bluuebunny/crossref_metadata_2025 huggingface dataset:
Improvement in Relative Survival of Patients with Vasculitis: Study of 101 Cases Compared to the General Population
Authors: Stratta P. , Marcuccio C. , Campo A. , Sandri L. , Messuerott A. , Colla L. , Rosso S. , Mazzucco G. , Mariani L. , Ciccone G.
Published: January 2017

Immunosuppressive treatment has changed the prognosis of renal vasculitis over time, but improvement in prognosis is difficult to analyze in different historical periods, and can be better demonstrated by comparison with life expectancy of sex- and age-matched people. Long-term survival of 101 patients diagnosed with systemic vasculitis at our center from 1975 to 2002 was retrospectively evaluated in comparison with that of the Region's age- and sex-matched population. Patient and kidney survival significantly increased over time. Multivariate analyses showed that risks of patient and renal death decreased by 10% and 7%, respectively, at each year of follow-up, and increased by 6.3% and 5.2% for each year of age. Relative survival significantly improved over time, approaching that of the general population for cases diagnosed after 1993, mainly in women <60 years (from 0.671 at 5-years in the first period to 0.916 in the last period), while 5-year-relative-survival was still 0.530 and 0.682 in men and women > 60 years, respectively. Poisson-based multinomial analyses confirmed the significant risk of the first periods of diagnosis and of dialysis in worsening of the relative survival of patients compared to that of the general population. Life expectancy in patients with renal vasculitis has improved over time, paralleling a significant increase in steroid pulse/cyclophosphamide association therapy and an earlier diagnosis due to the introduction of the ANCA test. Relative survival has considerably improved, and now approaches that expected in the general population for women, but not for men.
Serum Cytokines and Bioumoral Immunological Characterization of Psoriatic Patients in Long Term Etanercept Treatment
Authors: Cordiali-Fei P. , Ardigò M. , Mastroianni A. , Giuliani A. , D'Agosto G. , Bordignon V. , Trento E. , Vento A. , Berardesca E.
Published: January 2017

The purpose of this study is to evaluate blood cytokines and immunological parameters in psoriatic patients during long-term treatment with etanercept. Forty-five subjects of both sexes affected by psoriasis with or without arthritis entered the study and were treated with etanercept according to international standard protocols. Biochemical blood analysis was carried out at baseline and during follow-up every second month. In particular, the following parameters were kept under control: antinuclear antibodies, anti-nDNA antibodies, anti-histone antibodies, blood cell count, circulating lymphocyte subtypes (CD3, CD4, CD8, CD16, CD19) and IgE. Cytokine profiles (IL-1-α, IL-1-β, IL-6, IL-8, IL-10, IL-12, INF, TNF- α) were also evaluated in blood samples during the treatment up to 1 year of follow-up. A significant decrease in PASI score (p<0.01) and in several cytokine levels was observed, particularly in IL-1, IL-6, IFN-γ (p<0.01) and to a lesser extent in TNF-α (p<0.05). No statistically significant changes were recorded after 1 year of follow-up in blood immunological parameters, in particular in ANA titre, CD4/CD8 ratio, IgE levels, CD16, CD19 and eosinophils count. In conclusion, long-term treatment with etanercept leads not only to a significant improvement in PASI score, but also to significant changes (reduction) in several proinflammatory and modulatory cytokines involved in the pathogenesis of the disease; on the other hand, there are no effects on immunological or bioumoral parameters showing that etanercept modulates rather than suppresses the physiological responses during psoriasis treatment.
Evaluation by Real-Time PCR of the Expression of S. Flexneri Virulence-Associated Genes ospB and phoN2 under Different Genetical Backgrounds
Authors: Nicoletti M. , Santino I. , Petrucca A. , Del Chierico F. , Cannavacciuolo S. , Casalino M. , Sessa R. , Cipriani P.
Published: January 2017

Under conditions of activated type III secretion Shigella flexneri up-regulates the expression of numerous genes, including the virulence plasmid (pINV)-encoded ospB and phoN2 genes. ospB and phoN2 are virulence-associated genes which are part of a bicistronic transcriptional unit encoding OspB, a protein (effector) of unknown function secreted by the type III secretion (TTS) apparatus, and PhoN2 (apyrase or ATP-diphosphohydrolase), a periplasmic protein involved in polar IcsA localization on the surface of S. flexneri. In this work we used real-time PCR to measure transcription of ospB and phoN2 of wild-type S. flexneri strain M90T as well as of derivative mutants impaired in definite virulence traits. The results obtained confirmed and extended previous reports indicating that the expression of ospB and phoN2 genes is modulated in a virB-dependent, mxiE-independent manner under conditions of non-activated secretion, while their expression is considerably induced in a mxiE-dependent manner under conditions of activated secretion. That the expression of the ospB-phoN2 operon is up-regulated in condition of activated secretion, indicates that probably the expression of these two genes might be important, especially during the later stages of infection of S. flexneri.
Photo Allergic Contact Dermatitis: The 15-Year Experience of a Tertiary Reference Center in a Sunny Mediterranean City
Authors: Katsarou A. , Makris M. , Zarafonitis G. , Lagogianni E. , Gregoriou S. , Kalogeromitros D.
Published: January 2017

Photoallergic contact dermatitis (PACD) represents an important entity of photodermatoses while photopatch testing is the main diagnostic tool. The main goal of this study is to evaluate retrospectively the prevalence of photoallergic reactions and the offending agents in Athens during a 15-year period. The medical records of all patients with possible PACD between 1992 and 2006 were examined. All patients included in the analysis had undergone patch testing and photo-testing. Contact reactions were detected in 86 out of 207 participants (41.54%), while photocontact reactions were identified in 28/207 (13.52%) patients. The most common offending photoallergen was promethazine (25%), while chlorpromazine and oxybenzone were both detected in 12.5% of cases. PACD represents a unique proportion of photodermatoses in a sunny Mediterranean city such as Athens.
Cryptococcal Lymphadenitis as a Manifestation of Immune Reconstitution Inflammatory Syndrome in an HIV-Positive Patient: A Case Report and Review of the Literature
Authors: Putignani L. , Antonucci G. , Paglia M.G. , Vincenzi L. , Festa A. , De Mori P. , Loiacono L. , Visca P.
Published: January 2017

Cryptococcus neoformans infections are typically associated with T-cell deficiencies, including acquired immunodeficiency syndrome (AIDS). Although highly active antiretroviral therapy (HAART) has strongly reduced AIDS-related opportunistic infections, the restoration and reactivation of CD4+ cells can induce an immune reconstitution inflammatory syndrome (IRIS), consisting in a deregulated inflammatory response to latent infectious pathogens and/or to their residual antigens. Cryptococcal lymphadenitis has occasionally been documented in IRIS. Here we report a case of histology- and culture-negative cryptococcal lymphadenitis associated with IRIS in an adult AIDS patient with a history of disseminated cryptococcosis, after the start of fully adherent HAART. Appropriate diagnosis was established on nested-PCR and sequence analysis of the interspacer region 2 of C. neoformans ribosomal DNA, and detection of slow-growing blastospores in enrichment cultures of fine-needle lymph node aspirate. Review of recent literature and our case findings suggest that IRIS-associated cryptococcal lymphadenitis is more likely the flare up of a latent infection rather than an immunopathological response to residual antigen of unviable cryptococci.
The Immunosuppressor ST1959, a 3,5-Diaryl-S-Triazole Derivative, Inhibits T Cell Activation by Reducing NFAT Nuclear Residency
Authors: Lindstedt R. , Ruggiero V. , D'Alessio V. , Manganello S. , Petronzelli F. , Stasi M.A. , Vendetti S. , Assandri A. , Carminati P. , De Santis R.
Published: January 2017

3-(2-ethylphenyl)-5-(3-methoxyphenyl)-1H-1,2,4-triazole (ST1959) has shown therapeutic effects in several animal models of autoimmune diseases. In this study the effects of ST1959 were further investigated in a murine model of colitis. The evidence obtained indicates that the beneficial effects exerted by ST1959 rely upon a decreased local immunological response. The cellular effects of ST1959 were additionally investigated on human peripheral blood mononuclear cells and Jurkat T cells by measuring cytokine production, cell proliferation and activation of a set of transcription factors. ST1959 decreases human T cell proliferation and inhibits cytokine expression at the transcriptional level. Moreover, at doses inhibiting cytokine production, ST1959 blocks phorbol 12-myristate 13-acetate (PMA) and ionomycin-induced nuclear factor protein of activated T cell (NFAT1) activity, without impairing AP-1-and NF-κB-dependent transcription. Immunofluorescence data show that ST1959 inhibits the nuclear residency of NFAT1 in both Jurkat and human peripheral blood mononuclear cells activated with PMA/ionomycin. leptomycin B, an inhibitor of CRM1/exportinlα-dependent nuclear export, reverted the inhibitory effect of ST1959 on NFAT1 nuclear localization. This indicates that ST1959 may increase the nuclear export of NFAT1, downregulating NFAT1 activity via a mechanism different from that of cyclosporin A, since it does not affect NFAT phosporylation/dephosphorylation steps. These findings provide new insights into the molecular mechanisms underlying the immunomodulatory activity of ST1959.
Immunomodulatory Effects of 1,25-Dihydroxyvitamin D3 on TH1/TH2 Cytokines in Inflammatory Bowel Disease: An in Vitro Study
Authors: Ardizzone S. , Cassinotti A. , Trabattoni D. , Manzionna G. , Rainone V. , Bevilacqua M. , Massari A. , Manes G. , Maconi G. , Clerici M. , Porro G. Bianchi
Published: January 2017

Crohn's disease (CD) is associated with a higher type-1-helper T cell (Th1) cytokine expression, whereas ulcerative colitis (UC) appears to express a modified Th2 response. In addition to its classic role in calcium homeostasis, calcitriol, the hormonal active form of vitamin D, exerts immunoregulatory effects such as modulation of Th1/Th2 cytokines. Therefore, calcitriol administration could modify immune dysfunction in CD and UC. Nine patients with UC [M/F: 5/4; mean age 47 years, remission(R)/active(A) disease: 7/2], 8 patients with CD [M/F: 2/6; mean age 36, R/A 5/3] and 6 healthy controls (HC) [M/F: 3/3, mean age 46] were enrolled. Peripheral blood was collected after a drug-washout of 15 days and peripheral blood mononuclear cells were stimulated with mitogens alone or in the presence of physiological concentrations of calcitriol (100 pg/ml). Type 1 (IL-2, TNF-α, IFN-γ) and type 2 (IL-10) cytokine production was assayed on supernatants by ELISA. Compared to HC, TNF-α production was significantly higher both in UC (p=0.0002) and CD (p=0.0001) patients, at baseline and after incubation with calcitriol (UC p=0.0003, CD p=0.0009). The effects of calcitriol incubation were: 1) reduced IFN-γ (p=0.024) and increased IL-10 (p=0.06) production in UC patients; 2) reduced TNF-α production in CD (p=0.032); 3) no significant effects in HC. Calcitriol increased, albeit not significantly, IL-10 production in UC compared to CD patients (p=0.09). These results suggest an important modulatory role of vitamin D in the Th1/Th2 immune response. The observation that the effect of this modulation was different in CD compared to UC patients provides an interesting area of research into the pathogenesis and treatment of these inflammatory conditions.
Selection and Characterization of a Novel Agonistic Human Recombinant Anti-Trail-R2 Minibody with Antileukemic Activity
Authors: Secchiero P. , Sblattero D. , Chiaruttini C. , Melloni E. , Macor P. , Zorzet S. , Tripodo C. , Tedesco F. , Marzari R. , Zauli G.
Published: January 2017

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising natural anticancer therapeutic agent because through its “death receptors”, TRAIL-R1 and TRAIL-R2, it induces apoptosis in many transformed tumor cells, but not in the majority of normal cells. Hence, agonistic compounds directed against TRAIL death receptors have the potential of being excellent cancer therapeutic agents, with minimal cytotoxicity in normal tissues. Here, we report the selection and characterization of a new single-chain fragment variable (scFv) to TRAIL-R2 receptor isolated from a human phage-display library, produced as minibody (MB), and characterized for the in vitro anti-leukemic tumoricidal activity. The anti-TRAIL-R2 MB2.23 efficiently and specifically bound to membrane-associated TRAIL-R2 on different leukemic cell lines and could act as a direct agonist in vitro, initiating apoptotic signaling as well as complement-dependent cytotoxicity and antibody-dependent cell cytotoxicity, providing a rationale for further investigations of MB2.23 in anticancer therapy.
Osteoblast Apoptosis in Periodontal Disease: Role of TNF-Related Apoptosis-Inducing Ligand
Authors: Mori G. , Brunetti G. , Colucci S. , Oranger A. , Ciccolella E , Sardone F. , Pignataro P. , Mori C. , Karapanou V. , Ballini A. , Mastrangelo F. , Tetè S. , Grassi F. R. , Grano M.
Published: January 2017

Periodontal disease (Pd) is characterized by an increased osteoclast resorption and a decreased osteoblast (OB) bone formation. OBs obtained from alveolar bone of Periodontitis patients (Pp) undergo apoptosis in the presence of TNF-related apoptosis-inducing ligand (TRAIL). We studied the intracellular apoptotic pathway induced by TRAIL; TRAIL death (DR4, DR5) and decoy (DcR1, DcR2) receptors expression in Periodontitis patients' OBs (PpOBs), and we measured the concentration of TRAIL in the serum of Pp. We demonstrated that DNA fragmentation and activation of caspase-8 and caspase-3 in PpOBs, following TRAIL stimulation, occurred in shorter time; moreover, a higher amount of both caspases was activated in order to direct OBs. Down-regulation of DcR2 in PpOBs was demonstrated and high TRAIL levels were detected in the serum of Pp. In conclusion, our data suggest that PpOBs are more sensitive to TRAIL-induced apoptosis when compared to the control group. The down-regulation of DcR2 possibly leads to an imbalanced ratio between death and decoy receptors. Our findings highlight a role of TRAIL in the pathogenesis of Pd.
Necrotic Cell Death in Human Amniotic Cells Infected by Listeria Monocytogenes
Authors: Ammendolia M.G. , Superti F. , Bertuccini L. , Seganti L. , Cipriani D. , Longhi C.
Published: January 2017

Listeria monocytogenes can cause a placental-foetal infection that results in spontaneous abortion, premature labour, stillbirth, or neonatal sepsis and meningitis. Bacteria cross the maternofoetal barrier at the villous syncytiotrophoblast level and subsequently spread from the placenta to the fetus. L. monocytogenes is able to induce different kinds of death in a variety of cells. Murine hepatocytes, murine T and human B lymphocytes, and murine dendritic cells die by apoptosis, whereas bacterial infection of murine and human macrophages leads mainly to necrotic cell death. As we previously described the efficient infection and growth of L. monocytogenes in a human amniotic cell line, we investigated the fate of these cells in order to analyse the mode of cell death. Our results provide biochemical and morphological evidence of necrotic death induced by L. monocytogenes infection.
Efficacy of Cyclosporine in the Treatment of a Case of Infliximab-Induced Erythrodermic Psoriasis
Authors: Bruzzese V. , Pepe J.
Published: January 2017

A clinical case is described of infliximab-induced erythrodermic psoriasis resistant to conventional topical therapy and high-dose corticosteroids. Cyclosporine therapy for some months resolved the severe psoriasis picture. Pathogenetic mechanisms are considered through which anti-TNF agents could induce psoriasis. An activation of T lymphocytes with cutaneous overexpression of a CXCR3 subset and, mainly, an increase in IFN-α due to the blockage of TNF-α could be the causes for this paradoxical adverse event of biological agents. Cyclosporine can work in psoriasis induced by biological agents thanks to its peculiar suppressive activity on T lymphocytes and the concomitant specific action on keratocytes and angiogenesis.
Strategies for Successful Vaccination against Hepatocellular Carcinoma
Authors: Rinaldi M. , Iurescia S. , Fioretti D. , Ponzetto A. , Carloni G.
Published: January 2017

Current therapies against hepatocellular carcinoma (HCC) are not curative in the majority of patients. In the past, immunotherapy approaches aimed to non-specifically stimulate immune response were quite ineffective. New treatments based on stimulation of specific anti-tumor immune response are currently proposed and appear more promising. Tumor-specific antigens identified in HCC demonstrated immunogenicity both in preclinical and clinical trials. Effectiveness in animal studies raised interest in the clinical applicability of non-specific adoptive immunotherapy that prevented disease recurrence after tumor resection. Dendritic cell (DC)-based tumor vaccines achieved encouraging results, and cellular vaccines based on DCs have already entered clinical trials. Preventive and therapeutic DNA vaccination have been proposed, all based on tumor-associated antigens (TAAs), either modified or not, an example being alpha-fetoprotein (AFP). The concomitant expression of co-stimulatory molecules and cytokines was used to increase tumor immunogenicity. Syngeneic or nude mice models indicated that immunotherapy for HCC could stimulate an anti-tumor T-cell response leading to clinical benefit devoid of significant toxicity. The use of DNA-based vaccination raises exciting possibilities in preventing HCC in high-risk individuals such as those with cirrhosis. Novel immunotherapy strategies may contribute in the future to prevention and treatment of HCC.
In Vitro Evaluation of the Synergistic Activity of Neomycin-Polymyxin B Association against Pathogens Responsible for Otitis Externa
Authors: Tempera G. , Mangiafico A. , Genovese C. , Giudice E. , Mastrojeni S. , Nicolosi D. , Furneri P.M.
Published: January 2017

The most recent guidelines recommend, for otitis externa antibiotic therapy, the use of topical formulations in that they are very safe, have a quicker effect and do not induce bacterial resistance compared to systemic therapy. The choice of the class of antibiotics in empiric therapy of otitis externa must take into consideration the polymicrobic nature of the infection that includes both bacteria (Gram-positive and Gram-negative) and mycetes. For this reason, in this study we evaluated the synergic activity of neomycin in association with polymyxin B against the pathogens commonly responsible for otitis externa, compared to that of a single antibiotic (ciprofloxacin). The polymyxinB/neomycin association shows clear synergic effects with values of both Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) reduced by 3–4 times with respect to the single antibiotic; and in P. aeruginosa the synergistic effect of the neomycin/polymyxin B association with respect to neomycin was more evident (5–6 times), with an intrinsic in vitro activity constantly higher than that of ciprofloxacin alone or in association with hydrocortisone. From the analysis of the data obtained in vitro, we can conclude that the possibility of using a topical formulation containing a synergistic association of antibiotics, such as neomycin-polymyxin B, in such a way as to obtain the maximum effect in the minimum time with an increase in the spectrum of action of non-bacterial pathogens, is an optimal choice for the clinician for the empiric therapy of otitis externa.
Clinical Data and Inflammation Parameters in Patients with Cypress Allergy Treated with Sublingual Swallow Therapy and Subcutaneous Immunotherapy
Authors: Ventura M.T. , Carretta A. , Tummolo R.A. , Buquicchio R. , Arsieni A. , Murgia N.
Published: January 2017

The clinical efficacy of immunotherapy, either by high dose sublingual-swallow therapy (SLIT) or subcutaneous immunotherapy (SCIT), has been demonstrated in patients with pollinosis but few studies have been carried out analysing differences in these treatments in terms of an improvement of clinical and allergic phlogosis parameters. The aim of this double-blind placebo-controlled study is to investigate the efficacy of high dose SLIT and SCIT using a purified standardized Juniperus ashei extract in a population of allergic patients monosensitized to cypress. Forty patients with cypress-allergic rhino conjunctivitis were administered therapeutic or placebo SLIT or SCIT for 12 months. Laboratory parameters were studied, namely the eosinophil cationic protein (ECP) level in nasal lavage and in serum, as well as the number of eosinophils (EOS) in peripheral blood and in nasal lavage and the level of eosinophil chemotactic activity (ECA). These parameters were correlated with clinical symptoms, evaluated by means of the clinical symptoms score (CSS). After SCIT and SLIT the levels of ECP and ECA were reduced in nasal lavage. We also observed a significant reduction in the values of ECP in serum in the patients treated with SLIT. EOS were unchanged in peripheral blood, but significantly reduced in nasal lavage. These data were in accordance with the improvement of clinical symptoms, supported by the close correlation between CSS and laboratory parameters. Our data confirm a clinical improvement correlated with a decline in inflammation parameters after one year of immunotherapy, supporting the hypothesis that treatment with a major allergen of cypress is able to change the course of allergic rhinitis.
Structure and Function of the Nicotinic Arm of Acetylcholine Regulatory Axis in Human Leukemic T Cells
Authors: Chernyavsky A.I. , Arredondo J. , Galitovskiy V. , Qian J. , Grando S.A.
Published: January 2017

Although acetylcholine (ACh) is widely known as a neurotransmitter, it also functions as a local humoral factor translating environmental stimuli into alterations in T cell development and function. The cholinergic components present in neurons are expressed in T cells where they constitute an independent cholinergic system. Both non-immunologic and immunologic stimulations can alter expression and function of cholinergic elements in T cells. Recent studies have convincingly demonstrated regulation of immune system by auto/paracrine ACh, which provides a basis for development of new immunomodulatory therapies with nicotinic agonists. The purpose of our research is to integrate information about the structure and activity of the ACh regulatory axis with the phenotypic and functional alterations of T cells during their development and commitment. In this study, we used the ACh producing human leukemic T cell line CCRF-CEM (CEM) to investigate auto/paracrine mechanisms of T cell regulation through the nicotinic class of ACh receptors (nAChRs). The intact CEM expressed α3, α5, α6, α7, α9, β2 and β4 nAChR subunits. Stimulation of CEM with 10 μg/ml of phytohemagglutinin (PHA) for 16 h upregulated expression of the α3, α5, α7, α9 and β2 and downregulated that of α6 and β4 subunits, indicating that TCR activation leads to overexpression of high Ca2+-permeable ACh-gated ion channels. Activation of α7- and α3 nAChRs predominantly abrogated PHA-dependent upregulation of the pro-inflammatory cytokine TNF-α and IFN-γ receptors, respectively, at the mRNA and protein levels. Signaling through α7 and α3 nAChRs also significantly (p<0.05) altered expression of the cell state regulators p21 and Bcl-2, respectively, suggesting that downregulation of inflammation via nAChRs includes effects on the T cell cycle progression and apoptosis. These findings indicate that constant stimulation of α7 and α3 nAChRs by endogenously released ACh controls T cell activation and that signaling downstream of distinct nAChR subtypes targets specific inflammatory and cell cycle genes. Learning the cholinergic pharmacology of inflammation should allow to regulate specific types of immune reactions by selectively activating or blocking the types of nAChRs expressed by the immune cells mediating specific immune reactions.
Expression of Basic Fibroblast Growth Factor, its Receptors and Syndecans in Bladder Cancer
Authors: Marzioni D. , Lorenzi T. , Mazzucchelli R. , Capparuccia L. , Morroni M. , Fiorini R. , Bracalenti C. , Catalano A. , David G. , Castellucci M. , Muzzonigro G. , Montironi R.
Published: January 2017

Basic Fibroblast Growth Factor (bFGF) is a heparin-binding cationic protein involved in a variety of pathological conditions including angiogenesis and solid tumour growth. The basic Fibroblast Growth Factor Receptor (FGFR) family comprises at least 4 high affinity tyrosine kinase receptors that require syndecans for their function. Mounting evidence indicates that syndecans, that bind both bFGF and their FGFRs, will act as stimulators, whereas syndecans that only bind bFGF will act as inhibitors of signaling by sequestering the growth factor. Recent findings have highlighted the importance of syndecans in urological cancers. The aim of this study is to investigate the expression of bFGF, its receptors (R1 and R2) and syndecans (1-4) in invasive urothelial carcinoma and normal-looking urothelium by Western blotting, RT-PCR, and immunohistochemistry analyses. Interestingly, bFGF, FGFR1 and FGFR2 protein levels statistically increased in bladder cancer tissues. mRNA of FGFR1 and syndecans (1-4), showed a statistically significant increase while an mRNA increase in the other molecules analysed was not significant. bFGF, its receptors and syndecan immunostaining were mainly present in the urothelium both in normal-looking tissues and urothelial neoplastic cells. In conclusion, our data report that the bFGF, FGFR and syndecan expressions are altered in bladder tumours.
Simultaneous Characterization of Phospho-Proteins and Cell Cycle in Activated T Cell Subsets
Authors: Lanuti P. , Fuhrmann S. , Lachmann R. , Marchisio M. , Miscia S. , Kern F.
Published: January 2017

Multi-colour flow cytometry is the only technological platform that can analyse the highly complex cellular composition of the immune system in parallel and at single cell resolution. Analysis of the T cell compartment, in particular, requires the simultaneous measurement of multiple markers in order to account for lineage, phenotype and function. Flow cytometry also enables the analysis of intracellular signalling events. By combining the expression of surface markers, intracellular cytokines, phosphorylated versus unphosphorylated kinases, cell proliferation and DNA profile, mechanistic and kinetic information of subset-specific signalling may be obtained: this has not previously been achieved. Here we present a protocol which permits all of these aspects to be explored simultaneously. By comparing basic procedures previously described we were able to optimise different variables, including the choice of antibody/fluorochrome pairs, permeabilisation, fixation and labelling time, to obtain the best DNA staining of different cell types. We applied this method to study subset-specific signalling related to cytokine production and DNA synthesis in T cells responding to specific antigens.
N-Acetylcysteine Infusion Improves Hepatic Perfusion in the Early Stages of Systemic Sclerosis
Authors: Rosato E. , Zardi E.M. , Barbano B. , Menghi G. , Cianci R. , Amoroso A. , Afeltra A. , Pisarri S. , Salsano F.
Published: January 2017

The aim of our study is to evaluate portal and hepatic hemodynamic changes after N-acetylcysteine infusion in patients with systemic sclerosis. In an open-label study 40 patients with Systemic Sclerosis (SSc) were treated with 15mg/kg/hour intravenous N-acetylcysteine for 5 consecutive hours in a single day. Hepatic flow volume, congestion index, portal flow volume, resistance index and pulse rate index were measured in each subject before and after infusion. In all patients mean hepatic flow volume (HFV) and mean portal flow volume (PFV) values after the five-hour infusion with NAC increased not significantly. In 22 selected patients with active capillaroscopic pattern, modified Rodnan Total Skin Score (mRTSS) <18 and mild-moderate score to vascular domain of disease severity scale (DSS), mean HFV increased significantly when compared with mean HFV of 18 SSc patients with late capillaroscopic pattern, mRTSS >18 and severe-end stage score to vascular domain of DSS. The results of our study demonstrate that NAC is able to increase HFV and total liver perfusion after a single infusion in SSc patients with low disease activity and severity scores.
Herpes Simplex Virus Infection and Pemphigus
Authors: Marzano A.V. , Tourlaki A. , Merlo V. , Spinelli D. , Venegoni L. , Crosti C.
Published: January 2017

Pemphigus is a group of autoimmune blistering diseases of skin and/or mucous membranes caused by the presence of antibodies against adhesion molecules on the cell surface of keratinocytes. In genetically predisposed patients, several factors, including drugs, physical agents, neoplasms, hormones, and viruses, notably herpes simplex virus (HSV), have been hypothesized to trigger or exacerbate the disorder. To clarify whether HSV infection represents an aetiopathogenetic factor for pemphigus or a consequence of the immunosuppressive treatment, skin and/or mucosal swabs from 35 patients with pemphigus vulgaris or pemphigus foliaceus were tested for HSV by polymerase chain reaction. Twenty-three of these patients were newly diagnosed, while the remaining 12 had had a previous diagnosis and were under treatment with low-dosage oral corticosteroids. Repeat swabs were taken two weeks after starting intensive immunosuppressive therapy in 8 HSV-negative patients. All skin swabs (n = 27) resulted negative for both HSV-1/2, while oral swabs (n = 30) were positive for HSV-1 in 5 out of the 12 patients who were being treated with oral corticosteroids, but in none (n = 19) of the non-treated group (p = 0.0067, χ2 test). Five out of the 8 patients with repeat swabs became positive for HSV-1, prompting us to start antiviral therapy. In conclusion, HSV is unlikely to be a triggering factor for pemphigus, but its presence in pemphigus lesions seems to be a frequent and early complication of immunosuppression.
Immunoglobulin Production Pattern is Allergen-Specific in Polysensitized Patients
Authors: Ciprandi G. , De Amici M. , Tosca M.A. , Negrini S. , Puppo F. , Marseglia G.L.
Published: January 2017

Allergic rhinitis (AR) is characterized by Th2 polarized immune response, such as increased IL-4 and reduced IFN-γ production, and by a functional defect of T regulatory cells. This impaired immune response profile influences the pattern of immunoglobulin production in allergic patients. Therefore, the aim of this study is firstly to investigate the allergen-specific IgE, IgG, IgG4, and IgA serum level pattern in polysensitized AR patients with the same skin prick test positivity to some pollen allergens. Secondly, this study aims at relating immunoglobulin (Ig) values with some clinical and immunological parameters. Eighty polysensitized patients with AR were enrolled. Serum allergen-specific IgE, IgG, IgG4, and IgA for mites, Parietaria, grasses, and birch, TGF-β and sHLA-G were determined by the ELISA method. Allergic symptoms and drugs use were also assessed. Allergen-specific IgE, IgG, IgG4, and IgA serum levels were significantly different for each tested allergen (p=0.000T). There was a significant correlation between IgE levels and allergy severity, whereas IgA had an antagonistic behaviour, considering mite-specific immunoglobulins. In conclusion, the present study provides the first evidence that immunoglobulin production pattern depends on the specificity of the allergenic response.
Frailty Syndrome is Associated with Altered Circulating Redox Balance and Increased Markers of Oxidative Stress
Authors: Serviddio G. , Romano A.D. , Greco A. , Rollo T. , Bellanti F. , Altomare E. , Vendemiale G.
Published: January 2017

Frailty Syndrome (FS) is a condition described in aging and characterized by physical vulnerability to stress and lack of physiological reserve. In this study we aim to define whether circulating oxidative stress correlates to frailty in terms of glutathione balance and oxidative protein damage. In 62 elderly outpatients, classified as frail patients according to Fried's criteria, evaluation of reduced Glutathione (GSH), Oxidized Glutathione (GSSG), Tumor Necrosis Factor-α (TNF-α), Malonaldehyde- (MDA) and 4-hydroxy-2,3-nonenal- (HNE) protein plasma adducts were performed. A significant increase in the GSSG was observed in patients with FS when compared to non-frail. No difference was shown in the GSH amount, suggesting a glutathione oxidation more than impairment of the synthesis. TNF-α, MDA- and HNE- adducts, were significantly higher in FS as compared to non-frail patients. A logistic regression model correlating FS with redox balance showed a close relationship between glutathione ratio (OR= 1.8, 95% CI=1.2–2.5) and MDA adducts (OR= 2.8, 95% CI= 1.6–4.7) to frailty. Our findings show an association between oxidative imbalance and Frailty Syndrome. GSSG/GSH ratio and plasma protein adducts strongly predict the frailty conditions and seem to be reliable and easily measurable markers in the context of the multidimensional analysis of elderly patients.
Psoriasis Induced by Infliximab in a Patient Suffering from Crohn's Disease
Authors: Manni E. , Barachini P.
Published: January 2017

We describe the case of a 30-year-old female with no family history of psoriasis and suffering from Crohn's disease successfully treated with infliximab at the dosage of 5 mg/kg. At the 15th week from the start of therapy, the patient developed a palmoplantar pustular psoriasis, which spread to the arms, trunk and scalp with erythematosquamous plaques. Deeming the dermatitis onset due to the anti-TNF-α, we decided to discontinue infliximab, while starting with a topical therapy with emollients and corticosteroids and a systemic therapy with cyclosporine. These treatments achieved a clear improvement of psoriasis after 2 months and a complete regression of skin lesions after 4 months. Several cases have been reported of psoriasis induced by anti-TNF-α, which have shown to exert an effective therapeutic action on this disease. The pathogenic mechanism of such a paradoxical effect has not yet been explained, though a number of hypotheses were proposed, among which one of the most intriguing is that the rapid and strong blockade of TNF-α could result in an enhancement of INF-α activity with consequent induction of psoriasis.
The Use and Alpha-Lipoic Acid (ALA), Gamma Linolenic Acid (GLA) and Rehabilitation in the Treatment of Back Pain: Effect on Health-Related Quality of Life
Authors: Ranieri M. , Sciuscio M. , Cortese A.M. , Santamato A. , Di Teo L. , Ianieri G. , Bellomo R.G. , Stasi M. , Megna M.
Published: January 2017

The aim of this trial was to evaluate the effects of α-lipoic acid (ALA) and γ-linolenic acid (GLA) and the beneficial effect of physical exercise on positive sensory symptoms and neuropathic pain in patients with compressive radiculopathy syndrome from disc-nerve root conflict. Often these painful syndromes after the acute event, tend to recurrence becoming subacute or chronic syndromes that become for the period of interest disabiling is an event very important in these cases proper prevention, based on a maintenance drug therapy and the strengthening exercises of paravertebral muscles, flexibility exercises on the spine and when needed on the reduction of body weight. In this Observational Cohort, two-arm trial, 203 patients were enrolled and divided into two groups, the first, “ALA and GLA group”, (n = 101) received oral dose of 600 mg of α-lipoic acid (ALA) and 360 mg of γ-linolenic acid (GLA) and a rehabilitation program for six weeks, the second (n = 102) treated with only rehabilitation program. Patients were recruited at the centre of Physical Medicine and Rehabilitation, have undergone a physiatric examination at the primary outcome (t0) and secondary outcomes were recorded at monitoring visits scheduled at two weeks = t1, four weeks = t2, six weeks = t3, and at the same has been administered the following scale: VAS scale, SF-36, Oswestry Low Back Pain Disability Questionnaire, Aberdeen Back Pain Scale (ABPS), Revised Leeds Disability Questionnaire (LDQ), Roland and Morris Disability Questionnaire. Significant improvements has been noted in the “ALA and GLA group” for paresthesia, stabbing and burning pain, as showed by VAS (Visual Analogue Scale), Oswestry Low Back Pain Disability Questionnaire, Aberdeen Low Back Pain Scale; also, improvements of “quality of life” has been noted, in the same group, as showed by SF-36, LDQ (Revised Leeds Disability Questionnaire), Roland and Morris disability questionnaire. These all outcome measure showed decreases statistically significant. Oral treatment with α-lipoic acid (ALA) and γ-linolenic acid (GLA) for six weeks in synergy with rehabilitation therapy improved neuropathic symptoms and deficits in patients with radicular neuropathy.
Polylactide-Polyglycolide Resorbable Plates Stimulates Adipose Tissue-Derived Stem Cells towards Osteoblasts Differentiation
Authors: Sollazzo V. , Lucchese A. , Palmieri A. , Zollino I. , Iaccarino C. , Carnevali G. , Pezzetti F. , Brunelli G. , Carinci F.
Published: January 2017

Polylactide, polyglycolide materials or devices have been utilized routinely during maxillofacial, craniofacial, and orthopaedic reconstructive surgical procedures.(1) These materials combine the benefits of rigid fixation with the advantages of biodegradation, avoiding the need for implant removal and minimizing the risk of other complications.(2) To study how polylactide, polyglycolide acids plates (PLPG plates) can induce osteoblast differentiation and proliferation in mesenchymal stem cells, the expression levels of bone related genes (RUNX2, SP7, ALPL, SPP1, COL1A1, COL3A1 and FOSL1) and mesenchymal stem cells marker (ENG) were measured in adipose derived stem cells (ADSCs) and normal osteoblast (NO) cultivated on PLPG plates after 15 and 30 days of treatment using real time Reverse Transcription-Polymerase Chain Reaction. Significantly differentially expressed genes among ADSCs and NO were SP7, ENG, FOSL1, RUNX, ALPL and SPP1 in the first 15 days of treatment and SP7, ENG FOSL1, COL3A1 COL1A1, SPP1 and ALPL after 30 days. The present study demonstrated that PLPG plates strongly influences the behavior of ADSCs in vitro by enhancing proliferation, differentiation and deposition of matrix.
Double Demonstration of Oncogenic High Risk Human Papilloma Virus DNA and HPV-E7 Protein in Oral Cancers
Authors: Pannone G. , Santoro A. , Carinci F. , Bufo P. , Papagerakis S. M. , Rubini C. , Campisi G. , Giovannelli L. , Contaldo M. , Serpico R. , Mazzotta M. , Lo Muzio L.
Published: January 2017

Oncogenic HPVs are necessarily involved in cervical cancer but their role in oral carcinogenesis is debated. To detect HPV in oral cancer, 38 cases of formalin fixed-paraffin embedded OSCC were studied by both DNA genotyping (MY09/11 L1 consensus primers in combination with GP5-GP6 primer pair followed by sequencing) and immunohistochemistry (monoclonal Abs against capsid protein and HPV-E7 protein, K1H8 DAKO and clone 8C9 INVITROGEN, respectively). HPV-16 tonsil cancer was used as positive control. The overall prevalence of HPV infection in OSCCs was 10.5%. Amplification of DNA samples showed single HPV DNA infection in 3 cases (HPV16; HPV53; HPV70) and double infection in one case of cheek cancer (HPV31/HPV44). The overall HR-HPV prevalence was 7.5%. E-7 antigen was immunohistochemically detected in all HPV-positive cases. HPV+ OSCC cases showed an overall better outcome than HPV negative oral cancers, as evaluated by Kaplan-Meier curves. HPVs exert their oncogenic role after DNA integration, gene expression of E5, E6 and E7 loci and p53/pRb host proteins suppression. This study showed that HPV-E7 protein inactivating pRb is expressed in oral cancer cells infected by oncogenic HPV other than classical HR-HPV-16/18. Interestingly HPV-70, considered a low risk virus with no definite collocation in oncogenic type category, gives rise to the expression of HPV-E7 protein and inactivate pRb in oral cancer. HPV-70, as proved in current literature, is able to inactivates also p53 protein, promoting cell immortalization. HPV-53, classified as a possible high risk virus, expresses E7 protein in OSCC, contributing to oral carcinogenesis. We have identified among OSCCs, a subgroup characterized by HPV infection (10.5%). Finally, we have proved the oncogenic potential of some HPV virus types, not well known in literature.
Immunological Modifications Induced from Products Used during the Perioperative Period
Authors: Cardinale F. , Mastrototaro M. F. , Cappiello A. , Caffarelli C. , Caimmi S. , Barberi S. , Bernardini R.
Published: January 2017

Anesthetics and other products used during the perioperative period may influence immune function not only merely by reducing the HPA-axis stress response but also by directly modulating innate and adaptive immune responses. Most of the literature on the immune effects of anesthetics has been derived from in vitro or animal studies, due to the number of confounding variables in “real life” surgical settings. These immunosuppressive effects might not normally have clinical consequences for an immune-competent patient, but may act as important modifiers in postoperative morbidity and mortality. Furthermore, some inhibitory effects on neutrophil functions may provide a therapeutically beneficial effect under specific surgical clinical conditions, such as ischemia-reperfusion injury.
Effects of Electromagnetic Stimulation on Osteogenic Differentiation of Human Mesenchymal Stromal Cells Seeded onto Gelatin Cryogel
Authors: Saino E. , Fassina L. , Van Vlierberghe S. , Avanzini M.A. , Dubruel P. , Magenes G. , Visai L. , Benazzo F.
Published: January 2017

Bone tissue engineering typically uses biomaterial scaffolds, osteoblasts or cells that can become osteoblasts, and biophysical stimulations to promote cell attachment and differentiation. In this study, we investigated the effects of an electromagnetic wave on mesenchymal stromal cells isolated from the bone marrow and seeded upon gelatin cryogel disks. In comparison with control conditions without electromagnetic stimulus, the electromagnetic treatment (magnetic field, 2 mT; frequency, 75 Hz) increased the cell proliferation and differentiation and enhanced the biomaterial surface coating with bone extracellular matrix proteins. Using this tissue-engineering approach, the gelatin biomaterial, coated with differentiated cells and their extracellular matrix proteins, may be used in clinical applications as an implant for bone defect repair.
Stimulation of Bone Formation and Fracture Healing with Pulsed Electromagnetic Fields: Biologic Responses and Clinical Implications
Authors: Chalidis B. , Sachinis N. , Assiotis A. , Maccauro G. , Graziani Filippo
Published: January 2017

Pulsed electromagnetic fields (PEMF) have been used for several years to supplement bone healing. However, the mode of action of this non-invasive method is still debated and quantification of its effect on fracture healing is widely varied. At cellular and molecular level, PEMF has been advocated to promote the synthesis of extracellular matrix proteins and exert a direct effect on the production of proteins that regulate gene transcription. Electromagnetic fields may also affect several membrane receptors and stimulate osteoblasts to secrete several growth factors such as bone morphogenic proteins 2 and 4 and TGF-beta. They could also accelerate intramedullary angiogenesis and improve the load to failure and stiffness of the bone. Although healing rates have been reported in up to 87% of delayed unions and non-unions,.the efficacy of the method is significantly varied while patient or fracture related variables could not be clearly associated with a successful outcome.
The Clinical Diagnosis of Meniscal Tear is Not Easy. Reliability of Two Clinical Meniscal Tests and Magnetic Resonance Imaging
Authors: Rinonapoli G. , Carraro A. , Delcogliano A.
Published: January 2017

Aim: to clarify the reliability of two clinical meniscal tests, McMurray's and Apley's and the MRI imaging, in order to establish how to reduce unjustified arthroscopies. Methods: 102 patients were selected out of 160. All patients were submitted to a triple clinical examination (by a young surgeon and two skilled surgeons), MRI and an arthroscopic procedure. The investigated clinical tests were McMurray's and Apley's test. The positivity or negativity of the tests and MRI were compared to arthroscopic findings. Arthroscopy is considered the gold standard for the diagnosis of meniscal lesions. We measured the length of the meniscal lesions in order to correlate it to the clinical findings. Results: From the clinical examination, we got the following data: McMurray's test: sensitivity 79.7%, specificity 78.5%, accuracy 79.4%, positive likelihood ratio 3.7, negative likelihood ratio 0.2. Apley's test: sensitivity 83.7%, specificity 71.4%, accuracy 80.3%, positive likelihood ratio 2.9, negative likelihood ratio 0.2. The composite assessment is strictly dependent on how the discordance of the two tests is evaluated. The assessment of the clinical tests was done even in relation to medial or lateral meniscal lesion. No statistical difference was found about the length of the meniscal tear. MRI gave the following results: sensitivity 78.3%, specificity 85.7%, accuracy 80.3%. Conclusions: If we use, as diagnostic means, McMurray's and Apley's clinical tests and MRI as imaging procedure, we have an accuracy of about 80%. It is important to keep in mind that it is not possible to have the absolute certainty of make a correct diagnosis in case of meniscal lesions. Patients, too, have to be informed about the risk of a negative arthroscopy.
Fabrication, Characterization and Cell Cultures on a Novel Composite Chitosan-Nano-Hydroxyapatite Scaffold
Authors: Palazzo B. , Gallo A. , Casillo A. , Nitti P. , Ambrosio L. , Piconi C.
Published: January 2017

This paper deals with the characterizations made during the development of a nano-HAp loaded chitosan scaffold, obtained by the freeze-drying technique combined with a novel “in situ” crystal growth method. The nano-composites were characterized by a highly porous and interconnected structure. The XRD patterns and calculated domain sizes of the HAp nano-crystals nucleated on the chitosan scaffolds are very similar to the ones recorded for deproteinated bone apatite. Both osteoblasts (MG63) and mesenchimal cells (hMSC) were showing good proliferation and adhesion onto the scaffolds. The presence of extensive filopodia and excellent spreading in and around the interconnected porous structure, indicated a strong cellular adhesion and growth. Moreover a good hMSC osteogenic differentiation has been verified. The observations related to well-developed structure morphology, physicochemical properties and high cytocompatibility suggest that the obtained chitosan–nHA porous scaffolds are potential candidate materials for bone regeneration.