Massimo Gulisano

Exposure to global system for mobile communication (GSM) cellular phone radiofrequency alters gene expression, proliferation, and morphology of human skin fibroblasts.

Human skin fibroblasts were exposed to global system for mobile communication (GSM) cellular phone radiofrequency for 1 h. GSM exposure induced alterations in cell morphology and increased the expression of mitogenic signal transduction genes (e.g., MAP kinase kinase 3, G2/mitotic-specific cyclin G1), cell growth inhibitors (e.g., transforming growth factor-beta), and genes controlling apoptosis (e.g., bax). A significant increase in DNA synthesis and intracellular mitogenic second messenger formation matched the high expression of MAP kinase family genes. These findings show that these electromagnetic fields have significant biological effects on human skin fibroblasts.

Efficacy of eradicative radiotherapy for limited nodal metastases detected with choline PET scan in prostate cancer patients.

Aims and background In patients with recurrent prostate cancer, discriminating local or systemic recurrence is critical to decide second-line treatment. We investigated the capability of stereotactic body radiotherapy to treat limited nodal recurrences, detected using choline PET scan. Methods and study design Seventy-one patients with biochemical failure were studied after prostate cancer treatment: prostatectomy (28), radiotherapy (15) or both (28). Following computed tomography and choline PET imaging, stereotactic body radiotherapy was delivered on pathological lymphatic areas by 6 MV Linac, using dynamic micromultileaf collimation and intensity-modulated arc therapy optimization. Sixty days post-treatment, choline PET/CT imaging was carried out. Results Median follow-up was 29 months (range, 14.4–48). Choline PET detected recurrences in 39 of 71 patients. Median PSA velocity was 0.40 ng/ml/year in PET-negative patients and 2.88 ng/ml/year in PET-positive subjects (P <0.05). Twenty-five patients with limited nodal recurrences, out of the 71 submitted to choline PET, received eradicative radiotherapy. Persistent regression was recorded in 13; early spread to bone was found in 2 cases; lymph node recurrences in 8, all in sites outside the irradiated areas; 2 patients were lost to follow-up. At the 3-year follow-up, overall survival, disease-free survival and local control rates were 92%, 17% and 90%, respectively. In patients with a complete regression, PSA fell from 5.65 to 1.40 ng/ml (median). PSA nadir value (median 1.06 ng/ml) was maintained for 5.6 months (median). Conclusions Stereotactic body radiotherapy was effective in disease eradication of limited nodal recurrences from prostate cancer, saving patients from, or at least postponing, systemic treatments. Free full text available at www.tumorionline.it

EFFECT OF 0.2 T STATIC MAGNETIC FIELD ON HUMAN NEURONS : REMODELING AND INHIBITION OF SIGNAL TRANSDUCTION WITHOUT GENOME INSTABILITY

We describe the effect of the static magnetic field generated by a 0.2 T magnetic resonance tomograph on a normal human neuronal cell culture (FNC-B4). After 15 min exposure cells showed dramatic changes of morphology: they formed vortexes of cells and exposed branched neurites featuring synaptic buttons. At the same time, thymidine incorporation and inositol lipid signaling were significantly reduced. Control (sham exposed) or non-neuronal cells (mouse leukemia, and human breast carcinoma cells) did not show any alteration following exposure. Endothelin-1 release from FNC-B4 cells was also dramatically reduced after 5 min exposure. However, PCR analysis of 12 DNA microsatellites selected as gauges of genome instability, did not reveal any alteration following exposure, thus ruling out a direct effect of the magnetic field on DNA stability. These data can be interpreted as a specific effect of the static magnetic field on human neuronal cells and are consistent with the induction of remodeling and differentiation; they demonstrate that fields below 0.5 T have significant biological effects on human neurons.

Effects of 0.2 T static magnetic field on human skin fibroblasts

Human skin fibroblasts were exposed to 0.2 T static magnetic field generated by a magnetic resonance tomograph. After 1h exposure, cell morphology was modified in association with a concomitant decrease in the expression of some sugar residues of glycoconjugates. Study of cell proliferation and mitogenic signal transduction showed a decrease of thymidine incorporation and of second messenger formation. However, cell viability, assessed by colony forming assay, was unaffected. These results demonstrate that the static magnetic field generated by routinely used magnetic resonance tomograph induces alterations on human skin fibroblasts.

Proteomic analysis and protein carbonylation profile in trained and untrained rat muscles

Understanding the relationship between physical exercise, reactive oxygen species and skeletal muscle modification is important in order to better identify the benefits or the damages that appropriate or inappropriate exercise can induce. Unbalanced ROS levels can lead to oxidation of cellular macromolecules and a major class of protein oxidative modification is carbonylation. The aim of this investigation was to study muscle protein expression and carbonylation patterns in trained and untrained animal models. We analyzed two muscles characterized by different metabolisms: tibialis anterior and soleus. Whilst tibialis anterior is mostly composed of fast-twitch fibers, the soleus muscle is mostly composed of slow-twitch fibers. By a proteomic approach we identified 15 protein spots whose expression is influenced by training. Among them in tibialis anterior we observed a down-regulation of several glycolitic enzymes. Concerning carbonylation, we observed the existence of a high basal level of protein carbonylation. Although this level shows some variation among individual animals, several proteins (mostly involved in energy metabolism, muscle contraction, and stress response) appear carbonylated in all animals and in both types of skeletal muscle. Moreover we identified 13 spots whose carbonylation increases after training.

Transdermal Delivery of Heparin Using Pulsed Current Iontophoresis

PurposeIn clinical practice heparin has to be administered by injection with obvious disadvantages; thus, transdermal delivery by electrically assisted methods have been studied. In this study we evaluated the efficacy of a Food and Drug Administration-approved pulsed current iontophoresis system in delivering heparin through living rat skin.MethodsFluorescent and radioactive heparin as well as a commercial heparin preparation were delivered through rat skin via a pulsed current iontophoresis system.ResultsPulsed current iontophoresis allowed fluorescent heparin to cross the stratum corneum localizing in epidermis and dermis. Unfractionated, high-, and low molecular weight fraction pools, obtained by fractionating [35S]-unfractionated heparin on a molecular weight sieve, were then separately tested. Pulsed current iontophoresis elicited the transdermal delivery of low molecular weight heparin, but not that of high molecular weight heparin. Finally, pulsed current iontophoresis of an unfractionated pharmaceutical heparin preparation significantly decreased plasmatic factor Xa activity.ConclusionsWe hypothesize that this technique could be used to administer low molecular weight heparin in a cost-efficient and safe manner without the need for syringes and needles.

Cadmium modulates proliferation and differentiation of human neuroblasts

Cadmium is an environmental pollutant inducing numerous pathological effects, including neurological disorders and brain diseases. However, little is known about the molecular mechanisms of cadmium in affecting neurons and in inducing neurotoxicity in the development of the human brain. We have recently established, cloned, and propagated in vitro a primary long‐term cell culture (FNC‐B4) obtained from the human fetal olfactory neuroepithelium. In the present study, we show that different concentrations of cadmium chloride (CdCl2) induced dose‐dependent biological effects in FNC‐B4 cells. A low concentration (10 μM) of CdCl2 stimulated neuroblast growth, whereas a high concentration (100 μM) inhibited the growth and the viability of neuroblasts inducing morphological and cytoskeletal alterations as well as apoptotic cell death. We also observed that CdCl2 affected, in a dose‐dependent manner, the differentiation of FNC‐B4 neuroblasts, with increased mRNA and protein levels of differentiation markers and decreased expression levels of neuronal stem markers. Furthermore, differentiated cells co‐expressed glial and neuronal markers. We suggest that CdCl2 in FNC‐B4 neuroblasts might represent a selective cue by which, in a heterogeneous primary culture, the more differentiated mature cells die, whereas the undifferentiated cells, at the same time glial and neuronal progenitors, are forced to access a state of differentiation.

Internal jugular vein valves: an assessment of prevalence, morphology and competence by Color Doppler Echography in 240 healthy subjects

The presence of valves in the head and neck veins is known since classical anatomical dissections. Previous studies have investigated whether jugular veins show constant valves at their ostium and whether these valves are physiologically competent, but, unluckily, these studies have reported conflicting results. Further, recent observations have raised the question whether the incompetence of jugular vein valves might play a pivotal role in neurological disorders related to venous engorgement of the brain. In this study we examined 462 internal jugular veins by using an echocolorodoppler apparatus. In particular, we assessed the presence, morphology and competence of valves at their ostium. Unilateral jugular vein valves were present in 406 cases (88%), mainly on the right side. The most frequently observed morphology (75%, 305 cases) was the two-leaflet valve, and jugular vein valves were incompetent in the huge majority of cases (365 cases, 90%). Our findings confirm the anatomical variability predicted from classical anatomical studies but, unluckily, do not provide additional evidence on the possible role of jugular vein valves in physiology and pathology. Further studies are strongly needed to determine whether these valves actually play an important role in counteracting chest venous pressure and in preventing reflux towards the brain.

A Novel Role for a Major Component of the Vitamin D Axis: Vitamin D Binding Protein-Derived Macrophage Activating Factor Induces Human Breast Cancer Cell Apoptosis through Stimulation of Macrophages

The role of vitamin D in maintaining health appears greater than originally thought, and the concept of the vitamin D axis underlines the complexity of the biological events controlled by biologically active vitamin D (1,25(OH)(2)D3), its two binding proteins that are the vitamin D receptor (VDR) and the vitamin D-binding protein-derived macrophage activating factor (GcMAF). In this study we demonstrate that GcMAF stimulates macrophages, which in turn attack human breast cancer cells, induce their apoptosis and eventually phagocytize them. These results are consistent with the observation that macrophages infiltrated implanted tumors in mice after GcMAF injections. In addition, we hypothesize that the last 23 hydrophobic amino acids of VDR, located at the inner part of the plasma membrane, interact with the first 23 hydrophobic amino acids of the GcMAF located at the external part of the plasma membrane. This al1ows 1,25(OH)(2)D3 and oleic acid to become sandwiched between the two vitamin D-binding proteins, thus postulating a novel molecular mode of interaction between GcMAF and VDR. Taken together, these results support and reinforce the hypothesis that GcMAF has multiple biological activities that could be responsible for its anti-cancer effects, possibly through molecular interaction with the VDR that in turn is responsible for a multitude of non-genomic as well as genomic effects.

Iron Indices and Vitamin D Receptor Polymorphisms in Hemodialysis Patients

Cardiovascular disease caused by accelerated atherosclerosis is the major determinant of morbidity and mortality in chronic kidney disease patients. Vitamin D and its analogs provide survival benefit for hemodialysis (HD) patients. Vitamin D exerts its effects through the vitamin D receptor (VDR) that is coded for by a gene showing several polymorphisms that, in turn, are associated with a variety of diseases and differential responses to vitamin D. In this study, we evaluated the association between 4 VDR polymorphisms (ie, those identified by the restriction enzymes BsmI, ApaI, TaqI, and FokI) and iron indices (serum iron, transferrin, transferrin saturation, and ferritin) in 88 hemodialysis patients routinely treated with vitamin D. The absence or presence of the BsmI, ApaI, TaqI, and FokI restriction sites were denominated B and b, A and a, T and t, F and f, respectively. Our results show that in HD patients with transferrin saturation <20%, the F allele was more frequent than in HD patients with transferrin saturation >20% (P = .03). This relationship may provide a link between VDR alleles and iron and nutritional markers, which are highly predictive variables of cardiovascular morbidity and mortality in hemodialysis patients.