Nanostructures for SERS in living cell

Surface-enhanced Raman spectroscopy (SERS) has received renewed interest in recent years in fields such as trace analysis, biorelated diagnosis, and living cell study. However, the interference of impurities left on the surface from the preparation process of substrates limits to some extent the application of SERS. In the present paper, we propose a method to prepare clean SERS substrates by a combined method of hydrothermal green synthesis and thermal treatment to obtain a clean and impurity-free surface for SERS measurements, suitable for cells growth. The goal of such activity was the study of the membrane proteome, with special attention to prion protein (PrPC), in its physiological ambient. SERS has been used to evidence the PrPC-Cu(II) interaction in a rat neuroblastoma cell line (B104), known to overexpress the cellular prion protein PrPC

A HGF/cMET Autocrine Loop Is Operative in Multiple Myeloma Bone Marrow Endothelial Cells and May Represent a Novel Therapeutic Target

Purpose: The aim of this study was to investigate the angiogenic role of the hepatocyte growth factor (HGF)/cMET pathway and its inhibition in bone marrow endothelial cells (EC) from patients with multiple myeloma versus from patients with monoclonal gammopathy of undetermined significance (MGUS) or benign anemia (control group). Experimental Design: The HGF/cMET pathway was evaluated in ECs from patients with multiple myeloma (multiple myeloma ECs) at diagnosis, at relapse after bortezomib- or lenalidomide-based therapies, or on refractory phase to these drugs; in ECs from patients with MGUS (MGECs); and in those patients from the control group. The effects of a selective cMET tyrosine kinase inhibitor (SU11274) on multiple myeloma ECs' angiogenic activities were studied in vitro and in vivo. Results: Multiple myeloma ECs express more HGF, cMET, and activated cMET (phospho (p)-cMET) at both RNAand protein levels versus MGECs and control ECs. Multiple myeloma ECs are able to maintain the HGF/cMET pathway activation in absence of external stimulation, whereas treatment with anti-HGF and anti-cMET neutralizing antibodies (Ab) is able to inhibit cMET activation. The cMET pathway regulates several multiple myeloma EC activities, including chemotaxis, motility, adhesion, spreading, and whole angiogenesis. Its inhibition by SU11274 impairs these activities in a statistically significant fashion when combined with bortezomib or lenalidomide, both in vitro and in vivo. Conclusions: An autocrine HGF/cMET loop sustains multiple myeloma angiogenesis and represents an appealing new target to potentiate the antiangiogenic management of patients with multiple myeloma

Parvovirus B19 Infection Localized in the Intestinal Mucosa and Associated with Severe Inflammatory Bowel Disease▿

Infection by human parvovirus B19 is widespread and can be associated with a wide range of different pathologies and clinical manifestations. We provide the first evidence of localization of an active parvovirus B19 infection in the intestinal mucosa and its association with a severe inflammatory bowel disease, characterized by duodenal villous atrophy with increased intraepithelial lymphocytes and inflammatory infiltrates in the colonic mucosa. Virus in the intestinal mucosa was detected in cells of the inflammatory infiltrate, identified as T lymphocytes and selectively localized in sites of active tissue degeneration

Role of the Cellular Prion Protein in the neuron adaptation strategy to copper deficiency

CTR1 (Copper Transporter 1), PrPC (Cellular Prion Protein), NRAMP2 (Natural Resistance - Associated Macrophage Protein 2) and ATP7A proteins control the cell absorption and efflux of copper (Cu) ions in nervous tissues upon physiological conditions. Little is known about their regulation under reduced Cu availability, a condition underlying the onset of diffused neurodegenerative disorders. In the current study, rat neuron-like cells were exposed to Cu starvation for 48 h. The activation of Caspase-3 enzymes and the impairment of Cu,Zn Superoxide Dismutase (Cu,Zn SOD) activity depicted the initiation of a pro-apoptotic program, preliminary to the appearance of the morphological signs of apoptosis. The transcriptional response related to Cu transport proteins has been investigated. Notably, PrPC transcript and protein levels were consistently elevated upon Cu deficiency. The CTR1 protein amount was stable, despite a two-fold increase in the transcript amount, meaning the activation of post-translational regulatory mechanisms. NRAMP2 and ATP7A expression was unvaried. The up-regulated PrPC has been demonstrated to enhance the cell Cu uptake ability by about 50% with respect to the basal transport, and so sustain the Cu delivery to the Cu,Zn SOD cuproenzymes. Conclusively, the study suggests a pivotal role for PrPC in the cell adaptation to Cu limitation through a direct activity of ion uptake. In this view, the PrPC accumulation observed in several cancer cell lines could be interpreted as a molecular marker of cell Cu deficiency and a potential target of therapeutic interventions against disorders caused by metal imbalances

Metabolic phenotype of CD4+ T cells in Multiple Sclerosis subjects

Multiple sclerosis (MS) is a chronic inflammatory autoimmune demyelinating disease of the central nervous system. There are four clinical forms of MS, the most common of which is characterized by a relapsing remitting course (RRMS). The etiology of MS is unknown, but many studies suggested that genetic, environmental and infectious agents may contribute to the development of this disease. In experimental autoimmune encephalomyelitis (EAE), the animal model for MS, it has been shown that CD4+ T cells play a key role in MS pathogenesis. In fact, these cells are able to cross the blood-brain barrier and cause axonal damage with neuronal death. T cell activation critically depends on mitochondrial ATP synthesis and reactive oxygen species (ROS) production. Interestingly, lots of studies linked the oxidative damage arising from mitochondrial changes to neurodegenerative disorders, such as MS. Based on these evidences, this work focused on the metabolic reprogramming of CD4+ T cells in MS subjects, being this cell population directly implicated in pathogenesis of disease, paying attention to mitochondrial function and response to oxidative stress. Such aspects, once clarified, may open new opportunities for a therapeutic metabolic modulation of MS disorder

Molecular and functional expression of high conductance Ca 2+ activated K+ channels in the eel intestinal epithelium.

none8Several types of K+ channels have been identified in epithelial cells. Among them high conductance Ca2+- activated K+ channels (BK channels) are of relevant importance for their involvement in regulatory volume decrease (RVD) response following hypotonic stress. The aim of the present work was to investigate the functional and molecular expression of BK in the eel intestine, which is a useful experimental model for cell volume regulation research. In the present paper using rat BK channel-specific primer, a RT-PCR signal of 696 pb cDNA was detected in eel intestine, whole nucleotide sequence showed high similarity (83%) to the alpha subunit of BK channel family. BK channel protein expression was verified by immunoblotting and confocal microscopy, while the functional role of BK channels in epithelial ion transport mechanisms and cell volume regulation was examined by electrophysiological and morphometric analysis on the intact tissue. BKCa channels appeared to be localized along all the plasma membrane of the enterocytes; the apical part of the villi showed the most intense immunostaining. These channels were silent in basal condition, but were activated on both membranes (apical and basolateral) by increasing intracellular Ca2+ concentration with the Ca2+ ionophore ionomycin (1 µM). BKCa channels were also activated on both membranes by hypotonic swelling of the epithelium and their inhibition by 100 nM iberiotoxin (specific BKCa inhibitor) abolished the Regulatory Volume Decrease (RVD) of the intestinal cells after hypotonic swelling. In conclusion, our results demonstrated the molecular and functional expression of high conductance Ca2+ -activated K+ channels in eel intestine; the physiological role of these channels is mainly related to the RVD response of the epithelial cells following hypotonic swelling.I.F.(2007) = 3.557; Category = CELL BIOLOGY, Quartile in category = Q2 (from ISI Web of Knowledge). Category = PHYSIOLOGY, Quartile in category = Q2 (from ISI Web of Knowledge). Recentemente citato da Physiological Reviews (Physiol. Rev. 89: 193-277, 2009)M.G. LIONETTO; RIZZELLO A; GIORDANO M.E; M. MAFFIA; DE NUCCIO F; NICOLARDI G; HOFFMANN E.K; SCHETTINO TLionetto, Maria Giulia; Rizzello, Antonia; Giordano, Maria Elena; Maffia, Michele; DE NUCCIO, Francesco; Nicolardi, Giuseppe; HOFFMANN E., K; Schettino, Trifon

Sclerosi multipla. Le alterazioni metaboliche linfocitarie come potenziale strumento di follow-up della patologia

La Sclerosi Multipla (SM) è una malattia infiammatoria cronica demielinizzante, a carattere autoimmune, che colpisce il Sistema Nervoso Centrale (SNC). Nonostante siano stati effettuati una serie di studi allo scopo di chiarire le cause della patologia, alcuni meccanismi patogenetici restano ancora oscuri e l’eziologia rimane sconosciuta. Il modello murino della malattia, l'encefalopatia autoimmune, ha tuttavia permesso di ricondurre il processo di demielinizzazione all'azione dei linfociti T CD4+ che, una volta attivati, sarebbero in grado di penetrare attraverso la barriera ematoencefalica e di causare danno assonale con conseguente morte di cellule neuronali. L’attivazione delle cellule T è accompagnata da uno shift metabolico che prevede il passaggio da un metabolismo ossidativo energeticamente efficiente ad un programma prettamente glicolitico; nelle malattie autoimmuni i dati in letteratura concernenti il fenotipo metabolico dei linfociti T sono piuttosto esigui e, tuttora, non vi sono lavori condotti a tal riguardo nella SM