Giorgio Scarì

Growth factors, CD34 positive cells, and fibrin network analysis in concentrated growth factors fraction.

An interesting clinical option for optimizing healing tissue is the use of platelet concentrate. Platelets contain high quantities of growth factors, among these TGF‐β1 and VEGF, which are known to be implicated in tissue regeneration. CGF is produced by processing blood samples with a special centrifuge device; three layers are formed: top acellular plasma (PPP), middle CGF and bottom red blood cells (RBC) layers. Given that to date there are no data concerning the biological characteristic of CGF, the aim of this study was to evaluate the presence of TGF‐β1 and VEGF in CGF and also in PPP and RBC layers. In addition, since circulating stem cells are recruited from blood to injured tissue for healing we also evaluated the presence of CD34 positive cells. Our data show the presence of TGF‐β1 and VEGF in CGF and RBC layers. In addition, we show CD34 positive cells in CGF. Microsc. Res. Tech., 2011.

Phagocytosis of Biocompatible Gold Nanoparticles

We report the evidence for the cellular uptake of gold nanoparticles via the phagocytosis mechanism in murine macrophage cells strongly supported by TEM and optical microscopy. Nanoparticles were prepared using several biocompatible molecules of choice (5-aminovaleric acid, l-DOPA, melatonin, and serotonin hydrochloride) as stabilizers for gold colloids. Their surface chemistry was fully characterized by UV-vis, ATR-FTIR, (1)H NMR, and HR-MAS (1)H NMR spectroscopies, and size distribution was determined by CPS disc centrifuge and TEM. Differences in coatings were evaluated against cellular uptake, and a preferential movement of macrophages toward 5-aminovaleric acid-modified gold nanoparticles was shown, leading to the fast accumulation of nanoparticles in the cytosol.

Gold nanoparticles capped by a GC-containing peptide functionalized with an RGD motif for integrin targeting.

Gold nanoparticles were obtained by reduction of a tetrachloroaurate aqueous solution in the presence of a RGD-(GC)(2) peptide as stabilizer. As comparison, the behavior of the (GC)(2) peptide has been studied. The (GC)(2) and RGD-(GC)(2) peptides were prepared ad hoc by Fmoc synthesis. The colloidal systems have been characterized by UV-visible, TGA, ATR-FTIR, mono and bidimensional NMR techniques, confocal and transmission (TEM) microscopy, ζ-potential, and light scattering measurements. The efficient cellular uptake of Au-RGD-(GC)(2) and Au-(GC)(2) stabilized gold nanoparticles into U87 cells (human glioblastoma cells) were investigated by confocal microscopy and compared with the behavior of (GC)(2) capped gold nanoparticles. A quantitative determination of the nanoparticles taken up has been carried out by measuring the pixel brightness of the images, a measure that highlighted the importance of the RGD termination of the peptide. Insight in the cellular uptake mechanism was investigated by TEM microscopy. Various important evidences indicated the selective uptake of RGD-(GC)(2) gold nanoparticles into the nucleus.

Observations on the settlement of phallusia mammillata larvae: Effects of different lithological substrata

Abstract In the sessile marine tunicates, the selection of a suitable substratum by the larvae is an important and critical factor determining the distribution of species. The present paper investigated, under laboratory conditions, the role played by mineral content of the substratum in settlement of the ascidian Phallusia mammillata, using siliceous and carbonaceous stones. Individuals that attached to the different substrata and metamorphosed were scored. The data indicate that larvae could discriminate between the substrata on the basis of their silica content. Under the same laboratory conditions, the larvae that attached to siliceous stones grew faster and had a wider area of contact with the substratum than those that grew on carbonaceous stones. The present study concludes that silica is a mineral factor that can be discriminated by chemosensory palps of ascidian larvae during the choice of substratum. It is suggested that the mineral composition of the habitat can contribute, with other environmental factors, to regulate the spatial distribution of tunicate communities.

proPO system of Allogamus auricollis (Insecta): Effects of various compounds on phenoloxidase activity

The phenoloxidase activity in the hemolymph cell-free fraction from Allogamus auricollis was studied in the presence of Escherichia coli lipopolysaccharides and Saccharomyces cerevisiae β-1,3-glucans. The proPO system seems to be strongly activated by lipopolysaccharides (LPS). The basic activation observed in this model appears not to be affected by the use of protease inhibitors (α2 macroglobulin, soybean trypsin inhibitor), and, in addition, the LPS-activated proPO system is not inhibited by their presence. Calcium ions at high concentrations inhibit the phenoloxidase activity, and EDTA chelation strongly enhances dopachrome formation. Analytical polyacrylamide gel electrophoresis (PAGE) of the hemolymph cell-free fraction showed two main components, with a molecular mass of 76 and 80 kDa. After electro-elution from a native PAGE of L-dihydroxy-phenylalanine positive bands, the analytical SDS-PAGE again showed the same two major bands.

Gordius villoti (Nematomorpha) life cycle in relation with caddis fly larvae

Abstract The relationships between Gordius villoti Rosa and Allogamus auricollis Pictet larvae are described. The horsehair worms and host life cycles have been investigated in laboratory and field conditions and aspects of their morphology and behaviour are discussed.

Doxorubicin affects actin assembly in vitro

In vitro experiments on actin polymerization in the presence of doxorubicin show that the rate of salt-induced actin assembly is negatively affected by the drug. The decreased amount of actin monomers keeping their ability to self-interact to give F actin (microfilaments) probably explains the reduction of assembly value. Drug action is dose-dependent and various discrepancies are explained by the limitations of the techniques used.

Biochemical evidence of phenoloxidase activity (pro-PO system) in larvae of Allogamus auricollis (Insecta, Trichoptera)

Abstract 1. 1. Allogamus auricollis cell-free hemolymph proteins were analyzed by SDS-PAGE. Under reducing conditions the gel pattern showed two main components (83 and 76 kDa) and some lesser bands. 2. 2. After native PAGE, a single band showed phenoloxidase activity by in situ enzymatic staining. 3. 3. Spectrophotometric analysis of the cell-free plasma fraction was carried out with substrate and PTU as inhibitor.

Thermal equilibrium and temperature differences among body regions in European plethodontid salamanders.

Information on species thermal physiology is extremely important to understand species responses to environmental heterogeneity and changes. Thermography is an emerging technology that allows high resolution and accurate measurement of body temperature, but until now it has not been used to study thermal physiology of amphibians in the wild. Hydromantes terrestrial salamanders are strongly depending on ambient temperature for their activity and gas exchanges, but information on their body temperature is extremely limited. In this study we tested if Hydromantes salamanders are thermoconform, we assessed whether there are temperature differences among body regions, and evaluated the time required to reach the thermal equilibrium. During summers of 2014 and 2015 we analysed 56 salamanders (Hydromantes ambrosii and Hydromantes italicus) using infrared thermocamera. We photographed salamanders at the moment in which we found them and 1, 2, 3, 4, 5 and 15min after having kept them in the hands. Body temperature was equal to air temperature; salamanders attained the equilibrium with air temperature in about 8min, the time required to reach equilibrium was longer in individuals with large body size. We detected small temperature differences between body parts, the head being slightly warmer than the body and the tail (mean difference: 0.05°C). These salamanders quickly reach the equilibrium with the environment, thus microhabitat measurement allows obtaining accurate information on their tolerance limits.

Au– thymine , thymidine and thymidine 5′-monophosphate nanoparticles : chemical characterisation and cellular uptake studies into U87 cancer cells

Gold nanoparticles capped by thymine, thymidine and thymidine 5′-monophosphate were prepared and characterised by spectroscopy and microscopy techniques. AuNPs incubated with U87 cancer cells were taken up and showed a particular ability to overcome intracellular boundaries, being present in various cell compartments.