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L'orticoltura può influire positivamente sul benessere delle persone anziane, contrastando il declino fisico e cognitivo e migliorando la qualità della vita. Le moderne tecnologie fornisco-no un valido supporto per incoraggiare lo svolgimento di tali attività e promuovere uno stile di vita attivo. Il presente studio esplorativo si propone di valutare gli effetti di attività di orticultura comunitaria supportate da una piattaforma digitale in un campione misto di partecipanti over 60 durante il periodo della pandemia da COVID-19. I partecipanti di età superiore ai sessanta anni hanno preso parte ad un percorso della durata di sei mesi, diviso in due periodi. Durante i primi tre mesi i partecipanti si sono limitati a rispondere ai test che indagavano le seguenti va-riabili psicologiche: autostima, solitudine, depressione, qualità della vita, affetti, supporto sociale e funzionamento cognitivo. Nei successivi tre mesi i partecipanti sono stati coinvolti in attività di orticoltura, supportate da applicazioni digitali accessibili tramite smartphone, cui ha fatto seguito la rilevazione delle stesse variabili psicologiche. Gli effetti dell'orticultura sono stati valutati confrontando le due serie di rilevazioni. I risultati mostrano un incremento della qualità della vita degli anziani, delle funzioni cognitive e, in misura minore, del benessere soggettivo. Gli anziani che risultano spo-sati o conviventi manifestano in genere una migliore qualità della vita e a seguito dell'intervento sono meno propensi ad esprimere stati affettivi negativi. Non sono stati evidenziati segni di depressione. L'orticultura assistita digitalmente può migliorare la qualità della vita degli anziani durante la pandemia da COVID-19.

In this study, transmission electron microscopy atomic force microscopy, and surface enhanced Raman spectroscopy are combined through a direct imaging approach, to gather structural and chemical information of complex molecular systems such as ion channels in their original plasma membrane. Customized microfabricated sample holder allows to characterize Na

The effect of direct or indirect binding of intercalant molecules on DNA structure is of fundamental importance in understanding the biological functioning of DNA. Here we report on self-suspended DNA nanobundles as ultrasensitive nanomechanical resonators for structural studies of DNA-ligand complexes. Such vibrating nanostructures represent the smallest mechanical resonator entirely composed of DNA. A correlative analysis between the mechanical and structural properties is exploited to study the intrinsic changes of double strand DNA, when interacting with different intercalant molecules (YOYO-1 and GelRed) and a chemotherapeutic drug (Cisplatin), at different concentrations. Possible implications of our findings are related to the study of interaction mechanism of a wide category of molecules with DNA, and to further applications in medicine, such as optimal titration of chemotherapeutic drugs and environmental studies for the detection of heavy metals in human serum., Intercalating molecules can significantly change the conformation of DNA. Here, the authors fabricated resonators fully composed of DNA forming bundles between microfabricated pillars to study the vibration property of the DNA bundles with/without intercalant molecules.

Superhydrophobic surfaces display an extraordinary repulsion to water and water-based solutions. This effect emerges from the interplay of intrinsic hydrophobicity of the surface and its morphology. These surfaces have been established for a long time and have been studied for decades. The increasing interest in recent years has been focused towards applications in many different fields and, in particular, biomedical applications. In this paper, we review the progress achieved in the last years in the fabrication of regularly patterned superhydrophobic surfaces in many different materials and their exploitation for the manipulation and characterization of biomaterial, with particular emphasis on the issues affecting the yields of the fabrication processes and the quality of the manufactured devices.

Methods to produce protein amyloid fibrils, in vitro, and in situ structure characterization, are of primary importance in biology, medicine, and pharmacology. We first demonstrated the droplet on a super-hydrophobic substrate as the reactor to produce protein amyloid fibrils with real-time monitoring of the growth process by using combined light-sheet microscopy and thermal imaging. The molecular structures were characterized by Raman spectroscopy, X-ray diffraction and X-ray scattering. We demonstrated that the convective flow induced by the temperature gradient of the sample is the main driving force in the growth of well-ordered protein fibrils. Particular attention was devoted to PHF6 peptide and full-length Tau441 protein to form amyloid fibrils. By a combined experimental with the molecular dynamics simulations, the conformational polymorphism of these amyloid fibrils were characterized. The study provided a feasible procedure to optimize the amyloid fibrils formation and characterizations of other types of proteins in future studies., Zhang et al present an integrated real-time imaging and flow field control platform based on water droplet evaporation on super-hydrophobic substrate (SHS) to enable amyloid fibril aggregation. They apply this methodology to observe structural polymorphism in PHF6 peptide and full length Tau441.

In this work, micro-fabrication, super-hydrophobic properties and a physiologically compatible preparation step are combined and tailored to obtain background free biological samples to be investigated by Transmission Electron Microscopy (TEM) diffraction technique. The validation was performed evaluating a well-known parameter such as the DNA interbases value. The diffraction spacing measured is in good agreement with those obtained by HRTEM direct metrology and by traditional X-Ray diffraction. This approach addresses single molecule studies in a simplified and reproducible straightforward way with respect to more conventional and widely used techniques. In addition, it overcomes the need of long and elaborated samples preparations: the sample is in its physiological environment and the HRTEM data acquisition occurs without any background interference, coating, staining or additional manipulation. The congruence in the results reported in this paper makes the application of this approach extremely promising towards those molecules for which crystallization remains a hurdle, such as cell membrane proteins and fibrillar proteins.

The deep-sea brines of the Red Sea are remote and unexplored environments characterized by high temperatures, anoxic water, and elevated concentrations of salt and heavy metals. This environment provides a rare system to study the interplay between halophilic and thermophilic adaptation in biologic macromolecules. The present article reports the first DNA polymerase with halophilic and thermophilic features. Biochemical and structural analysis by Raman and circular dichroism spectroscopy showed that the charge distribution on the protein's surface mediates the structural balance between stability for thermal adaptation and flexibility for counteracting the salt-induced rigid and nonfunctional hydrophobic packing. Salt bridge interactions via increased negative and positive charges contribute to structural stability. Salt tolerance, conversely, is mediated by a dynamic structure that becomes more fixed and functional with increasing salt concentration. We propose that repulsive forces among excess negative charges, in addition to a high percentage of negatively charged random coils, mediate this structural dynamism. This knowledge enabled us to engineer a halophilic version of Thermococcus kodakarensis DNA polymerase.-Takahashi, M., Takahashi, E., Joudeh, L. I., Marini, M., Das, G., Elshenawy, M. M., Akal, A., Sakashita, K., Alam, I., Tehseen, M., Sobhy, M. A., Stingl, U., Merzaban, J. S., Di Fabrizio, E., Hamdan, S. M. Dynamic structure mediates halophilic adaptation of a DNA polymerase from the deep-sea brines of the Red Sea.

This review looks at the different approaches, techniques, and materials devoted to DNA studies. In the past few decades, DNA nanotechnology, micro-fabrication, imaging, and spectroscopies have been tailored and combined for a broad range of medical-oriented applications. The continuous advancements in miniaturization of the devices, as well as the continuous need to study biological material structures and interactions, down to single molecules, have increase the interdisciplinarity of emerging technologies. In the following paragraphs, we will focus on recent sensing approaches, with a particular effort attributed to cutting-edge techniques for structural and mechanical studies of nucleic acids.

In bio-imaging by electron microscopy, damage of the sample and limited contrast are the two main hurdles for reaching high image quality. We extend a new preparation method based on nanofabrication and super-hydrophobicity to the imaging and structural studies of nucleic acids, nucleic acid–protein complexes (DNA/Rad51 repair protein complex) and neuronal ion channels (gap-junction, K+ and GABAA channels) as paradigms of biological significance and increasing complexity. The preparation method is based on the liquid phase and is compatible with physiological conditions. Only in the very last stage, samples are dried for TEM analysis. Conventional TEM and high-resolution TEM (HRTEM) were used to achieve a resolution of 3.3 and 1.5 Å, respectively. The EM dataset quality allows the determination of relevant structural and metrological information on the DNA structure, DNA–protein interactions and ion channels, allowing the identification of specific macromolecules and their structure.

Super-hydrophobic micro-patterned surfaces are ideal substrates for the controlled self-assembly and substrate-free characterization of biological molecules. In this device, the tailored surface supports a micro-volume drop containing the molecules of interest. While the quasi-spherical drop is evaporating under controlled conditions, its de-wetting direction is guided by the pillared microstructure on top of the device, leading to the formation of threads between the neighboring pillars. This effect has been exploited here to elucidate the mechanism triggering the formation of amyloid fibers and oligomers in tau related neurodegenerative diseases. By using Raman spectroscopy, we demonstrate that the fiber bridging the pillars contains β-sheets, a characteristic feature of amyloid aggregation. We propose that the combination of laminar flow, shear stress and molecular crowding taking place while the drop is evaporating on the SHMS, induces the reorganization of the tau protein secondary structure and we suggest that this effect could in fact closely mimic the actual mechanism occurring in the human brain environment. Such a straightforward technique opens up new possibilities in the field of self-assembly of biomolecules and their characterization by different methods (SEM, AFM, Raman spectroscopy, TEM), in a single device.

In cystic fibrosis, deletion of phenylalanine 508 (F508del) in the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel causes misfolding and premature degradation. One possible approach to reducing the detrimental health effects of cystic fibrosis could be the identification of proteins whose suppression rescues F508del-CFTR function in bronchial epithelial cells. However, searches for these potential targets have not yet been conducted, particularly in a relevant airway background using a functional readout. To identify proteins associated with F508del-CFTR processing, we used a high-throughput functional assay to screen an siRNA library targeting 6,650 different cellular proteins. We identified 37 proteins whose silencing significantly rescued F508del-CFTR activity, as indicated by enhanced anion transport through the plasma membrane. These proteins included FAU, UBE2I, UBA52, MLLT6, UBA2, CHD4, PLXNA1, and TRIM24, among others. We focused our attention on FAU, a poorly characterized protein with unknown function. FAU knockdown increased the plasma membrane targeting and function of F508del-CFTR, but not of wild-type CFTR. Investigation into the mechanism of action revealed a preferential physical interaction of FAU with mutant CFTR, leading to its degradation. FAU and other proteins identified in our screening may offer a therapeutically relevant panel of drug targets to correct basic defects in F508del-CFTR processing.

The shear flow generated at the rim of a drop evaporating on a micro-fabricated super-hydrophobic surface has been used to suspend and orient single/few lysozyme amyloid fibrils between two pillars for substrate-free characterization. Micro Raman spectroscopy performed on extended fibers evidenced a shift of the Amide I band main peak to the value attributed to β-sheet secondary structure, characteristic of the amyloid fibers. In addition, given the orientation sensitivity of the anisotropic molecule, the Raman signal of the main secondary structure was nicely enhanced for a fiber alignment parallel to the polarization direction of the laser. The substrate-free sample generated by this suspending technique is suitable for other structural analysis methods, where fiber crystals are investigated. It could be further employed for generation of arrays and patterns in a controllable fashion, where bio-compatible material is needed.

The A- and B-form are two of the most common structural conformations of double strand DNA present in nature and they can interchange on the basis of the helices hydration [1,2]. Herein we demonstrate that the use of non-destructive techniques such as Raman spectroscopy coupled with the use of a super-hydrophobic device, allows the clear identification of the DNA hydration state, of the backbone (phosphate+deoxyribose sugar) conformation and of the nucleotides. There is a wide prospect for an increase of knowledge in biomolecules using this combined approach resulting in a significant impact in the study of more complex supramolecular assemblies and of fine chemical variation along the genomic loci undergoing to epigenetic variations. Display Omitted Non-destructive Raman spectroscopy technique coupled with super-hydrophobic devices for biological fibers investigationSuper-hydrophobic surfaces comprising micro-pillars for DNA localizationDiscrimination between A- and B-form of dsDNA suspended between micro-pillars by means of Raman analysis.Electron Microscopy preliminary investigation of suspended dsDNA molecules

This study presents a simple and rapid fabrication technique involving injection molding and particle leaching (IM/PL) to fabricate the porous scaffold for tissue engineering applications. Sodium Chloride (NaCl) and Sucrose are separately mixed with the poly-epsilon-caprolactone (PCL) granules using a screwed thermo-regulated extruder, then the biocompatible scaffolds are fabricated through injection molding. The micro/nano-structure of the samples and their different grade of porosity were characterized by scanning electron microscopy and mercury intrusion porosimetry. Bone marrow-derived mesenchymal stem cells are chosen for cell culture and Hoechst 33342 staining was used to verify the biocompatibility of the polymeric porous surfaces. We concluded that, by using the same fast solvent free injection/leaching process, the use of Sucrose as porogen, instead of NaCl, allowed the obtainment of biocompatible scaffolds with a higher grade of porosity with suitable cell adhesion capacity for tissue engineering purpose. PCL/NaCl, PCL/Sucrose composites were injection molded.Particulate leaching confers to the scaffolds different grades of porosity.Human hMSCs have attached, survived and proliferated well on PCL/Sucrose scaffolds.Molded PCL/Sucrose scaffolds could be good candidates for tissue engineering. Display Omitted

For decades, lipid droplets have been considered as the main cellular organelles involved in the fat storage, because of their lipid composition. However, in recent years, some new and totally unexpected roles have been discovered for them: (i) they are active sites for synthesis and storage of inflammatory mediators, and (ii) they are key players in cancer cells and tissues, especially in cancer stem cells. In this review, we summarize the main concepts related to the lipid droplet structure and function and their involvement in inflammatory and cancer processes.

Nanowire arrays and networks with precisely controlled patterns are very interesting for innovative device concepts in mesoscopic physics. In particular, DNA templates have proven to be versatile for the fabrication of complex structures that obtained functionality via combinations with other materials, for example by functionalisation with molecules or nanoparticles, or by coating with metals. Here, the controlled motion of the a three-phase contact line (TCL) of DNA-loaded drops on superhydrophobic substrates is used to fabricate suspended nanowire arrays. In particular, the deposition of DNA wires is imaged in situ, and different patterns are obtained on hexagonal pillar arrays by controlling the TCL velocity and direction. Robust conductive wires and networks are achieved by coating the wires with a thin layer of gold, and as proof of concept conductivity measurements are performed on single suspended wires. The plastic material of the superhydrophobic pillars ensures electrical isolation from the substrate. The more general versatility of these suspended nanowire networks as functional templates is outlined by fabricating hybrid organic-metal-semiconductor nanowires by growing ZnO nanocrystals onto the metal-coated nanowires.

Nature Communications 5: Article number: 3513 (2014); Published: 29 April 2016; Updated: 31 October 2016 This article was published without any competing financial interests statement. A revised declaration that lists potentially competing financial interests is provided below: A.D.M., M.M. and M.A.are affiliated with Bcs Biotech S.

Objective To assess the effects of Day Care (DC) on older subjects with dementia and their caregivers. Methods Thirty patients with dementia, consecutively admitted to a DC, were compared with 30 patients, matched for age and cognitive function, who received usual home care (HC). Primary caregivers were compared as well. At baseline and after 2 months, patients were assessed for cognitive and functional status, behavioral and psychological symptoms [Neuropsychiatric Inventory (NPI)] and psychotropic drugs use, and caregivers were evaluated for care burden [Caregiver Burden Inventory (CBI)] and depressive symptoms. Results After adjusting for potential confounders, NPI score significantly decreased in DC group, with a reduction of psychotropic drugs prescription, whereas it increased in HC. No significant between-group difference was observed for cognitive and functional change. CBI significantly decreased in DC, but not HC, caregivers, with no significant between-group difference in depressive symptoms change. Conclusions A 2-month period of DC assistance is effective in reducing behavioral and psychological symptoms of dementia patients and in alleviating caregivers' burden. Copyright © 2008 John Wiley & Sons, Ltd.

Cystic fibrosis (CF) is caused by mutations in the CFTR chloride channel. Deletion of phenylalanine 508 (F508del), the most frequent CF mutation, impairs CFTR trafficking and gating. F508del-CFTR mistrafficking may be corrected by acting directly on mutant CFTR itself or by modulating expression/activity of CFTR-interacting proteins, that may thus represent potential drug targets. To evaluate possible candidates for F508del-CFTR rescue, we screened a siRNA library targeting known CFTR interactors. Our analysis identified RNF5 as a protein whose inhibition promoted significant F508del-CFTR rescue and displayed an additive effect with the investigational drug VX-809. Significantly, RNF5 loss in F508del-CFTR transgenic animals ameliorated intestinal malabsorption and concomitantly led to an increase in CFTR activity in intestinal epithelial cells. In addition, we found that RNF5 is differentially expressed in human bronchial epithelia from CF vs. control patients. Our results identify RNF5 as a target for therapeutic modalities to antagonize mutant CFTR proteins.

In order to develop surfaces with improved cell culture biocompatibility, we optimized a solvent-casting and particulate-leaching fabrication technique to create porous three-dimensional polycaprolactone scaffolds. These biocompatible porous surfaces were realized by means of NaCl particles as porogen; salt leaching by immersion in distilled water created porosity and pore interconnectivity in the material. Scanning electron microscopy and mercury intrusion porosimetry were used for the measurement of porosity, pore size distribution, permeability and compressibility. To evaluate scaffold biocompatibility, fibroblasts were cultured on the porous surfaces and confocal immunofluorescence characterization indicated that they were effective for in vitro cell culture and practical tissue engineering applications.

The DNA helix and its internal structures were directly imaged; characteristic lengths and inner components were measured and reported., The structure of DNA was determined in 1953 by x-ray fiber diffraction. Several attempts have been made to obtain a direct image of DNA with alternative techniques. The direct image is intended to allow a quantitative evaluation of all relevant characteristic lengths present in a molecule. A direct image of DNA, which is different from diffraction in the reciprocal space, is difficult to obtain for two main reasons: the intrinsic very low contrast of the elements that form the molecule and the difficulty of preparing the sample while preserving its pristine shape and size. We show that through a preparation procedure compatible with the DNA physiological conditions, a direct image of a single suspended DNA molecule can be obtained. In the image, all relevant lengths of A-form DNA are measurable. A high-resolution transmission electron microscope that operates at 80 keV with an ultimate resolution of 1.5 Å was used for this experiment. Direct imaging of a single molecule can be used as a method to address biological problems that require knowledge at the single-molecule level, given that the average information obtained by x-ray diffraction of crystals or fibers is not sufficient for detailed structure determination, or when crystals cannot be obtained from biological molecules or are not sufficient in understanding multiple protein configurations.

A strategy for an innovative, continuous and reversible LSPR tuning using DNA origami actuation to modulate the nanometric separation of two gold nanoparticles has been developed. The actuation mechanism is based on DNA hybridization, in particular three different DNA sequences were shown to induce resonance shift of up to 6 nm.

TMEM16A/ANO1 is a calcium-activated chloride channel expressed in several types of epithelia and involved in various physiological processes, including proliferation and development. During mouse embryonic development, the expression of TMEM16A in the olfactory epithelium is dynamic. TMEM16A is expressed at the apical surface of the entire olfactory epithelium at embryonic day E12.5 while from E16.5 its expression is restricted to a region near the transition zone with the respiratory epithelium. To investigate whether TMEM16A plays a role in the development of the mouse olfactory epithelium, we obtained the first immunohistochemistry study comparing the morphological properties of the olfactory epithelium and nasal glands in TMEM16A-/- and TMEM16A+/+ littermate mice. A comparison between the expression of the olfactory marker protein and adenylyl cyclase III shows that genetic ablation of TMEM16A did not seem to affect the maturation of olfactory sensory neurons and their ciliary layer. As TMEM16A is expressed at the apical part of supporting cells and in their microvilli, we used ezrin and cytokeratin 8 as markers of microvilli and cell body of supporting cells, respectively, and found that morphology and development of supporting cells were similar in TMEM16A-/- and TMEM16A+/+ littermate mice. The average number of supporting cells, olfactory sensory neurons, horizontal and globose basal cells were not significantly different in the two types of mice. Moreover, we also observed that the morphology of Bowman’s glands, nasal septal glands and lateral nasal glands did not change in the absence of TMEM16A. Our results indicate that the development of mouse olfactory epithelium and nasal glands does not seem to be affected by the genetic ablation of TMEM16A.

In this work we review novel strategies and new physical effects to achieve compositional and structural recognition at single molecule level. This chapter is divided in two main parts. The first one introduces the strategies currently adopted to investigate matter at few molecules level. Exploiting the capability of surface plasmon polaritons to deliver optical excitation at nanoscale, we introduce a technique relying on a new transport phenomenon with chemical sensitivity and nanometer spatial resolution. The second part describes how micro and nanostructured superhydrofobic textures can concentrate and localize a small number of molecules into a well-defined region, even when only an extremely diluted solution is available. Several applications of these devices as micro- and nano-systems for high-resolution imaging techniques, cell cultures and tissue engineering applications are also discussed.

Item does not contain fulltext Nail-patella syndrome (NPS), an autosomal dominant disorder characterized by nail dysplasia, absent or hypoplastic patellae, iliac horns, and often associated with nephropathy and, less frequently, with open angle glaucoma, is caused by mutations in the LMX1B gene. Inter-familial and intra-familial phenotypic variability raises the question whether modifier genes can be identified to explain differences in the expression and severity of clinical features of NPS. Genes encoding proteins that interact with the LMX1B protein are good candidates and, therefore, methods to search for interactions can be used to this purpose. By the yeast two-hybrid screening we detected the CLIM2 gene as a LMX1B interactor, confirming previous reports which described the same interaction by biochemical methods. Sequencing of the CLIM2 coding region in seven NPS cases in which no LMX1B mutation had been found, did not detect any molecular variant in these patients. Moreover, by genotyping a polymorphic dinucleotide repeat close to the CLIM2 gene in affected members of a large Dutch NPS family with high incidence of nephropathy, we were unable to find a correlation between the presence of a specific allele and the expression of nephropathy. In conclusion, although the results of this study could not provide any proof of CLIM2 involvement in the pathogenesis of NPS or in determination of the clinical phenotype, we suggest that the CLIM2 gene can be considered as a good candidate for further studies on normal and disturbed kidney development associated with NPS or other hereditary glomerulopathies.

We report on an Italian boy with the Meier-Gorlin syndrome (ear-patella-short stature syndrome). This rare autosomal recessive disorder comprises the triad of microtia, absent patellae, and growth retardation with prenatal onset. The patient had also an acute torsion of his left spermatic cord, a condition related to a congenital defect of the tunica vaginalis. Because this syndrome had been suggested as the human equivalent of the short ear mouse [Lacombe et al., 1994: Ann. Genet. 37:184-191], a mutation analysis of the BMP5 gene was performed and found normal. The LMX1B and the SHOX genes were also evaluated considering the absent patellae and short stature, respectively, and were found normal as well.

A prototype for a DNA origami nanorobot is designed, produced, and tested. The cylindrical nanorobot (diameter of 14 nm and length of 48 nm) with a switchable flap, is able to respond to an external stimulus and reacts by a physical switch from a disarmed to an armed configuration able to deliver a cellular compatible message. In the tested design the robot weapon is a nucleic acid fully contained in the inner of the tube and linked to a single point of the internal face of the flap. Upon actuation the nanorobot moves the flap extracting the nucleic acid that assembles into a hemin/G-quadruplex horseradish peroxidase mimicking DNAzyme catalyzing a colorimetric reaction or chemiluminescence generation. The actuation switch is triggered by an external nucleic acid (target) that interacts with a complementary nucleic acid that is beard externally by the nanorobot (probe). Hybridization of probe and target produces a localized structural change that results in flap opening. The flap movement is studied on a two-dimensional prototype origami using Förster resonance energy transfer and is shown to be triggered by a variety of targets, including natural RNAs. The nanorobot has potential for in vivo biosensing and intelligent delivery of biological activators. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Graphical abstractDisplay Omitted Superhydrophobic surface constituted with micropillars.Single strand DNA and dsDNA spectroscopic analysis.Low concentrated biomolecules Raman analysis.Non-enhanced Raman spectroscopy for DNAs molecules.Distinction of ss and dsDNAs via Raman spectroscopy. A novel approach for the study of low concentrated DNAs (60pM) using microRaman spectroscopy is reported. A superhydrophobic substrate with array of microPillars is fabricated over which the sample was drop casted. The substrate concentrates the molecules in a very small area with higher molecular density, enabling to carry out the microRaman measurements. Two different DNAs (single strand and double strand) were used to investigate through Raman technique. A spectral Raman difference was found to distinguish the ssDNA and dsDNAs. The approach can be of interest for a wide variety of applications ranging from biological materials interactions characterization to the biomedical field.

Superhydrophobicity refers to surfaces on which drops assume a quasi-spherical shape and a high contact angle (more than 150°). This well-known phenomenon occurs in nature and we can take advantage of this principle fabricating bio-inspired superhydrophobic surfaces constituted of ordinate arrays of silicon micro-pillars.

Recent advances in the single-molecule detection and manipulation provided unexpected solutions for the understanding of the physio-pathological behavior of individual biological macromolecules. Modern techniques of patterning at the micro- and nanometer scale combined with chemical treatments are being used to create surfaces that stretch the hydrophobic behavior to the limit. The ability to create surfaces with high static water contact angles (usually greater than 150°) is essential for a variety of applications, ranging from the development of biosensors to the implementation of sensitive and reliable single-molecule collection and sample preparation methods. Thus, superhydrophobic devices could be considered as nano-biotechnological single-molecule detection tools that can be applied to a wide range of high-resolution studies. To outline the paper, single-molecule detection topics and theoretical principles of superhydrophobicity are first introduced. A comprehensive overview is then given, describing how different types of devices with superhydrophobic surfaces are realized. Finally, the usefulness of the presented devices for a wide range of applications and the concluding comments are proposed.

Tetralogy of Fallot (TOF) (OMIM #187500) is the most frequent conotruncal congenital heart defect (CHD) with a range of intra- and extracardiac phenotypes. TBX5 is a transcription factor with well-defined roles in heart and forelimb development, and mutations in TBX5 are associated with Holt-Oram syndrome (HOS) (OMIM#142900). Here we report on the screening of 94 TOF patients for mutations in TBX5, NKX2.5 and GATA4 genes. We identified two heterozygous mutations in TBX5. One mutation was detected in a Moroccan patient with TOF, a large ostium secundum atrial septal defect and complete atrioventricular block, and features of HOS including bilateral triphalangeal thumbs and fifth finger clinodactyly. This patient carried a previously described de novo, stop codon mutation (p.R279X) located in exon 8 causing a premature truncated protein. In a second patient from Italy with TOF, ostium secundum atrial septal defect and progressive arrhythmic changes on ECG, we identified a maternally inherited novel mutation in exon 9, which caused a substitution of a serine with a leucine at amino acid position 372 (p.S372L, c.1115C>T). The mother's clinical evaluation demonstrated frequent ventricular extrasystoles and an atrial septal aneurysm. Physical examination and radiographs of the hands showed no apparent skeletal defects in either child or mother. Molecular evaluation of the p.S372L mutation demonstrated a gain-of-function phenotype. We also review the literature on the co-occurrence of TOF and HOS, highlighting its relevance. This is the first systematic screening for TBX5 mutations in TOF patients which detected mutations in two of 94 (2.1%) patients. © 2014 Wiley Periodicals, Inc

From single molecule to suspended DNA nanowires How DNA can be suspended in ordered nanometric bundles Andrea Falqui, Monica Marini, Marco Francardi and Enzo di Fabrizio 1 Biological and Environmental Sciences and Engineering Division, King Abdullah University for Science and Technology, Thuwal 23955-6900, Saudi Arabia 2 Physical Sciences and Engineering Division, King Abdullah University for Science and Technology, Thuwal 23955-6900, Saudi Arabia * Corresponding author: Falqui, A. (andrea.falqui@kaust.edu.sa)

Background Deletion of phenylalanine 508 is the most frequent mutation causing cystic fibrosis. It causes multiple defects: 1) misfolding of the protein causing retention at the ER (processing defect); 2) reduced channel activity (gating defect); 3) reduced plasma membrane residency time due to increased internalization rate and defective recycling. Methods Druggability of F508del-CFTR was demonstrated by several studies. Correctors are molecules able to improve maturation and trafficking to the membrane of F508del- CFTR. Correctors could act as pharmacological chaperones or as proteostasis regulators. Pharmacological chaperones act directly on mutant CFTR, while proteostasis regulators modify the proteostasis environment leading to beneficial effects on CFTR maturation. Results The use of a single compound is not sufficient to promote a therapeutically relevant F508del-CFTR rescue. Drug therapy for CF will require combinations of correctors exploiting different mechanisms of action, i.e. pharmacological chaperones combined together or with a proteostasis regulator. Conclusion Development of more effective CF drugs could therefore rely on a better understanding of the molecular events underlying CFTR processing/degradation. This review will focus on most promising pathways and related targets for the development of novel CF pharmacotherapies.

Background. Benzodiazepines are among the most commonly prescribed drugs in Italy and they are often used inappropriately according to guidelines for their rational use.The aim of this study was to investigate the prevalence and pattern of use of benzodiazepine amongst the general population aged 65-84 years in the Friuli-Venezia Giulia Region, in North-East Italy. Methods. A total of 40 general practitioners participated in the study. Two data sources were used in the research. The first was the Health Search Database, the second was a short questionnaire administered by the general practitioners to the 65 to 84 year old patients attending their surgeries for any reason during the study period. Data on the use of benzodiazepines between 1st February and 31st July 2001 were extracted from both the Health Search Database using drug prescriptions and the questionnaires.Results. Of the 10,468 patients aged 65-84 years with complete demographical data in the general practitioners’ patient lists, 2,369 subjects used benzodiazepines, hypnotics and over the counter drugs. Overall prevalence of benzodiazepine use was 21.5% (95% confidence interval: 19.8-23.1%). Of the benzodiazepine users, 66.9% consumed a short-intermediate half-life and 33.1% a long half-life benzodiazepine. Most patients took benzodiazepines at night (68.2%), less frequently in the daytime and at night (23.7%), or in the daytime only (8.1%). Most users (89.2%) said they had been taking benzodiazepine for years.Conclusions. Benzodiazepine use was associated with patient characteristics, such as being female, using analgesics or antidepressants and the presence of a chronic disease especially cancer or chronic heart failure.

Summary Cankers and dieback on London plane caused by Diaporthe scabra have been observed in urban plantings in Catania, Italy. Symptoms were delayed spring flush, trunk cankers, small chlorotic leaves associated with dead terminal or lateral branches and an early defoliation. Isolations from infected wood on PDA led to the isolation of the anamorph Phomopsis scabra with α-conidia production. The infected wood placed in a moist chamber developed pycnidia with α and β conidia and perithecia of the teleomorph D. scabra. Sequence analysis of the internal transcribed spacers ITS1 and ITS2, the ribosomal intragenic spacer beta-tubulin 1 and beta-tubulin 2 genes, congruently indicated that the D. scabra isolate is phylogenetically related to two strains of D. helianthi isolated in Italy from sunflower and to strains of D. eres. Inoculations on young plants of London plane growing in pots confirmed the pathogenicity of the isolated fungus. To our knowledge, this is the first report of the disease in Italy.