bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Phytoncide from Trees on Human Natural Killer Cell Function
Authors: Li Q. , Kobayashi M. , Wakayama Y. , Inagaki H. , Katsumata M. , Hirata Y. , Hirata K. , Shimizu T. , Kawada T. , Park B.J. , Ohira T. , Kagawa T. , Miyazaki Y.
Published: January 2017

We previously reported that the forest environment enhanced human natural killer (NK) cell activity, the number of NK cells, and intracellular anti-cancer proteins in lymphocytes, and that the increased NK activity lasted for more than 7 days after trips to forests both in male and female subjects. To explore the factors in the forest environment that activated human NK cells, in the present study we investigate the effect of essential oils from trees on human immune function in twelve healthy male subjects, age 37–60 years, who stayed at an urban hotel for 3 nights from 7.00p.m. to 8.00a.m. Aromatic volatile substances (phytoncides) were produced by vaporizing Chamaecyparis obtusa (hinoki cypress) stem oil with a humidifier in the hotel room during the night stay. Blood samples were taken on the last day and urine samples were analysed every day during the stay. NK activity, the percentages of NK and T cells, and granulysin, perforin, granzyme A/B-expressing lymphocytes in blood, and the concentrations of adrenaline and noradrenaline in urine were measured. Similar control measurements were made before the stay on a normal working day. The concentrations of phytoncides in the hotel room air were measured. Phytoncide exposure significantly increased NK activity and the percentages of NK, perforin, granulysin, and granzyme A/B-expressing cells, and significantly decreased the percentage of T cells, and the concentrations of adrenaline and noradrenaline in urine. Phytoncides, such as α-pinene and β-pinene, were detected in the hotel room air. These findings indicate that phytoncide exposure and decreased stress hormone levels may partially contribute to increased NK activity.
Immunomodulatory Activity of a Plant Extract Containing Human Papillomavirus 16-E7 Protein in Human Monocyte-Derived Dendritic Cells
Authors: Di Bonito P. , Grasso F. , Mangino G. , Massa S. , Illiano E. , Franconi R. , Fanales-Belasio E. , Falchi M. , Affabris E. , Giorgi C.
Published: January 2017

This study reports the immunomodulatory activity on human Monocyte Derived Dendritic Cells (MDDCs) of a vaccine preparation shown to be effective against an HPV16-related tumour in an animal model. The vaccine is composed of extract from Nicotiana benthamiana leaves containing HPV16 E7 protein expressed by a potato virus X-derived vector (NbPVX-E7). The effect of the extract was evaluated on MDDC differentiation and maturation by monitoring the phenotypic expression of specific markers. The results show that NbPVX-E7 does not induce monocyte differentiation to dendritic cells, but does induce MDDC maturation. Plant extract does not influence MDDC-uptake of E7-FITC while it significantly improves the Ovalbumin-FITC uptake, considered as a model antigen. Importantly, NbPVX-E7-pulsed MDDCs/PBMCs are able to prime human blood-derived lymphocytes from healthy individuals to induce HPV16 E7-specific cytotoxic activity. This is a propaedeutic study for a possible use of E7-containing plant extract in human immunotherapy of HPV-related lesions.
HLA-A*01 is Associated with Late Onset of Alzheimer's Disease in Italian Patients
Authors: Guerini F.R. , Tinelli C. , Calabrese E. , Agliardi C. , Zanzottera M. , De Silvestri A. , Franceschi M. , Grimaldi L.M.E. , Nemni R. , Clerici M.
Published: January 2017

In this study, the distribution of HLA-A alleles was analyzed in Italian Alzheimer's Disease (AD) patients. Interaction between HLA alleles, APOE genotypes, age of onset, and gender were also analyzed. The results were compared to those obtained in healthy controls (HC). One hundred-seventy-three AD patients and 258 age-and-sex-matched healthy controls were enrolled in the study. AD patients were classified according to age at the onset of disease using quartiles of the distribution. HLA-A genotyping was performed by PCR-SSP; APOE genotyping was performed by RFLP. A correlation between late disease onset and HLA-A*01 was observed. Thus, HLA-A*01, calculated as number of alleles, was significantly more present in patients with age of onset > 74 years than in HC (20% vs 10.5%; p=0.014); the distribution of this allele was skewed also in patients 68.1–74.0 years of age (16.3%), even if the difference did not reach statistical significance. The relative risk ratio (RRR) of AD onset calculated by a multinomial logistic regression adjusted for sex and presence of APOE-4 confirmed a significant association of HLA-A*01 with AD onset > 74.0 years of age (RRR=2.2; 95%CI: 1.1 – 4.6; p=0.033). A high RRR (2.04) was also present in patients 68.1 – 74 years (p=0.064). Lower age of disease onset did not correlate with HLA-A*01. Data herein suggest that the presence of HLA-A*01 results in delayed AD development, even in patients carrying APOE-4. These results could offer new insights into the etiopathogenesis of Alzheimer's disease.
Does Spleen Volume Play a Role in Patients with HCV-Related Chronic Hepatitis?
Authors: Tarantino G. , Conca P. , Tarantino M. , Di Minno M.N.D. , Grimaldi E. , Chianese D. , Riccio A. , Scopacasa F. , Capone D.
Published: January 2017

As the lymphotropism of hepatitis C virus (HCV) has already been ascertained, and in the light of the fact that the immune defense system is an organized network composed of functionally interrelated tissues, this study was carried out to verify the possible involvement of spleen in HCV-related chronic hepatitis. In this cross-sectional study we measured spleen longitudinal diameter by ultrasound, beta2-microglobulin serum levels and splenic artery resistivity index (SARI) by Doppler in 51 patients treated with standard combined (Peg-Interferon plus Ribavirin) antiviral therapy. Thirty-three patients (17 females) completed the regimen and were compared to 31 controls (16 females). The mean basal values of spleen longitudinal diameter were higher in patients with chronic hepatitis than in controls, i.e., 116 mm (9.4) versus 102.7 mm (9.3), P = 0.0001. In the same patients a significant trend towards increased spleen longitudinal diameter was found after antiviral therapy, independently of the stage of HCV-related chronic hepatitis. The median values of the beta2-microglobulin concentrations were not significantly higher in the patients with HCV-related chronic hepatitis compared to controls, i.e., 1.3 (0.5 – 2.6) versus 1 (0.6 – 1.4), P = 0.16, although during the course of therapy they were significantly increased. SARI values of HCV-related chronic hepatitis patients were different from those of controls, but were unvaried compared to values at the end of treatment. Neither spleen measurements nor serum beta2-microglobulin levels were able to predict therapeutic response to antiviral therapy. A stimulation/ expansion of lymphoid tissue was found in patients with HCV-related chronic hepatitis. Such evidence raises the question whether physicians should continue to prescribe antiviral therapy in non-responders and supports the use of a new scheme (SLD plus β2-MG) to diagnose this ongoing, apparently reversible but nevertheless dangerous immunologic process.
Exhaled Nitric Oxide and Nitric Oxide Synthase Expression in Hodgkin's Disease
Authors: Guida G. , Culla B. , Scirelli T. , Bellone G. , Sciascia S. , Brussino L. , Novero D. , Palestro G. , Heffler E. , Gavarotti P. , Rolla G. , Bucca C.
Published: January 2017

Hodgkin's disease (HD) is a malignant lymphoma with frequent mediastinal involvement, characterized by a significant inflammatory infiltration. Exhaled Nitric Oxide (FENO), is present in healthy humans, and has been proven to be increased in eosinophilic diseases such as allergic asthma. We investigated whether FENO is increased in mediastinal HD and whether NO is produced by lymphoma tissue. To this aim FENO was measured in 56 HD patients, 17 with and 39 without bulky mediastinal involvement, in the period from January 2007 to December 2008. Thirty-seven patients were reassessed after remission. Lymph node biopsies of 10 patients were evaluated for inducible (iNOS) and constitutive (eNOS) nitric oxide synthase expression by immunohistochemistry. FENO resulted significantly related to the mediastinal mass maximum diameter (p=0.009) and was significantly higher in patients with as compared to those without bulky mediastinal disease (38.7 ppb, CI95% 19.3–58.0, versus 20.7 ppb, CI95% 16.6–24.7; p=0.009). iNOS and eNOS immunoreactivity was observed in tumour and inflammatory cells (eosinophils and histiocytes). Only in patients with bulky mediastinal HD there was a significant decrease in FENO (from 50.4 ppb CI95% 18.0–82.8 to 11.1 ppb CI95% 4.4–17.8, p=0.011). In conclusion, high FENO and NOS expression in lymph-nodes indicate that NO is a component of the inflammatory network of HD. FENO may be proposed for the assessment and follow up of bulky mediastinal HD patients.
Verbena Officinalis Essential Oil and its Component Citral as Apoptotic-Inducing Agent in Chronic Lymphocytic Leukemia
Authors: De Martino L. , D'Arena G. , Minervini M.M. , Deaglio S. , Fusco B.M. , Cascavilla N. , De Feo V.
Published: January 2017

We evaluated the pro-apoptotic activity of Verbena officinalis essential oil and of its main component citral, on lymphocytes collected from normal blood donors and patients with chronic lymphocytic leukemia (CLL). The number of apoptotic cells was greater in CLL patients than in healthy subjects at all different times of incubation (4, 8 and 24 hours) for samples treated with Verbena officinalis essential oil (A) and citral (B) as well vs controls at different concentrations (0.1% and 0.01%). The greater pro-apoptotic ability was showed by both essential oil of Verbena officinalis and citral at lower concentrations (after 4 h A 0.1%: 17.8% vs 37.1%; A 0.01%: 15.8% vs 52%; B 0.1%: 18.4% vs 46.4%; B 0.01%: 15.8% vs 54.2%; after 8 h A 0.1%: 23% vs 38%; A 0.01%: 22.2% vs 55%; B 0.1%: 32% vs 42.2%; B 0.01%: 22% vs 54.3%; after 24 h A 0.1%: 5% vs 20.7%; A 0.01%: 25.8% vs 47.2%; B 0.1%: 18.4% vs 46.4%; B 0.01%: 15.8% vs 54.2%). Patients carrying deletion 17p13 (p53 mutation) showed a reduced ability to undergo apoptosis with respect to patients with other genomic aberrations or normal karyotype. The proapoptotic activity of Verbena officinalis essential oil and citral is thought to be due to a direct procaspase 3 activation. These data further support evidence that indicate natural compounds as a possible lead structure to develop new therapeutic agents.
Analysis on the Co-Localization of Asbestos Bodies and Fas or CD163 Expression in Asbestos Lung Tissue by in-Air Micro-Pixe
Authors: Matsuzaki S. , Shimizu Y. , Dobashi K. , Nagamine T. , Satoh T. , Ohkubo T. , Yokoyama A. , Ishii Y. , Kamiya T. , Arakawa K. , Makino S. , Utsugi M. , Ishizuka T. , Tanaka S. , Shimizu K. , Mori M.
Published: January 2017

To prevent and control diseases caused by exposure to various agents, it is necessary to determine the harmful level for intervention and to establish a method for measuring that level. In-air microparticle-induced X-ray emission (in-air micro-PIXE) analysis is based on irradiation of specimens with a proton ion microbeam, and has been modified for biological application. Two-dimensional analysis and quantitative analysis using the system confirmed that asbestos induced apoptosis by upregulating Fas expression and also revealed the accumulation of CD163-expressing macrophages in the lungs of patients with asbestosis. By quantitative comparison of the area of Fas or CD163 expression and the Fas- or CD163-negative area in asbestos lung tissue, the harmful levels which caused the expression of Fas or CD163 could be estimated on Silica, Ferrous iron, and Magnesium (the components of asbestos) deposition. These results indicate that the system could be useful for investigating the pathogenesis of inhaled particle-induced immune reactions and for determining harmful levels of exogenous agents.
Beta2-Glycoprotein I is a Target of T Cell Reactivity in Patients with Advanced Carotid Atherosclerotic Plaques
Authors: Profumo E. , Buttari B. , Alessandri C. , Conti F. , Capoano R. , Valesini G. , Salvati B. , Riganò R.
Published: January 2017

Evidence in animal models that beta2-glycoprotein I (β2GPI), the principal target of autoimmune antiphospholipid antibodies, is involved in the initiation and progression of atherosclerosis, prompted us to investigate the possible role of this self protein as a target autoantigen of immune reactions in patients with carotid atherosclerosis. Plaque-infiltrating T lymphocytes from patients, and circulating T lymphocytes from patients and healthy subjects were tested by cell proliferation assay and by flow cytometry for intracellular cytokine expression in response to β2GPI. ELISA was used to detect cytokine production in culture supernatants and anti-β2GPI/anti-cardiolipin antibodies in serum samples. Eight of 35 PBMC samples and 1 of 5 plaque-infiltrating T lymphocyte samples from patients proliferated in response to β2GPI, whereas PBMC from healthy subjects did not. Patients' PBMC samples that proliferated in response to β2GPI produced significantly higher IFN-γ and TNF-α than non-proliferating PBMC. β2GPI-specific plaque-derived T lymphocytes expressed IFN-γ, TNF-α and IL-4, suggesting concomitant Th1 and Th2 activation. Only one patient's serum was positive for anti-β2GPI and anti-cardiolipin IgM antibodies. These new findings indicate that β2GPI induces a cellular immune response in a subpopulation of patients with carotid atherosclerosis thus contributing to the inflammatory responses involved in carotid atherosclerotic disease.
Essential Amino Acids Improve Insulin Activation of Akt/mTOR Signaling in Soleus Muscle of Aged Rats
Authors: Flati V. , Caliaro F. , Speca S. , Corsetti G. , Cardile A. , Nisoli E. , Bottinelli R. , D'Antona G.
Published: January 2017

Essential amino acids (EAA) improve basal muscle protein synthesis in the elderly. Nevertheless, in settings of prolonged supplementation, putative signal pathways of EAA are currently unknown. The purpose of this study was to test the effects of prolonged supplementation of EAA enriched mixture (12-L-Amin) on Insulin/Insulin-like Growth Factor-1 (IGF1) pathway by measuring total and phosphorylated Akt (Ser473) and its upstream (IRS1 at Ser636) and downstream (mTOR at Ser2448, p70S6K at Thr389) targets in basal conditions and following acute insulin (0.1 U/L) incubation in vitro. To this aim, soleus muscles were dissected from male Wistar rats divided in three groups of 7 each: adults (AD, 10 mo of age), elderly (EL, 22 mo of age) and elderly supplemented (EL-AA, 12-L-Amin 1.5gr/Kg die in drinking water for 3mo). EL showed reduced basal and post-insulin mTOR and p70S6K activation and reduced post-insulin IRS1 degradation relative to AD. EL-AA showed an increase of post-insulin Akt activation, no change in basal and post-insulin phospho-mTOR, lower reduction of phospho-p70S6K and increased post-insulin IRS1 degradation relative to AD. These results demonstrate that chronic 12-L-Amin administration exerts anti-ageing effects on the activation/inactivation of the Insulin/IGF1/mTOR pathway which is identified as putative target of EAA in the elderly.
Identification of SARS-COV Spike Protein-Derived and HLA-A2-Restricted Human CTL Epitopes by Using a New Muramyl Dipeptide-Derivative Adjuvant
Authors: Chen Y.-Z. , Liu G. , Senju S. , Wang Q. , Irie A. , Haruta M. , Matsui M. , Yasui F. , Kohara M. , Nishimura Y.
Published: January 2017

Severe acute respiratory syndrome (SARS) spread during the winter of 2003, and attempt have been made to develop vaccines against SARS corona virus (SARS-CoV). The present study provides a strategy to rapidly identify SARS-CoV-derived antigenic peptides recognized by HLA-A2-restricted cytotoxic T lymphocytes (CTLs). Forty-three candidate peptides having HLA-A2-binding motifs were selected in silico and HLA-A2/Db chimeric MHC class I-transgenic mice were immunized with these peptides and a new derivative of muramyl dipeptide that can induce upregulation of HLA-DR, CD80, CD86, and CD40 in human CD14+ antigen presenting cells, was administered as an adjuvant. Six HLA-A2-restricted mouse CTL epitopes were identified, including two new epitopes which have never been reported before. One of the novel peptides was naturally processed and successfully induced HLA-A2-restricted specific CTLs in both HLA transgenic mice and healthy donors. The method was useful, convenient and efficient for rapid identification of CTL epitopes derived from SARS-CoV proteins and will be possibly applicable for other pathogens to develop a peptide-based vaccine.
Effect of Temperature on the Shift of Pseudomonas Fluorescens from an Environmental Microorganism to a Potential Human Pathogen
Authors: Donnarumma G. , Buommino E. , Fusco A. , Paoletti I. , Auricchio L. , Tufano M.A.
Published: January 2017

Pseudomonas fluorescens is a Gram-negative bacterium generally considered of scarce clinical significance. However, in the last few years, the isolation of P. fluorescens as causative agent of nosocomial infections has rapidly increased. P. fluorescens is a psychrophile microorganism which grows at an optimal temperature of 25–30°C. In spite of this constraint, it has recently been reported that the human physiological temperature does not appear to be a barrier for this microorganism. In this study we examined the ability of P. fluorescens, grown at 28°C or at 37°C, to adhere to cultured human A549 pulmonary cells and to form biofilm. The ability of P. fluorescens to induce expression of proinflammatory cytokines, beta-defensin 2 and the intercellular adhesion molecule-1 was also investigated. Our results clearly indicate that inflammatory mediators are induced when the microorganism is grown at a lower temperature, while biofilm is formed only at 37°C. The results presented are consistent with previous reports indicating P. fluorescens as an opportunistic pathogen and underscore the urgent need for further studies to better characterize the virulence of this microorganism.
Clinical Outcome Evaluation following Cyclosporine a Treatment in Moderate to Severe Psoriasis: A Retrospective Study
Authors: Colombo D. , Flori L. , Altomare G. , Aste N. , Sgarbi S.
Published: January 2017

This retrospective study was conducted on 193 patients treated in three Italian Psoriasis Units with the aim of evaluating the evolution of psoriasis severity and the safety of cyclosporin A (Sandimmun Neoral®) in moderate to severe psoriasis, at the regimens usually employed in common clinical practice. Cyclosporin A (CyA) was administered for a mean period of 14 months, the mean number of treatment courses was 1.6 (range 1–4), and the mean dosage ranged from 1.5 to 3.1 mg/kg/die. Ninety percent of patients obtained complete therapeutic success or clinical remission, defined as complete clearance of lesions or clearance of lesions with residual minor pigmentations respectively, when treated with CyA in monotherapy. The mean Psoriasis Area and Severity Index (PASI) decreased from 23.31 before CyA administration to 5.64 at the end of treatment. The clinician's judgement on CyA tolerability was good/very good in 90% of cases. Adverse events occurred in 36% of patients, with hypertension being the most commonly reported (17.6%). The results of this study indicate that in the common clinical practice CyA in moderate to severe psoriasis is usually employed at low doses, resulting both safe and effective.
Comparison of Anti-Bacterial Prophylactic Properties of Two Different Vascular Grafts: Action of Anti-Bacterial Graft Coating and Systemic Antibiotic Treatment
Authors: Artini M. , Scoarughi G.L. , Papa R. , Cellini A. , Avanzolini C. , Mileto E. , Rizzo L. , Selan L.
Published: January 2017

The main problem associated with artificial vascular devices is the risk of bacterial infections, mostly sustained by coagulase negative staphylococci or Staphylococcus aureus. Many efforts have been made to identify materials refractory to bacterial adhesion. The aim of our study is to verify the antimicrobial properties of two kinds of vascular prosthesis to prevent early onset infections and the efficacy of the concomitant action of a systemic antibiotic treatment. Adult male Wistar rats were used. We subcutaneously implanted in four groups a silver-coated prosthesis fragment, and a rifampicin-soaked prosthesis fragment in the remaining four groups. We inoculated in the site of implant a high bacterial burden of S. aureus in four groups and a low burden in the remaining groups. Systemic levofloxacin was administered for seven days in four groups representing the two kinds of prosthesis; after 21 days the rats were sacrificed, prosthesis fragments were sonicated and the corresponding supernatants were plated for bacterial counts. The rifampicin-soaked prostheses explanted from rats treated with levofloxacin were sterile, regardless of the bacterial inoculum. In other groups some prostheses were colonized. In the experimental rat model used, the action of local and systemic antibiotic treatment was able to reduce colonisation of artificial prostheses.
Efforts to Counteract Locally the Effects of Systemic Immunosupression: A Review on the Use of Imiquimod, a Topical Immunostimulator in Organ Transplant Recipients
Authors: Trakatelli M. , Katsanos G. , Ulrich C. , Kalabalikis D. , Sotiriadis D. , Stockfleth E.
Published: January 2017

The potent systemic immunosuppression therapy necessary to sustain a life-saving solid organ transplant is associated with an increased incidence of various infections including human papillomavirus infection and skin cancers in organ transplant recipients. Imiquimod, a topical agent that functions through local induction of a specific anti-viral or anti-tumor immune response, appears to be a promising therapeutic option that could potentially counteract in situ the effects of systemic immunosupression in this vulnerable group. Up-to-date studies using this local immune-response modifier in transplanted patients have yielded reassuring and encouraging results regarding its safety and efficacy in this population. However, in order to establish the use of imiquimod as a standard treatment option for organ transplant recipients, additional research and clinical trials are required.
TNFR1 mRNA Expression Level and TNFR1 Gene Polymorphisms are Predictive Markers for Susceptibility to Develop Invasive Pulmonary Aspergillosis
Authors: Sainz J. , Salas-Alvarado I. , López-Fernández E. , Olmedo C. , Comino A. , García F. , Blanco A. , Gómez-Lopera S. , Oyonarte S. , Bueno P. , Jurado M.
Published: January 2017

Tumour necrosis factor (TNF) is primarily secreted by monocytes/macrophages and activated T lymphocytes in response to fungal infections. TNF acts through TNF receptor 1 (TNFR1) triggering a pro-inflammatory response, and therefore plays a pivotal role in immune regulation and host immune responses. We hypothesized that single nucleotide polymorphisms (SNPs) in TNFR1 gene may influence the innate immune response against Aspergillus. Three SNPs were genotyped in 275 individuals (144 immunocompromised haematological patients with high-risk of developing IPA and 131 healthy controls): TNFR1-383(A/C) (rs2234649) and TNFR1-609(G/T) (rs4149570) in the 5′ UTR region, and TNFR1+36(A/G) SNP (rs767455) in the first exon of the gene. Of the 144 haematological patients, 77 patients developed Invasive Pulmonary Aspergillosis (IPA) infection and the remaining 67 patients were not infected. TNFR1+36(A/G) and TNFR1-609(G/T) were associated with IPA susceptibility (p=0.033 and p=0.018, respectively). A role of TNFR1 genetic variants in the susceptibility of patients to develop IPA was also supported by the significantly lower TNFR1 mRNA expression level in IPA than in IPA-resistant patients and the strong correlation between the TNFR1-609 genetic variant and the expression levels of TNFR1. There was also a tendency for a higher frequency of galactomannan (GM) positivity in patients with TNFR1-609G/G genotype than in patients with TNFR1-609G/T (p=0.0909) or TNFR1-609T/T (p=0.0913) genotype. Predictive sequence analysis of the effects of TNFR1-609 promoter polymorphism revealed that this SNP might play a critical role in modifying the affinity of ICSBP/IRF-8, a transcription factor that is involved in the TNFR1-mediated activation of NFκB signalling pathway. Taken together, these data suggest that TNFR1 polymorphisms influence the risk of IPA disease and might be useful for risk stratification strategies. These findings need to be confirmed in validation studies with larger samples of haematological patients.
Continuous Treatment of Plaque-Type Psoriasis with Etanercept: An Observational Long-Term Experience
Authors: Esposito M. , Giunta A. , Mazzotta A. , Babino G. , Talamonti M. , Chimenti M.S. , Chimenti S.
Published: January 2017

To assess the long-term efficacy and safety profile and the patient-reported outcomes (PRO) in patients with moderate-to-severe plaque-type psoriasis receiving continuous etanercept treatment. An open-label study was conducted to evaluate etanercept as long-term treatment for moderate-to-severe plaque psoriasis. Continuous therapy was administered at a dose of 50 mg subcutaneously twice weekly for 12 weeks followed by a continuous treatment with 50 mg subcutaneously once weekly or 25 mg twice weekly throughout a 96-week study. The primary measure of efficacy was the proportion of patients with PASI 75 at week 24, 48 and 96. Patient-reported outcomes (PRO) were also assessed during the study, at week 24, 48 and 96, including the Dermatology Life Quality Index (DLQI) and the Psoriasis Disability Index (PDI). At baseline, mean PASI score, DLQI and PDI for patients eligible to initiate treatment with etanercept showed significant disease severity, quality-of-life impairment and psoriasis-related disability. At week 96, patients showed statistically significant and meaningful improvements. The continuous etanercept regimen provided a consistent improvement in both clinical disease parameters and PRO measures.
Apoptosis and Autoimmunity Induced by Clodronate in Systemic Lupus Erythematosus Mononuclear Circulating Cells
Authors: Cacciapaglia F. , Spadaccio C. , Gregorj C. , Margiotta D. , Coccia R. , De Marco F. , Chello M. , Picardi A. , Amoroso A. , Afeltra A.
Published: January 2017

The aim of this study is to evaluate the effect of clodronate on apoptosis of human systemic lupus erythematosus circulating mononuclear cells and to analyze possible correlations with changes in autoantibody production in vitro. Lympho-monocytes from 20 SLE patients were isolated and incubated with or without addition of 1 μM clodronate for 72 hours. Apoptosis and release of genomic material was assessed by immunofluorescent detection of cleaved caspase-3 and by Cell-Death-Detection ELISAPLUS kit (Roche). Anti-Nucleosome IgG and anti-dsDNA IgM and IgG autoantibody levels were determined in supernatants by commercially available ELISA kits. Clodronate induced apoptosis in monocytes as confirmed by cleaved caspase-3 immunostaining and by quantification of cleaved nucleosome in the supernatants (treated 0.22±0.05 O.D. vs untreated 0.09±0.04 O.D.; P<0.001). This finding was coupled with a significant increasing in supernatants of IgG anti-Nucleosome (treated 6.5±1.1 vs untreated 5.5±0.6 IU/mL; p=0.001) and IgM (treated 3.0±1.3 vs 2.2±0.9 IU/mL; p=0.02) and IgG (treated 4.0±1.8 vs untreated 2.8±1.5 IU/mL; p=0.02) anti-dsDNA autoantibody levels. Our findings stressed the proapoptotic activity of clodronate, as well as its potential autoimmunity induction in SLE mononuclear circulating cells. Clinical studies could clarify the role of bisphosphonates on autoantibody production and worsening of disease activity.
Antiproliferative, Protective and Antioxidant Effects of Artichoke, Dandelion, Turmeric and Rosemary Extracts and Their Formulation
Authors: Menghini L. , Genovese S. , Epifano F. , Tirillini B. , Ferrante C. , Leporini L.
Published: January 2017

Artichoke, dandelion, turmeric extracts and rosemary essential oil are commonly used as ingredients in many herbal preparations to treat hepatic and gallbladder disorders. In the present work we compare the activity of each single extract with a commercial mixture for antiproliferative, antiradical and protective effects against induced oxidant stress effect. In ABTS and DPPH tests, turmeric extract is the most active, followed by artichoke and dandelion. All samples exhibited antiproliferative activity in a dose-dependent manner against HepG2 cells. In the same cell lines, the protective effect of pre-treatment with the extracts were detected by evaluating the prostaglandin E2 release, a marker of oxidative stress induced by hydrogen peroxide. The treatments with the extracts were efficient in reducing the release of PGE2 induced by oxidative stimulus. The positive results of the cell viability test, together with the protective and antiradical activity confirm the rationale for the use of these ingredients in commercial formulations as a health aid tool in modern phytotherapy.
Effect of Inhibition of the Ubiquitin-Proteasome-System and IκB Kinase on Airway Inflammation and Hyperresponsiveness in a Murine Model of Asthma
Authors: El-Hashim A.Z. , Renno W.M. , Abduo H.T. , Jaffal S.M. , Akhtar S. , Benter I.F.
Published: January 2017

The current treatment of asthma is far from optimal and there is a need for novel therapeutic approaches. NFκB has recently been highlighted as an important pro-inflammatory transcriptional factor and its blockade is believed to represent a new therapeutic approach for asthma. The purpose of this study is to investigate the effects of blocking the actions of NFκB, through inhibition of the ubiquitin-proteasome system (UPS) or IκB kinase (IKK), in a murine model of asthma. Treatment with the UPS inhibitor, MG-132 (0.03 and 0.1 mg/kg), did not significantly affect the ovalbumin-induced increase in total and differential cell numbers, histological changes such as perivascular and peribronchial inflammatory cell infiltration, perivascular and peribronchial fibrosis or the increased Penh to methacholine. In contrast, treatment of mice with the IKK inhibitor, BAY 11-7085, (3 and 10 mg/kg) dose-dependently inhibited the ovalbumin-induced increase in airway leukocyte influx and decreased the percentage of airway lymphocytes, neutrophils and eosinophils. Also, BAY 11-7085-treated (10 mg/kg) mice showed a significant decrease in the histologically assessed inflammatory indices as well as a significant reduction in the ovalbumin-induced increase in Penh to inhaled methacholine. Furthermore, BAY 11-7085 significantly inhibited the ovalbumin-induced increase in the level of phosphorylation of IκBα and extracellular regulated kinases (ERK) 1/2, whilst MG-132 significantly increased the phosphorylation of (ERK) 1/2. These findings confirm the critical role that NFκB plays in airway inflammation, highlight the importance of IKK in regulating the pro-inflammatory activity of NFκB and also suggest that UPS may not be a useful drug target for asthma treatment.
Do DNA-Methylation and Histone Acetylation Play a Role in Clear Cell Renal Carcinoma? Analysis of Radical Nephrectomy Specimens in a Long-Term Follow-up
Authors: Minardi D. , Lucarini G. , Filosa A. , Zizzi A. , Milanese G. , Polito M. , Polito M. , Di Primio R. , Montironi R. , Muzzonigro G.
Published: January 2017

We investigated global methylation and histone acetylation in 50 conventional clear cell renal carcinomas (RCC), treated with radical nephrectomy, to assess their possible role as diagnostic biomarkers. The features considered in this study were patient age, tumor size and grade, percentage and intensity of 5-methylcytosine (5mc) and Acetyl-Histone (Lys 9) expression in tumor tissue. All considered parameters were correlated with patient specific survival. The mean percentage of global cellular methylation in tumoral tissue was significantly higher compared to normal peritumoral tissue (p<0.0001), while the intensity of cellular methylation was significantly higher in normal tissue than in tumoral tissue (p=0.001). The mean percentage of histone cellular acetylation in tumoral tissue was significantly lower compared to normal peritumoral tissue (p=0.0005), while the intensity of mean acetylation in neoplastic tissue was similar to the normal tissue. The percentage of global DNA methylation was significantly higher in grades 3 and 4 tumors (p=0.033). Global DNA methylation and histone acetylation in tumoral tissue did not correlate with survival. Fuhrman grade was statistically significant for prognosis (p=0.031). In conclusion, global hypermethylation and histone hypoacetylation play an important role in RCC carcinogenesis; Fuhrman grade is still considered the most important factor for patient survival; 5mc can have a role as markers of aggressiveness.
Therapeutic Effectiveness of Minimal Doses of Rituximab in a Patient with Rheumatoid Arthritis
Authors: Bruzzese V.
Published: January 2017

We report the case of a man suffering from rheumatoid arthritis, resistant to common DMARDs and anti-TNF-alpha, who received an excellent response, in terms of effectiveness and depletion of CD20 positive B-lymphocytes, to minimal doses of anti-CD20 monoclonal antibody (rituximab). The dose used was only 100 mg, repeated after 2 weeks. Already after the first infusion of rituximab, a profound depletion of CD20 B-lymphocytes and an improvement of clinical symptoms were evident. The patient, after 4 months from the first two infusions, maintained an accentuated lymphocyte depletion and obtained a low disease activity, passing from an initial DAS28 of 6.3 to a DAS28 of 2.8. The possible practical implications of this observation are taken into consideration.
Pathogenesis and Clinical Approaches to Anticonvulsant Hypersensitivity Syndrome: Current State of Knowledge
Authors: Scaparrotta A. , Verrotti A. , Consilvio N.P. , Cingolani A. , Di Pillo S. , Di Gioacchino M. , Verini M. , Chiarelli F.
Published: January 2017

Anticonvulsant hypersensitivity syndrome (AHS) is a rare, but severe and potentially fatal, adverse reaction that occurs in patients who are treated with commonly used older anticonvulsant drugs (phenytoin, carbamazepine and phenobarbital) and/or with some newer agents (lamotrigine). Paediatric patients are at an increased risk for the development of AHS for the higher incidence of seizure disorder in the first decade of life. Hypersensitivity reactions range from simple maculopapular skin eruptions to a severe life-threatening disorder. AHS is typically associated with the development of skin rash, fever and internal organ dysfunctions. Recent evidence suggests that AHS is the result of a chemotoxic and immunologically-mediated injury, characterized by skin and mucosal bioactivation of antiepileptic drugs and by major histocompatibility complex-dependent clonal expansion of T cells. Early recognition of AHS and withdrawal of anticonvulsant therapy are essential for a successful outcome. In vivo and vitro tests can be helpful for the diagnosis that actually depends essentially on clinical recognition.
Use of Complementary and Alternative Medicine in Children with Recurrent Acute Otitis Media in Italy
Authors: Marchisio P. , Bianchini S. , Galeone C. , Baggi E. , Rossi E. , Albertario G. , Torretta S. , Pignataro L. , Esposito S. , Principi N.
Published: January 2017

Controlling environmental factors, chemoprophylaxis, immunoprophylaxis and surgery are considered possible means of preventing recurrent acute otitis media (RAOM), but there are no available data concerning the paediatric use of complementary and alternative medicine (CAM). We evaluated the uses of CAM (homeopathy and/or herbal medicine) as means of preventing AOM in children with a history of RAOM. Eight hundred and forty Italian children with RAOM (⩾3 episodes in six months) aged 1–7 years were surveyed in 2009 using a face-to-face questionnaire, filled by parents or caregivers, that explored the prevalence, determinants, reasons, cost, and perceived safety and efficacy of CAM. About one-half (46%) of the children used CAM, significantly more than the number who used immunoprophylaxis (influenza vaccine 15%; p<0.05), PCV-7 34%; p< 0.05) or chemoprophylaxis (2%; p<0.001). Use of CAM in the family was the only important factor positively associated with the use of CAM in children (adjusted OR 7.94; 95% CI: 5.26–11.99). The main reasons for using CAM were a fear of the adverse effects of conventional medicine (40%) and to increase host defences (20%). CAM was widely seen as safe (95%) and highly effective (68%). CAM prescribers were paediatricians in 50.7% of cases; self-initiation was reported by 23% of respondents. CAM expenditure was between €25 and €50/month in 27.6% of cases and ⩾ €50/month in 16%. Children with RAOM should be considered among the categories of subjects likely to be using CAM. Together with the fact that paediatricians are the main prescribers, this is worrying because of the current lack of evidence regarding the efficacy, safety and cost-effectiveness of CAM in the prevention of RAOM.
Inflammatory Cells, Cytokines and Matrix Metalloproteinases in Amicrobial Pustulosis of the Folds and other Neutrophilic Dermatoses
Authors: Marzano A.V. , Cugno M. , Trevisan V. , Lazzari R. , Fanoni D. , Berti E. , Crosti C.
Published: January 2017

Amicrobial pustulosis of the folds (APF) is a rare cutaneous disease characterized by relapsing sterile pustules frequently associated with autoimmune disorders. Although APF pathophysiology is still undefined, scattered reports suggest involvement of neutrophils. The aim of the present study is to evaluate the role of the skin inflammatory infiltrate, selected multifunctional cytokines and effectors of tissue damage in APF and other neutrophilic dermatoses. We studied, by immunohistochemical methods, inflammatory cell markers (CD3, CD163, myeloperoxidase), cytokines (TNF-α, IL-8, IL-17), metalloproteinases (MMP-2, MMP-9) and vascular-endothelial-growth-factor (VEGF) in lesional skin from six patients with APF, 11 with pyoderma gangrenosum (PG), 7 with Sweet's syndrome, and in 20 normal skin samples. Immunoreactivities of CD3, CD163, myeloperoxidase, TNF-α, IL-8, IL-17, MMP-2, MMP-9 and VEGF were significantly higher in APF, PG and Sweet's syndrome than in controls (p=0.0001). IL-8 was more expressed in PG than in APF (P=0.002) and Sweet's syndrome (p=0.001). In APF, MMP-9 reactivity was higher than in Sweet's syndrome, (p=0.035), but less intense than in PG (p=0.020). Our study supports the role of proinflammatory cytokines/chemokines and MMPs as important effectors for the tissue damage in APF similarly to classic neutrophilic dermatoses.
Immunogenicity of An Interferon-β1a Product
Authors: Kauffman M.A. , Sterin-Prync A. , Papouchado M. , González E. , Vidal A.J. , Grossberg S.E. , Chuppa S. , Odoriz B. , Vrech C. , Diez R.A. , Ferro H.H.
Published: January 2017

In order to determine whether Blastoferon®, a biosimilar interferon (IFN)-β1a formulation, shares epitopes with other known IFN-β products, a series of neutralization bioassays were performed with a set of well-characterized anti-IFN-β monoclonal antibodies and human sera (World Health Organization Reference Reagents). The bioassay was the interferon-induced inhibition of virus cytopathic effect on human cells in culture (EMC virus and A-549 cells). Computer-calculated results were reported as Tenfold Reduction Units (TRU)/ml. To further assess Blastoferon® immunogenicity, in vivo production of anti-IFN β antibodies was determined in sera of patients included in the pharmacovigilance plan of Blastoferon® by the level of IFN-β1a binding antibodies (by enzyme immunoassay -EIA) and neutralizing antibodies (in the Wish-VSV system). The highly characterized neutralizing monoclonal antibodies A1 and A5 that bind to specific regions of the IFN-β molecule reacted positively with the three β1a IFNs: Blastoferon®, Rebif®, and the IFN-β WHO Second International Standard 00/572. As expected, the non-neutralizing monoclonal antibodies B4 and B7 did not neutralize any of the IFN-β preparations. The commercially available monoclonal antibody B-02 reacted essentially equally with Rebif® and Blastoferon®. The WHO Reference Reagent human serum anti-IFN-β polyclonal antibody neutralized all the IFN-β products, whereas the WHO Reference Reagent human serum anti-IFN-α polyclonal antibody G037-501-572 appropriately failed to react with any of the IFN-β products. On the basis of in vitro reactivity with known, well-characterized monoclonal and polyclonal antibody preparations, Blastoferon® shares immunological determinants with other human interferon-β products, especially IFN-β1a. In vivo antibodies were detected by EIA in 72.9% of 37 chronically treated multiple sclerosis patients, whereas neutralizing antibodies were found in 8.1% of them. Blastoferon® appears to have immunological characteristics comparable to other IFN-β1a products.
Exaggerated Imiquimod Application Site Reactions in the Context of Systemic Tumor Necrosis Factor-Alpha Inhibition: More Than a Coincidental Occurrence?
Authors: Saggini A. , Saraceno R. , Chimenti S.
Published: January 2017

Topical imiquimod and tumor necrosis factor (TNF)-α inhibitors have gained wide acceptance as safe and effective treatments for non-melanoma skin cancer (NMSC) and moderate to severe psoriasis, respectively. While mild to moderate application site reactions (ASRs) are a well-known and common phenomenon associated with imiquimod, the potential of TNF-α blockers to elicit cutaneous inflammatory reactions has only recently been recognized. We present two cases of severe, atypical ASRs which occurred during NMSC treatment with imiquimod in the context of infliximab administration for psoriasis, and consider the grounds supporting a non-fortuitous association. To date, no relation has ever been suggested between TNF-α inhibitors and increased susceptibility to developing exaggerated ASRs with imiquimod. We believe that this subject deserves further analysis; meanwhile, increased attention should be drawn to the possibility of this adverse interaction, as simultaneous treatment with TNF-α blockers and imiquimod is becoming increasingly frequent in daily practice.
Ulerythema Ophryogenes, a Rare and Often Misdiagnosed Syndrome: Analysis of an Idiopathic Case
Authors: Dianzani C. , Pizzuti A. , Gaspardini F. , Bernardini L. , Rizzo B. , Degener A.M.
Published: January 2017

Keratosis pilaris (KP) is a follicular hyperkeratosis disorder which is frequently detected in the adult population (44%), mostly in female adolescents (80%). It is a genetic autodominant dermatosis with variable penetrance, but no specific gene association has been determined, even though association to the presence of chromosome 18p deletion has been reported in some cases. We report the case of a 51-year-old Caucasian woman affected by keratosis pilaris gradually progressing with age and with a story of multiple abortions. Standard karyotype and CGH array analyses did not reveale any genetic abnormality. Virological analyses detected the presence of HPV 36 DNA inside the dorsum biopsy, leading to hypothesize its involvement in the evolution of the lesion. Clinical history and patient examination led the diagnosis of an idiopathic case of Ulerythema ophryogenes. The analysis of more cases could be useful to verify the involvement of cutaneous HPV in the progression of the clinical manifestation of the KP variants.
Atherosclerosis: A Classic Inflammatory Disease
Authors: Anogeianaki A. , Angelucci D. , Cianchetti E. , D'alessandro M. , Maccauro G. , Saggini A. , Salini V. , Caraffa A. , Teté S. , Conti F. , Tripodi D. , Shaik-Dasthagirisaheb Y.B.
Published: January 2017

Atherosclerosis is an inflammatory disease due to a diet high in saturated fat, hypercholesterolemia, obesity, hypoglycemia, etc. mainly mediated by the infiltration of macrophage and T cells into the vascular wall. Once the endothelial is damaged monocytes penetrate the tissue and are transformed in scavenger cells. Upon stimulation of Th1 cells, a group of cytokines is released and contributes to the inflammatory response of atherosclerotic tissue. When macrophages proliferate they amplify inflammatory response through the secretion of growth factors and cytokines such as TNF and IL-1. In addition, chemokines such as RANTES and other C-C chemokines are generated, and matrix metalloprotinease 9 (MMP-9) are produced by activated monocytes. However, the immune system in atherosclerosis still remains unclear. Here, in this study we revisited the inter-relationship between atherosclerosis and inflammation.
Effects of Paraquat and Capsaicin on the Expression of Genes Related to Inflammatory, Immune Responses and Cell Death in Immortalized Human HaCat Keratinocytes
Authors: Paolillo N. , Piccirilli S. , Giardina E. , Rispoli V. , Colica C. , Nisticò S.
Published: January 2017

The present experiments were aimed to characterize in immortalized human HaCat keratinocytes the gene expression induced by paraquat and capsaicin, two agents known to induce cell death or to affect inflammatory and pain pathways, respectively. In particular, the following set of genes were analysed by qRealtime PCR: CXCL10,CXCL11, IL-10 (inflammatory and immune responses), TP73, BCL2, (apoptotic and anti-apoptotic genes), MMP9 (proteolysis), SOD-1, BAK-1 and CAT (peroxysomal and microsomal oxidation pathways). In this way, we were able to differentiate the two toxins since they had a different profile of gene expression. In fact, paraquata was found to activate set of genes involved in inflammatory (CXL10,CXL11 and IL-10), and cell death (BCL2, BAK-1, MMP9) pathways. Another specific site of action of paraquat was represented by an activation of the gene involved in SOD-1 transcription. On the contrary, capsaicin was found to produce only an up-regulation of BCL2, an anti-apoptotic gene and MMP9, whereas no significant changes were reported in genes involved in inflammatory and immune responses. Finally, in comparison to previous experiments carried out with TNF-α and IL-1β, we have shown that paraquat produced a similar pattern of activation of set of genes involved both in inflammation and apoptosis.
Time-Related Dynamics of Variation in Core Clock Gene Expression Levels in Tissues Relevant to the Immune System
Authors: Mazzoccoli G. , Sothern R.B. , Greco A. , Pazienza V. , Vinciguerra M. , Liu S. , Cai Y.
Published: January 2017

Immune parameters show rhythmic changes with a 24-h periodicity driven by an internal circadian timing system that relies on clock genes (CGs). CGs form interlocked transcription-translation feedback loops to generate and maintain 24-h mRNA and protein oscillations. In this study we evaluate and compare the profiles and the dynamics of variation of CG expression in peripheral blood, and two lymphoid tissues of mice. Expression levels of seven recognized key CGs (mBmall, mClock, mPer1, mPer2, mCry1, mCry2, and Rev-erbα) were evaluated by quantitative RT- PCR in spleen, thymus and peripheral blood of C57BL/6 male mice housed on a 12-h light (L)-dark (D) cycle and sacrificed every 4 h for 24 h (3–4 mice/time point). We found a statistically significant time-effect in spleen (S), thymus (T) and blood (B) for the original values of expression level of mBmal1 (S), mClock (T, B), mPer1 (S, B), mPer2 (S), mCry1 (S), mCry2 (B) and mRev-Erbα (S, T, B) and for the fractional variation calculated between single time-point expression value of mBmal1 (B), mPer2 (T), mCry2 (B) and mRev-Erbα (S). A significant 24-h rhythm was validated for five CGs in blood (mClock, mPer1, mPer2, mCry2, mRev-Erbα), for four CGs in the spleen (mBmal1, mPer1, mPer2, mRev-Erbα), and for three CGs in the thymus (mClock, mPer2, mRev-Erbα). The original values of acrophases for mBmal1, mClock, mPer1, mPer2, mCry1 and mCry2 were very similar for spleen and thymus and advanced by several hours for peripheral blood compared to the lymphoid tissues, whereas the phases of mRev-Erbα were coincident for all three tissues. In conclusion, central and peripheral lymphoid tissues in the mouse show different sequences of activation of clock gene expression compared to peripheral blood. These differences may underlie the compartmental pattern of web functioning in the immune system.