M. Chiarpotto

Transient state kinetic investigation of ferritin iron release

Increased iron concentration in tissues appears to be a factor in the genesis and development of inflammatory and degenerative diseases. By means of real-time small angle x-ray scattering measurements, we studied the kinetics of iron release from the ferritin inorganic core as a function of time and distance from the iron core centre. Accordingly, the iron release process follows a three step model: (i) a defect nucleation in the outer part of the mineral core, (ii) the diffusion of the reducing agent towards the inner part of the core, and (iii) the erosion of the core from the inner to the outer part.

Self-assembling of large ordered DNA arrays using superhydrophobic patterned surfaces

In this paper we present a simple and robust method to realize highly ordered arrays of stretched and suspended DNA molecules over the millimeter length scale. To this end we used an ad hoc designed superhydrophobic surface made of high aspect-ratio silicon pillars, where we deposited a droplet containing genomic DNA. A precise positioning of DNA strands was achieved by shaping the silicon pillars so that sharpened features resembling tips were included. Such features allowed us to accurately control the droplet de-wetting dynamics, pinning DNA strands in a well-defined position above pillars. The proposed technique has the potential to positively impact on the development of novel DNA chips for genetic analysis.

Quantitative analysis of autophagic flux by confocal pH-imaging of autophagic intermediates

Although numerous techniques have been developed to monitor autophagy and to probe its cellular functions, these methods cannot evaluate in sufficient detail the autophagy process, and suffer limitations from complex experimental setups and/or systematic errors. Here we developed a method to image, contextually, the number and pH of autophagic intermediates by using the probe mRFP-GFP-LC3B as a ratiometric pH sensor. This information is expressed functionally by AIPD, the pH distribution of the number of autophagic intermediates per cell. AIPD analysis reveals how intermediates are characterized by a continuous pH distribution, in the range 4.5–6.5, and therefore can be described by a more complex set of states rather than the usual biphasic one (autophagosomes and autolysosomes). AIPD shape and amplitude are sensitive to alterations in the autophagy pathway induced by drugs or environmental states, and allow a quantitative estimation of autophagic flux by retrieving the concentrations of autophagic intermediates.

Mechanism of aluminium bio-mineralization in the apoferritin cavity

Many experimental evidences point out the correlation between the presence of aluminum-ferritin complex and neursopathological disorders. In these complexes, two different ranges of Aluminium (Al) atoms are usually found, i.e., just few atoms or several hundreds. Here, we investigated the in-vitro Al-apoferritin binding, with the aim to elucidate the mechanism behind the formation of Al-ferritin complexes in-vivo. To this purpose, we studied the mineralization of Al in its ionic and complexed form with citrate demonstrating that high Al levels found in clinical studies can be obtained only conveying Al by small physiological ligands.

Controlling the Cassie-to-Wenzel Transition: an Easy Route towards the Realization of Tridimensional Arrays of Biological Objects

In this paper we provide evidence that the Cassie-to-Wenzel transition, despite its detrimental effects on the wetting properties of superhydrophobic surfaces, can be exploited as an effective micro-fabrication strategy to obtain highly ordered arrays of biological objects. To this purpose we fabricated a patterned surface wetted in the Cassie state, where we deposited a droplet containing genomic DNA. We observed that, when the droplet wets the surface in the Cassie state, an array of DNA filaments pinned on the top edges between pillars is formed. Conversely, when the Cassie-to-Wenzel transition occurs, DNA can be pinned at different height between pillars. These results open the way to the realization of tridimensional arrays of biological objects.

Controlling DNA Bundle Size and Spatial Arrangement in Self-assembled Arrays on Superhydrophobic Surface

The use of superhydrophobic surfaces (SHSs) is now emerging as an attractive platform for the realization of one-dimensional (1D) nanostructures with potential applications in many nanotechnological and biotechnological fields. To this purpose, a strict control of the nanostructures size and their spatial arrangement is highly required. However, these parameters may be strongly dependent on the complex evaporation dynamics of the sessile droplet on the SHS. In this work, we investigated the effect of the evaporation dynamics on the size and the spatial arrangement of self-assembled 1D DNA bundles. Our results reveal that different arrangements and bundle size distributions may occur depending on droplet evaporation stage. These results contribute to elucidate the formation mechanism of 1D nanostructures on SHSs.