Bisperoxovanadium, a phospho-tyrosine phosphatase inhibitor, reprograms myogenic cells to acquire a pluripotent, circulating phenotype.

Satellite cells are the main source of myogenic progenitors in postnatal skeletal muscle, but their use in cell therapy for muscle disorders is limited because these cells cannot be delivered through circulation and they are rapidly exhausted in severe myopathies. The search for alternative donor cells is ongoing, but none of the candidates so far show all the features required for successful colonization and repair of diseased muscle. In this study, we show that bisperoxovanadium, a phospho-tyrosine phosphatase inhibitor, induces myogenic cells to acquire a gene expression profile and a differentiation potential consistent with the phenotype of a circulating precursors, while maintaining their myogenic potential. These effects are mediated, at least in part, by NF-kappaB activation through the Tyr42-IkappaB-alpha phosphorylation, as shown by the expression of the dominant negative mutant form of the p50 NF-kappaB subunit. Moreover, when bisperoxovanadium-treated cells are injected into the femoral artery of alpha-sarcoglican null dystrophic mice, they are able to circulate and to reach muscle tissue; importantly, they contribute to muscle regeneration, as shown by the expression of alpha-sarcoglican in some fibers. Our observations indicate that bisperoxovanadium, or similar compounds, may prove very valuable to obtain and to expand, from committed cells, multipotent cell populations suitable for gene-cell therapy applications and may help to understand the molecular basis of genome reprogramming and "stem-ness."

Engineering of FePt nanoparticles by e-beam co-evaporation.

Fe(50)Pt(50) nanoparticles were deposited on thermally oxidized Si substrates by electron-beam co-evaporation of Fe and Pt, at substrate temperatures T(s) between 300 and 700 degrees C. The co-deposition led to the formation of drop-like, coalesced nanoparticles, chain-like structures or continuous films, the morphology being dependent on T(s) or the nominal thickness of the layer, f. The nanoparticles have a mean diameter D(p) between 3 and 45 nm, which increases with increasing f. The degree of crystallization in the ordered face centred tetragonal (fct) phase of the samples depends strongly on the growth conditions and increases with increasing T(s) and f. Nanoparticles with a higher proportion of the fct phase exhibit higher coercivity, with a maximum value of approximately 10.3 kOe (for the specimens prepared at 600 degrees C with f = 8.5 nm). Conversely, samples with a high proportion of the cubic phase are either superparamagnetic or ferromagnetically soft. The thermal annealing performed on selected samples resulted in structural transformation as well as magnetic hardening that depended on f and D(p).

Analysis of DNA oxidative damage related to cell proliferation.

In vivo and in vitro cell populations exhibit a different sensitivity and a heterogeneous response to many genotoxic agents. Several studies have been carried out to evaluate the possibility that the different sensitivity of the cells is related to their proliferative status. In this study, the sensitivity of proliferating (P) and quiescent (Q) C3H10T1/2 cells to oxidative damage and their repair capability has been investigated by single cell gel electrophoresis (SCGE) and micronucleus test. Furthermore the possibility to simultaneously detect DNA damage and cell cycle position has been evaluated. Our results showed a dose-related increase of DNA damage in exponential and plateau phase cells treated with hydrogen peroxide (doses ranging between 2.5 and 100 microM). DNA damage was almost completely repaired within 2 h after treatment in both culture conditions. The percentage of cells in the various phases of the cell cycle has been determined by comet assay and by flow cytometry, and a good agreement between the results of the two techniques was found. Untreated exponentially growing cells in G1 phase showed a lower tail moment than S and G2/M cells. The same cell cycle dependence was evidenced in cells treated with low doses of H(2)O(2), while, at the higher doses, all cells showed a similar level of damage. These results confirm the sensitivity of the Comet Assay in assessing DNA damage, and support its usefulness in evaluating cell cycle-related differential sensitivity to genotoxic agents.

Integrated techniques in the identification of ovarian tumors.

This work is intended to be an effort to standardise the diagnostic iter of ovarian pathology. This protocol is carried out with a clinical examination and later by USG and celioscopy. It tests the management of functional and endometriosic cysts and the other ovarian pathologies.

Echopatterns of the ovarian dermoid tumor.

The authors report the ovarian dermoid echopatterns observed in 189 cases. The ultrasonic key of the diagnosis is a particular "pearly-gray" colour. The early diagnosis by USG gives the Gynecologist some advantages in: a) avoiding the operation in those cases of small mass; b) making a pfammenstiel incision in the surgical operation; c) trying a conservative surgery.

FePt and CoPt nanoparticles co-deposited on silicon dioxide-a comparative study.

We compare CoPt and FePt nanoparticles grown under identical conditions on oxidized Si substrates by electron beam co-evaporation. Growth was performed under high vacuum conditions at substrate temperatures of 1023 K and was immediately followed by an annealing step. This process forms CoPt and FePt nanoparticles with mean diameters between ∼17 and ∼22 nm. In particular, the annealing step results in grain size enlargement for all samples and in a progressive magnetic hardening of the nanoparticles which reach maximum perpendicular coercivities of ∼6.6 kOe (for the CoPt) and ∼10.2 kOe (for the FePt nanoparticles). We show that, during this annealing step, a progressive transition towards the hard magnetic L1(0) ordered phase takes place in both materials. In contrast to FePt, CoPt nanoparticles must be annealed in order to crystallize in this phase.

Protein kinase C theta co-operates with calcineurin in the activation of slow muscle genes in cultured myogenic cells.

Adult skeletal muscle fibers can be divided into fast and slow twitch subtypes on the basis of specific contractile and metabolic properties, and on distinctive patterns of muscle gene expression. The calcium, calmodulin-dependent protein phosphatase, calcineurin, stimulates slow fiber-specific genes (myoglobin (Mb), troponin I slow) in cultured skeletal muscle cells, as well as in transgenic mice, through the co-operation of peroxisome-proliferation-activator receptor gamma co-activator 1alpha (PGC1alpha) myocyte enhancer factor 2 (MEF2), and nuclear factor of activated T cells (NFAT) transcription factors. Specific protein kinase C isoforms have been shown to functionally co-operate with calcineurin in different cellular models. We investigated whether specific protein kinase C isoforms are involved in calcineurin-induced slow skeletal muscle gene expression. By pharmacological inhibition or exogenous expression of mutant forms, we show that protein kinase C theta (the protein kinase C isoform predominantly expressed in skeletal muscle) is required and co-operates with calcineurin in the activation of the Mb promoter, as well as in the induction of slow isoforms of myosin and troponin I expression, in cultured muscle cells. This co-operation acts primarily regulating MEF2 activity, as shown by using reporter gene expression driven by the Mb promoter mutated in the specific binding sites. MEF2 activity on the Mb promoter is known to be dependent on both PGC1alpha and inactivation of histone deacetylases (HDACs) activity. We show in this study that protein kinase C theta is required for, even though it does not co-operate in, PGC1alpha-dependent Mb activation. Importantly, protein kinase C theta regulates the HDAC5 nucleus/cytoplasm location. We conclude that protein kinase C theta ensures maximal activation of MEF2, by regulating both MEF2 transcriptional complex formation and HDACs nuclear export.