals via liquid antisolvent precipitation followed by immediate (on-line) spray drying

The aim of this study is to investigate the feasibility of producing pharmaceutical nanoformulations for enhanced oral or pulmonary delivery of poorly water-soluble drugs via liquid antisolvent precipitation followed by immediate (on-line) spray drying. A poorly water-soluble corticosteroid, budesonide, was chosen as the model drug. Budesonide nanoparticles were prepared through liquid antisolvent precipitation, and then processed into a powdered nanoformulation which consists of budesonide nanoparticles embedded in mannitol microcrystals by immediate (on-line) spray drying. The size of the freshly precipitated and the reconstituted budesonide particles was analyzed by dynamic light scattering (DLS). The spray-dried nanoformulation, together with budesonide and mannitol raw materials, their physical mixture and the spray-dried mannitol, were characterized by field emission scanning electron microscopy (FESEM), differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD). In vitro dissolution test and aerosol deposition study were conducted on the spray-dried nanoformulation and the physical mixture of budesonide and mannitol raw materials. It was found that the spray-dried nanoformulation, consisting of mannitol microcrystals comprising budesonide nanocrystals with z-average mean size of 520?±?11.4?nm, exhibited enhanced drug dissolution rate and higher fine particle fraction (FPF). The results of this study indicated the potential of the combined process of liquid antisolvent precipitation followed by immediate (on-line) spray drying to be used as a direct and continuous formulation process to produce powdered nanoformulations to achieve enhanced oral or pulmonary delivery of poorly water-soluble drugs.