bluuebunny/crossref_metadata_2025 huggingface dataset:
The Relationship of Atopy, Smoking, and Sensitization to Methyltetrahydrophthalic Anhydride
Authors: Yokota K. , Johyama Y. , Yamaguchi K. , Takeshita T. , Morimoto K.
Published: January 2017

We investigated the association of smoking, atopy and helper T (Th) cytokines with sensitization to methyltetrahydrophthalic anhydride (MTHPA) in occupationally exposed subjects. A population of 147 workers from two condenser plants using epoxy resin with MTHPA underwent a questionnaire survey and serologic investigations. Total and MTHPA-specific IgE levels were measured by the Pharmacia CAP system, and serum levels of interleukin (IL)-4, IL-13, and interferon-gammma (IFN-g) by enzyme immunoassay. The Pharmacia CAP-Phadiatop® test, which detects serum IgE specific to most common aeroallergens, was also used. Ninety-six (65%) of the currently exposed workers had positive MTHPA-specific IgE. A significant difference was found in the frequency of positive specific IgE between atopic and non-atopic subjects (P<0.01), but not between smokers and non-smokers. As for smoking, the frequency of positive specific IgE was significantly (P<0.005) higher in smokers than that in non-smokers in non-atopic subjects. Multiple logistic regression analysis also confirmed significant contributions of atopy and smoking to the development of specific IgE (odds ratio=3.2 and 10.5, respectively), suggesting that atopic subjects and non-atopic smokers may be at increased risk of developing sensitization to MTHPA. It is also possible that subjects who became sensitized to MTHPA may become sensitized to common aeroallergens. On the other hand, none of the Th cytokines contributed to the elevation of specific IgE levels. These results suggest that atopic subjects and non-atopic smokers are at increased risk of sensitization by MTHPA. However, to evaluate conclusively the effect of atopy on sensitization, further prospective studies are necessary.
Respiratory Risks in Tunnel Construction Workers
Authors: Arcangeli G. , Cupelli V. , Montalti M. , Pristera M. , Baldasseroni A. , Giuliano G.
Published: January 2017

Respiratory diseases are frequent in tunnel construction workers. A group of 144 subjects randomly selected from the population of 2516 workers engaged in the construction of the railway tunnel under the Appennine Mountains, from Bologna to Firenze was investigated. A group of 69 males comparable for age, living area and habits was studied as a control group. Assessment of air pollutants (NO, NO2, SO2, total dust, silica %) was carried out by means of fixed monitoring stations as well as personal monitors. All the subjects included in the study were examined with a standardised protocol which included physical examination, lung function tests (before and after shift work) and a questionnaire to collect respiratory symptoms. Low concentrations of environmental pollutants were evidenced. Significantly lower values of FEV, and PEF were determined in the worker group pared to controls. A significant decrease in respiratory parameters was shown after shift work. Variables capable of influencing the decrease in parameters include smoking habits, work activity, presence of cough and expectoration, period of the year (spirometries resulted worse in the winter time). Significantly lower values of FEV1 and PEF were evidenced in the workers compared to controls. In spite of the present low work environmental exposure conditions, some physiologic parameters appear altered in tunnel construction workers. This may depend on a variety of noxious agents present in the working environment.
Effect of Varying Types of Anti-Arthritic Drugs on TH1 and TH2 Immune Responses in Mice
Authors: Yamaki K. , Uchida H. , Li X. , Yanagisawa R. , Takano H. , Hayashi H. , Mori Y. , Yoshino S.
Published: January 2017

The present study was undertaken to study the effect of varying types of anti-arthritic drugs on Th1 and Th2 immune responses in mice. To immunize mice, ovalbumin (OVA) emulsified with complete Freund's adjuvant was injected s.c. at the base of the tail (day 0). Indomethacin (IND) as a non-steroidal antiinflammatory drug (NSAID), dexamethasone (DEX) as a steroidal antiinflammatory drug, methotrexate (MTX), auranofin (AUR), and D-penicillamine (D-PA) as anti-rheumatic drugs were orally administrated daily from days 0 to 20. On day 21, anti-OVA IgG2a and interferon (IFN)-γ as indicators of Th1 responses and anti-OVA IgG1 and interleukin (IL) −10 as those of Th2 responses were measured. Treatments with IND, DEX, MTX and AUR were followed by decreases in OVA-specific IgG and proliferation of spleen cells to the antigen. Treatments with IND, DEX, MTX and AUR inhibited both Th1 and Th2 immune responses, although the inhibitory effects of these drugs on the antigen-specific IgG2a and IFN-7 production appeared to be greater than those on IgG1 and IL-10 production. D-PA failed to influence anti-OVA IgG, IgG2a and IgG1 production as well as IFN-γ and IL-10 secretion. Administrations of all the drugs used resulted in suppression of antigen (OVA)-induced arthritis in mice which was associated with inhibition of anti-OVA IgG2a but not IgG1 production. These results suggest that anti-arthritic drugs including IND, DEX, MTX and AUR appear to suppress Th1 and, to a lesser extent, Th2 immune responses, and their antiinflammatory effects on human rheumatoid arthritis might be at least in part explained by downregulation by these drugs of Th1 responses involved in the disease.
Xaliproden (SR57746A) Induces 5-Ht1A Receptors-Mediated Map Kinase Activation in Pc12 Cells
Authors: Appert-Collin A. , Duong F. H. T. , Degrace P. Passilly , Bennasroune A. , Poindron P. , Warter J.-M. , Gies J.P.
Published: January 2017

Neurotrophic growth factors are involved in cell survival. However, natural growth factors have a very limited therapeutic use because of their short half-life. In the present study, we investigated the mechanism of action of a non peptidic neurotrophic drug, Xaliproden, a potential molecule for the treatment of motoneuron diseases, since the transduction pathways of this synthetic 5-HT1A agonist are very poorly understood. Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the ERK1 and ERK2 isoforms of MAP kinase, which then rapidly decrease to the basal level. We demonstrate that isoforms of the she adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced MAP kinases activation. The inhibitor of Ras farnesylation, FPT-1, and the protein kinase C inhibitors, GF 109203X and chelerythrine, inhibited the Xaliproden-induced MAP kinase activation, suggesting p21Ras and PKC involvement. Moreover, the observations that the 5-HT1A antagonist, pindobind, and pertussis toxin abolished the Xaliproden-induced ERK stimulation suggested that Xaliproden activates the MAP kinase pathways by stimulating the G-protein-coupled receptor, 5-HT1A. These results demonstrated clearly that the non peptidic compound, Xaliproden, exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins. These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by MAP Kinase pathway by a pertussis toxin-sensitive mechanism involving 5-HT1A receptors, p21 Ras and MEK-1 and by PKC and Akt pathways.
A Study of the Purine Derivative AIT-082 in G93A SOD1 Transgenic Mice
Authors: Jiang F. , Li W.P. , Kwiecien J. , Turnbull J.
Published: January 2017

AIT-082 is a purine derivative with neuroprotective and neurotrophic activity that is desirable in a candidate therapy for Amyotrophic Lateral Sclerosis. Consequently, we investigated the effect of AIT-082 in a transgenic mouse model of ALS. AIT-082 (0, 1, 3, 10, 30, 60, 100 mg/kg) was given to TgN(SOD1-G93A)1Gur transgenic mice from age 30 days until death. The age at onset of clinical signs of disease and the age at death were recorded for each animal. Disease progression was measured by the weekly average distance run in a running wheel. Analysis was made by the Kaplan Meier method with log rank statistics, log rank for trend and Cox regression. Neuropathological study of the brain, spinal cord, muscles and other organs was undertaken at death. In a second experiment we studied the effect of AIT-082 (30 mg/kg) at the onset of disease and during survival of transgenic G93A SOD1 mice, beginning dosing at different ages (20, 30, 40, 60, 80 days). Disease onset was mildly earlier (i.e. worse) at 1 and 10 mg/kg AIT-082 and mildly delayed at 30 mg/kg. This improvement did not reach the usual statistical significance. There was no difference in the age at death for any treatment dose. There was no difference in the neuropathology of treated and untreated G93A mice. However, there was an early improvement in the running wheel function at all tested doses. Using Cox regression, after adjustment for sex, the mice in the running wheels had slightly delayed onset of disease without change in survival and, after adjustment for exercise, the female mice had slightly improved survival. Consequently, AIT-082 would not be an attractive candidate for ALS clinical trials as monotherapy and justification for its use in combination therapy would require additional laboratory support. There was dissociation between the endpoints of disease progression (as judged by running wheel performance) and disease onset and survival. AIT-082 improved early running wheel performance yet led to accelerated late decline and had no impact on survival. It is possible that the drug facilitates early sprouting that leads to accelerated late decline.
A Clinical Trial of Oral Hyposensitization in Systemic Allergy to Nickel
Authors: Schiavino D. , Nucera E. , Alonzi C. , Buonomo A. , Pollastrini E. , Roncallo C. , De Pasquale T. , Lombardo C. , La Torre G. , Sabato V. , Pecora V. , Patriarca G.
Published: January 2017

Nickel allergy is the most common contact allergy. Some nickel-sensitive patients present systemic (cutaneous and/or digestive) symptoms related to the ingestion of high nickel-content foods, which significantly improve after a specific low nickel-content diet. The etiopathogenetic role of nickel in the genesis of systemic disorders is, furthermore, demonstrated by the relapse of previous contact lesions, appearance of widespread eczema and generalized urticaria-like lesions after oral nickel challenge test. The aim of this study is to investigate the safety and efficacy of a specific oral hyposensitization to nickel in patients with both local contact disorders and systemic symptoms after the ingestion of nickel-containing foods. Inclusion criteria for the recruitment of these patients were (other than a positive patch test) a benefit higher than 80% from a low nickel-content diet and a positive oral challenge with nickel. Based on the previous experiences, our group adopted a therapeutic protocol by using increasing oral doses of nickel sulfate associated to an elimination diet. Results have been excellent: this treatment has been effective in inducing clinical tolerance to nickel-containing foods, with a low incidence of side effects (gastric pyrosis, itching erythema).
Non-Invasive Assessment of Airway Inflammation in Ship-Engine Workers
Authors: Barreto M. , Villa M.P. , Corradi M. , Barberi S. , Monaco G. , Martella S. , Bohmerova Z. , Sabatino G. , Ronchetti R.
Published: January 2017

Smoking is harmful for respiratory function. In young to middle-aged men the damage is insidious and difficult to demonstrate. The respiratory impairment could increase under specific stressful conditions in the professional environment. On the hypothesis that exhaled markers are useful for assessing airway susceptibility to inhaled irritants, we measured exhaled markers and lung function in smoking and non-smoking engine-driver military coastguards before and after a patrol at sea. Eighteen men, mean age 39 yrs (range 23–58 yrs), 8 smokers, underwent spirometry, exhaled and nasal nitric oxide (eNO, nNO), exhaled carbon monoxide (CO) and exhaled breath condensate (EBC) for measures of hydrogen peroxide (H2O2), leukotriene B4 (LTB4), proteins (Prots), 8-isoprostanes (8-IsoPs), nitrite (NO2) and nitrosothiols (RS-NOs) at baseline and after an 8-hour patrol navigation on board small, high-speed diesel-powered ships. At baseline, the smokers showed higher middle flows and CO levels, lower eNO and nNO than non-smokers, but similar levels of EBC markers; geometric means (95% confidence interval), CO: 23.6 (14.5 to 38.3) vs. 3.5 (2.5 to 5.3) ppm; eNO: 7.9 (4.8 to 12.9) vs. 26.7 (15.7 to 45.5) ppb, p=0.000. After navigation, Prots, 8-IsoPs and RS-NOs (but not lung function variables or other markers) significantly increased only in smokers; baseline vs post-navigation RS-NOs: 0.27 (0.11 to 0.65) vs. 1.30 (0.58 to 2.89) μmol, p=0.012. The respiratory consequences of a stressing environment in engine-driver military coastguards who actively smoke are better assessed by measuring EBC markers than by eNO, nNO or lung function. By increasing airway inflammation from oxidative-stress, tobacco smoking appears to interact with other chemical or physical factors elicited during sea navigation. Precisely what these factors are deserves further investigation.
Evidence of Lower Oxidative Stress in the Air Spaces of Patients with Reversible Copd
Authors: Serviddio G. , Carpagnano G.E. , Rollo T. , Tamborra R. , Barbaro M.P. Foschino , Vendemiale G. , Altomare E.
Published: January 2017

The mechanism responsible for the reversibility of airflow limitation in stable chronic obstructive pulmonary disease (COPD) patients is unknown. The aim of this study is to assess the relationship between the reversibility of airflow limitation, the redox balance and the inflammatory cells in the sputum of patients with stable COPD. For this purpose we examined 15 normal healthy control subjects and 20 nonatopic COPD patients. The COPD patients were divided into two groups: reversible COPD (increase in FEV1>200 ml and/or ≥12% after 200 μg of inhaled salbutamol) or non-reversible COPD. GSH, GSSG were measured in induced sputum and blood. Protein carbonyls were evaluated by WB in sputum and IL-4 and IL-6 and TNF-α in plasma and sputum. GSH oxidation and protein oxidation were lower in reversible COPD patients than in those with no reversibility. The sputum eosinophil count was significantly higher in the reversible group than in the non-reversible group, and IL-4 concentration was higher in the same patients both in sputum and in plasma. In contrast, IL-6 and TNF-α were increased in non-reversible COPD patients in both biological samples. We conclude that airflow reversibility in COPD patients is associated with airway oxidative stress and activation of eosinophil inflammatory pattern in sputum and blood, suggesting that these patients could respond to specific pharmacological treatment.
Immunoinflammatory Activation during the Acute Phase of Lacunar and Non-Lacunar Ischemic Stroke: Association with Time of Onset and Diabetic State
Authors: Licata G. , Tuttolomondo A. , Corrao S. , Di Raimondo D. , Fernandez P. , Caruso C. , Avellone G. , Pinto A.
Published: January 2017

Several studies have stressed the involvement of inflammation in the pathophysiology of acute brain ischemia, but the role of immunoinflammatory activation in diabetic stroke patients has not yet been fully evaluated. The aim of our study was to evaluate immunoinflammatory activation of acute phase of stroke in relation to time of symptoms onset, diabetic state and diagnostic subtype. We enrolled 60 patients (32 diabetics; 28 non- diabetics) with acute ischemic stroke and 123 subjects without acute ischemic stroke, and measured levels of IL-1β, TNF-α, IL-6, IL-10, E-selectin, P-selectin, sICAM-1, sVCAM-1, VWF, 24–72 h and 7–10 days after stroke onset; TPA, PAI-1 plasma levels at 24–72h. Our stroke patients exhibited significantly higher plasma levels of cytokines, selectins, adhesion molecules and PAI-1, and diabetic stroke patients exhibited higher plasma levels of PAI-1 in comparison with non-diabetic ones. Lacunar strokes in comparison with those non-lacunar exhibited significantly lower levels of TNF-α and IL1-β, P-selectin and ICAM-1. Moreover, diabetic patients with lacunar strokes exhibited a minor grade of immunoinflammatory activation of the acute phase at 24–72h and 7–10 days after stroke onset. The minor grade of immunoinflammatory activation of patients with lacunar strokes, particularly diabetic ones, could be related to the minor extension of the infarct size, owing to the typical microvascular disease of diabetic subjects which could also explain the reported better outcome of this subtype of ischemic stroke.
Increased Expression of Specific Thioredoxin Family Proteins; A Pilot Immunohistochemical Study on Human Hepatocellular Carcinoma
Authors: Cunnea P. , Fernandes A.P. , Capitanio A. , Eken S. , Spyrou G. , Björnstedt M.
Published: January 2017

Hepatocellular Carcinoma (HCC) is one of the most frequent cancers worldwide, however, prognosis remains poor following its discovery. We investigate the Thioredoxin superfamily of proteins as diagnostic markers for HCC. Furthermore, we delineate possible roles of the endoplasmic reticulum member of the superfamily, ERdj5, in carcinogenesis. Using antibodies against Thioredoxin 1, Thioredoxin Reductase 1 and ERdj5, we performed immunohistochemistry on paraffin embedded liver biopsy sections from HCC patients. All three redox proteins exhibited elevated expression levels in tumor tissue compared to internal control, with ERdj5 showing a remarkable 3-fold increase. In vitro cell viability experiments using Hepatocellular Carcinoma HuH7 cells treated with ERdj5 small interfering RNA showed that ERdj5 knockdown cells exhibited less resistance to Doxorubicin (chemotherapy drug), but more resistance to Tunicamycin (Endoplasmic Stress inducer), compared to control cells. In conclusion, we introduce members of the Thioredoxin superfamily as possible immunohistochemical markers in the diagnostics of hepatocellular carcinoma and indicate a potential defensive role for ERdj5 in chemotherapeutic drug resistance.
Interleukin-6, Interleukin-10 and Heat Shock Protein-90 Expression in Renal Epithelial Neoplasias and Surrounding Normal-Appearing Renal Parenchyma
Authors: Cardillo M.R. , Ippoliti F.
Published: January 2017

Cytokines, notably the interleukins IL-6 and IL-10, have an important role in the development and progression of renal-cell carcinomas, acting in the host-tumor interaction and in tumor bulk. Heat shock proteins (HSP), in particular HSP-90, may have a regulatory role in cytokine biosynthesis and prognostic implication in some tumors. To define the roles of the cytokines IL-6 and IL-10 and HSP-90 in the progression of renal-cell carcinoma we analyzed immunohistochemical expression of these proteins in human renal-cell carcinomas from 95 total nephrectomies. IL-6, IL-10 and HSP-90 proteins were more strongly expressed in epithelium and stroma of the renal tumoral compartment than in adjacent normal peritumoral tissue. But the difference reached significance only for HSP-90 protein. The percentage of cells expressing IL-6, IL-10 and HSP-90 immunoreactivity was higher in benign epithelial tumors, than in normal peritumoral tissue, but lower than in renal-cell carcinomas. Whereas HSP-90 immunoreactivity seemed higher in more aggressive histological phenotypes (collecting-duct carcinoma) of renal-cell carcinomas, IL-10 protein levels were higher in more advanced TNM stage (pT3) tumors. Our observation suggests that IL-6 and IL-10 and HSP-90 may be useful markers associated with the development and progression of renal-cell carcinomas and have independent functional roles in this malignant condition.
Induction of Macrophage Glutamine: Fructose-6-Phosphate Amidotransferase Expression by Hypoxia and by Picolinic Acid
Authors: Manzari B. , Kudlow J.E. , Fardin P. , Merello E. , Ottaviano C. , Puppo M. , Eva A. , Varesio L.
Published: January 2017

We studied the expression of glutamine:fructose-6-phosphate amidotransferase (GFAT), the rate limiting enzyme in the hexosamine biosynthetic pathway controlling protein glycosylation. We obtained the first evidence that the GFAT mRNA and protein are constitutively expressed in murine mononuclear phagocytes (Mf) and inducible by picolinic acid (PA), a catabolite of tryptophan, hypoxia and desferrioxamine (DFX). These stimuli share the property to transactivate gene expression through the Hypoxia Responsive Element (HRE). The promoter of GFAT contains the consensus sequence of HRE in position −74/-65 (GFAT-HRE), and we studied the role of HRE on the activation of the promoter utilizing appropriate expression vectors. We found that GFAT-HRE is essential for the response to hypoxia, PA or DFX and that Hypoxia Inducible Factor-1α (HIF-1α) can augment this response. Finally, we demonstrate that iron chelation is part of the mechanism by which PA and DFX activate GFAT expression. Our results provide the first indication that hypoxia, PA or DFX induce the transcription of GFAT gene in murine Mf cell lines and that the HRE of the promoter is essential for this response.
Correlation between Serum IgE and Salivary Cortisol Levels in Subjects with Type I Allergic Disorders
Authors: Toda M. , Sakaguchi Y. , Morimoto K.
Published: January 2017

This study compares the relationship between serum IgE and salivary Cortisol levels in 42 normal and in 18 type I allergic subjects. Levels of serum total IgE and salivary Cortisol were determined with the UniCAP system and ELISA respectively. In the type I allergic subjects, there was a significant correlation between serum IgE and salivary Cortisol levels ( p < 0.01). In the normal subjects, on the other hand, no correlation was found. These findings suggest that there may be an association between stress and allergic disorders.
Poly-L-Lactic Acid β-Tricalcium Phosphate Screws: A Preliminary In Vivo Biocompatibility Study
Authors: Magarelli N. , Savastano M.A. , Palmieri D. , Zappacosta R. , Lattanzio G. , Salini V. , Orso C.A. , Guglielmi G. , Colosimo C.
Published: January 2017

The aim of this study is to assess the biocompatibility of two types of Poly-L-lactic acid (PLLA) screws (with either hydroxiapatite (HA) or β-tricalcium phosphate (β-TCP)) implanted in the left femur of four sheep euthanized at 42, 50, 57 and 84 days after surgery. Titanium screws were also implanted for comparison purposes. No signs of inflammation were seen in the 240 specimens. A rating of “+/-”for macrophages and “-”for neutrophils was assigned to all specimens. All specimens were assigned a rating which ranged from “+/-” to “+++” for fibroblasts and osteoblasts. The presence of macrophages, neutrophils and fibroblasts/osteoblasts was not statistically different for the four implantation periods. PLLA implants with β-TCP have a biocompatibility comparable to PLLA implants with HA.
Pentoxifylline Inhibits Vγ9/Vδ2 T Lymphocyte Activation of Patients with Active Behçet's Disease in Vitro
Authors: Accardo-Palumbo A. , Ferrante A. , Ciccia F. , Cadelo M. , Giardina A.R. , Impastato R. , Triolo G.
Published: January 2017

The aim of this study is to evaluate the in vitro effect of pentoxifylline (PTX) on T Vγ9/Vδ2 lymphocyte function in Behçet's disease (BD). We investigated the effect of PTX on Vγ9/Vδ2 T cell expansion and expression of TNFRII receptor and perforin content before and after PTX addition by means of FACS analysis lymphocyte cultures from patients with active and inactive BD and healthy subjects. The addition of PTX at a concentration of 1 mg/ml determined a significant inhibition of cell expansion, a down regulation of TNF receptor expression and inhibited the PMA-induced degranulation of perforin. Taken together these data indicate that PTX is capable of interfering with Vγ9/Vδ2 T cell function in BD, and although cell culture models cannot reliably predict all of the potential effects of the drug in vivo, our results encourage the possibility that this drug may find use in a range of immunological disorder characterized by dysregulated cell-mediated immunity.
Isolated Lichen Planus of the Lip
Authors: Petruzzi M. , De Benedittis M. , Pasture L. , Pannone G. , Grassi F.R. , Serpico R.
Published: January 2017

Oral lichen planus (OLP) is a relatively common disorder whose cause is still unknown. It occurs mostly on the buccal mucosa, but the gingivae, tongue, floor of the mouth and retromalar pads may also be affected. It rarely occurs on the lips and usually in association with oral lesions. We report a case series of ten patients with a history of isolated swelling of the lower and/or upper lip, erosions and crusting. General medical history, examination of the oral cavity and recording of signs and symptoms were carried out for each patient. Among the six different clinical variants of OLP described by Andreasen, the atrophic-erosive form was the most common in the course of isolated LP of the lip in our series. Five cases presented HCV hepatitis. A complete remission of lesions was observed in eight patients after topical treatment with clobetasol propionate 0.05% and tocopherol oil, while partial improvement was noted in those remaining. Isolated LP of the lip is unusual and presents a diagnostic challenge, however an appropriate differential diagnosis is fundamental. Lesions of the lips might represent a more or less precocious phase of oral involvement. Moreover the reasons for the unique localization on the lips need to be explored. Several variables, including age, duration of lesions, concomitance of other diseases, and genetic predisposition may be involved. Isolated LP of the lip is a well-known condition which responds well to topical treatment with corticosteroids. A thorough medical management and active early treatment are necessary to improve symptoms and might also be a relevant prevention strategy from squamous cell carcinoma risk, although data to fully support this statement still need investigation.
Synthetic Gel of Carotid Artery Plaque
Authors: Terzuoli L. , Felici C. , Ciari I. , Guerranti R. , Pagani R. , Marinello E. , Porcelli B.
Published: January 2017

Atherosclerosis is a complex disease that affects medium and large arteries, leading to the formation and progression of plaque. In this process the proteins play an essential role and as a consequence, proteomic-based strategies examining the protein content of cells or tissues could offer a useful approach for the study of plaque proteins. Due to the heterogeneous cell composition of plaque, proteome analysis of whole lesions is difficult, besides being also complicated by the presence of plasma proteins that cannot be completely eliminated. A good way to study variations in protein expression among series of gels is to construct a synthetic gel. This type of gel is obtained by averaging the positions, shapes and optical densities of spots in a given set of gels. To be included in the synthetic gel, spots must be found in at least three gels. To obtain a profile representative of the proteome of atherosclerotic plaque, canceling its high variability, we constructed a synthetic gel using an average of ten carotid plaque samples. We then compared it with an equivalent synthetic gel constructed using ten plasma samples from the same carotid surgery patients. For the comparison of two synthetic gels (plasma/plaque) we could discriminate plasma proteins from plaque proteins. Besides analysis of spots common to plasma, the synthetic gel is useful to detect spots exclusive to plaque, thus simplifying a very complex mixture.
Report of a Case of Discoid Lupus Erythematosus Localised to the Oral Cavity: Immunofluorescence Findings
Authors: Serpico R. , Pannone G. , Santoro A. , Mezza E. , Piccolo S. , Esposito V. , Busciolano M. , Ciavarella D. , Muzio L. Lo , Bufo P.
Published: January 2017

Discoid Lupus Erythematosus (DLE) is a chronic disease with a typical cutaneous involvement. This pathology rarely involves mucosa: oral cavity is interested in 20% of DLE patients. We describe a case of oral DLE in a 50-year-old woman with an anamnesis for autoimmune disorders. This study shows the helpful role of immunofluorescence in the diagnosis of autoimmune diseases. The first diagnostic step was the clinical observation of the oral mucosa: the lesion area was erythematous, athrophic and hyperkeratotic. The patient then underwent laboratory examination. We utilized human epithelial cells (Hep-2010) for Indirect Immuno-Fluorescence (IIF). Moreover, the biopsy site for Direct Immuno-Fluorescence (DIF) and histopathological analysis was the untreated oral lesion. IIF detected an increase of Anti-Nuclear Antibody (ANA) and positivity for SSA-RO. By DIF, we observed IgG/IgA/fibrinogen along basal layer. Multiple biopsies reported signs of chronic basal damage. Steroid systemic therapy induced a considerable lesion regression. We suggest the use of immunofluorescence with the integration of further data to improve diagnosis of rare diseases and to establish a suitable therapy.
An Optimised Data Analysis for the Balb/c3T3 Cell Transformation Assay and its Application to Metal Compounds
Authors: Ponti J. , Munaro B. , Fischbach M. , Hoffmann S. , Sabbioni E.
Published: January 2017

The Balb/c3T3 cell transformation assay (CTA) is an available in vitro system to detect the carcinogenic potential of chemicals. Currently, the European Centre for the Validation of Alternative Methods (ECVAM) is validating this test, assessing its reliability and relevance. Its endpoint is the formation of type III foci, which is, when using clone A31-1-1, a very rare event that usually does not occur at all for negative controls. The carcinogenic potential of a compound tested is assessed by comparing the number of foci in treated and untreated cells. The objective of the present work is to optimise the data analysis for this endpoint by applying the most commonly used approach by a t-test and the Fisher's exact test as an alternative approach. For this purpose selected metal compounds classified as carcinogenic (NaAsO2, CdCl2 cisPt), as suspected carcinogenic (C6H5)4AsCl, CH3HgCl), or as compounds without evidence of carcinogenic properties in humans ((NH4)2PtCl6, NaVO3) as well as a non-carcinogenic (AgNO3) were analysed. Our evaluation revealed that the t-test approach, which assumes normality of data, is not appropriate. The results demonstrated that the statistical analysis by Fisher's exact test better reflects the data properties and greatly facilitates the interpretation of Balb/c3T3 CTA data regarding carcinogenic potential.
Quantitative Immunohistochemical Analysis of Matrilysin 1 (MMP-7) in Various Renal Cell Carcinoma Subtypes
Authors: Patraki E. , Cardillo M.R.
Published: January 2017

The role of matrilysin 1 or matrix metalloproteinase-7 (MMP-7) in cancer is extremely complex and poorly understood. In this study we investigated differential expression of MMP-7 in the epithelium and stroma of 95 paraffin-embedded renal tumor samples by immunohistochemistry and compared tumoral with normal peritumoral renal tissue. We also determined a possible correlation of the immunohistochemical findings with histological subtype, tumor grade and stage of RCC. In all areas examined MMP-7 protein expression was significantly higher in epithelium than in stroma (P<.01). MMP-7 was more less expressed in peritumoral normal areas than in benign epithelial neoplasias (renal papillary and oncocytomas) and RCC carcinomas, reaching the highest immunopositive reaction in chromophobe RCC subtypes, followed by conventional clear-cell and chromophilic-papillary RCC histological subtypes and the lowest levels in more aggressive RCC histotypes (spindle-cell and collecting-duct RCCs). MMP-7 reached their highest levels in high-grade and high-stage RCCs. Our observation suggests an important role of MMP-7 in the development and progression of renal cancer. The differential expression of MMP-7 in the various histological RCC subtypes may reflect the malignant phenotype and more aggressive behavior of RCCs.
Inflammation, Oxidative Stress and Systemic Effects in Mild Chronic Obstructive Pulmonary Disease
Authors: Barbaro M.P. Foschino , Carpagnano G.E. , Spanevello A. , Cagnazzo M.G. , Barnes P.J.
Published: January 2017

Chronic obstructive pulmonary diseases (COPD) is a pulmonary disease characterized by systemic abnormalities. The aim of this study is to investigate inflammation and systemic effects in mild COPD. Twenty-seven mild stable smoking related COPD patients and 15 age-matched healthy subjects were enrolled in the study. IL-6, TNF-α and IL-4 in plasma, sputum and exhaled breath condensate were measured. We also measured exhaled nitric oxide (NO) and pH in sputum and in breath condensate. Moreover, fat-free mass, body mass index (BMI), respiratory muscle strength, plasma oxidative stress and C-reactive protein (CRP) were measured. Higher concentrations were found of CRP, of diacron reactive oxygen metabolites (DROMs) and of IL-6, TNF-α and IL-4 either in plasma or in supernatant of induced sputum or in exhaled breath condensate of COPD subjects compared to healthy controls. Furthermore, higher concentrations were observed of exhaled NO and lower exhaled pH in breath condensate of COPD when compared with healthy subjects. In the group of COPD patients, the subjects with airway reversibility showed an increase of sputum eosinophils and exhaled NO, whereas the subjects without airway obstruction reversibility showed an increase in sputum neutrophils, TNF-α and IL-6. We also found a trend towards a decrease in fat-free mass and respiratory muscle strength in COPD compared to healthy subjects and a negative correlation between these systemic indices (fat-free mass, maximal inspiratory pressure, maximal expiratory pressure) and TNF-α concentrations in the blood, sputum and breath condensate. We conclude that mild COPD subjects present an increase in inflammatory markers in blood and in airways of similar pattern and furthermore, the neutrophilic pattern of airway inflammation observed in the group of COPD subjects without an airway obstruction reversibility makes it more likely that systemic features are present.
The-374T/A Variant of the Rage Gene Promoter is Associated with Clinical Restenosis after Coronary Stent Placement
Authors: Falcone C. , Emanuele E. , Buzzi M.P. , Ballerini L. , Repetto A. , Canosi U. , Mazzucchelli I. , Schirinzi S. , Sbarsi I. , Boiocchi C. , Cuccia M.
Published: January 2017

Upregulation of the receptor for advanced glycation end products (RAGE) may play a crucial role in neointimal formation upon vessel injury. The −374T/A variant of the RAGE gene promoter, which has been associated with an altered expression of the cell-surface receptor, could exert a protective effect toward the development of vascular disease. The aim of this study is to determine the impact of this common genetic variant in the occurrence of clinical in-stent restenosis after coronary stent implantation. The −374T/A polymorphism of the RAGE gene promoter was evaluated by PCR-RFLPs in 267 patients with coronary artery disease who underwent coronary stent implantation and a subsequent coronary angiography 6–9 months later for suspected restenosis. In-stent restenosis was assessed by means of quantitative angiography. Carriers of the-374AA genotype showed a significantly reduced risk of developing restenosis after percutaneous transluminal intervention than non-carriers. To determine whether the protective effect of the homozygous AA genotype toward clinical restenosis was independent of potential confounders, we performed multivariable logistic regression analysis. After allowance for clinical and biochemical risk factors and stent length, the AA genotype remained significantly associated with a reduced prevalence of in-stent restenosis. No relation was evident between the RAGE genotype and established cardiovascular risk factors. In conclusion, the −374AA genotype of the RAGE gene promoter could be associated with a reduced risk of in-stent restenosis after coronary stent implantation.
CD4+CD25+ Lymphocyte Subsets in Chronic Graft versus Host Disease Patients Undergoing Extracorporeal Photochemotherapy
Authors: Rubegni P. , Sbano P. , Cevenini G. , Perari M.G. , Marotta G. , Risulo M. , Carcagnì M.R. , D'Ascenzo G. , De Aloe G. , Fimiani M.
Published: January 2017

Extracorporeal photochemotherapy (ECP) has been used successfully for the treatment of chronic Graft versus Host Disease (cGvHD). However, the mechanism by which ECP exerts its protective effects remains elusive. Some recent observations have suggested a possible role of certain subsets of T lymphocytes with immunosuppressive properties (T-regulatory cells) that coexpress CD4 and high levels of the interleukin-2 receptor chain: CD4+CD25+ T lymphocytes. We studied whether ECP affects the percentage of these cells in the peripheral blood of patients with cGvHD. The study population consisted of 14 patients with cGvHD refractory to systemic steroids. On enrolment in each cycle of ECP, patients underwent clinical examination, blood chemistry analysis and other instrumental procedures to document and assess involvement of the various organs and systems. For cytofluorimetric identification and phenotyping of CD4+CD25+ T lymphocytes, peripheral blood samples were collected in EDTA anticoagulant before ECP, after 48 hours, and after 6 and 12 months from the start of treatment. The 14 patients in this study received a total of more than 300 cycles of ECP, with only minor side effects. The clinical outcome was negative in 2 patients and positive in 12 patients. Within-subject analysis indicated that the percentage of CD4+CD25+ T lymphocytes before ECP and after 12 months of treatment was significantly increased. Our study confirms that changes in the percentage of CD4+CD25+ T cells induced by ECP could be a central aspect in the cascade of immune events leading to the immunological and clinical effects of this treatment in patients with cGvHD.
Intranasal Flunisolide Treatment in Children with Adenoidal Hypertrophy
Authors: Ciprandi G. , Varricchio A. , Capasso M. , Varricchio A.M. , De Lucia A. , Ascione E. , Avvisati F. , Capristo C. , Marseglia G.L. , Barillari U.
Published: January 2017

Adenoidal hypertrophy (AH) represents one of the most frequent indications for surgery in children and it has been proposed that treatment with intranasal corticosteroids can decrease the size of AH. Therefore, the aim of the study is to evaluate the effect of the use of intranasal flunisolide among children affected by AH. 178 children with AH were evaluated in this randomised and controlled study. Inclusion criteria for the study required that each patient had to have a HI or IV degree of AH on the initial endoscopic examination. Children were treated with intranasal flunisolide or isotonic saline solution for 8 weeks. After treatment, endoscopy was performed to re-evaluate AH degree. Flunisolide treatment was associated with significant (p<0.04) reduction of AH degree. There was moreover a consistent reduction of children (46 out of 58) proposed to adenoidectomy. No clinically important adverse events were reported. In conclusion, this preliminary study demonstrates that an 8-week treatment with intranasal flunisolide is significantly associated with reduction of AH, thus preventing the recurrence to adenoidectomy, and is safe.
Changes in Matrix Extracellular Phosphoglycoprotein Expression before and during in Vitro Osteogenic Differentiation of Human Dental Papilla Mesenchymal Cells
Authors: Tetè S. , Nargi E. , Mastrangelo F. , Zizzari V. , D'Apolito G. , Traini T. , Costanzo G. , Dadorante V. , D'Alimonte I. , Caputi S. , Caciagli F. , Ciccarelli R.
Published: January 2017

The purpose of this study is to characterise the expression of matrix extracellular phosphoglycoprotein (MEPE) in cultured mesenchymal cells isolated from human dental papilla (PaMCs) of impacted third molars either before or during differentiation of these cells into osteo/odontoblasts. PaMCs, like mesenchymal cells deriving from human dental pulp (DPMCs), resulted positive for a number of mesenchymal markers including CD146 and STRO-1. During the first week in culture they showed a faster proliferation rate than DPMCs, coupled to an earlier down-regulation of MEPE. Also when the cells were further cultured in osteogenic medium (containing β-glycerophosphate, ascorbic acid and dexamethasone) for 40 days, MEPE down-regulation coupled to an increased expression of osteogenic markers, such as osteocalcin and alkaline phosphatase, occurred earlier in PaMCs than in DPMCs. Thus, our data, indicating that also in PaMCs MEPE expression is higher when cells proliferate, whereas it is downregulated as cells differentiated, are in favour of a role of MEPE as an early regulator of odontogenic differentiation. We also confirm the superior proliferative potential of PaMCs in comparison with DPMCs, coupled to a more rapid induction of osteogenic differentiation. Therefore, these cells represent an optimal source to be conveniently used for dental tissue engineering and tooth regeneration.
Pampiniform Plexus and Oxidative Stress during Chronic Hypoxia and Hyperoxia
Authors: Verratti V. , Di Giulio C. , Berardinelli F. , Tiboni G.M. , Pellicciotta M. , Brunetti L. , Ferrante C. , Orlando G. , Bianchi G. , Cacchio M. , Vacca M. , Tenaglia R.
Published: January 2017

The aim of the present study is to investigate oxidative stress produced by experimental hypoxia and hyperoxia in young and old pampiniform plexus rats, in order to evaluate the oxidative role of oxygen. Oxidative stress causing molecular and cellular dysfunction increases in hypertension and can therefore be considered a state of oxidative stress. This consideration makes us reflect on the responsibility of oxidative stress in the veins of the pampiniform plexus, notoriously under high hydrostatic pressure. After experimental hypoxia and hyperoxia we studied the 8-iso-PGF2alpha release (a specific index of cellular oxidative stress) in young and old left pampiniform plexus rats. The basal 8-iso-PGF2alpha release showed a statistically significant difference P=0.0067 between young and old rats PP. After hypoxia and hyperoxia, the release was higher in young rats as compared to normoxia, respectively P=0.0001 and P=0.0002. After hypoxia the release was not modified in old rats P=0.544 while after hyperoxia the release was increased in old rats as compared to control P<0.0001. The results show how chronic hypoxia and hyperoxia represent two important causes of oxidative stress and lipid peroxidation in pampiniform plexus rats. In young rats an increase of oxidative stress suggests that pampiniform plexus is sensitive to variations of oxygen supply. In old rats the pampiniform plexus is liable to a reduction of oxygen-sensing mechanisms and it is possible that the missing oxidative answer to the hypoxia in old rats is attributable in all likelihood to adaption to a hypoxic condition typical of aging.
Total IL-12 Levels are Increased in Paediatric Atopic Dermatitis: Correlations with Age and Disease Severity
Authors: Piancatelli D. , Bellotta L. , Del Beato T. , Duse M. , Della Penna M.R.
Published: January 2017

Alterations in the immune response play an important role in the pathogenesis of atopic dermatitis (AD). We evaluated the role of Th1/Th2 cytokine imbalance in paediatric AD and its progression towards other allergic manifestations. The levels of total interleukin (IL)-12 (p70+p40), IL-12 p70 and soluble CD30 (sCD30) were measured in paediatric patients affected by AD and in age-matched, non-atopic subjects (controls). The serum levels of total IL-12 and sCD30 were higher in patients than in controls (p=0.003 and p=0.053, respectively). Total IL-12 and sCD30 were also particularly increased in patients with severe disease. Serum levels of total IL-12 were negatively correlated with patient age (p=0.001): they were significantly higher in patients younger than 30 months, as compared to age-matched controls and to older patients and controls. Total IL-12 and sCD30 production were not significantly correlated with disease outcomes (atopic march) or with a family history of atopy. Our data show that total IL-12 levels are strongly associated with Th2 activation and severe disease in children with AD. The increased production of total IL-12 at an early age might indicate a different immune modulation and suggests the possibility of new therapeutic approaches for allergic diseases, in particular AD, early in childhood.
No Evidence of Involvement of Chlamydia Pneumoniae in Lung Cancer by Means of Quantitative Real-Time Polymerase Chain Reaction
Authors: Sessa R. , Santino I. , Pietro Di M. , Schiavoni G. , Ripa C. , Galdiero M. , Iannone M. , Izzo L. , Mingazzini P.L. , Bolognese A. , Del Piano M.
Published: January 2017

Chlamydia pneumoniae, an obligate intracellular pathogen, is well-known as etiological agent of acute respiratory infections; the repeated or prolonged exposure to chlamydial antigens may promote the persistence of C. pneumoniae in the respiratory tract leading to chronic diseases, such as chronic obstructive pulmonary disease and asthma. The predilection of C. pneumoniae to cause respiratory tract infections combined with its persistent nature suggest that it might play a role in lung cancer. The aim of our study is to evaluate the involvement of C. pneumoniae in pathogenesis of lung cancer. We therefore investigated the presence of C. pneumoniae DNA in tumor lung tissues by using real-time PCR assay. Simultaneously, tumor and healthy tissues from the same patient with primary carcinoma lung were analyzed. C. pneumoniae DNA was not detected in a single lung tumor tissue by means of an highly sensitive, and specific real-time PCR assay based on FRET hybridization probes. In conclusion, this study does not support the involvement of C. pneumoniae in the pathogenesis of lung cancer, suggesting that further investigations are needed to clarify other potential causative factors for the development of this malignancy.
Real-Time Polymerase Chain Reaction and Laser Capture Microdissection: An Efficient Combination Tool for Chlamydophila Pneumoniae DNA Quantification and Localization of Infection in Atherosclerotic Lesions
Authors: Ciervo A. , Mancini F. , Sale P. , Russo A. , Cassone A.
Published: January 2017

Chlamydophila pneumoniae has been implicated in atherosclerosis, but the role of this obligate intracellular pathogen in the development of the above pathology is still unclear. In particular, its presence and quantitative distribution within lesional areas has not yet been defined. We studied 18 carotid biopsies obtained from patients undergoing endoartherectomy. By laser microdissection (LCM), two different sites (intra-plaque and plaque-adjacent areas) were taken from each lesion, and the presence and quantity of the pathogen DNA were determined by real-time polymerase chain reaction (Real-time PCR). A total of 8 plaques, exclusively, from patients with unstable angina, were positive in real-time PCR. The bacterial DNA was detected in both lesional areas of 3 plaques which contained the highest number of DNA copies (1,900 to 2,200 copy numbers), while C. pneumoniae DNA was detected only in the intra-plaque area of the other 5 positive (500 to 1,600 copy numbers). No C. pneumoniae DNA was found in the other 10 plaques of which 6 were from patients with unstable angina and 4 from stable angina patients. No DNA from Helicobacter pylori or Cytomegalovirus was found in any plaque. This is the first report where both the target lesion and an adjacent reference site were evaluated for the presence of C. pneumoniae DNA by the combination of LCM and Real-time PCR assays. The integration of these two methodologies offer an excellent tool for in situ studies and may help to elucidate the putative role of C. pneumoniae in atherosclerosis.