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International Journal of Immunopathology and Pharmacology
1. Vol. 21, no. 3
1. Chemokines and bone remodeling pp 485 - 492 E. Galliera1, M. Locati1,2, A. Mantovani1,2 and M.M. Corsi1,3 1 Institute of General Pathology, Medical Faculty, University of Milan, Milan, Italy 2 IRCCS Istituto Clinico Humanitas, Rozzano, Italy 3 Laboratory of Biotechnological Applications, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy
Bone remodeling is characterized by spatial and temporal coupling of bone resorption and formation and is necessary for skeletal growth and normal bone structure maintenance. Imbalance of this process is related to metabolic bone disorders such as osteoporosis or rheumatoid arthritis. For this reason, bone remodeling is under the control of several local and systemic factors, including molecules of the immune system. The importance of the interplay of both the skeletal and immune systems is reflected by the emerging interdisciplinary research field, called osteoimmunology, focused on common aspects of osteology and immunology. This review focuses on the role of inflammatory mediators, such as cytokines in bone remodeling and, in particular, a subfamily of chemotactic cytokines or chemokines which are involved not only in several aspects of physiological bone remodeling but also in pathological bone disorders, such as rheumatoid arthritis or osteoporosis. Understanding the role of inflammation and chemokines will provide new insights for the treatment of diseases affecting both skeletal and immune systems, by the development of new therapeutic strategies targeting common inflammatory mediators.
2. Strategies to overcome obstacles to successful immunotherapy of melanoma pp 493 - 500
F. Pandolfi, R. Cianci, S. Lolli, I.S. Dunn1, E.E. Newton2, T.J. Haggerty1,3, L.A. Boyle1,3 and J.T. Kurnick1,3
Institute of Internal Medicine, Catholic University, Rome, Italy 1CytoCure LLC, Beverly, MA, USA 2Boston University School of Medicine, Boston, USA 3Massachussetts General Hospital and Harvard Medical School, Boston, USA
The immunogenicity of malignant melanomas has been recognized by the observed recruitment of tumor-specific cytotoxic T-cells (CTL), leading to the identification of several melanoma associated antigen (MAA). However, numerous strategies to treat melanoma with immunotherapy have resulted in only partial success. In this editorial, we discuss recent data related to the ability of tumors to elude immune responses. We therefore discuss different strategies to induce a clinically effective immune response. These approaches include 1) immunostimulation: including peptide/protein based vaccines, dendritic cell vaccines, and adoptive cell transfer; and 2) overcoming immunosuppression, including targeting of checkpoint molecules such as CTLA-4, circumventing the activity of Tregs, and assuring antigen expression by tumor cells (thwarting antigen silencing). Finally, we discuss recent advances in gene therapy, including adoptive therapy with engineered T cell receptors (TCRs). These issues lead to the conclusion that successful immunotherapy in malignant melanoma requires a combination of strategies aimed at both inducing immunostimulation and blocking immunosuppression.
3. Antioxidative role of interleukin-6 in septic lung injury in mice pp 501 - 508
K. Inoue1, 2, H. Takano1, 2, R. Yanagisawa1, M. Sakurai1, A. Shimada3, M. Satoh4, S. Yoshino5, K. Yamaki5 and T. Yoshikawa2
1Environmental Health Sciences Division, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan 2Inflammation and Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan 3Department of Veterinary Pathology, Faculty of Agriculture, Tottori University, Tottori, Japan 4Department of Pharmacology, Aichi Gakuin University, Nagoya, Japan 5Department of Pharmacology, Kobe Pharmaceutical University, Kobe, Japan
We have previously demonstrated the protective role of interleukin (IL)-6 against septic lung injury induced by lipopolysaccharide (LPS) using IL-6 knock-out (-/-) mice. This protection is mediated, at least partly, through the inhibition of the enhanced local expression of proinflammatory cytokines. In the present study, we addressed whether IL-6 regulates oxidative stress in the lung generated by LPS exposure using IL-6 (-/-) and corresponding wild type (WT) mice. Intraperitoneal LPS (1mg/kg) challenge induced transcriptional expressions of inducible nitric oxide synthase and heme oxygenase -1 in the lung of mice with both genotypes. In the presence of LPS, these expressions were significantly greater in IL-6 (-/-) than in WT mice. Immunohistochemistry also showed that LPS induced a significant increase in 8-hydroxy-2’-deoxyguanosine formation in the lung as compared to vehicle. Furthermore, the formation was more intense in IL-6 (-/-) than in WT mice in the presence of LPS challenge. In the presence of LPS, lipid peroxidation in the lung was significantly greater in IL-6 (-/-) than in WT mice. These data suggest that the possible mechanisms in which endogenous IL-6 protects against septic lung injury induced by LPS involve, at least in part, its antioxidative properties.
4. A quercetin containing supplement reduces niacin-induced flush in humans pp 509-514 D. Kalogeromitros1, M. Makris1, C. Chliva1, X. Aggelides1, D. Kempuraj2 and T.C. Theoharides1,2
1Allergy Clinical Research Center, Allergy Section, Attikon Hospital, University of Athens Medical School, Athens, Greece 2Molecular Immunopharmacology and Drug Discovery Laboratory, Department of Pharmacology and Experimental Therapeutics, Tufts University School of Medicine, Medical Center, Boston, MA, USA
5. Interferon gamma-signature transcript profiling and IL-23 upregulation in response to Helicobacter pylori infection pp 515 -526
J.R. Vivas1,5, B. Regnault2, V. Michel1,6, F.I. Bussiere1,6, P. Ave3, M. Huerre3, A. Labigne1, M.M. D’Elios4 and E. Touati1,6
1Unite de Pathogenie Bacterienne des Muqueuses, Institut Pasteur, Paris, France 2Genopole DNA Chip Platform, Institut Pasteur, Paris, France 3Unite de Recherche et d’Expertises en Histotechnologie et Pathologie, Institut Pasteur, Paris, France 4Department of Internal Medicine, University of Florence, Florence, Italy 5Department of Molecular Microbiology, Centro de Investigaciones Biologicas, CIB-CSIC, Madrid, Spain 6Unite Postulante de Pathogenese de Helicobacter, Institut Pasteur, Paris, France
Helicobacter pylori infection is the major cause of gastroduodenal pathologies including gastric cancer. The long persistence of bacteria and the type of immune and inflammatory response determine the clinical issue. In this study, the global gene expression profile after 6 and 12 months of H. pylori infection was investigated in the mouse stomach, using the Affymetrix GeneChip Mouse Expression Array A430. Genes related to the inflammatory and immune responses were focused. Levels of selected transcripts were confirmed by reverse transcription polymerase chain reaction. Twenty- five and nineteen percent of the differentially expressed genes observed at 6 and 12 months post-infection respectively, were related to immune response. They are characterized by an interferon (IFN)gamma-dependent expression associated to a T helper 1 (Th1) polarised response. In-depth analysis revealed that an up-regulation of IL-23p19, took place in the stomach of H. pylori infected-mice. Strong IL-23p19 levels were also confirmed in gastric biopsies from H. pylori-infected patients with chronic gastritis, as compared to healthy subjects. Our microarray analysis revealed also, a high decrease of H+K+-ATPase transcripts in the presence of the H. pylori infection. Association of gastric Th1 immune response with hypochlorhydria through the down-regulation of H+K+-ATPase contributes to the genesis of lesions upon the H. pylori infection. Our data highlight that the up-regulation of IL-23 and of many IFN gamma signature transcripts occur early on during the host response to H. pylori, and suggest that this type of immune response may promote the severity of the induced gastric lesions.
6. Impact of porcine Orexin A on glucagon plasma concentrations in pigs pp527 - 538 P. Papakonstantinou, N. Tziris, I. Kesisoglou1, A. Gotzamani-Psarrakou2, Chr. Tsonidis3, M.N. Patsikas and L.G. Papazoglou
AHEPA University Hospital, 3rd Surgical Clinic, Aristotle University of Thessaloniki, Greece 1Department of Nuclear Medicine, Aristotle University of Thessaloniki, Greece 22nd Neurosurgical Clinic A.U.T., Aristotle University of Thessaloniki, Greece 3Department of Clinical Sciences, Veterinary Medicine, Aristotle University of Thessaloniki, Greece
In 1998, Orexin A was added to the long list of orexigenic neuropeptides of the brain’s physiology. Orexin A is involved in the central control of appetite and in energy homeostasis, as well as in the regulation of many other physiological functions. It is produced by a small cluster of the brain’s neurons, located mainly in and around the lateral hypothalamic area. This site is known to be involved in regulating feeding in mammals. An intracerebroventricular injection of Orexin A into the rat’s brain causes an impressive increase in the consumption of food, while an intravenous injection induces changes on glucagon plasma concentrations in rats. In addition, there are signs of changes on glucagon plasma concentrations when Orexin A acts on individual pancreatic islets of rats. In this study, we investigated the potential effects of the central administration of porcine Orexin A on glucagon plasma concentrations in pigs, and examined whether these changes are associated with the possible effect of the neuropeptide on the enteroinsular axis.
7. Involvement of cPLA2 inhibition in dexamethasone-induced thymocyte apoptosis pp 539 - 552 B. Cinque, D. Fanini, L. Di Marzio1, P. Palumbo, V. Donato2, C. La Torre, E. Velardi2, S. Bruscoli2, C. Riccardi2 and M.G. Cifone
Department of Health Science, University of L’Aquila, L’Aquila, Italy 1Department of Drug Science, University of Chieti, Chieti, Italy 2Department of Clinical and Experimental Medicine, Section of Pharmacology, University of Perugia, Perugia, Italy
Various molecular mechanisms have been suggested to be involved in dexamethasone induced thymocyte apoptosis. In this study we show that pharmacological inhibition of cytoplasmic PLA2 in mouse thymocytes for 18 h with arachidonyl trifluoromethyl ketone (AACOCF3) (10 mM) and palmitoyl trifluoromethyl ketone (PACOCF3) (10 mM) induced a drastic increase of thymocyte apoptosis comparable to that observed following Dex (10-7 M) treatment, while inhibition of secretory PLA2 with p-bromophenacyl bromide (pBPB) (20 mM) did not. AACOCF3-induced thymocyte apoptosis, similarly to Dex-induced thymocyte apoptosis, was eliminated by cell pre-treatment with the PI-PLCb inhibitor, U73122, but not by the PC-PLC inhibitor D609. These observations were corroborated by the ability of AACOCF3, like Dex, to induce a rapid and transient increase in DAG generation. In addition, AACOCF3-induced apoptosis involved the activation of the acidic sphingomyelinase (aSMase) but not of the neutral sphingomyelinase (nSMase), as evaluated by measurements of enzyme activity in cell extracts following thymocyte exposure to AACOCF3 and by the ability of monensin to inhibit AACOCF3-induced thymocyte apoptosis. In addition, the AACOCF3 apoptotic effect resulted in an early increase of ceramide levels. AACOCF3-induced thymocyte apoptosis involved the activation of caspase 3, and cell pre-treatment with a caspase 3 inhibitor prevented AACOCF3-induced apoptosis. These observations suggest that cPLA2 inhibition may have a role in Dex-induced thymocyte apoptosis and highlight the importance of cPLA2 activity in thymocyte survival.
8. Antibodies as predictors of complex autoimmune diseases and cancer pp 553 - 566
A. Vojdani
Immunosciences Lab., Inc., Beverly Hills, CA, USA
The pathologic role of autoantibodies in many autoimmune diseases is widely accepted. An enzyme immunoassay was used for measurement of antibodies against disease-specific antigens and etiologic agents for cross-reactive antigens associated with them. This antibody assay was applied to a panel of antigens for the detection of different neuroautoimmune diseases that included multiple sclerosis, motor peripheral neuropathies, multifocal motor neuropathy, amyotrophic lateral sclerosis, pediatric autoimmune neuropsychiatric disorder associated with streptococcal infection. We studied women with pregnancies complicated by neural tube defect, neuroborreliosis, autism and patients with possible somatic hypermutation. Antibodies were also measured against antigens and etiologic agents associated with primary biliary cirrhosis and chronic obstructive pulmonary disease. And, finally, antibodies were measured against several tumor antigens or peptides which are expressed in prostatic, breast and colon tissues. This panel of different autoantibodies was applied to 290 patients with neuroautoimmune disorders, cancer, and possible somatic hypermutation. The levels of these antibodies against different tissue-specific antigens and etiologic agents associated with them were significantly elevated in patients versus controls. We hope that this novel 96 antigen-specific ELISA will be used in additional studies that will prove its clinical efficacy, not only for the early diagnosis of many neuroautoimmune, liver and lung autoimmune disorders, but also for prognosis and the implementation of preventive steps for many complex diseases.
9. In-air micro-particle induced X-ray emission analysis of asbestos and metals in lung tissuePp 567 -576
Y. Shimizu, K. Dobashi1, T. Kusakbe2, T. Nagamine2, M. Oikawa2, T. Satoh3, J. Haga3, Y. Ishii3, T. Ohkubo3, T. Kamiya3, K. Arakawa3, T. Sano4, S. Tanaka5, K. Shimizu6, S. Matsuzaki, M. Utsugi and M. Mori
Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Gunma, Japan 1Gunma University Faculty of Health Science, Gunma, Japan 221st Century COE Program, Gunma University Faculty of Health Science, Gunma, Japan 3Japan Atomic Energy Agency, Takasaki Advanced Radiation Research Institute, Gunma, Japan 4Department of Tumor Pathology, Gunma University Graduate School of Medicine, Gunma, Japan 5Department of General Surgical Science, Gunma University Graduate School of Medicine, Gunma, Japan 6Divisiont of Thoracic and Visceral Organ Surgery, Gunma University Graduate School of Medicine, Gunma, Japan
Inhalation of asbestos increases the risk of lung cancer and pulmonary fibrosis. It is difficult to directly assess the distribution and content of inhaled particles in lung tissue sections. The purpose of this study is to employ an in-air micro particle induced X-ray emission (in-air micro-PIXE) system for assessment of the spatial distribution and content of asbestos and other metals in lung tissue. A proton ion-microbeam from this system was applied to irradiate lung tissue of patients with or without asbestosis, tumor tissue from both groups, and asbestos fibers (in vitro). The content of each element composing asbestos and those of other metals were calculated and their distribution was assessed from the characteristic X-ray pattern for each element obtained after irradiation. This in-air micro-PIXE system could identify the location of asbestos bodies composed of Si, Mg, and Fe in lung tissue sections. Macrophage and lymphocytes accumulated in that area. This new system also revealed deposits of titanium, nickel, and cobalt in the lung tissues, in addition to asbestos bodies. The Si and Fe content were higher in lungs with asbestosis than in lungs without asbestosis or in tumor tissue. Analysis of asbestos fibers composed of chrysotile, crocidolite, and amosite showed that the ratios of Si, Fe, and Mg corresponded with those for the chemical structures. In-air micro-PIXE analysis is useful for assessing the distribution and quantities of asbestos bodies and also other metals in lung tissue comparing to immune-related cell localizations, and is also useful for analysis of standard asbestos fibers.
10. Murine models of anaemia of inflammation: extramedullary haematopoiesis represents a species specific difference to human anaemia of inflammation that can be eliminated by splenectomy Pp 577 -584
T.E.O. Schubert1,5, F. Obermaier2, P. Ugocsai3, D.N. Mannel4, B. Echtenacher4, F. Hofstadter1 and P. Haerle2 1Department of Pathology, University of Regensburg, Regensburg, Germany2Department of Internal Medicine I, University of Regensburg, Regensburg, Germany3Department of Laboratory Medicine, University of Regensburg, Regensburg, Germany 4Department of Immunology, University of Regensburg, Regensburg, Germany5Laboratory of Pathology, Frankfurt, Germany
In contrast to humans, mice physiologically exhibit extramedullary haematopoiesis in the spleen. In spite of this crucial species specific difference not much is known about the contribution of extramedullary haematopoiesis to overall erythropoiesis in models of anaemia of inflammation (AI). The objective of this study is to characterize murine AI with respect to extramedullary haematopoiesis and to develop a model more closely resembling human AI. Three different models of AI [caecal ligation and puncture (CLP), collagen induced arthritis (CIA) and DSS induced chronic colitis (DSSC)] were characterized with respect to red blood parameters, iron metabolism and extramedullary haematopoiesis. Arthritic animals were splenectomised to prevent extramedullary haematopoiesis. Anaemia caused by systemic inflammation was found in all three models. Splenic extramedullary haematopoiesis was markedly increased as reflected by increment in spleen weights and increase of the red pulp resulting in increased reticulocyte counts. Splenectomised arthritic animals did not show increased reticulocyte counts indicating that most of the reticulocytes were produced in the spleen. Our results demonstrate that murine AI differs from human AI with respect to increased splenic extramedullary haematopoiesis. Our data demonstrate that induction of AI in splenectomised mice represents a good way to model human AI.
11. Quantification of sirolimus and everolimus by immunoassay techniques: test specificity and cross-reactivity evaluation Pp 585 - 594 M. Pieri, N. Miraglia, A. Gentile1, G. Polichetti1, L. Castiglia, S. Federico2, M. Sabbatini2, V. Basile1, G. Tarantino3, A. Acampora and D. Capone1
Department of Public Medicine and Social Health, Federico II University of Naples, Naples, Italy 1Department of Neurosciences, Unit of Clinical Pharmacology, Federico II University of Naples, Naples, Italy 2Chair of Nephrology, Section of Renal Transplantation, Federico II University of Naples, Naples, Italy 3Department of Clinical and Experimental Medicine, Section of Hepatology in Internal Medicine, Federico II University of Naples, Naples, Italy
The possible cross-reactivity of immunoassays with structurally-related drugs was investigated. Innofluor® Certican® (FPIA) calibrators were measured by using IMx® Sirolimus assay (MEIA) and MEIA Sirolimus calibrators were analysed by using FPIA Certican® assay. Drug concentrations were measured in 95 and 100 samples from renal transplanted patients (RTP) on sirolimus or everolimus treatment by using immunoassays and LC/ESI-MSMS. A high cross-reactivity was found both for MEIA and FPIA. High correlation degrees, confirmed by the Bland-Altman and the Eksborg tests, were found between drug concentrations measured in real samples by both immunoassays (r=0.909 and r=0.970, respectively). LC/ESI-MSMS analysis of samples containing sirolimus showed no positivity for everolimus. Similarly, samples from patients on treatment with everolimus resulted negative as far as regards sirolimus. MEIA and FPIA could be considered mutually reliable and accurate alternatives for the specific-drug immunoassay. It should be noticed that in patients switching from one drug to the other unreal overestimation of the blood levels of the current administered immunosuppressant can occur.
12. Potential role of culture mediums for successful isolation and neuronal differentiation of amniotic fluid stem cells Pp 595 – 602
M. Orciani, M. Emanuelli1, C. Martino2, A. Pugnaloni, A.L. Tranquilli 2 and R. Di Primio
Department of Molecular Pathology and Innovative Therapies, Histology Section, Marche Polytechnic University, Ancona, Italy 1Institute of Biochemical Biotechnologies, Marche Polytechnic University, Ancona, Italy 2Department of Obstetrics and Gynecology, Marche Polytechnic University, Salesi Hospital, Ancona, Italy
In recent years, the use of stem cells has generated increasing interest in regenerative medicine and cancer therapies. The most potent stem cells derive from the inner cell mass during embryonic development and their use yields serious ethical and methodological problems. Recently, a number of reports suggests that another suitable source of multipotent stem cells may be the amniotic fluid. Amniotic fluid mesenchymal stem cells (AFMSCs) are capable of extensive self-renewal, able to differentiate in specialized cells representative of all three germ layers, do not show ethical restriction, and display minimal risks of teratomas and a very low immunogenity. For all these reasons, amniotic fluid appears as a promising alternative source for stem cell therapy. Their recent discovery implies a lack of knowledge of their specific features as well as the existence of a protocol universally recognized as the most suitable for their isolation, growth and long-term conservation. In this study, we isolated stem cells from six amniotic fluids; these cells were cultured with three different culture mediums [Mesenchymal Stem Cell Medium (MSCGM), PC-1 and RPMI-1640], characterized by cytofluorimetric analysis, and then either frozen or induced to neuronal differentiation. Even if the immunophenotype seemed not to be influenced by culture medium (all six samples cultured in the above-mentioned mediums expressed surface antigens commonly found on stem cells), cells showed different abilities to differentiate into neuron-like cells and to re-start the culture after short\long-term storage. Cells isolated and cultured in MSCGM showed the highest proliferation rate, and formed neuron-like cells when sub-plated with neuronal differentiation medium. Cells from PC-1, on the contrary, displayed an increased ability to re-start culture after short\long term storage. Finally, cells from RPMI-1640, even if expressing stem cells markers, were not able to differentiate in neuron-like cells. Further studies are still needed in order to assess the effective role of culture medium for a successful isolation, growth, differentiation and storage of AFMSCs, but our data underline the importance of finding a universally accepted protocol for the use of these cells.
13. C-reactive protein changes in the uncomplicated course of arthroscopic anterior cruciate ligament reconstruction
Pp 603 - 608 V. Calvisi and S. Lupparelli Department of Orthopaedic Surgery, University of L’Aquila, L’Aquila, Italy
The diagnosis of septic arthritis following arthroscopic anterior cruciate ligament (ACL) reconstruction is often elusive and can only be confirmed by joint aspiration, although arthrocentesis carries a risk for superinfection. C-reactive protein (CRP) may prove a useful laboratory test to substantiate clinical suspicion. The present study investigated the post-operative variations of CRP in 58 patients (age range 15-52, median age 25) undergoing ACL reconstruction with either bone-patellar tendon-bone (BPTB) or hamstring tendon (HT) who did not develop infection at 6 months follow-up. CRP titre was determined on the 1st, 3rd, 7th, 15th, and 30th post-operative day by immunoprecipitation in patients divided according to the type of autograft (BPTB: 13 patients; HT: 45 patients). Mean CRP significantly increased on the 1st post-operative day, peaked on the 3rd day and decreased on the 7th day, while levels on the 15th and 30th days did not differ from baseline. The trend of CRP changes did not differ in relation to the type of autograft. The results of our study suggest that close clinical surveillance may be advisable when CRP levels deviate from the reference values 2 weeks after surgery. In these circumstances, suspicion of septic arthritis warrants aspiration and culturing in order to avert a diagnostic delay.
14. Lack of inflammatory cells in the oral mucosa of subjects undergoing sublingual immunotherapy Pp 609- 614
F. Marcucci1, C. Incorvaia2, L. Sensi1, G. Di Cara1, G. Cadario3, A. Cavaliere4, P. Moingeon5, P. Puccinelli6, M. Di Gioacchino6 and F. Frati1,6
1Pediatrics, University Department of Medical and Surgical Specialties and Public Health, Perugia, Italy Allergy/Pulmonary Rehabilitation, ICP Hospital, Milan, Italy 3Allergy and Clinical Immunology, San Giovanni Battista Hospital, Turin, Italy 4Department of Pathological Anatomy, University of Perugia, Italy 5Medical and Scientific Department, Stallergenes, Antony, France 6Medical and Scientific Department, Stallergenes, Milan, Italy 7Department of Medicine and Science of Ageing, Section of Allergy, Clinical Immunology and Occupational Medicine, G. d’Annunzio University, Chieti, Italy
The cells involved in allergic inflammation, such as mast cells, basophils, and eosinophils, have been thoroughly studied in the nose, the lungs and the skin, demonstrating an evident increase in response to the introduction of the specific allergen, while little is known in the mucosal system and particularly in the oral mucosa. We investigated such tissue by using the model of sublingual immunotherapy (SLIT), by which high doses of the specific allergen enter the mouth. Oral biopsies were carried out on seven subjects allergic to grass pollen and treated with SLIT by a grass extract. In biopsies carried out before SLIT there was a very low number of mast cells and eosinophils both in the epithelium and subepithelium layers, and insignificant changes were detected after SLIT. These findings show the lack of allergic inflammation in the oral mucosa upon contact with the specific allergen and confirm the role of the mouth as a tolerogenic site, which is conceivable considering the different attitude of the mouth, where the antigens transit to undergo digestion, in respect to the airways or the skin, where the antigen absorption is potentially dangerous.
15. Immunohistochemical evaluation of global DNA methylation and histone acetylation in papillary urothelial neoplasm of low malignant potential pp 615-624 F. Barbisan, R. Mazzucchelli, A. Santinelli, D. Stramazzotti, M. Scarpelli, A. Lopez-Beltran1, L. Cheng2 and R. Montironi
Section of Pathological Anatomy, Polytechnic University of the Marche Region, School of Medicine, United Hospitals, Ancona, Italy 1Department of Pathology, Reina Sofia University Hospital and Faculty of Medicine, Cordoba, Spain 2Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, Indianapolis, IN, USA
A preceding study has shown that karyometry detected subvisual differences in chromatin organization status between non-recurrent and recurrent papillary urothelial neoplasm of low malignant potential (PUNLMP). The status of chromatin organization depends on epigenetic events, such as DNA methylation and histone acetylation. The aim of this study is to explore global DNA methylation and global histone acetylation in non-recurrent and recurrent PUNLMP. 5-methylcytosine (5MeC) and acetylated histone H3 lysine 9 (AcH3K9) were investigated by immunohistochemistry (IHC) in 20 PUNLMP cases (10 non-recurrent and 10 recurrent), in 5 cases of normal urothelium (NU) and in 5 cases of muscle invasive pT2 urothelial carcinoma (UC). For global DNA methylation, the mean percentage of positive nuclei in the cells adjacent to the stroma increased from NU (79%) through non-recurrent and recurrent PUNLMP (86% and 93%, respectively) to UC (97%). The percentages of positive nuclei in the intermediate cell layers and in the superficial cells in the four groups were similar to those adjacent to the stroma. The proportion of nuclei with weak-to-moderate intensity was far greater than that of those strongly stained and increased steadily from NU to UC. For global histone acetylation, the mean percentage of positive nuclei was highest in non-recurrent PUNLMP (i.e. 90%) and lowest in recurrent PUNLMP (i.e. 81%). In NU and UC the mean percentages of positive nuclei were 84% and 86%, respectively. The percentage of positive nuclei decreased from the cell layer adjacent to the stroma to the superficial cell layer. The proportion of nuclei with weak-to-moderate intensity was slightly greater than that of those strongly stained. In comparison with global DNA methylation, the proportion of strongly stained nuclei was much higher. In conclusion, there are differences in global DNA methylation and histone acetylation patterns between non-recurrent and recurrent PUNLMP. Further studies are needed to elucidate the complex interplay between chromatin structure, its modifications and recurrence of PUNLMP.
16. Inhaled thiamphenicol and acetilcysteine in children with acute bacterial rhinopharyngitis Pp 625-630 A. Varricchio, M. Capasso1, M. Di Gioacchino2 and G. Ciprandi3 ENT Unit, San Gennaro Hospital, Naples, Italy 1Paediatric Unit, Piedimonte Matese Hospital, Caserta, Italy 2Allergy Related Disease Unit, G. d’Annunzio University Foundation, Chieti, Italy 3Department of Internal Medicine, Azienda Ospedaliera-Universitaria San Martino, Genova, Italy
Antibiotic abuse for treating rhinopharyngitis induces the occurrence of resistant bacteria. As topical drugs might reduce this phenomenon, the aims of our study are to evaluate inhaled thiamphenicol associated with acetylcysteine in children with acute bacterial rhinopharyngitis and to compare it with the use of saline solution. The trial was conducted as randomized, parallel group, and single blind. Children, aged 3-6 years, with acute bacterial rhinopharyngitis were treated with aerosolized thiamphenicol associated with acetylcysteine (250 mg: ½ vial in the morning and ½ vial in the evening) (Group A) or saline solution twice daily (Group B), both of them for 5 days. Both treatments were administered using a new device: Rinowash. The following parameters were assessed: nasal obstruction, mucopurulent rhinorrhea, post-nasal drip, cough, sore throat, fever, and cultures. Of 104 patients screened, 90 children, median age 3.7 years (44 females and 46 males), completed the study: 60 in Group A and 30 in Group B. Actively-treated children achieved a significant improvement of all parameters, but fewer than the control group. In conclusion, inhaled thiamphenicol associated with acetylcysteine may represent a valid treatment for acute bacterial rhinopharyngitis in children, as it is effective, safe, economic, and simple to use.
17. Improvement in relative survival of patients with vasculitis: study of 101 cases compared to the general population Pp 631- 642 P. Stratta, C. Marcuccio1, A. Campo2, L. Sandri3, A. Messuerott1, L. Colla1, S. Rosso4, G. Mazzucco5, L. Mariani6 and G. Ciccone7
Department of Clinical and Experimental Medicine, Nephrology and Transplantation & IRCAD Amedeo Avogadro University, Maggiore Hospital, Novara; Department of Internal Medicine, 1Section Nephrology of the University of Turin, Molinette Hospital, Turin, Italy 2Renal Care Units of S. Lazzaro Hospital, Alba, Italy 3Pinerolo Hospital, Pinerolo, Italy 4CPO-Piemonte Cancer Registry, Biomedical Science and Human Oncology, University of Torino, Italy 5Sections Pathology, University of Turin, Italy 6Gynaecological Oncology, IRCC, University of Turin, Italy 7Epidemiology Division, University of Turin, Italy
Immunosuppressive treatment has changed the prognosis of renal vasculitis over time, but improvement in prognosis is difficult to analyze in different historical periods, and can be better demonstrated by comparison with life expectancy of sex- and age-matched people. Long-term survival of 101 patients diagnosed with systemic vasculitis at our center from 1975 to 2002 was retrospectively evaluated in comparison with that of the Region’s age- and sex-matched population. Patient and kidney survival significantly increased over time. Multivariate analyses showed that risks of patient and renal death decreased by 10% and 7%, respectively, at each year of follow-up, and increased by 6.3% and 5.2% for each year of age. Relative survival significantly improved over time, approaching that of the general population for cases diagnosed after 1993, mainly in women <60 years (from 0.671 at 5-years in the first period to 0.916 in the last period ), while 5-year-relative-survival was still 0.530 and 0.682 in men and women > 60 years, respectively. Poisson-based multinomial analyses confirmed the significant risk of the first periods of diagnosis and of dialysis in worsening of the relative survival of patients compared to that of the general population. Life expectancy in patients with renal vasculitis has improved over time, paralleling a significant increase in steroid pulse/cyclophosphamide association therapy and an earlier diagnosis due to the introduction of the ANCA test. Relative survival has considerably improved, and now approaches that expected in the general population for women, but not for men.
18. Serum cytokines and bioumoral immunological characterization of psoriatic patients in long term etanercept treatment Pp 643-650 P. Cordiali-Fei, M. Ardigo, A. Mastroianni, A. Giuliani, G. D’Agosto, V. Bordignon, E. Trento, A. Vento and E. Berardesca
San Gallicano Dermatological Institute, Rome, Italy
The purpose of this study is to evaluate blood cytokines and immunological parameters in psoriatic patients during long-term treatment with etanercept. Forty-five subjects of both sexes affected by psoriasis with or without arthritis entered the study and were treated with etanercept according to international standard protocols. Biochemical blood analysis was carried out at baseline and during follow-up every second month. In particular, the following parameters were kept under control: antinuclear antibodies, anti-nDNA antibodies, anti-histone antibodies, blood cell count, circulating lymphocyte subtypes (CD3, CD4, CD8, CD16, CD19) and IgE. Cytokine profiles (IL-1-α, IL-1-β, IL-6, IL-8, IL-10, IL-12, INF, TNF- α) were also evaluated in blood samples during the treatment up to 1 year of follow-up. A significant decrease in PASI score (p<0.01) and in several cytokine levels was observed, particularly in IL-1, IL-6, IFN-γ (p<0.01) and to a lesser extent in TNF-α (p<0.05). No statistically significant changes were recorded after 1 year of follow-up in blood immunological parameters, in particular in ANA titre, CD4/CD8 ratio, IgE levels, CD16, CD19 and eosinophils count. In conclusion, long-term treatment with etanercept leads not only to a significant improvement in PASI score, but also to significant changes (reduction) in several proinflammatory and modulatory cytokines involved in the pathogenesis of the disease; on the other hand, there are no effects on immunological or bioumoral parameters showing that etanercept modulates rather than suppresses the physiological responses during psoriasis treatment.
19. Sucralfate modulates uPAR and EGFR expression in an experimental rat model of cervicitis Pppp 651-658 C. Mannari, S. Santi, M. Migliori, C. Filippi, N. Origlia1, M. Sanso2, E. Boldrini2 and L. Giovannini
Department of Neuroscience, Pharmacology section, University of Pisa, Pisa, Italy 1Institute of Neuroscience (CNR), Pisa, Italy 2Farmigea SpA, Pisa, Italy
Sucralfate is a drug used in the treatment of gastric and duodenal ulcer; it is cytoprotective and able to increase the bioavailability of several growth factors, modulating the wound healing process. In this study we tested the possible therapeutic effect of Sucralfate in the treatment of ulcerative lesion occurring in uterine cervix; to investigate such effect we used an experimental rat model of cervicitis in which the uPAR and EGFR expression were evaluated. Cervicitis was induced in wild and ovariectomized wistar female rats by an acetic acid-soaked tampon. The animals were divided into two main groups (4 and 7 days) and Sucralfate was administered topically until the day they were sacrificed. In order to distinguish physiological and drug-induced healing, quantitative and qualitative uPAR and EGFR expression were evaluated by using Western Blot and Immunohistochemistry techniques. Western Blot analysis demonstrated an increased expression of both receptors after 4 days from wounding in wild and ovariectomized animals. In particular in ovariectomized animals the expression of uPAR and EGFR increased after 4 days while it reduced following the administration of Sucralfate. In wild rats the same was observed for uPAR expression, while EGFR was different; in fact, its expression increased significantly at day 4 in the animals treated with the drug and only at day 7 in those untreated. Immunohistochemistry highlighted a noteworthy epithelial colocalization of EGFR and uPAR after 4 days in the animals treated with Sucralfate. We conclude that Sucralfate can promote the healing of ulcerative cervicitis and moreover, it reduces the normal healing time because of its modulatory property on uPAR and EGFR expression.
20. Antibodies to carbonic anhydrase in patients with connective tissue diseases: relationship with lung involvement Pp 659-668 D. Caccavo, A. Afeltra1, A. Rigon1, M. Vadacca1, B.B. Zobel1, D. Zennaro1, L. Arcarese1, F. Buzzulini1, N.M. Pellegrino and A. Amoroso2
Department of Internal Medicine, Immunology and Infectious Diseases, University of Bari, Bari, Italy 1University Campus Bio-Medico, Rome, Italy 2Department of Clinical Medicine, Sapienza University, Rome, Italy
The aim of this study is to evaluate the presence of antibodies to carbonic anhydrase I and/or II (ACAI and ACAII) in patients affected by connective tissue diseases (CTD) and to investigate their association with lung involvement evaluated by High resolution CT scan (HRCT). Ninety-six patients affected by CTD were studied, i.e. 33 rheumatoid arthritis (RA), 8 psoriatic arthritis (PA), 8 ankylosing spondilitis (AS), 23 Systemic Lupus Erythematosus (SLE), 10 Sjogren Syndrome (SS), and 14 Systemic Sclerosis (SSc). ACA were detected by ELISA. The lung involvement was evaluated by means of a previously described HRCT score. According to a receiver operator characteristic curve, patients were divided into those with HRCT score ≥ 10 and those with HRCT score < 10, where HRCT score ≥ 10 was predictive of interstitial lung disease. ACAI and/or ACAII were detected in 30/96 patients (31.2%) (P<0.0001 in comparison with controls). In particular, the prevalence of ACAI and/or ACAII was significantly higher in patients with RA (P = 0.002), PA (P < 0.0001), SLE (P = 0.0003) and SSc (P< 0.0001). A positive correlation was found between HRCT scores and CRP or ACAI levels (P=<0.0001 and P=0.004, respectively). Thirty-nine of 96 patients (40.6%) showed a HRCT score ≥ 10 and both their CRP and ACAI levels were significantly higher when compared with patients showing a HRCT score < 10 (P<0.0006 and P = 0.0009, respectively). Moreover, C3 and C4 complement fractions inversely correlated with HRCT scores (P = 0.0004 and P<0.0001, respectively) and lower values of C3 and C4 complement fractions were found in patients with HRCT score ≥ 10 than in those with HRCT score < 10 (P = 0.014 and P = 0.007, respectively). Due to the lower levels of complement fractions detected in patients with HRCT score ≥ 10, a possible immune-complex-mediated pathogenic mechanism of lung involvement could be suggested. 21. Sublingual desensitization in patients with wasp venom allergy: preliminary resultsG. Patriarca, E. Nucera, C. Roncallo, A. Aruanno, C. Lombardo, M. Decinti, L. Pascolini, M. Milani1, A. Buonomo and D. SchiavinoDepartment of Allergology, Catholic University, Rome, Italy1Alk-Abello Medical Department, Lainate, Milan, ItalyThe aim of this paper is to assess in an open prospective pilot case-control study the tolerability, safety and efficacy of an ultra-rush sublingual immunotherapy (SLIT) protocol with Vespula venom in wasp allergic patients compared to subcutaneous immunotherapy (SCIT). Forty-one wasp allergic patients were treated with sublingual (SLIT group) or subcutaneous (SCIT group) ultrarush immunotherapy with Vespula venom extract. All patients underwent skin tests and serum specific IgE and IgG4 detection before enrolment and after 6, 12 and 24 months of immunotherapy. The SLIT group consisted of 21 (6 females and 15 males) patients who received increasing doses of Vespula venom (Aquagen, ALK-Abelló) until the final dose of 30 drops of extract in 3 hours, containing 100,000 SQ-U/ml. The maintenance dose was of 10 drops of pure venom extract 3 times a week, for a total dose of 100,000 SQ-U weekly (corresponding to 100 μg of venom extract). The SCIT group consisted of 20 patients (16 males and 4 females) who were treated with subcutaneous ultrarush immunotherapy with Vespula venom extract (Pharmalgen, Alk-Abelló). Patients received 101.1 mg of Vespula venom in 3 hours and were treated with 100 mg of wasp venom monthly. During the ultrarush sublingual treatment 2 patients (9.5%) experienced mild side-effects. Specific IgE and specific IgG to wasp venom did not show any significant modification. Four patients were field-stung by a wasp during the treatment (for a total of 6 stings). Two patients (3 stings), with a previous clinical history of a grade III and IV reaction, did not experience any reaction. One patient, with a previous grade II reaction, showed a large local reaction. The fourth patient, with a previous grade III reaction, was re-stung twice (after 12 and 24 months) with two systemic reactions (SR) (mild throat constriction). During the ultrarush SCIT phase, 3 (15%) patients experienced side-effects: 2 of them showed a large local reaction and 1 had headache and stomach ache. Specific IgE showed a significant (P=0.001) increase after 6 months of treatment and then returned to baseline levels while specific IgG showed a significant (P=0.001) increase after 6, 12 and 24 months in comparison with baseline. Nine patients were field-stung during the treatment: 8 of them experienced large local reactions; one patient (11%) experienced an SR (dizziness). Our results, even if in a small number of patients, suggest that in patients with Hymenoptera sting allergy SLIT could be efficacious with a good tolerability profile when compared to SCIT. Larger studies are needed to assess efficacy, safety and tolerability profile of wasp venom SLIT.22. Comparison of thermal infrared and laser doppler imaging in the assessment of cutaneous tissue perfusion in scleroderma patients and healthy controls
A. Merla1,2, L. Di Donato1,2, G.L. Romani1,2, M. Proietti3 and F. Salsano3 1Department of Clinical Sciences and Bioimaging, G. d Annunzio University, Chieti, Italy 2ITAB, Institute for Advanced Biomedical Technologies, G. d’Annunzio University Foundation, Chieti, Italy 3Department of Clinical Immunology and Allergy, Sapienza University, Rome, Italy
In this study we propose a non-invasive method to calculate blood flow by means of thermal infrared imaging and bio-heat transfer modeling. The method is able to provide high time-resolution series of cutaneous blood flow images with the same spatial resolution of the thermal images. The method was tested against a standard laser Doppler imaging system, which to date is considered the gold standard for non-invasive assessment of cutaneous blood flow, on both healthy subjects and patients suffering from systemic sclerosis (SSc; a pathological condition with microvessel endothelium injury). Twenty healthy subjects and twenty SSc patients simultaneously underwent laser Doppler and thermal imaging of the dorsum of the hand. A linear correlation between perfusion values obtained with the two methods was found for the healthy control group (R = 0.85, Pearson Product Moment Correlation). A significant correlation was not observed for the SSc patient group. The results of this study suggest that combined laser Doppler, thermal imaging and bio-heat transfer modeling could effectively discriminate between healthy vs. impaired conditions of the cutaneous tissue thermal properties and cutaneous vasculature. Such method, in addition to providing a potential effective imaging-based tool for a variety of biomedical and clinical applications ranging from diagnostics to treatment follow-up, may help the understanding of the morphological and functional impairment secondary to the disease. The thermal imaging-based method provided faster and better time-resolved imaging of cutaneous perfusion than standard laser Doppler techniques as the thermal cameras can provide up to 100 complete 524x524 pixel images per second, thus allowing real time monitoring of tissue perfusion rates.
23. Histological and clinical survey of polylactic-polyglycolic acid and dextrane copolymer in maxillary sinus lift: a pilot in vivo study
R. Martuscelli, M.C. Maltarello1, N.M. Maraldi2, C. Sbordone3 and L. Sbordone4
Department of Oral Science and Maxillofacial Surgery, Federico II University of Napoli, Naples, Italy 1Lab. Cell Biology and Electron Microscopy-Istituti Ortopedici Rizzoli (IOR), Bologna, Italy 2Lab. Cell Biology and Electron Microscopy-IOR, Bologna, Department of Anatomical Sciences, University of Bologna, Bologna, Italy 3Department of Maxillo-Facial Surgery, Federico II University of Naples, Naples, Italy 4Implantology and Periodontology, Department of Surgery, University of Pisa, Italy
Of various proposed alternatives to autogenous bone, a synthetic, degradable copolymer of PLA-GLA and dextrane seems to be a promising biomaterial for maxillary sinus lift. Consecutive partially edentulous patients showing severe monolateral posterior maxillary atrophy were treated via sinus lift using PLA-GLA-dextrane copolymer as the sole filler. Delayed implant positioning was performed and cores of regenerated tissues and native bone controls were retrieved and evaluated by light and electron microscopy, histomorphometry, microhardness and qualitative X-ray analysis. Seven sinuses in 7 patients were augmented with PLA-GLA-dextrane copolymer. Six to nine months after the copolymer ‘graft’, 17 bone cores were retrieved: all histological sections contained newly synthesized, mineralized material and new bone in various stages of development. Histomorphometry revealed average Trabecular Bone Volume (TBV) values ranging from 51% (6 months) to 77% (9 months). Backscattered scanning electron microscopy (BSE) in experimental and control samples confirmed histology findings. Microhardness values suggested newly formed bone at nine months was not as hard as native bone. Ca and P content was similar in 9-month regenerated and native bone. Seventeen implants were inserted in the second stage of surgery: resulting Implant Success (SR) and Cumulative Success (CSR) up to 3 years were 100% following Albrektsson’s criteria. Sinus lift augmentation using PLA-GLA-dextrane copolymer as the sole filler resulted in uneventful surgeries. New bone formation was evident histologically and its maturation was still in progress after 9 months. Successful, staged implant positioning was achieved in regenerated tissue.
24. The increase of endothelial progenitor cells in the peripheral blood: a new parameter for detecting onset and severity of sepsis
C. Becchi, S. Pillozzi1, L.P. Fabbri, M. Al Malyan, C. Cacciapuoti, C. Della Bella2, M. Nucera, M. Masselli1, S. Boncinelli, A. Arcangeli1 and A. Amedei2,3
Department of Medical and Surgical Critical Care, Section of Anaesthesia and Intensive Care, Florence, Italy 1Department of Experimental Pathology and Oncology, Florence, Italy 2Department of Internal Medicine, University of Florence, Florence, Italy 3Department of Biomedicine, Azienda Ospedaliera Universitaria Careggi, Florence, Italy
Sepsis is a clinical syndrome characterized by non-specific inflammatory response with evidence of profound changes in the function and structure of endothelium. Recent evidence suggests that vascular maintenance, repair and angiogenesis are in part mediated by recruitment from bone marrow (BM) of endothelial progenitor cells (EPCs). In this study we were interested in whether EPCs are increasingly mobilized during sepsis and if this mobilization is associated with sepsis severity. Our flow cytometry data demonstrate that in the CD34+ cell gate the number of EPCs in the blood of patients with sepsis had a four-fold increase (45 ± 4.5% p<0.001) compared to healthy controls (12 ± 3.6%) and that this increase was already evident at 6 hours from diagnosis (40.6 ± 4.2%), reaching its maximum at 72 hours. Also the percentage of cEPCs identified in the patients with sepsis (35 ± 4.6% of the CD34+ cell) was statistically different (p<0.001) compared to that found in the blood of patients with severe sepsis (75 ± 4.9%). In addition, we proved that at six hours after sepsis diagnosis, VEGF, CXCL8 and CXCL12 serum levels were significantly higher in septic patients compared to healthy volunteers 559 ± 82.14 pg/ml vs 2.9 ± 0.6 (p<0.0001), 189.8 ± 67.3 pg/ml 15 vs 11.9 ± 1.6 (p=0.014) and 780.5 ± 106.5 pg/ml; vs 190.2 ± 71.4 (p < 0.001). Our data suggest that the cEPC evaluation in peripheral blood, even at early times of diagnosis, in patients with sepsis can be envisaged as a valuable parameter to confirm diagnosis and suggest further prognosis.
25. Evaluation by real-time PCR of the expression of S. flexneri virulence-associated genes ospB and phoN2 under different genetical backgrounds
M. Nicoletti1, I. Santino2, A. Petrucca1,3, F. Del Chierico1, S. Cannavacciuolo2, M. Casalino4, R. Sessa2 and P. Cipriani2,3
1Dipartimento di Scienze Biomediche, University G. d Annunzio, Chieti, Italy 2Dipartimento di Scienze di Sanita Pubblica, Sapienza Universita di Roma, Rome, Italy 3Laboratorio di Microbiologia Clinica, II Facolta di Medicina e Chirurgia, Ospedale Sant’Andrea, Rome, Italy 4Dipartimento di Biologia, Università di Roma Tre, Rome, Italy
Under conditions of activated type III secretion Shigella flexneri up-regulates the expression of numerous genes, including the virulence plasmid (pINV)-encoded ospB and phoN2 genes. ospB and phoN2 are virulence-associated genes which are part of a bicistronic transcriptional unit encoding OspB, a protein (effector) of unknown function secreted by the type III secretion (TTS) apparatus, and PhoN2 (apyrase or ATP-diphosphohydrolase), a periplasmic protein involved in polar IcsA localization on the surface of S. flexneri. In this work we used real-time PCR to measure transcription of ospB and phoN2 of wild-type S. flexneri strain M90T as well as of derivative mutants impaired in definite virulence traits. The results obtained confirmed and extended previous reports indicating that the expression of ospB and phoN2 genes is modulated in a virB-dependent, mxiE-independent manner under conditions of non-activated secretion, while their expression is considerably induced in a mxiE-dependent manner under conditions of activated secretion. That the expression of the ospB-phoN2 operon is up-regulated in condition of activated secretion, indicates that probably the expression of these two genes might be important, especially during the later stages of infection of S. flexneri.
26. Virulence traits in Escherichia coli strains isolated from outpatients with urinary tract infections
C. Longhi, A. Cossu, V. Iebba, M.R. Massaro, D. Cipriani, F. Chiarini, M.P. Conte, L. Seganti, J. Osborn and S. Schippa
Department of Public Health Sciences, Sapienza University, Rome, Italy This study aims to characterize phenotypic and genotypic virulence traits in Escherichia coli strains, isolated from outpatients with urinary tract infections, comparing with those obtained from inpatients. Information on the pathogenic behavior of the uropathogenic strains was obtained by monitoring different biological properties, such as autoagglutination, hemagglutination, adhesiveness to and invasion of human bladder (HT1376) cells, biofilm formation, phylogenetic grouping, and virulence-related genes. The results show similar behavior in the two groups concerning autoagglutination, hemagglutination, and biofilm formation. None of the strains examined was invasive. However, in strains from outpatients there was an increased adhesion to HT1376 cells compared with clinical strains, a significant higher presence of genes codifying for adhesins and cell protection factors, and a lower proportion of strains belonging to B1 group. These findings add further information on the pathogenic traits of community E. coli, since strains isolated from the outpatients’ group were differently armed in comparison with those of clinical cases, and more suitable to infect healthy individuals.
27. Photoallergic contact dermatitis: the 15-year experience of a tertiary reference center in a sunny Mediterranean city
A. Katsarou, M. Makris1, G. Zarafonitis, E. Lagogianni, S. Gregoriou and D. Kalogeromitros1
Department of Dermatology, A. Sygros Hospital, Medical school, University of Athens, Athens, Greece 1Allergy Unit, 2nd Dept of Dermatology and Venereology, Attikon University Hospital, Medical school, University of Athens, Athens, Greece
Photoallergic contact dermatitis (PACD) represents an important entity of photodermatoses while photopatch testing is the main diagnostic tool. The main goal of this study is to evaluate retrospectively the prevalence of photoallergic reactions and the offending agents in Athens during a 15-year period. The medical records of all patients with possible PACD between 1992 and 2006 were examined. All patients included in the analysis had undergone patch testing and photo-testing. Contact reactions were detected in 86 out of 207 participants (41.54%), while photocontact reactions were identified in 28/207 (13.52%) patients. The most common offending photoallergen was promethazine (25%), while chlorpromazine and oxybenzone were both detected in 12.5% of cases. PACD represents a unique proportion of photodermatoses in a sunny Mediterranean city such as Athens.
28. Blood cholesterol concentration measured by CR3000: fingerstick versus venous sampling
V. Sblendorio, B. Palmieri and G. Riccioni1
General Surgery and New Technologies Laboratory Research, University of Modena-Reggio Emilia, Italy 1Cardiology Unit, San Camillo de Lellis Hospital, Manfredonia, Foggia, Italy
Currently, the clinical practice of desktop or Point of Care (PoC) analyzers for lipid measurements has gained wide popularity. Designed to quickly perform measurements on microlitre (µL) quantities of blood, these instruments can be used in non-laboratory settings, such as physicians’ offices or field-testing sites and can provide measurements in whole blood, serum, or plasma, using either venous or capillary blood samples. The aim of this study is to examine the relationship between cholesterol determinations in venous and capillary samples using the CR3000 PoC system. The study was performed on 21 unselected adult volunteers, and no exclusion criteria was adopted. The mean cholesterol concentration for the venous blood samples measured was 164 mg/dL. The values obtained in the capillary blood samples averaged 168 mg/dL, which is only slightly higher (e.g., 2.87%) than the venous sample measurements. Moreover, the total variance was statistically similar for venous and capillary measurements (F value=1.199, where the upper critical value of the F distribution is 2.124, p<0.05). The results of our study support the concept that CR3000 total cholesterol testing can be performed safely and accurately in either venous or capillary specimens.
29. Prevalence of atopic symptoms among blood donor carriers of mannose-binding lectin variant alleles
F. Cardinale, I. Chinellato, G.L. Marseglia1, E. Nettis2, A. Polizzi, M.S. Loffredo, T. Santostasi, R. Tesse, P. Trerotoli3, D. Di Monte4 and L. Armenio
Department of Pediatrics, School of Medicine, University of Bari, Bari, Italy 1Department of Pediatrics, School of Medicine, University of Pavia, Pavia, Italy 2Department of Allergy and Clinical Immunology, School of Medicine, University of Bari, Bari, Italy 3Department of Internal and Public Medicine, School of Medicine, University of Bari, Bari, Italy 4Transfusion Unit, Department of Internal Medicine, Azienda Ospedaliera Policlinico, Bari, Italy
Mannose-binding lectin (MBL) is a C-type soluble collectin involved in the innate immune response. Carriers of MBL gene variant alleles (MBLva) have decreased plasma concentrations of MBL and increased susceptibility to bacterial and viral infections. The aim of the present study is to test the hypothesis that carriers of MBLva could have a different frequency of atopic symptoms as compared to wild-type carriers. A total of 385 consecutively enrolled Caucasian blood donors were studied. Blood specimens underwent genomic analysis and genotyping for MBLva by polymerase chain reaction (PCR). MBLva carrier status was associated with a reduced frequency of allergic rhinitis (OR 0.41 [95% CI 0.2 to 0.8], c2= 6.98, p=.008). No relationship was found between MBLva carrier status and asthma or atopic skin symptoms. MBLva might be one of the host-related genetic factors involved in atopic disorders, namely allergic rhinitis.
30. Visceral leishmaniasis revealing chronic granulomatous disease in a child
A. Finocchi1,2, P. Palma1,2, G. Di Matteo1, M. Chiriaco1, L. Lancella2, A. Simonetti1,2, I. Rana3, S. Livadiotti2 and P. Rossi1,2
1Division of Paediatrics, Dept. of Public Health, University of Tor Vergata, Rome, Italy 2Division of Immunology and Infectious Diseases Children’s Hospital Bambino Gesu, Rome, Italy 3Division of Haematology Children’s Hospital Bambino Gesu, Rome, Italy
We report the first description of visceral leishmaniasis (VL) infection as a harbinger of chronic granulomatous disease (CGD) in a 3-year old child. Although VL is not frequently suspected in CGD patients, our case emphasises the importance of a complete evaluation of the immune system in children presenting with VL in order to exclude underlying immunodeficiency states. As the prognosis of CGD is poor, with high morbidity and mortality, establishing an early diagnosis has important practical implications in the successful treatment of these patients. Following the diagnosis, the patient received Human Leukocyte Antigen (HLA) identical sibling bone marrow transplantation (BMT). The child is now 2 years post-transplant and is in good general conditions with normal blood counts, and evidence of full-donor chimerism in repeated fluorescence in situ hybridization (FISH) studies.
31. Characterization of a Staphylococcus aureus strain showing high levels of biofilm formation isolated from a vascular graft: case report
D. Petrelli, A. Repetto1, M.C. Di Luca, B. Parente2, V. Tavolini2, P. Cao2, C. Ripa3, M. Prenna and L.A. Vitali
Dipartimento Biologia MCA, Universita di Camerino, Camerino, Italy 1Struttura Complessa di Microbiologia, Ospedale S. Maria della Misericordia, Perugia, Italy 2Struttura Complessa di Chirurgia Vascolare e Endovascolare, Universita degli Studi di Perugia, Perugia, Italy 3U.O. Cardiologia, UTIC, Istituto Scientifico INRCA, Ancona, Italy
A methicillin-susceptible Staphylococcus aureus strain, SA-DZ1, was isolated from an infected bypass crossover graft. Its general microbiological features were reminiscent of those previously described for the wound Wiley strain. Removal of the prosthetic device was necessary to resolve the infection. SA-DZ1 grown under different conditions showed a very strong and distinctive biofilm-producing phenotype, which was also visualized by confocal laser scanning microscopy. The biofilm extracellular matrix was essentially polysaccharidic, as determined by differential growth and physicochemical tests. By Multi Locus Sequence Typing (MLST), SA-DZ1 was classified as st94, a single locus variant of st8. Several other genetic traits assayed by PCR, such as agr-type and the presence of gene encoding proteins involved in adhesion and virulence (e.g. ica operon), confirmed the identifying features of this clinical isolate.
32. Cryptococcal lymphadenitis as a manifestation of immune reconstitution inflammatory syndrome in an HIV-positive patient: a case report and review of the literature
L. Putignani1,2, G. Antonucci1, M.G. Paglia1, L. Vincenzi1, A. Festa1, P. De Mori1, L. Loiacono1 and P. Visca1,3
1National Institute for Infectious Diseases Lazzaro Spallanzani I.R.C.C.S., Rome, Italy 2Children’s Hospital and research Institute Bambino Gesù, Rome, Italy 3Department of Biology, University of Roma Tre, Rome, Italy
Cryptococcus neoformans infections are typically associated with T-cell deficiencies, including acquired immunodeficiency syndrome (AIDS). Although highly active antiretroviral therapy (HAART) has strongly reduced AIDS-related opportunistic infections, the restoration and reactivation of CD4+ cells can induce an immune reconstitution inflammatory syndrome (IRIS), consisting in a deregulated inflammatory response to latent infectious pathogens and/or to their residual antigens. Cryptococcal lymphadenitis has occasionally been documented in IRIS. Here we report a case of histology- and culture-negative cryptococcal lymphadenitis associated with IRIS in an adult AIDS patient with a history of disseminated cryptococcosis, after the start of fully adherent HAART. Appropriate diagnosis was established on nested-PCR and sequence analysis of the interspacer region 2 of C. neoformans ribosomal DNA, and detection of slow-growing blastospores in enrichment cultures of fine-needle lymph node aspirate. Review of recent literature and our case findings suggest that IRIS-associated cryptococcal lymphadenitis is more likely the flare up of a latent infection rather than an immunopathological response to residual antigen of unviable cryptococci.
33. Extragenital lichen sclerosus and atrophicus treated with topical steroids and retinoids in a child with vitiligo
C. Guerriero, S. Manco, A. Paradisi, R. Capizzi, B. Fossati and G. Fabrizi1
Department of Dermatology, Catholic University of the Sacred Heart, Rome,Italy 1Department of Dermatology, University of Molise School of Medicine, Campobasso, Italy
Lichen sclerosus and atrophicus (LSA) most commonly affects the anogenital region. Extragenital involvement is rare, and women are reported to be affected 6 to 10 times more often than men. The aetiology of LSA is unclear, but genetic, physiological and environmental factors are thought to be involved. Several lines of evidence support the hypothesis of an autoimmune basis for LSA; an increased incidence of tissue-specific antibodies and an association with autoimmune disorders such as vitiligo, alopecia areata, thyroid disease and pernicious anaemia have been reported. We describe a paediatric patient with extragenital LSA associated with vitiligo who was successfully treated with topical steroids and retinoids.
34. Consequences of long-lasting persistent allergic rhinitis in adolescents
G.L. Marseglia, M. Grignani, P. Civallero, B. Colombo1, M. Di Gioacchino2, A. Marchi, A. Perrone2 and G. Ciprandi1
Department of Pediatric Science, Pavia University, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy 1Department of Internal Medicine, Azienda Ospedaliera Universitaria San Martino, Genoa, Italy 2Allergy Related Disease Unit, Chieti University Foundation, Chieti, Italy
While it is well known that asthma is characterized by airway remodelling, only few studies have investigated this issue in patients affected by allergic rhinitis (AR). The aim of the present study is to investigate functional and structural consequences of long-lasting persistent AR (PER) in a cohort of adolescents. Eighty patients, forty with short-lasting and forty with long-lasting PER were prospectively and consecutively evaluated both clinically and by performing skin prick test, nasal cytology, and rhinomanometry. Eosinophils were significantly higher in patients presenting with long-lasting PER rather than in those with short-lasting PER (P<0.0001). The degree of inflammation was significantly associated with impaired nasal airflow (rs= -0.81). This study provides evidence that adolescents with long-lasting PER may show a progressive worsening of nasal function depending on the inflammation.
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