atherosclerosis: a classic inflammatory atherosclerosis: a classic inflammatory disease a....

INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY

0394-6320 (2011)Copyright © by BIOLIFE, s.a.s.

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Atherosclerosis is the condition in which the inner layers of the artery wall, known as intima, become thick and irregular due to the deposits of fats (mainly in the form of cholesterol and another fat called phospholipid) and other substances. When these deposits occur in the arteries that supply blood to the heart the condition is called coronary atherosclerosis. As the build-up grows, the artery narrows and the flow of blood to the heart muscle is reduced. Like any other muscle, the heart needs

blood to provide it with oxygen. When that blood flow is reduced or completely blocked, some of the cells in the heart muscle can die. Scientists divide the telltale sign of atherosclerosis into two major types: the earliest deposits called fatty streaks, and advance deposits called plaques. Fatty streaks which contain about 25% fat can be seen in the arteries even in children. During the development of fatty streaks, the cells that line the arteries are stimulated to take in more cholesterol than they can handle.

EDITORIAL

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.

ATHEROSCLEROSIS: A CLASSIC INFLAMMATORY DISEASE

A. ANOGEIANAKI1, D. ANGELUCCI2, E. CIANCHETTI3, M. D’ALESSANDRO4, G. MACCAURO5, A. SAGGINI6, V. SALINI7, A. CARAFFA8, S. TETÉ9, F. CONTI10,

D. TRIPODI9 and Y.B. SHAIK-DASTHAGIRISAHEB11

1Physiology Department, School of Medicine, Aristotle University of Thessaloniki, Greece;2AnatomoPathology Division, University of Chieti-Pescara, Chieti, Italy; 3Division of Senology,

Ortona Hospital, Italy; 4Radiology Division, Santo Spirito Hospital, Pescara, Italy; 5Department of Orthopaedics, Catholic University of Rome, Rome, Italy; 6Department of Dermatology, University

of Rome Tor Vergata, Rome, Italy; 7Orthopaedics Division, University of Chieti, Chieti, Italy; 8Orthopaedics Division, University of Perugia, Perugia, Italy; 9Departmentof Oral, Nano and

Biotechnologies, University G. d’Annunzio, Chieti, Italy; 10Department of Gyneacology, “Santo Spirito” Hospital, Pescara, Italy; 11Department of Medicine, Boston University School of Medicine,

Boston, MA, USA

Received May 4, 2011 – Accepted July 26, 2011

Mailing address: Dr. Antonia Anogeianaki,Physiology Department,Faculty of Medicine,Aristotle University of Thessaloniki,Thessaloniki, GreeceTel: ++30 6937001080 Fax: ++30 2310 999079e-mail: [email protected]

Key words: atherosclerosis, inflammation, cholesterol, phospholipid

Vol. 24, no. 4, 817-825 (2011)






The family of autoinflammatory diseases, an expanding group of rare conditions characterized by spontaneously relapsing bouts of inflammation, especially localized in one or more areas, joints, skin, central nervous system, etc., in the absence of specific autoantibodies or antigen-specific T cells, encompasses cryopyrin-associated periodic syndromes (CAPS), also known as “cryopyrinopathies”. These syndromes were characterized at a molecular level with the identification in 2001 of mutations in the NLRP3 gene (first called cold induced autoinflammatory

syndrome 1 or CIAS1), located on the long arm of chromosome 1q44 and containing nine exons (1). The CAPS include three conditions along a spectrum from the mildest to the most severe: (a) familial cold autoinflammatory syndrome (FCAS, MIM 120100), (b) Muckle-Wells syndrome (MWS, MIM 191900) and (c) chronic infantile neurological cutaneous articular syndrome (CINCAs, MIM 607115, also known as “neonatal onset multisystem inflammatory disease or NOMID), all caused by different dominantly inherited or de novo mutations in the same above-cited gene, and in which severity

BRIDGING THE GAP BETWEEN THE CLINICIAN AND THE PATIENT WITH CRYOPYRIN-ASSOCIATED PERIODIC SYNDROMES

L. CANTARINI1, O.M. LUCHERINI2, B. FREDIANI1, M.G. BRIZI1, B. BARTOLOMEI3, R. CIMAZ4, M. GALEAZZI1 and D. RIGANTE3

1Interdepartmental Research Center of Systemic Autoimmune and Autoinflammatory Diseases, Unit of Rheumatology, Policlinico “Le Scotte”, University of Siena, Siena;

2Department of Evolutionary Biology, University of Siena, Siena; 3Department of Pediatric Sciences, Università Cattolica Sacro Cuore, Policlinico “A. Gemelli”, Rome;

4Department of Pediatrics, Rheumatology Unit, Anna Meyer Children’s Hospital and University of Florence, Italy

Received May 16, 2011 – Accepted September 27, 2011

EDITORIAL

Cryopyrin-associated periodic syndromes are categorized as a spectrum of three autoinflammatory diseases, namely familial cold auto-inflammatory syndrome, Muckle-Wells syndrome and chronic infantile neurological cutaneous articular syndrome. All are caused by mutations in the NLRP3 gene coding for cryopyrin and result in active interleukin-1 release: their rarity and shared clinical indicators involving skin, joints, central nervous system and eyes often mean that correct diagnosis is delayed. Onset occurs early in childhood, and life-long therapy with interleukin-1 blocking agents usually leads to tangible clinical remission and inflammatory marker normalization in a large number of patients, justifying the need to facilitate early diagnosis and thus avoid irreversible negative consequences for tissues and organs.

Mailing address: Luca Cantarini, MD, PhD. Rheumatology Unit, Policlinico “Le Scotte”, University of Siena, Viale Bracci 1, 53100 Siena, Italy Tel: ++39 347 9385457Fax: ++39 57740450. e-mail: [email protected]

Key words: Cryopyrin-associated periodic syndromes, familial cold autoinflammatory syndrome, Muckle-Wells syndrome, chronic infantile neurological cutaneous articular syndrome, neonatal onset multisystem inflammatory disease

Vol. 24, no. 4, 827-836 (2011)

836 INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY





Ethyl tertiary butyl ether (ETBE) is an oxygenate added to gasoline to reduce the emissions of carbon monoxide and volatile organic compounds and has been proposed as a potential substitute for methyl tertiary butyl ether. The use of ETBE as a gasoline additive, at an amount of up to 17% weight, indicates that it could be produced in very large amounts depending on how widespread its use becomes within the gasoline supply chain. The main source of exposure to ETBE is anticipated to be via inhalation and the contact with the skin while fueling automobiles and potentially via

ingestion through drinking water (1). The use of ETBE as a gasoline additive is increasing rapidly in Europe, as a result of the highly debated European Biofuels Directive which set a minimum market share for biofuels of 5.75% in 2010 (2). In Japan, it is calculated that the amount of ETBE mixed in gasoline will be 7% and total consumption of ETBE will be 840,000 kL (630,000 tons) per year after 2010. Based on this consumption, it is calculated that approximately 2,100 tons/year, 4,700 tons/year and 6,300 tons/year of ETBE will be emitted from oil refineries, gas

EFFECTS OF SUBCHRONIC INHALATION EXPOSURE TO ETHYL TERTIARY BUTYL ETHER ON SPLENOCYTES IN MICE

Q. LI1, M. KOBAYASHI1, H. INAGAKI1, Y. HIRATA1, K. HIRATA1, T. SHIMIZU1, R-S. WANG2, M. SUDA2, T. KAWAMOTO3, T. NAKAJIMA4 and T. KAWADA1

1Department of Hygiene and Public Health, Nippon Medical School, Tokyo; 2National Institute of Occupational Safety and Health, Kawasaki; 3Department of Environmental Health, University

of Occupational and Environmental Health, Kitakyushu; Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya, Japan

Received June 16, 2011 – Accepted October 6, 2011

Ethyl tertiary-butyl ether (ETBE) is a motor fuel oxygenate used in reformulated gasoline. The current use of ETBE in gasoline or petrol is modest but increasing. To investigate the effects of ETBE on splenocytes, mice were exposed to 0 (control), 500 ppm, 1750 ppm, or 5000 ppm of ETBE by inhalation for 6 h/day for 5 days/wk over a 6- or 13-week period. Splenocytes were harvested from the control and exposed mice, and the following cell phenotypes were quantified by flow cytometry: (1) B cells (PerCP-Cy5.5-CD45R/B220), (2) T cells (PerCP-Cy5-CD3e), (3) T cell subsets (FITC-CD4 and PE-CD8a), (4) natural killer (NK) cells (PE-NK1.1), and (5) macrophages (FITC-CD11b). Body weight and the weight of the spleen were also examined. ETBE-exposure did not affect the weight of the spleen or body weight, while it transiently increased the number of RBC and the Hb concentration. The numbers of splenic CD3+, CD4+, and CD8+ T cells, the percentage of CD4+ T cells and the CD4+/CD8+ T cell ratio in the ETBE-exposed groups were significantly decreased in a dose-dependent manner. However, ETBE exposure did not affect the numbers of splenic NK cells, B cells, or macrophages or the total number of splenocytes. The above findings indicate that ETBE selectively affects the number of splenic T cells in mice.

Mailing address: Qing Li, MD, PhDDepartment of Hygiene and Public Health, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, JapanTel: ++81 3 3822 2131 Fax: ++81 3 5685 3065e-mail: [email protected]

Key words: B cells, ethyl tertiary-butyl ether (ETBE), flow cytometry, NK, splenocytes, T cells

Vol. 24, no. 4, 837-847 (2011)






IDENTIFICATION OF DIFFERENTIALLY-EXPRESSED PROTEINS BETWEEN EARLY SUBMUCOSAL NON-INVASIVE AND INVASIVE COLORECTAL CANCER USING

2D-DIGE AND MASS SPECTROMETRY

J. ZHANG, M-Q. SONG, J-S. ZHU, Z. ZHOU, Z-P. XU, W-X. CHEN and N-W. CHEN

Department of Gastroenterology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China


Early detection and diagnosis of colorectal cancer (CRC) are closely related to a better therapeutic outcome, and the five-year survival rate of early CRC is over 90%. Though endoscopic minimally invasive treatment has become a quick and effective therapy for early CRC, endoscopic biopsies are usually not deep enough to obtain tissues from the submucosal layer and it is difficult to determine whether early CRC has infiltrated into the submucosa. Therefore, in the present study, we constructed tumor models of early submucosal non-invasive CRC (SNICRC) and submucosal invasive CRC (SICRC) in Fischer-344 rats induced by N-methyl-N-nitrosourea (MNU). The differentially-expressed proteins were analyzed and identified in SNICRC, SICRC and normal control (NC) tissues using highly sensitive two-dimensional differential gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS). Proteomic data revealed 132 protein spots between SNICRC and SICRC, 162 protein spots between SICRC and NC and 154 protein spots between SNICRC and NC which were found differentially expressed. These differential spots were picked, in-gel digested and peptide mass fingerprints were obtained by MALDI-TOF-MS/MS. Finally, five differentially-expressed proteins in SNICRC, SICRC and NC were identified, and increases in Transgelin, peptidylprolyl isomerase A (PPIA) and tropomyosin alpha isoform d were observed, while decreases in carbonic anhydrase 2 (CAII) and an unnamed protein were detected in SICRC compared with SNICRC and NC. Furthermore, Fluorescence-based quantitative polymerase chain reaction (FQ-PCR), Western blotting and immunohistochemistry assays also revealed significant upregulation of Transgelin expression and down-regulation of CAII expression in SICRC tissues. In conclusion, 2D-DIGE is confirmed to be an efficient strategy that enables us to identify differentially-expressed proteins between early SNICRC and SICRC. The potential biomarkers such as Transgelin and CAII may be used for the detection of early SICRC

Mailing address: Prof. Jin-Shui Zhu, Department of Gastroenterology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200233, China Tel: ++86 21 64369181 exit58970 Fax: ++86 21 64837019e-mail: [email protected]; [email protected]

Key words: colorectal cancer, proteomics, 2D-DIGE, mass spectrometry, transgelin, carbonic anhydrase 2

Colorectal cancer (CRC) is a major cause of cancer incidence and mortality, and there are nearly one million new cases of CRC diagnosed worldwide each year and half a million deaths (1). The past two decades have brought major advances in CRC

screening and therapeutics including advances in colonoscopy therapy. However, it is hard for endoscopic examination or other means to detect early CRC lesions.

Thus, research focuses on specific strategies

Vol. 24, no. 4, 849-859 (2011)






For a long time microarray technology has been successfully used in cultured cells, including human epidermal keratinocytes, to characterize specific modifications of gene expression and transcriptional changes (1-3).

Paraquat dichloride (1,1′-dimethyl-4,4′-bipyridilium dichloride), a herbicide that has been used widely throughout the world over the past 40 years, is known to produce pulmonary lesions (4)

and a variety (5) of skin disease including contact dermatitis, premalignant lesions and squamous cell carcinoma (6-7). In addition, paraquat has been shown to produce evident neurotoxic effects after intracerebroventricular or intracerebral microinjection in experimental animals; however, it seems that the herbicide does not exhibit a selective neurotoxicity towards the dopaminergic nigro-striatal system since potent behavioral and

EFFECTS OF PARAQUAT AND CAPSAICIN ON THE EXPRESSION OF GENES RELATED TO INFLAMMATORY, IMMUNE RESPONSES AND CELL DEATH IN

IMMORTALIZED HUMAN HaCat KERATINOCYTES

N. PAOLILLO1, S. PICCIRILLI2, E. GIARDINA3, V. RISPOLI4, C. COLICA4 and S. NISTICÒ5

1IRCCS “C. Mondino Institute of Neurology” Foundation, Pavia; 2Center of Pharmaceutical Biotechnology, University of Rome Tor Vergata; 3Department of Biopathology and Diagnostic

Imaging, School of Medicine, Tor Vergata University, Rome; 4Department of Pharmacobiological Sciences, University Magna Graecia of Catanzaro; 5Department of Dermatology, School of

Medicine, University of Rome Tor Vergata, Italy

Received April 1, 2011 – Accepted July 28, 2011

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.

Mailing address: Prof. Steven Nisticò, Dermatology Unit, Department of Internal Medicine, University of Rome Tor Vergata, Via Montpellier, 1, 00133 Rome, Italy Tel: ++39 06501703159 Fax: ++39 06501703307e-mail: [email protected].

Key words: paraquat, capsaicin, keratinocytes, inflammation, apoptosis, transcriptional response, gene expression

Vol. 24, no. 4, 861-868 (2011)






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 (mBmal1, 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.

TIME-RELATED DYNAMICS OF VARIATION IN CORE CLOCK GENE EXPRESSION LEVELS IN TISSUES RELEVANT TO THE IMMUNE SYSTEM

G. MAZZOCCOLI1, R.B. SOTHERN2, A. GRECO3, V. PAZIENZA4, M. VINCIGUERRA5,S. LIU6 and Y. CAI6

1Department of Internal Medicine and Chronobiology Unit, Scientific Institute and Regional General Hospital “Casa Sollievo della Sofferenza”, San Giovanni Rotondo; 2The Rhythmometry Laboratory, College of Biological Sciences, Biological Sciences Center, University of Minnesota,

St. Paul, Minnesota, USA; 3Geriatrics Unit and Gerontology-Geriatrics Research Laboratory, Department of Medical Sciences, Scientific Institute and Regional General Hospital “Casa Sollievo

della Sofferenza”, San Giovanni Rotondo, Italy; 4Research Laboratory of Gastroenterology Unit, Scientific Institute and Regional General Hospital “Casa Sollievo della Sofferenza”, San Giovanni Rotondo, Italy; 5Institute of Hepatology, Birkbeck College, London, United Kingdom; 6Department of Neurology and Neurobiology, Xuanwu Hospital of Capital Medical University, Key Laboratory

for Neurodegenerative Diseases of Ministry of Education, Beijing, P.R. China

Received May 24, 2011 – Accepted July 25, 2011

Mailing address: Gianluigi Mazzoccoli, MD, Department of Internal Medicine and Chronobiology Unit, Scientific Institute and Regional General Hospital, “Casa Sollievo della Sofferenza”, San Giovanni Rotondo (FG), Italy Tel: ++39 0882 835321 Fax: ++39 0882 410563 e-mail: [email protected]

Key words: clock gene, circadian rhythm, immune system

Vol. 24, no. 4, 869-879 (2011)


881

Lactic acid bacteria (LAB) have been proposed as adjuvant for SIT, as various studies conducted on humans and animals converge to define the effect of LAB as anti-Th2 modulators (1) and Treg inducers (2). LABs have been shown to be effective in the prevention and control of allergy in children

with atopic dermatitis (3-4), although some studies have shown the association of atopic disorders with specific strains of intestinal microflora at birth (5). As a mechanism of action, LAB can bind to toll-like receptors (TRL) leading to dendritic cell (DCs) activation and production of IL-12 or IL-10

LACTOBACILLUS PARACASEI LP6 FAVORS IMMUNE MODULATION INDUCED BY ALLERGOID TREATMENT IN RAGWEED SENSITIZED MICE

C. PETRARCA1,3, F. LAZZARIN1,3, P. LANUTI2,3, M. MARCHISIO2,3, S. MISCIA2,3, C. ROSSI4, M. BRAGA5, G. MISTRELLO6 and M. DI GIOACCHINO1,3

1Unit of Allergy and Immunotoxicology, Center for Ageing Sciences, “Università G. d’Annunzio” Foundation, Chieti; 2Unit of Cytomorphology, Center for Ageing Sciences,

“Università G. d’Annunzio” Foundation, Chieti; 3Department of Medicine and Ageing Science (DMSI), University “G. d’Annunzio” of Chieti-Pescara, Chieti; 4Animal Facility, Center for Ageing

Sciences, “Università G. d’Annunzio” Foundation, Chieti; 5Allergy and Clinical Immunology, School of Specialization, Spedali Civili, Brescia; 6Research Center, Lofarma SpA, Milano, Italy

Received May 12, 2011 – Accepted August 2, 2011

It has been hypothesized that lactic acid bacteria (LAB) could be used as adjuvant for specific immunotherapy (SIT), as various studies conducted on humans and animals converge to define LAB as anti-Th2 modulators and Treg inducers. In the present study we evaluated the effects of LAB, in particular Lactobacillus paracasei Lp6 (Lp6), in a mouse model of ragweed (RW) allergy. Groups of Balb/c mice, experimentally sensitized towards ragweed, were treated by viable Lp6 or by RW-allergoid with or without co-administration of Lp6. A control group was sham-sensitized with PBS and sham-treated with water and a group was sensitized with RW and treated with water. Serum IgE, RW-induced release of IFN-γ, IL-4 and IL-10 from splenocytes and the frequency of CD4CD25 regulatory T cells (Tregs) expressing Foxp3 or IL-10 were evaluated in various groups. RW-allergoid treatment induced a reduction of serum IgE, with a decrease in RW-induced release of IL-4, and an increase in IL-10 and IFN-γ, along with a significant change in the frequency of Tregs, both CD25+ and -. The joint RW-allergoid+Lp6 treatment induced the highest degree of suppression of allergen-driven IL-4, the greatest reduction of IL-4/IFN-γ and IL-4/IL-10 ratios and the most significant increase of Foxp3 and IL-10 expressing Tregs. The study shows that Lp6 strengthens the immune modulation induced by allergoid-SIT in RW-sensitized mice, essentially characterized by a differential induction of Tregs associated to a reduction of IL-4; data converge to define a role of SIT adjuvant for Lp6.

Mailing address: Prof. Mario Di Gioacchino (MD),Unit of Allergy and Immunotoxicology,Center of Ageing Science, “Università G. d’Annunzio” Foundation, Via Colle dell’Ara, 66100 Chieti, ItalyTel: ++39 0871 54 1291 Fax: ++39 0871 54 13 00e-mail: [email protected]

Key words: probiotic, ragweed, specific immunotherapy, T regulatory cells, Foxp3



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Vol. 24, no. 4, 881-893 (2011)






Trichinellosis is caused by the parasitic nematode Trichinella, after ingestion of raw or undercooked meat by the host. This parasite has the peculiarity of having an intracellular localization both at intestinal and muscle level where the L1 larvae live and grow in a modified skeletal muscle fibre cell, called ‘nurse cell’ that induces, in encapsulating Trichinella species, the formation of a collagen capsule which protects the parasite from effector cells of the immune system (1). The immune response of the

host at muscle level has received increasing attention in recent years (2).

IgE and eosinophils derived from the activation of Th2 cells have been considered protective against helminths for some time although, to date, in vivo results have provided controversial results (3-4).

T helper type 2 (Th2) cells producing cytokines such as interleukin (IL)-4, IL-5, IL-9, IL-10 and IL-13 (2, 5), are not only involved in helminth infections but also in the development of pathological

STIMULATION OF TH1 RESPONSE BY HELICOBACTER PYLORI NEUTROPHIL ACTIVATING PROTEIN DECREASES THE PROTECTIVE ROLE OF IgE AND

EOSINOPHILS IN EXPERIMENTAL TRICHINELLOSIS

L. CHIUMIENTO1, G. DEL PRETE2,3, G. CODOLO4,5, M. DE BERNARD4,5,A. AMEDEI2,3, C. DELLA BELLA2, M. PIAZZA1, S. D’ELIOS2, L. CAPONI1,

M.M. D’ELIOS2,3 and F. BRUSCHI1

1Department of Experimental Pathology, M.B.I.E., Pisa University; 2Department of Internal Medicine, Florence University, Florence; 3Deparment of Biomedicine, Azienda Ospedaliero-

Universitaria Careggi, Florence; 4Department of Biology, University of Padua, Padua; 5Venetian Institute of Molecular Medicine, Padua, Italy

Received April 8, 2011 – Accepted September 1, 2011

Th2 responses seem to play an important role in defence against Trichinella spiralis (Ts). The Neutrophil Activating Protein of Helicobacter pylori (HP-NAP), that induces IL-12, and IL-23 expression and shifts to Th1 allergen-specific Th2 cells in vitro was used as an anti-Th2 agent in BALB/c mice infected with T. spiralis. The muscle larvae (ML) burden was lower (p < 0.02) in untreated infected animals than those infected treated with HP-NAP. In both groups there was an inverse relationship between ML burden of each animal and total IgE level (controls: r -0.617, p = 0.0013 and HP-NAP-treated: r -0.678, p = 0.0001) or eosinophil count, evaluated in the same mouse on day 42 (r -0.390, p = 0.0592 and r -0.803, p = 0.0001, respectively). Inflammatory response around the nurse cell-parasite complex was significantly higher in HP-NAP-treated infected animals than in those untreated infected, on the contrary the number of eosinophils, counted around each complex was significantly lower in the first animal group. This study provides evidence of a powerful anti-Th2 activity in vivo by HP-NAP and for the partial protective effect of Th2 responses in T. spiralis infection.

Mailing address: Dr. Mario M. D’Elios, Department Internal Medicine, University of Florence, viale Morgagni 85, 50134 Florence, Italy Tel: ++39055 4271 026 Fax: ++39055 4271 494 e-mail: [email protected]

Key words: experimental trichinellosis, mouse, Th1 adjuvant, Helicobacter pylori neutrophil activating protein

Vol. 24, no. 4, 895-903 (2011)






The inflammatory process is actively involved in atherosclerosis and underlies all phases of atherosclerotic plaque development: the beginning, the progression and the plaque rupture (1). In recent years, mediators and effectors of this cascade have been deeply studied in order to better define the mechanism that leads to acute clinical events, and systemic approaches are pursued to discover serum

biomarkers useful to identify patients with plaque at risk of future vascular events (2-3).

Galectin-3 is a 26 kD β-galactosidase-binding protein belonging to the Galectin family which contains more than ten members. This protein has a carboxyl terminal carbohydrate recognition domain (CRD) and an amino-terminal tandem repeat (4). It is expressed in the epithelia of many organs and

GALECTIN-3 PLASMA LEVELS AND CORONARY ARTERY DISEASE: A NEW POSSIBLE BIOMARKER OF ACUTE CORONARY SYNDROME

C. FALCONE1,2,3, S. LUCIBELLO2, I. MAZZUCCHELLI2, S. BOZZINI2, A. D’ANGELO2,S. SCHIRINZI1,2, R. TOTARO1,2, R. FALCONE2, M. BONDESAN1,2 and G. PELISSERO3

1Department of Cardiology, Istituto di Cura Città di Pavia University Hospital, Pavia; 2Interdepartimental Center for Research in Molecular Medicine (CIRMC), University of Pavia,

Pavia; 3IRCCS San Donato Hospital, San Donato Milanese, Italy

Received March 24, 2011 – Accepted September 26, 2011

Inflammation plays a key role in atherosclerosis. Galectin-3 is a macrophage- and endothelium-derived mediator actively involved in the regulation of many aspects of inflammatory cell behaviour. The aim of this study is to quantify plasma Galectin-3 in patients with coronary artery disease (CAD) and different clinical manifestation at the moment of observation in order to verify whether Galectin-3 could be a useful biomarker of atherosclerotic state. We enrolled 125 patients affected by CAD, angiographically documented (70 stable, 55 unstable). They underwent accurate examinations and anamnestic data was collected. The most important traditional risk factors, such as age, hypertension, and body mass index, were reported. Plasma Galectin-3 was quantified using an ELISA kit. Unstable patients (n = 55) had a higher plasma Galectin-3 levels in respect to the stable subjects (27.75 ng/mL (19.27-39.09) vs 6.48 ng/ml (4.88-8.83), p<0.001. A trend in correlation between plasma Galectin-3 levels and number of vessels compromised seems to be present: CAD patients with three-vessel disease had higher levels of Galectin-3 than patients with one-or two-vessel disease (17.39 ng/ml (10.75-29.82) vs 9.18 ng/ml (5.56-23.22), p= 0.058. The significantly higher plasma Galectin-3 levels in patients with unstable angina in respect to the stable angina confirm the involvement of Galectin-3 in promoting macrophage activation and monocyte attraction. Despite the distribution of CAD in patients with acute and chronic coronary disease being similar, we may hypothesize that Galectin-3 could be a useful biomarker of atherosclerotic plaque and in particular of its destabilization.

Mailing address: Prof. Colomba Falcone,Department of Cardiology, Istituito di Cura Città di Pavia University Hospital, Via Parco Vecchio 27, Pavia, ItalyTel: ++39 0382 433650 Fax: ++39 0382 576821e-mail: [email protected]

Key words: Galectin-3, CAD, stable and unstable angina

Vol. 24, no. 4, 905-913 (2011)






More than 5000/µL circulating clonal B-cells are required to diagnose chronic lymphocytic leukemia (CLL), the commonest form of leukemia in Western countries (1). However, a number of clonal B-cells less than 5000/µL, in absence of other features of

lymphoproliferative disorders, characterizes a new disease entity, the so-called monoclonal B-cell lymphocytosis (MBL) (2-4). Because the B-cell threshold chosen is based on the risk of developing clinical complications and/or dying from the disease,

CIRCULATING REGULATORY T CELLSIN “CLINICAL” MONOCLONAL B-CELL LYMPHOCYTOSIS

G. D’ARENA1, G. ROSSI2, M.M. MINERVINI2, L. SAVINO2, F. D’AURIA1, L. LAURENTI3, M.I. DEL PRINCIPE4, S. DEAGLIO5, A. BIAGI4,

L. DE MARTINO6, V. DE FEO6, T. STATUTO4, P. MUSTO1 and G. DEL POETA4

1Department of Onco-Hematology, IRCCS “Centro di Riferimento Oncologico della Basilicata” (CROB), Rionero in Vulture; 2Hematology and Stem Cell Transplantation Unit,IRCCS “Casa

Sollievo della Sofferenza” Hospital, San Giovanni Rotondo; 3Hematology Chair, Catholic University of Sacred Heart, Rome; 4Hematology Chair, University of Tor Vergata, Rome;

5Laboratory of Immunogenetics, University of Turin, Turin; 6Department of Pharmacology, University of Salerno, Salerno, Italy

Received July 21, 2011 – Accepted October 10, 2011

Regulatory T-cells (Tregs) constitute a small subset of cells involved in antitumour immunity and are generally increased in patients with chronic lymphocytic leukemia (CLL). No data is available on Tregs in monoclonal B-cell lymphocytosis (MBL), a disease entity characterized by less than 5000/µL circulating clonal B-cells in absence of other features of lymphoproliferative disorders. We used multicolour flow cytometry to evaluate the number of circulating Tregs in 56 patients with “clinical” MBL, 74 patients with previously untreated CLL and 40 healthy subjects. MBL patients showed a lower absolute number of Tregs, compared to CLL patients, but slightly higher than controls. Moreover, the absolute cell number of Tregs directly correlated both with more advanced Rai/Binet clinical stages and peripheral blood B-cell lymphocytosis. Of note, the absolute number of Tregs was found lower in MBL patients than in CLL patients staged as 0/A Rai/Binet. The study showed that Tregs increase gradually from normal subjects to “clinical” MBL patients and are significantly higher in CLL patients as compared to MBL patients. Moreover, a significant direct relationship was found between higher Treg values and a higher tumor burden expressed by B-lymphocytosis or more advanced clinical stages. In light of this data, MBL seems to be a preliminary phase preceding CLL. The progressive increase of Treg numbers might contribute both to the clinical evolution of MBL to overt CLL and to CLL progression.

Mailing address: Giovanni D’Arena, MDDepartment of Onco-Hematology, IRCCS “Centro Riferimento Oncologico della Basilicata” (CROB), Via Padre Pio n. 1, Rionero in Vulture (Pz), ItalyTel: ++39 0972 726729 Fax: ++39 0972 726217 e-mail: [email protected]

Key words: regulatory T cells, monoclonal B-cell lymphocytosis, chronic lymphocytic leukemia

Vol. 24, no. 4, 915-923 (2011)






Lung epithelial cells are normally well oxygenated but may be exposed to hypoxia in many pathological conditions, including chronic obstructive pulmonary diseases (COPD) (1). A phosphodiesterase (PDE) inhibitor sildenafil, also a cGMP-enhancer, attenuated chronic airway inflammation and

pulmonary hypertension (2). Targeting the vascular endothelial growth factor (VEGF) receptor is not merely limited to cancer therapy but also as an adjunct to existing inflammation treatment (3). KMUP-1 stimulated endothelial NO and displayed cGMP-dependent inhibition of Rho kinase (ROCK)

KMUP-1 INHIBITS H441 LUNG EPITHELIAL CELL GROWTH, MIGRATION AND PROINFLAMMATION VIA INCREASED NO/CGMP AND INHIBITED RHO KINASE/

VEGF SIGNALING PATHWAYS

B.N. WU1, H.Y. CHEN1, C.P. LIU2, 3, L.Y. HSU1 and I.J. CHEN1

1Department of Pharmacology, School of Medicine, Kaohsiung Medical University, Kaohsiung;2Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung;

3Division of Cardiology, Department of Internal Medicine, Yuan’s General Hospital, Kaohsiung, Taiwan

Received May 6, 2011 – Accepted October 3, 2011

This study investigates whether KMUP-1 protects soluble guanylate cyclase (sGC) and inhibits vascular endothelial growth factor (VEGF) expression in lung epithelial cells in hypoxia, therapeutically targeting epithelial proinflammation. H441 cells were used as a representative epithelial cell line to examine the role of sGC and VEGF in hypoxia and the anti-proinflammatory activity of KMUP-1 in normoxia. Human H441 cells were grown in hypoxia for 24-72 h. KMUP-1 (1, 10, 100 μM) arrested cells at the G0/G1 phase of the cell cycle, reduced cell survival and migration, increased p21/p27, restored eNOS, increased soluble guanylate cyclase (sGC) and PKG and inhibited Rho kinase II (ROCK-II). KMUP-1 (0.001-0.1 μM) concentration dependently increased eNOS in normoxia and did not inhibit phosphodiesterase-5A (PDE-5A) in hypoxic cells. Hypoxia-induced factor-1α (HIF-1α) and VEGF were suppressed by KMUP-1 but not by L-NAME (100 μM). The PKG inhibitor Rp-8-CPT-cGMPS (10 μM) blunted the inhibition of ROCK-II by KMUP-1. KMUP-1 inhibited thromboxane A2-mimetic agonist U46619-induced PDE-5A, TNF-α (100 ng/ml)-induced iNOS, and ROCK-II and associated phospho-p38 MAPK, suggesting multiple anti-proinflammatory activities. In addition, increased p21/p27 by KMUP-1 at higher concentrations might contribute to an increased Bax/Bcl-2 and active caspase-3/procaspase-3 ratio, concomitantly causing apoptosis. KMUP-1 inhibited ROCK-II/VEGF in hypoxia, indicating its anti-neoplastic and anti-inflammatory properties. KMUP-1 inhibited TNF-α-induced iNOS and U46619-induced PDE-5A and phospho-p38 MAPK in normoxia, confirming its anti-proinflammatory action. KMUP-1 could be used as an anti-proinflammatory agent to reduce epithelial inflammation.

Mailing address: Ing-Jun Chen PhD, Dept and Graduate Institute of Pharmacology, School of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan Fax: ++886 7 3234686 e-mail: [email protected]

Key words: lung epithelial cell, migration, proliferation, Rho kinase, vascular endothelial growth factor

Vol. 24, no. 4, 925-940 (2011)






ACTIVATION OF HUMAN MONOCYTE-DERIVED DENDRITIC CELLS IN VITRO BY THE BIOLOGICAL RESPONSE MODIFIER ARABINOXYLAN RICE BRAN

(MGN-3/BIOBRAN)

M. GHONEUM1 and S. AGRAWAL2

1Department of Otolaryngology, Charles Drew University of Medicine and Science, Los Angeles, CA; 2University of California at Irvine, Division of Basic and Clinical Immunology, Irvine, CA, USA

Received April 15, 2011 – Accepted October 3, 2011

Both authors contributed equally to this work

Arabinoxylan rice bran (MGN-3/Biobran) is a potent biological response modifier (BRM) that activates natural killer (NK) cells, T cells and monocytes. Currently, little is known regarding the effects of MGN-3 on dendritic cells (DCs), the cell type that bridges innate and adaptive immunity. Therefore, we examined the stimulatory effects of MGN-3 on DCs. Human monocyte-derived DCs were treated with MGN-3 at different concentrations (5-20 μg/ml) for 24 hours in vitro. Activation of DCs was determined by assessing the expression of co-stimulatory and maturation markers (CD40, CD80, CD83, CD86 and HLA-DR) by flow cytometry, and production of cytokines by ELISA. DC function was determined by assessing their ability to activate naïve T cells. Activation of T cells was assessed by measuring cell proliferation and cytokine production. MGN-3 treatment, in a dose-dependent manner, resulted in: 1) up-regulation of the surface expression of CD83 and CD86, on DCs; 2) an increase in the production of pro-inflammatory and immuno-regulatory cytokines (IL-1β, IL-6, IL-10, TNF-α, IL-12p40 and low levels of IL-12p70 and IL-2) by DCs; and 3) MGN-3 stimulated DC induced CD4+T cell proliferation and their production of cytokines, IFN-γ, IL-10, IL-17. Results suggest that MGN-3 functions as a natural adjuvant for DC activation and thus may be used in DC-based vaccine strategies against infections and cancer.

Mailing address: Mamdooh Ghoneum, Ph.D., Charles Drew University of Medicine and Science, Department of Otolaryngology, 1621 E. 120th Street, Los Angeles, California, 90059, U.S.A. Tel: ++1 323 563 5953, Fax: ++1 310 474 6724. e-mail: [email protected]

Key words: MGN-3, dendritic cells, CD4+T cells, in vitro, biological response modifiers

Rice bran was obtained from milling rice. It is composed of pericarp and seed coat together as a major portion of the aleurone layer, a part of the endosperm, and some part of the embryo or germ. Accordingly, several health benefits have been identified with rice bran. It is a rich source of proteins, lipids, vitamins, and trace minerals (1), and is well-known as a great source of dietary fiber (2). Earlier studies have shown that rice bran lowers serum cholesterol, and additionally reduces

the postprandial lipemia in humans (3-4). It is of great interest to note that rice bran derivatives possess anti-cancer activity, in vivo and in vitro. Recent studies by our laboratory and that of others have demonstrated the anti-cancer activity of rice bran derivatives. Arabinoxylan rice bran (MGN-3/Biobran) exerts in vivo tumor inhibitory effects against Ehrlich carcinoma-bearing mice (5-6), and rice bran polyphenol, cycloartenyl ferulate, induces apoptosis in human colorectal adenocarcinoma in

Vol. 24, no. 4, 941-948 (2011)






The subtypes of CD2 Ig superfamily, are essential for the cross-talk between immune cells and co-stimulation (1-3). 2B4 (CD244) receptor together with CD48, Ly-

9, NTB-A/Ly-108A and CRACC form the members of SLAM family (4-7). Characteristically these receptors possess single transmembrane immunoglobulin-like

2B4 (CD244) IS INVOLVED IN EOSINOPHIL ADHESION AND CHEMOTAXIS, AND ITS SURFACE EXPRESSION IS INCREASED IN ALLERGIC RHINITIS

AFTER CHALLENGE

A.E. EL-SHAZLY1, M. HENKET2, P.P. LEFEBVRE1 and R. LOUIS2

1Department of Oto-Rhino-Laryngology and Head and Neck Surgery, GIGA-Research, Liege University Hospitals (Centre hospitalier Universaitaire-C.H.U.), Liege; 2Department of

Pulmonology, GIGA-Research, Liege University Hospitals (Centre Hospitalier Universaitaire-C.H.U.), Liege, Belgium


A role for the subtypes of CD2 Ig superfamily receptors has been recently demonstrated in eosinophilic inflammation in experimental asthma and atopic asthmatics. We investigated the functions of 2B4 (CD244) molecules in eosinophil adhesion and chemotaxis, and correlated the results to the pathophysiology of allergic rhinitis (AR). Herein, we show that agonistic stimulation of 2B4 by C1.7, the anti-human 2B4 functional grade purified antibody, resulted in significant increase of eosinophils and eosinophil cell line (Eol-1 cells) adhesion to collagen type IV, and random migration. These functions were associated with tyrosine kinase phosphorylation of several protein residues of low molecular weight. Flow cytometry (FACS) experiments demonstrated that Eol-1 cells, normal peripheral blood eosinophils and eosinophils from AR patients, express surface 2B4 molecules. In vitro AR model demonstrated that the CC-chemokine receptor CCR3 stimulation by eotaxin induced significant increase in the expression of surface 2B4 in eosinophils and Eol-1 cells. Immunofluorescence confocal microscopy images showed that eotaxin induces also redistribution of 2B4 molecules towards the pseudopods in eosinophils and Eol-1 cells, changing their shape. Blocking of 2B4 molecules by the corresponding neutralizing antibody inhibited eotaxin induced Eol-1-adhesion, chemotaxis and the cytoskeleton changes. Pretreatment of Eol-1 cells with 1 μM genistein blocked eotaxin-induced Eol-1 adhesion, chemotaxis and 2B4 up-regulated expression. In vivo correlation demonstrated the expression of 2B4 molecules in eosinophils from AR patients to be significantly increased, after nasal provocation challenge. These results identify a novel role for 2B4 molecules in eosinophil functional migratory response and may point to a novel tyrosine kinase-mediated ligation between CCR3 receptor and 2B4 co-receptor in eosinophil chemotaxis. If so, then 2B4 molecules would be a novel target for therapeutic modalities in diseases characterized by eosinophilia such as AR.

Mailing address: Prof. Dr. A. E. El-Shazly,Department of Otolaryngology and Head and Neck Surgery, Rhinology unit, Liege University Hospital-(C.H.U.), 4000-Liege, BelgiumFax: ++32 43667525e-mail: [email protected]

Key words: eosinophils, Eol-1, chemotaxis, adhesion, allergic rhinitis, 2B4 (CD244), CD2, tyrosine kinase

Vol. 24, no. 4, 949-960 (2011)






INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY Vol. 24, no. 3, 0-0 (2011)



Unauthorized reproduction may result in financial and other penalties603

EXPRESSION OF IL-23, VEGF AND TLR2/TLR4 ON MONONUCLEAR CELLS AFTER EXPOSURE TO PSEUDOMONAS AERUGINOSA

R. PAOLILLO, C. ROMANO CARRATELLI, S. SORRENTINO, N. MAZZOLA, L. MITA and A. RIZZO

Department of Experimental Medicine, Section of Microbiology and Clinical Microbiology, Faculty of Medicine and Surgery, Second University of Naples, Naples, Italy

Received December 23, 2010 – Accepted May 9, 2011

Pseudomonas aeruginosa is a Gram-negative, aerobic bacillus causing infections of the respiratory and other organ systems in susceptible hosts. Although it does not cause pulmonary infections in immunocompetent individuals, P. aeruginosa causes chronic lung infection in individuals with cystic fibrosis and nosocomial pneumonia resulting in significant morbidity and mortality. Exogenous administration of an important P. aeruginosa virulence factor, lipase, present in P. aeruginosa culture supernatant, induces potent mononuclear cell activation leading to the production of numerous proinflammatory cytokines. In particular, P. aeruginosa culture supernatant stimulated increased proliferation of THP-1 cells and monocytes (MN). The addition of culture supernatant to THP-1 cells and MN also induced Interleukin (IL)-23 and vascular endothelial growth factor (VEGF) release in a time-dependent manner. To investigate whether any compounds present in the supernatant lipase contributed to releasing IL-23 and VEGF, the culture supernatant from P. aeruginosa containing lipase was treated with hexadecylsulfonylfluoride (AMSF). The AMSF-treated culture supernatant (CS) did not show any induction on the IL-23 and VEGF release compared to the cells treated with CS without AMSF. We also showed that Toll-like receptors (TLR)2/TLR4 are expressed in THP-1 cells and MN treated with P. aeruginosa CS in a time-dependent fashion. Flow cytometry analysis revealed a higher TLR4 and a lower TLR2 expression at 48 and 72 h of treatment. The treatment of cells with TLR4 neutralizing antibody, and to a lesser extent with TLR2 neutralizing antibody, resulted in a decrease in P. aeruginosa CS-induced IL-23 and VEGF production.

Mailing address:Prof. C. Romano Carratelli,Via Santa Maria di Costantinopoli, 1680138 – Napoli, ItalyTel: ++39 081566 5658 Fax: ++39 081566 56 68e-mail: [email protected]

Key words: Pseudomonas aeruginosa, lipase, IL-23, VEGF, human mononuclear cells, TLR2/TLR4

The Gram-negative bacterium Pseudomonas aeruginosa is an opportunistic pathogen that causes a wide range of acute and chronic infections and disease associated with significant inflammatory responses. The course and severity of the disease are determined by the efficiency of the local or systemic immunological and non-immunological defenses of the host. A wide variety of cells including

lymphocytes, monocytes and respiratory tract cells, like respiratory epithelial cells, produce cytokines in response to Gram-negative bacteria or their products (1-2). As a primary interface between pathogens and the host, mononuclear cells are crucial cells for the innate immune response (3), and appear to be the primary source of proinflammatory cytokines during infections. Moreover, it is becoming clearer that

INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY Vol. 24, no. 3, 0-0 (2011)



Unauthorized reproduction may result in financial and other penalties603

EXPRESSION OF IL-23, VEGF AND TLR2/TLR4 ON MONONUCLEAR CELLS AFTER EXPOSURE TO PSEUDOMONAS AERUGINOSA

R. PAOLILLO, C. ROMANO CARRATELLI, S. SORRENTINO, N. MAZZOLA, L. MITA and A. RIZZO

Department of Experimental Medicine, Section of Microbiology and Clinical Microbiology, Faculty of Medicine and Surgery, Second University of Naples, Naples, Italy

Received December 23, 2010 – Accepted May 9, 2011

Pseudomonas aeruginosa is a Gram-negative, aerobic bacillus causing infections of the respiratory and other organ systems in susceptible hosts. Although it does not cause pulmonary infections in immunocompetent individuals, P. aeruginosa causes chronic lung infection in individuals with cystic fibrosis and nosocomial pneumonia resulting in significant morbidity and mortality. Exogenous administration of an important P. aeruginosa virulence factor, lipase, present in P. aeruginosa culture supernatant, induces potent mononuclear cell activation leading to the production of numerous proinflammatory cytokines. In particular, P. aeruginosa culture supernatant stimulated increased proliferation of THP-1 cells and monocytes (MN). The addition of culture supernatant to THP-1 cells and MN also induced Interleukin (IL)-23 and vascular endothelial growth factor (VEGF) release in a time-dependent manner. To investigate whether any compounds present in the supernatant lipase contributed to releasing IL-23 and VEGF, the culture supernatant from P. aeruginosa containing lipase was treated with hexadecylsulfonylfluoride (AMSF). The AMSF-treated culture supernatant (CS) did not show any induction on the IL-23 and VEGF release compared to the cells treated with CS without AMSF. We also showed that Toll-like receptors (TLR)2/TLR4 are expressed in THP-1 cells and MN treated with P. aeruginosa CS in a time-dependent fashion. Flow cytometry analysis revealed a higher TLR4 and a lower TLR2 expression at 48 and 72 h of treatment. The treatment of cells with TLR4 neutralizing antibody, and to a lesser extent with TLR2 neutralizing antibody, resulted in a decrease in P. aeruginosa CS-induced IL-23 and VEGF production.

Mailing address:Prof. C. Romano Carratelli,Via Santa Maria di Costantinopoli, 1680138 – Napoli, ItalyTel: ++39 081566 5658 Fax: ++39 081566 56 68e-mail: [email protected]

Key words: Pseudomonas aeruginosa, lipase, IL-23, VEGF, human mononuclear cells, TLR2/TLR4

The Gram-negative bacterium Pseudomonas aeruginosa is an opportunistic pathogen that causes a wide range of acute and chronic infections and disease associated with significant inflammatory responses. The course and severity of the disease are determined by the efficiency of the local or systemic immunological and non-immunological defenses of the host. A wide variety of cells including

lymphocytes, monocytes and respiratory tract cells, like respiratory epithelial cells, produce cytokines in response to Gram-negative bacteria or their products (1-2). As a primary interface between pathogens and the host, mononuclear cells are crucial cells for the innate immune response (3), and appear to be the primary source of proinflammatory cytokines during infections. Moreover, it is becoming clearer that

Vol. 24, no. 4, 961-974 (2011)






in the body. The symptoms of this disturbance include weakness, nausea and vomiting, impaired breathing, hypotension, hypocalcaemia, arrhythmia and asystole. Hypermagnesemia is defined by a serum magnesium concentration > 2.1 mEq/L (> 1.05 mmol/L), and the clinical consequences vary by serum concentration (>4.0 mEq/L hyporeflexia, >5.0 mEq/L prolonged atrioventricularconduction, >10.0 mEq/L complete heart block,>13.0 mEq/L cardiac arrest) (6). Serum magnesium levels are regulated by the kidneys, and a normal plasma magnesium concentration is approximately 1.4-1.7 mEq/L.

Hypermagnesemia is often iatrogenic and is commonly associated with excess magnesium intake by patients with renal insufficiency or renal failure. Concentrated sources of magnesium include

A well-composed diet provides an organism with all the essential nutrients (1). Uncontrolled dietary supplementation of magnesium or use of magnesium therapy may cause disruptions in metabolic processes, which, in turn, may result in a disruption of homeostasis (2, 3). An analysis of the daily consumption of magnesium is highly important (4, 5). The increased intake of foods containing magnesium in combination with unjustified dietary supplementation with widely available solid, oral forms containing magnesium salts or multi-component preparations may lead to hypermagnesemia. Hypermagnesemia is an electrolyte disturbance in which there is an abnormally elevated level of magnesium in the blood. Usually this results in an excess of magnesium

Magnesium is one of the most important cations for an organism. The aim of our study is to evaluate whether the use of a magnesium formulation as a diet supplement or medical treatment is necessary. The 24-hour recall method was used to obtain information regarding the daily magnesium consumption of 949 people. The results were compared with the Estimated Average Requirement (EAR) and Recommended Daily Allowance (RDA) values. The average daily requirement for magnesium was exceeded by 292 (183 women and 109 men) of the 949 respondents. This research confirmed excessive magnesium intake by both men and women that exceeded both the EAR and the RDA. Uncontrolled, excessive dietary supplementation or medical treatment with magnesium by this group may constitute a health threat.

MAGNESIUM CONTENT IN DAILY FOOD PORTIONS AND THE INFLUENCE OF SUPPLEMENTATION

E. NIEDWOROK1, M. MUC-WIERZGOŃ2, E. NOWAKOWSKA-ZAJDEL2,L. DUL3 and K. KLAKLA2

1Department of Human Nutrition, Medical University of Silesia, Bytom; 2Department of Internal Diseases, Medical University of Silesia, Bytom; 3Department of Biostatistics, Medical University of

Silesia, Bytom, Poland

Received March 23, 2011 – Accepted November 4, 2011

Mailing address: Prof. MałgorzataMuc-Wierzgoń,Dept. of Internal Diseases,Medical University of Silesia,Żeromskiego 7 st,41-902 Bytom, PolandTel/Fax:48/32 2812122e-mail:[email protected]

Key words: hypermagnesemia, magnesium, nutrition, 24-recall method

Vol. 24, no. 4, 975-981 (2011)






Class-switch recombination defects also known as hyper-IgM syndromes (HIGMs) are rare primary immune deficiencies characterized by normal or increased serum immunoglobulin (Ig)M and low or normal IgG levels (1). Since it was first described nearly 50 years ago, seven different forms of HIGMs have been identified with either X-linked or autosomal (recessive - dominant) inheritance and a broad range of clinical manifestations

mainly represented by increased susceptibility to opportunistic infections and autoimmunity (2-3).

The most common HIGM form (HIGM1) is characterized by mutations of the CD40L, whereas the CD40 (HIGM3) defect is more rare. CD40/CD40L pathway is essential for Ig Class Switch Recombination (CSR) and Somatic Hypermutation (SHM); patients suffer from recurrent infections caused by bacteria, viruses and fungi, and may

HYPER-IgM, NEUTROPENIA, MILD INFECTIONS AND LOW RESPONSE TO POLYCLONAL STIMULATION: HYPER-IgM SYNDROME OR COMMON VARIABLE

IMMUNODEFICIENCY?

M.M. ROSADO1, A. PICCHIANTI DIAMANTI2, S. CASCIOLI1, S. CECCARELLI1, S. CAPORUSCIO2, R. D’AMELIO2, R. CARSETTI1 and B. LAGANÀ2

1Research Center, Ospedale Pediatrico Bambino Gesù, Laboratory of Flow-cytometry and B cell development, IRCSS, Rome; 2”Sapienza” University of Rome, II School of Medicine, S. Andrea

University Hospital, Rome, Italy

Received February 23, 2011 – Accepted August 1, 2011

M.M.R., A.P.D., R.C. and B.L. contributed equally to this work

Mailing address: Prof. B. Laganà, Sapienza University of Rome, II School of Medicine, S. Andrea University Hospital, Via di Grottarossa 1039, 00189 Rome, Italy Tel: ++39 06 33776043 Fax: ++39 06 33775331 e-mail: [email protected]

Key words: intravenous immunoglobulin, hyper-IgM syndrome,common variable immunodeficiency, class switch recombination

A young woman presenting respiratory infections, polyarthritis, severe neutropenia, and increased serum IgM was treated with Intravenous Immunoglobulin (IVIG) with good clinical and laboratory outcome followed by a loss of efficacy. The increased serum IgM associated to recurrent infections and autoimmune manifestations suggested the diagnosis of a hyper–IgM syndrome (HIGMs). The frequency of peripheral T cells, the expression of CD40 on the patients’ B cells and CD40L on T cells and the Activation-Induced cytidine Deaminase (AID) and Uracyl-DNA glycosylase (UNG) at mRNA level was comparable to controls. In contrast, the frequency of B cells was one half of the healthy control and all cells showed an atypical phenotype. Although AID and UNG were normal, class-switch recombination was not very efficient because circulating switched memory were reduced and, once stimulated with CpG, generated less antibody-secreting cells than controls. An increase in serum B Lymphocytes stimulator (BLyS) was also found. The patient presented a peculiar clinical and immunological phenotype fitting for many aspects of both HIGM4 and Common Variable Immunodeficiency (CVID). These findings underline the need to better explore the complex link between these two diseases.

Vol. 24, no. 4, 983-991 (2011)






Opioids play an important role in the clinical management of pain. The µ-opioid receptor, encoded by the gene of human opioid receptor µ−1 (OPRM1), is the primary site of action for the most commonly used opioids (1). In literature a wide interindividual variation in sensitivity to opioids has been demonstrated,

suggesting potential variability in the gene (OPRM1) and in the µ opioid receptor protein (MOR). Several studies tried to relate OPRM1 genotyping in the context of pain therapy, to identify patient populations that will benefit from a pharmacogenetically-guided pain therapy (2).

Single-nucleotide polymorphisms (SNPs) are

MICRO OPIOID RECEPTOR A118G POLYMORPHISM AND POST-OPERATIVE PAIN: OPIOIDS’ EFFECTS ON HETEROZIGOUS PATIENTS

A. DE CAPRARIS1, G. CINNELLA2, A. MAROLLA3, P. SALATTO1, S. DA LIMA1,P. VETUSCHI1, L. CONSOLETTI1, L. GESUALDO4 and M. DAMBROSIO2

1Unit of Anesthesia and Intensive Care, University Hospital of Foggia, Foggia; 2Department of Surgical Science, Unit of Anesthesia and Intensive Care, University Hospital of Foggia, Foggia;

3Department of Surgical Sciences, University Hospital of Foggia, Foggia; 4Department of Biomedical Sciences, Unit of Nephrology, University Hospital of Foggia, Foggia, Italy

Received February 4, 2011 – Accepted September 5, 2011

The Single-Nucleotide-Polymorphism (SNP) 118A>G in the µ-1 Opioid Receptor gene (OPRM1) is associated with a decrease in the analgesic effects of opioids. The aim of this study is to assess whether 118A>G polymorphism could influence the analgesic response to opioid-based postoperative pain (POP) therapy. The study consisted of two parts: section α, observational, included 199 subjects undergoing scheduled surgical procedures with pain management standardized on surgery invasiveness and on expected level of postoperative pain; section β, randomized, included 41 women undergoing scheduled caesarean delivery with continuous intra-operative epidural anesthesia and post-operative analgesia (CEA). In both sections, POP was measured over 48 h (T6h-T24h-T48h) by the visual analogue scale (VAS). In section β we also tested the responsiveness of hypothalamic-pituitary-adrenal axis (HPA) expressed by cortisol levels. In section α, with cluster analysis, subjects were analyzed according to their genotype: a group (#1) of 34 patients reporting VAS score >3 at every time lapse was identified and included only A118G carriers, while wild-type (A118A - absence of 118A>G polymorphism) patients were unevenly distributed between those with cluster #2 (VAS score <3 at every study steps) and those with cluster #3 (VAS score progressively reducing from T6h). In section β, A118G carriers receiving epidural sufentanil had the lowest VAS scores at T24h; also in these patients, cortisol levels remained more stable, with a mild decrease at T6h. This study shows that the OPRM1 118A>G polymorphism affects postoperative pain response in heterozygous patients: they have a different postoperative pain response than patients with wild-type genes, which may affect the efficacy of the analgesic therapy.

Mailing address: Dr. Antonella De Capraris, Istituto di Anestesia e Rianimazione Universitaria,Ospedali Riuniti, Università degli Studi di Foggia,Viale L. Pinto, 171100 Foggia, ItalyTel/Fax ++39 0881 732387e-mail: [email protected]

Key words: postoperative pain; μ opioid receptor; 118A>G polymorphism, cortisol

Vol. 24, no. 4, 993-1004 (2011)






NEO-ADJUVANT CHEMO/IMMUNOTHERAPY IN THE TREATMENT OF STAGE III (N2) NON-SMALL CELL LUNG CANCER:

A PHASE I/II PILOT STUDY

G.B. RATTO1, R. COSTA2, P. MAINERI3, A. ALLOISIO1, M.T. PIRAS1, A. D’AGOSTINO4, G. TRIPODI5, L. RIVABELLA5, B. DOZIN6, P. BRUZZI6

and G. MELIOLI4

1U.O.C Chirurgia Toracica, Istituto Nazionale per la Ricerca sul Cancro, Genoa; 2U.O.S Antonio e Biagio e C. Arrigo Hospital, Alessandria; 3U.O.C Chirurgia Toracica, Ospedale Santa Corona,

Pietra Ligure, Savona; 4U.O.C Laboratorio Centrale di Analisi, Istituto G. Gaslini, Genoa; 5 U.O.C Centro Trasfusionale, Istituto G. Gaslini, Genoa; 6U.O.C Epidemiologia Clinica, Istituto Nazionale

per la Ricerca sul Cancro, Genoa, Italy

Received February 23, 2011 – Accepted September 1, 2011

In a previous randomized study, we showed that adjuvant immunotherapy with tumor-infiltrating lymphocytes and recombinant interleukin-2 (rIL-2) significantly improved survival in resected N2-Non Small Cell Lung Cancer (NSCLC) patients. The present study assesses feasibility, safety and potential efficacy of combined neo-adjuvant chemotherapy and immunotherapy with peripheral blood mononuclear cells (PBMC) and rIL-2 in resectable N2-NSCLC patients. Eighty-two consecutive N2-NSCLC patients underwent neo-adjuvant chemotherapy with cisplatin and gemcitabine. Out of the 82 patients, 23 were also subjected to leukapheresis prior to neo-adjuvant chemotherapy while the remaining 59 did not. Collected PBMC were analyzed for viability and phenotype and then stored frozen in liquid nitrogen. Thawed PBMC were infused intravenously, 5 days before surgery. After the infusion, rIL-2 was administered subcutaneously until surgery. Only patients with a partial or complete response to neo-adjuvant chemotherapy underwent surgery: 13 patients in the experimental immunotherapy group (A) and 32 in the reference group (B). The two groups were homogeneous for all major prognostic factors. Median leukapheresis yield was 10 billion PBMC, (range 3-24 billions). Two to six billion PBMC were infused. The phenotypic analysis showed that similar proportions of CD4 and CD8 cells were present in leukapheresis products, and thawed PBMC, as well as in T lymphocytes isolated from the removed tumours. No severe adverse effects were observed following immunotherapy. No significant differences in overall survival (OS) and event-free survival (EFS) were seen between the two groups. However, the 5-year OS in group A was almost twice as much compared to group B (59% vs 32%). After adjustment for major prognostic factors, a statistically significant 66% reduction in the hazard of death was seen in patients receiving immunotherapy. The OS benefit was more evident in patients with adenocarcinoma than in those with squamous cell carcinoma. This study supports the favorable toxicity profile and potential efficacy of combining neo-adjuvant chemotherapy and immunotherapy with PBMC and rIL-2 in the treatment of N2-NSCLC patients.

Mailing address: Giovanni Melioli, MDLaboratorio Centrale di AnalisiIstituto Giannina GasliniLargo Gerolamo Gaslini 5, 16147 Genova, ItalyTel: ++39 010 5636557 Fax: ++39 010 3994168e-mail: [email protected]

Key words: non-small cell lung cancer, neo-adjuvant immunotherapy, neo-adjuvant chemotherapy, surgery, survival

Vol. 24, no. 4, 1005-1016 (2011)






Chronic Fatigue Syndrome (CFS) is still an unclear clinical entity, characterized by an unexplained disabling fatigue, immunological alterations and aspecific accompanying symptoms: sleep disorders, neurocognitive dysfunctions and multiple somatic complains, such as muscle pain,

multi-joint pain and unusual headache (1-2). The Centers for Disease Control (CDC) criteria (3) exclude CFS when subjects are affected by bipolar disorders and major depressive disorders with psychotic or melancholic features. Major depressive disorders without psychotic or melanchonic features,

PSYCHOPHYSICAL DISTRESS AND ALEXITHYMIC TRAITS IN CHRONIC FATIGUE SYNDROME WITH AND WITHOUT COMORBID DEPRESSION

G. SEPEDE1, D. RACCIATTI2, V. GORGORETTI2, M. NACCI3, E. PIZZIGALLO2,M. ONOFRJ1, M. DI GIANNANTONIO1,4, C. NIOLU5, R.M. SALERNO1 and F. GAMBI1

1Department of Neuroscience and Imaging, “G. d’Annunzio” University, Chieti; 2Clinic of Infectious Diseases, Department of Medicine and Aging, “G. d’Annunzio" University, Chieti;

3Department of Mental Health, National Health Trust, Taranto; 4Center of Mental Health, National Health Trust, Chieti, Italy; 5Psychiatric Unit, Tor Vergata University, Rome, Italy

Received March 15, 2011 – Accepted October 3, 2011

Patients with Chronic Fatigue Syndrome (CFS) often report a comorbid depressive disorder. Comorbid depression may negatively influence the long-term outcome of CFS therefore it must be correctly diagnosed and treated. The aim of the present study is to provide a clinical and psychometric assessment of CFS patients with and without depressive features. A comparative analysis between 57 CFS subjects (CDC, 1994), 17 of whom with a comorbid depression, and 55 matched healthy volunteers was assessed to evaluate the presence of any psychophysical distress and alexithymic traits, by means of Symptom Checklist-90-R (SCL-90R) and Toronto Alexithymia Scale (TAS-20). The severity of fatigue was also assessed in all CFS patients using the Fatigue Impact Scale (FIS). With regard to psychiatric comorbidity, the SCL-90R scores showed higher levels of somatic complaints in CFS patients than in healthy subjects, whereas augmented depressive and obsessive-compulsive symptoms were observed only in the depressed CFS subgroup. When comparing the TAS-20 scores, we observed a selective impairment in the capacity to identify feelings and emotions, as measured by the Difficulty in Identifying Feelings subscale (DIF), non-depressed CFS patients showing an intermediate score between depressed CFS and healthy controls. Finally, in terms of FIS scores, a statistical trend versus a higher fatigue severity in depressed CFS patients, with respect to non-depressed ones, was observed. In conclusion, comorbid depression in CFS significantly increased the level of psychophysical distress and the severity of alexithymic traits. These findings suggest an urgent need to address and treat depressive disorders in the clinical care of CFS cases, to improve social functioning and quality of life in such patients.

Mailing address: Gianna Sepede, MD, PhD. Department of Neuroscience and Imaging, ITAB - Institute for Advanced Biomedical Technologies, University “G. d’Annunzio” of Chieti, Via dei Vestini 33, 66013 Chieti (CH), Italy Tel: ++39 0871 3556901 Fax: ++39 0871 3556930 e-mail: [email protected]

Key words: alexithymia, Chronic Fatigue Syndrome (CFS), comorbidity, depression

Vol. 24, no. 4, 1017-1025 (2011)






The pathogenesis of acute exacerbations in chronic obstructive pulmonary disease (COPD) is still poorly understood and the role of bacteria is highly controversial. Various infectious agents, including bacteria, viruses and atypical bacteria, are known to increase airway inflammation and are implicated in up to 80% of acute exacerbations, while bacterial infections are involved in more than half of the cases (1-2). As for other chronic lung diseases such as cystic fibrosis, even in the case of COPD, bacteria have been

hypothesized to persist for months in respiratory tract growing as a biofilm and being probably responsible for exacerbations (AECOPD) (3). A biofilm is a structured community of bacteria enveloped in a self-produced polymeric matrix and adherent to an inert or living surface (4). Whereas planktonic bacteria are generally susceptible to host-defense mechanisms and to antibiotic treatments, bacteria in biofilms are substantially resistant and concentrations of antibiotics required to achieve bactericidal activity against

MODULATION OF BIOFILM OF STRAINS ISOLATED FROM PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE BY LEVOFLOXACIN,

MOXIFLOXACIN, CIPROFLOXACIN, AMOXICILLIN/CLAVULANIC ACID AND CEFTRIAXONE

L. DRAGO1,2, R. MATTINA3, D. LEGNANI4, C.L. ROMANÒ5, E. VIANELLO6,C. RICCI7 and E. DE VECCHI1

1Laboratory of Clinical Chemistry and Microbiology, IRCCS Galeazzi Institute, Milan;2Laboratory of Clinical Microbiology, Department of Clinical Sciences L. Sacco, University of

Milan; 3Department of Public Health, Microbiology and Virology, University of Milan; 4Institute of Respiratory Disease, L. Sacco Teaching Hospital, University of Milan; 5C.R.I.O. Unit, IRCCS

Galeazzi Institute Milan; 6Dept Human Morphology and Biomedical Sciences “Città Studi”, University of Milan; 7Clinical Epidemiology and Biometry Unit, IRCCS Policlinico San Donato,

Milan, Italy

Received March 25, 2011 – Accepted September 5, 2011

The ability of levofloxacin, moxifloxacin, ciprofloxacin, amoxicillin/clavulanic acid and ceftriaxone to interfere on biofilm produced by Pseudomonas aeruginosa, Haemophilus influenzae and Streptococcus pneumoniae isolated from patients with chronic obstructive pulmonary disease was evaluated. The effects of antibiotics were evaluated on formation of biofilm (at 1/2, 1/4 and 1/8 X MIC) and on pre-formed biofilm (at epithelial lining fluid peak concentrations) by means of a spectrophotometric method. Levofloxacin was the most active compound followed by ciprofloxacin, moxifloxacin and amoxicillin/clavulanic acid and ceftriaxone. Levofloxacin may contribute to clear the reservoir of pathogens involved in chronic obstructive pulmonary disease, thus leading to decreased occurrence of acute exacerbations.

Mailing address: Dr L. Drago,Laboratory of Clinical Chemistry and Microbiology, IRCCS Galeazzi Orthopaedic Institute, via Riccardo Galeazzi 4, 20161 Milan, ItalyTel: ++39 0266214839Fax: ++39 0250319651 e-mail: [email protected]

Key words: fluoroquinolones, β-lactams, cephalosporins, Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa

Vol. 24, no. 4, 1027-1035 (2011)






Atopic dermatitis (AD) is a clinically defined chronic inflammatory skin disease affecting 10%-20% of children in Western societies (1). AD is characterized by a complex genetic background, a disturbed immune system, environmental trigger factors and typical skin manifestations (2).

The most important pathogenetic aspect of AD

is the presence of defective skin and gut barrier functions that play a critical role in inducing abnormal inflammation and activation of the immune system upon exposure to ubiquitous environmental allergens and infectious pathogens (3).

Probiotics have been defined as “live microorganisms which, when administered in

EFFECTS OF LACTOBACILLUS SALIVARIUS LS01 (DSM 22775) TREATMENT ON ADULT ATOPIC DERMATITIS: A RANDOMIZED PLACEBO-CONTROLLED STUDY

L. DRAGO1, E. IEMOLI2, V. RODIGHIERO1, L. NICOLA1, E. DE VECCHI1

and S. PICONI2

1Laboratory of Clinical Microbiology, Department of Clinical Sciences Luigi Sacco, University of Milan, and Laboratory of Clinical Chemistry and Microbiology, IRCCS Galeazzi Orthopaedic

Institute, Milan; 2Allergy and Clinical Immunology Unit “Luigi Sacco” Hospital, Milan, Italy


Atopic dermatitis (AD) is a common inflammatory skin disease characterized by xerosis, pruritus and eczema. The role of probiotics in the prevention and the treatment of AD have been extensively studied in children with controversial results while there are few studies on an adult population. The aim of this randomized, double-blind, placebo-controlled study is to evaluate the clinical efficacy of the intake of a probiotic strain (Lactobacillus salivarius LS01) in the treatment of adult patients with AD. A group of 38 patients was treated with probiotics or placebo (maltodextrin) for 16 weeks. The study was performed from January (T0) to May, 2009 (T16). The assessment of efficacy was based on change in SCORAD (SCORing Atopic Dermatitis) index, dermatology life quality index (DLQI) improvement, cytokine production by PBMCs and ability to modify faecal microbial flora. No significant adverse events were recorded during the study. Patients treated with probiotics showed a statistically improvement of both clinical parameters (SCORAD p< 0.0001 and DLQI p= 0.021) at the end of treatment (T16) compared with the placebo group. Furthermore, after four months of treatment there was a significant reduction of Th1 cytokines (IL-12+IFNγ) (p= 0.03) and Th1/Th2 ratio (IL-12+IFNγ/IL-4+IL-5) (p= 0.019) only in placebo-treated patients. A statistically relevant decrease of staphylococci in faeces of the probiotic-treated group was also observed at the end of treatment. In our study, the administration of L. salivarius LS01 was well tolerated and was associated with a significant improvement of clinical manifestation and QoL. This probiotic strain could have an important role in modulating Th1/Th2 cytokine profiles and could be considered as an important adjunctive therapy in the treatment of adult AD.

Mailing address: Dr Lorenzo Drago,Clinical-Chemistry and Microbiology Lab,IRCCS Galeazzi Institute, University of Milan,Via R.Galeazzi, 4, 20166 Milano, ItalyTel: ++390266214718Fax: ++390266214774e-mail: [email protected]

Key words: probiotics, adults, atopic dermatitis, Th1 cells, Th2 cells

Vol. 24, no. 4, 1037-1048 (2011)






Exposure to allergens induces an immunologic response that involves both serum allergen specific IgE and IgG4 (1) which have been suggested to represent soluble markers of sensitization and allergic disease (2).

Both IgG4 and IgE are regulated by Th2 lymphoid cells (3-4). IgG4, in particular, have been hypothesized to act as blocking antibodies capable of preventing IgE-mediated effector cell triggering (5), possibly by an allergen-blocking effect at the

level of mast cells and/or antigen-presenting cells (6). Furthermore, IgG4 antibodies have been shown to be expressed after prolonged immunization and they have been hypothesized to be involved in the development of tolerance (7-10).

A number of studies performed in an Alpine sanatoria (11-17) have demonstrated that, in asthmatic children, staying at a high altitude is associated with a global improvement of the disease, including clinical respiratory symptoms, lung

A COMPARISON BETWEEN IgE AND IgG4 AS MARKERS OF ALLERGY IN CHILDREN: AN EXPERIMENTAL TRIAL IN A MODEL

OF NATURAL ANTIGEN AVOIDANCE

G.L. PIACENTINI1, S. GUERRESI1, A. KANTAR2, L. LUBRANO1, F. OLIVIERI1, A.L. BONER1 and D.G. PERONI1

1Pediatric Section, Department of Life and Reproduction Sciences, University of Verona, Verona;2Istituto Pio XII, Misurina (BL), Italy

Received May 17, 2011 – Accepted October 21, 2011

IgG4 have been hypothesized to act as blocking antibodies capable of preventing IgE-mediated effector cell triggering. This study aims to evaluate the changes in IgG4 in children during a period of natural antigen avoidance. Serum IgE and IgG4 were evaluated in a group of asthmatic children, aged between 7 and 17 years, admitted to the residential house Istituto Pio XII (Misurina, BL, Italy), located at 1,756 m, in a natural model of antigen avoidance. All the patients were skin prick test positive to at least two of the following allergens: Dermatophagoides pteronissynus, Dermatophagoides farinae, cat epithelium, timothy grass pollen and Parietaria pollen. During the 180 days of hospitalization, serum specific IgE and IgG4 were measured six times. A significant decrease (p≤0.05) in serum specific IgE to house dust mite and pollen allergens was observed; by contrast, no significant variations were shown by IgG4 and IgG4/IgE ratio. No significant relationship was found between serum specific IgE, IgG4 and IgG4/IgE ratio variations and the re-exposure to house dust mite allergens during the Christmas holidays. A positive correlation between specific IgE and specific IgG4 was observed at each considered time (T0: r=0.57, p=0.08; T1: r=0.85, p=0.001; T3: r=0.76, p=0.01). The positive correlation between specific IgE and specific IgG4, enduring throughout the entire time of study, suggests a relationship between these classes of immunoglobulins.

Mailing address: Diego G. Peroni, MDPaediatric Department, Department of Life and Reproduction Sciences, University of Verona, Policlinico G.B. Rossi,P.le L.A. Scuro, 37134 Verona, ItalyFax: ++39 045 8124790e-mail: [email protected]

Key words: antigen avoidance, asthma, IgE, IgG4

Vol. 24, no. 4, 1049-1056 (2011)






Cystic fibrosis (CF) is an autosomal recessive disease characterized by dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR) channel (1) leading to abnormalities in ion transport across the epithelial cells. Particularly relevant is the malfunctioning of respiratory tract, and high iron concentrations are found in airway secretions (2-3).

LACTOFERRIN DECREASES INFLAMMATORY RESPONSE BY CYSTIC FIBROSIS BRONCHIAL CELLS INVADED WITH BURKHOLDERIA CENOCEPACIA IRON-

MODULATED BIOFILM

P. VALENTI1, A.CATIZONE2, F. PANTANELLA1, A. FRIONI1, T. NATALIZI1,M. TENDINI1 and F. BERLUTTI1

1Department of Public Health and Infectious Disease, Sapienza University of Rome, Rome; 2Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, Sapienza University of

Rome, Rome, Italy

Received February 17, 2011 – Accepted July 26, 2011

In cystic fibrosis (CF) high iron concentration in airway secretion plays a pivotal role in bacterial multiplication and biofilm formation as well as in inflammatory response. Burkholderia cenocepacia, an opportunistic facultative pathogen responsible for chronic lung infections and cepacia syndrome, recurrently infects CF patients. Lactoferrin (Lf), an iron binding multifunctional glycoprotein synthesized by exocrine glands and neutrophils, has been found at higher concentration in the airway secretions of infected CF patients than in healthy subjects. Here the influence of milk derivative bovine lactoferrin (bLf), an emerging important regulator of iron and inflammatory homeostasis, on invasiveness of B. cenocepacia iron-modulated biofilm, as well as on inflammatory response by infected CF bronchial (IB3-1) cells, is reported. bLf did not significantly affect invasion efficacy by biofilm-forming B. cenocepacia clinical strains. Conversely, the addition of bLf to cell monolayers during infection significantly decreased the pro-inflammatory Interleukin (IL)-1β and increased the anti-inflammatory IL-11 expression compared to that observed in cells infected in the absence of bLf. The bLf ability to modulate genes expressed following B. cenocepacia infection seems related to its localization to the nucleus of infected IB3-1 cells. These results provide evidence for a role of bLf in the protection of infected CF cells from inflammation-related damage, thus extending the therapeutic potential of this multifunctional natural protein.

Mailing address: Dr Francesca Berlutti,Department of Public Health and Infectious Disease, Sapienza University of Rome, p.le A. Moro, 500185 Rome, ItalyTel: ++39 06 49914638Fax: ++39 06 49914626 e-mail: [email protected]

Key words: lactoferrin, iron-modulated biofilm, cystic fibrosis bronchial cell infection, inflammatory response

A hallmark of CF is a chronic lung infection due to opportunistic bacteria inducing sustained neutrophil-mediated inflammation (4-5). Aggressive antibiotic and anti-inflammatory treatments may ameliorate CF patient symptoms in the short-term (6), but they do not consistently reduce the bacterial load, which may be related to persistently increased airway iron concentrations (3). As a matter of fact, high iron

Vol. 24, no. 4, 1057-1068 (2011)






Bronchial asthma is a chronic inflammatory disease of the airways characterized by airflow obstruction, usually reversible either spontaneously or after therapy, bronchial hyperreactivity (BHR) and accelerated deterioration of lung function that can sometimes lead to irreversible airway obstruction. Currently, the up-to-date GINA guidelines classify asthma according to 4 levels of increasing severity: 1) Intermittent 2) Mild Persistent, 3) Moderate Persistent, 4) Severe Persistent, which correspond to different treatments (1).

Intermittent asthma is generally characterized by occasional symptoms that may occur over long periods of time or by symptomatic periods, even relevant and sustained, alternated with long periods of remission. Asthma can be triggered by many factors (physical exercise, allergens, etc.), it occurs especially during early childhood, can remain stable for a long time and usually requires no control therapy, but only as-needed or for short periods (2). Bronchial reactivity should also be monitored in subjects with allergic rhinitis, due to

LETTER TO THE EDITOREVALUATION OF BRONCHIAL HYPERREACTIVITY WITH MANNITOL

DRY POWDER CHALLENGE TEST IN A PAEDIATRIC POPULATION WITH INTERMITTENT ALLERGIC ASTHMA OR ALLERGIC RHINITIS

F. DECIMO, C. CAPRISTO, R. AMELIO, N. MAIELLO, A.F. CAPRISTOand M. MIRAGLIA DEL GIUDICE

Department of Paediatrics, II University of Naples, Italy

Received March 3, 2011 – Accepted July 22, 2011

We evaluated the bronchial hyperreactivity (BHR) with a new bronchial challenge test, mannitol dry powder, in a paediatric population with intermittent allergic asthma or allergic rhinitis who did not respond to an exercise challenge test. We selected 50 children, aged 9-16 years, with intermittent allergic bronchial asthma (Group 1) or allergic rhinitis without clinical manifestation of asthma for at least 12 months (Group 2). All patients performed the following tests in three different days (≥ 48 hours apart): Day 1: exhaled nitric oxide (FeNO) determination followed by baseline spirometry and reversibility to inhaled β2-agonists; Day 2: exercise challenge test followed by FeNO determination; Day 3: mannitol challenge test followed by FeNO determination. Forty children completed the study. Eighteen subjects of Group 1 (90%) and 5 subjects of Group 2 (25%) resulted positive to the mannitol test. Positive mannitol challenge subjects showed no statistically significant differences compared to negative subjects as regard baseline spirometry, reversibility to salbutamol and response to the exercise challenge test, but they had significantly higher FeNO values. In conclusion, the mannitol challenge test can be a diagnostic tool more useful than the exercise challenge test to identify BHR in a paediatric population with intermittent allergic asthma or allergic rhinitis because it is better reproducible, quick and easy to perform and well tolerated.

Mailing address: Fabio Decimo, MD,Seconda Università di Napoli, Dipartimento di Pediatria, Via L. de Crecchio 4, 80138 Napoli, ItalyTel: ++39 0815665468 Fax: ++39 0815665467e-mail: [email protected]

Key words: intermittent asthma, rhinitis, bronchial hyperresponsiveness, aspecific bronchial provocation test, mannitol dry powder

Vol. 24, no. 4, 1069-1077 (2011)

1074 INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY Vol. 24, no. 4, 0-0 (2011)





Anetoderma is a benign condition characterized by round or oval macular lesions with focal loss of dermal elastic tissue resulting in localized areas of flaccid or herniated sac-like skin (1). In literature, a primary and secondary anetoderma are described.

Primary anetoderma is an idiopathic manifestation while secondary anetoderma is related to various conditions. The term primary anetoderma implies that the lesions occur in clinically normal skin although there could be an association with other dermatological or systemic diseases or conditions, without a well-established relationship. In the secondary anetoderma the lesion occurs on the same site as another skin lesion (2).

We report our experience of two women affected by primary anetoderma. The first 54-year-old patient, affected by myasthenia gravis, antiphospholipid antibody positivity and autoimmune hemolytic anemia, was referred to our department for the presence of pink papules, which were soft to the touch, less than 2 cm in diameter, localized in the spine and

limbs (Fig. 1A). These lesions had appeared about 10 years before the diagnosis of myasthenia gravis had been made at the age of 20 years, the diagnosis of autoimmune hemolytic anemia was made at the age of 37 and the antiphospholipid antibody presence had been found 5 years previously.

A skin biopsy on the upper arm was performed, showing loss of normal elastic tissue throughout the papillary and upper reticular dermis that allows the skin to bulge (Fig. 1B). The ultrastructural examination of the dermis showed abnormal elastic fibers characterized by heterogeneous low density of the amorphous substance. Macrophages were seen at the edge of the fragmented elastic fibers (Fig. 2).

The second patient is a 65-year Italian old woman suffering from multiple sclerosis for 15 years, observed by us for a pink papule, soft to the touch, localized on the torso. Histopathologic examination revealed loss of normal elastic tissue throughout the papillary and upper reticular dermis that allows the skin to bulge.

The anetoderma is a rare skin pathology caused

LETTER TO THE EDITORANETODERMA: EVIDENCE OF THE RELATIONSHIP WITH AUTOIMMUNE DISEASE

AND A POSSIBLE ROLE OF MACROPHAGES IN THE ETIOPATHOGENESIS

S. PERSECHINO1, C. CAPERCHI1, G. CORTESI1, F. PERSECHINO1, S. RAFFA2, E. PUCCI3,A. TAMMARO1 and M.R. TORRISI2

1Dermatology Unit, S. Andrea Hospital, University of Rome “Sapienza”; 2Cellular Diagnostics Unit, S. Andrea Hospital, University of Rome “Sapienza”; 3Department of Histopathology, S.

Andrea Hospital, University of Rome “Sapienza”, Italy

Received April 1, 2011 – Accepted August 31, 2011

Anetoderma is a benign condition characterized by round or oval macular lesions with focal loss of dermal elastic tissue resulting in localized areas of flaccid or herniated saclike skin. Often, the anetoderma is associated with immuno-mediated pathogenetic mechanism. In this article, we describe the association between anetoderma and autoimmune diseases, by underlining the role and the action of macrophages as a possible etiopathogenesis.

Mailing address: Giorgia Cortesi, MDU.O.C. Dermatology, University of Rome ‘Sapienza’Via di Grottarossa, 103500189 Rome, ItalyTel: ++39-06-33775822 Fax: ++39-06-33775378e-mail: [email protected]

Key words: anetoderma, autoimmune diseases, macrophages, electron microscopy






A 36-year-old woman was admitted to our Department in May, 2006 for a necrotic lesion on the left foot, which had appeared two months previously during a trip to the Caribbean. The patient did not remember any traumatic event to her foot. After two days she started to have fever, arthralgia and asthenia until being admitted to our hospital. Her personal history was remarkable only for asthma, that had been diagnosed 8 years previously and treated in the last 3 years with Montelukast, and for chronic sinusitis confirmed by a paranasal computed tomography. Laboratory examinations showed an increased white blood cell count (22.600/µL) with predominance of eosinophils (57%), erythrocyte sedimentation rate 21 mm/h, albumin 3,53 g/dl and pANCA negative. All the laboratory examinations carried out in order to detect a parasitologic etiology (Toxocara, Toxoplasma, Echinococcus) were normal.

A 12-lead ECG showed sinus tachycardia at heart

rate of 110 bpm, negative T waves in all the leads. The patient underwent an echocardiogram, a chest x-ray and other laboratory examinations. The ECG demonstrated a normal-sized left ventricle with a normal ejection fraction and a pericardial effusion. The chest x-ray revealed diffuse interstizial infiltrate. We extended the diagnostic workup by high resolution computed tomography (HRCT), that showed bilateral confluent patchy lung infiltrates associated with a ground glass pattern. Other laboratory examinations revealed troponin I 0.07 ng/ml (0.00-0.06), proBNP 2296 pg/ml (woman <153), lactate dehydrogenase 729 U/L (313-618) and perinuclear antineutrophilic cytoplasmic antibody positive. Bronchoalvear lavage revealed 78% eosinophilis. Electromiography of the lower extremities was notable for right sural neuropathy consistent with mononeuritis. The patient also underwent a bone marrow biopsy, that showed an eosinophilic expansion and, suspecting

CHURG-STRAUSS AND MONTELUKAST

M. CARLESIMO, E. MARI, E. PALESE, G. PRANTEDA and G. CAMPLONE

U.O.C. Dermatology Sant’Andrea Hospital, II School University of Rome “Sapienza”, Rome, Italy

Received March 29, 2011 – Accepted July 22, 2011

All the authors contributed equally to the paper

LETTER TO THE EDITOR

Mailing address: Elena Mari, MDU.O.C. Dermatology II Unit, University of Rome “Sapienza”Via di Grottarossa, 1039 00189 Rome, ItalyTel: ++39 06 33775378Fax: ++39 06 33775081e-mail: [email protected]

Churg-Strauss syndrome (CSS) is a systemic small vessel vasculitis involving lungs, skin, heart, gastrointestinal tract and peripheral nerves. We report the case of a 36-year-old woman with a necrotic lesion on the left foot of two months duration, associated with hypereosinophilia, patchy lung infiltrates, cardiac damage and a mononeuritis. The personal history was remarkable only for an asthma, treated with Montelukast, a leukotriene receptor antagonist (LRA). Clinical symptoms, laboratory exams and instrumental examinations led us to the diagnosis of CSS. In recent years several studies have reported the possible relationship between use of leukotriene receptor antagonist (LRA) and CSS expression. We report this case to underline the possible relationship between LRA and CSS and its etiopathogenetic mechanism.

Key words: Churg-Strauss, leukotriene receptor antagonist, montelukast, leukotriene B4, vasculitis

Vol. 24, no. 4, 1079-1082 (2011)






Systemic Lupus Erythematosus (SLE) is associated with an increased risk of Non-Hodgkin’s Lymphoma (NHL) (1). The risk of hematological malignancies is approximately threefold higher in patients with SLE than in the general population (standardized incidence ratio 2.75, 95% CI 2.13-3.49), particularly for NHL (standardized incidence ratio 3.64, 95% CI 2.63-4.93) (2). It is uncertain whether these lymphomas are due to immunologic derangements, genetic factors, viruses, or medication. Recent investigations have reported a predominance of aggressive histological subtypes, in particular diffuse large B cell lymphoma (3). Similarities

of a variety of immunological disturbances that characterize both rheumatic conditions and lymphomas have been suggested as a linkage between these disorders and immunosuppressive drugs or viral infections, especially Epstein-Barr Virus (EBV) (4). The role of EBV infection in the pathogenesis of NHL has already been described. However, association between EBV and SLE remains unclear (5-6). Many hypotheses have been proposed on the role of EBV in SLE etiology. EBV initially infects epithelial and B-cells, and then a latent infection of B-cells with occasional reactivation and productive cycles of viral replication develops.

AGGRESSIVE LARGE B-CELL LYMPHOMA IN A SYSTEMIC LUPUS ERYTHEMATOSUS PATIENT WITH CHRONIC ACTIVE EPSTEIN-BARR VIRUS

INFECTION: A CASE REPORT

L. ABENAVOLI1, I. PENNACCHIA2, E. STIGLIANO2, A. CARBONE2, F.M. VECCHIO2

and V. ARENA2

1Department of Health Sciences, University “Magna Græcia”, Catanzaro;2Pathologic Anatomy Unit, Catholic University, Rome, Italy

Received March 22, 2011 – Accepted August 5, 2011

The first two authors contributed equally to this manuscript

A link between Epstein-Barr Virus (EBV) infection, Systemic Lupus Erythematosus (SLE) and Non-Hodgkin’s Lymphoma (NHL) has been recently reported in literature. Here we report a case of diffuse Large B-Cell Lymphoma (DLBCL) with a particularly aggressive clinical course in an SLE patient with EBV infection. A 49-year-old woman with a long history of SLE was admitted to the Department of Experimental and Clinical Medicine and dramatically died a few hours later. The autopsy described no evidence of active lymphoproliferative disorder. Instead, histological examination demonstrated an atypical lymphocitic proliferation in lymph node, kidneys, pericardium and uterus. Immunoistochemically, the lymphomatous cells were positive with CD19, CD20, CD22 and CD79a, which was consistent with a DLBCL. The cells were also reactive to EBV markers, indicating the possible role of previous EBV infection in DLBCL pathogenesis.

Mailing address: Ludovico Abenavoli, M.D.Department of Experimental and Clinical Medicine, University “Magna Græcia”, Viale Europa, 88100 Catanzaro, Italy Tel: ++39 0961 3694387 Fax: ++39 0961 754220e-mail: [email protected]

Key words: Epstein-Barr virus, systemic lupus erythematosus, non-Hodgkin’s lymphoma, lymphoproliferative disorders, B cell lymphoma

LETTER TO THE EDITOR

Vol. 24, no. 4, 1083-1086 (2011)






Alopecia Areata (AA) is a chronic inflammatory skin disease affecting about 2–3% of new patients attending Dermatologic Clinics and it can affect any age without a specific preponderance of race or sex (1). About 20% of patients affected by AA has a family history of disease, indicating a genetic predisposition: major histocompatibility complex, cytokines and immunoglobulin genes have been

emphasized, suggesting a genetic multifactorial involvement (2). The aetiology still remains unknown, although evidence supports the hypothesis that it is a tissue-specific autoimmune disease (2).

AA is clinically characterized by a non-scarring hair loss involving anagen stage hair follicles of the scalp and/or of any hair-bearing body surface with a possible nail localization in 10% of patients.

LETTER TO THE EDITORVIDEOCAPILLAROSCOPIC PATTERN OF ALOPECIA AREATA BEFORE AND AFTER

DIPHENYLCICLOPROPENONE TREATMENT

G. GANZETTI, A. CAMPANATI, O. SIMONETTI, I. CATALDI, K. GIULIODORIand A.M OFFIDANI

Dermatological Clinic, Department of Medical Sciences, Ancona Hospital,Polytechnic University of Marche, Ancona, Italy


The first two authors contributed equally to this paper

Alopecia areata (AA) is an inflammatory skin disease the most effective therapy for which is diphenylcyclopropenone (DPCP). Videodermatoscopy and intra-vital capillaroscopy (IVCP) are two non-invasive techniques that help in the differential diagnosis of alopecias. It is known that, after DPCP therapy, there is a histologically proven significant increase of VEGF in hair follicle keratinocites and a consequent increase in capillary vessels in the dermis of the same follicles. The aim of our study is to emphasize any clinical and videodermatoscopic-videocapillaroscopic changes after DPCP treatment in 20 patients affected by alopecia areata. Videodermatoscopic images and an intravital videocapillaroscopic analysis were performed at T0, T12 and T24 to emphasize clinical modifications and microscopic changes in vascular pattern before and after DPCP treatment. At T0, videodermatoscopy showed the presence of “exclamation point” hairs, hair follicles filled with hyperkeratotic plugs (“yellow dots”), hair follicles containing cadaverized hairs (“black dots”) and broken hairs. IVCP highlighted a pale scalp, and vessels were not visible. At 24 weeks (T24), videodermatoscopy revealed the disappearance or a statistically significant reduction of AA hallmarks and an increase of number of vellus hairs. Videocapillaroscopy showed a statistically significant increase of new vessels and, where neoangiogenesis were more marked, a major hair regrowth was evident. Our study emphasizes that, after DPCP therapy, neoangiogenesis is detectable by videocapillaroscopy and these new capillaries could be considered an initial positive attempt to compensate capillary loss of T0 alopecia areata images.

Mailing address: Dr Giulia Ganzetti, via conca 71, 60020 Ancona, ItalyTel: ++39 3473699154Fax: ++39 0715963446e-mail: [email protected]

Key words: alopecia areata, DPCP, videocapillaroscopy

Vol. 24, no. 4, 1087-1091 (2011)






Clinical outcomes of influenza A (H1N1) virus are reported as ranging from self-limited illness to respiratory failure or death (1-3). Bacterial pneumonia, usually caused by Staphylococcus aureus (often methicillin-resistant), Streptococcus pneumoniae, Streptococcus pyogenes, and sometimes other bacteria, has been suspected or diagnosed in about one third of hospitalized children (4) and in 20-24% of patients admitted to ICU for pandemic H1N1 influenza (5). The first case of necrotizing pneumonia due to community-acquired methicillin-resistant S. aureus during influenza A (H1N1) has been recently described in a previously healthy adolescent (6). To our knowledge, only one case of pneumococcal necrotizing pneumonia complicating pandemic H1N1 influenza has been reported to date (7).

We herein report a case of necrotizing pneumonia due to S. pneumoniae that was diagnosed within a series of 18 children with pandemic influenza A (H1N1) admitted to a tertiary care pediatric hospital in Italy.

Case reportA 3-year-old previously healthy Caucasian girl

was admitted to a district hospital because of a seven-day history of flu-like symptoms and fever despite treatment with ibuprofen. She had no history of recent travel abroad, and nobody was positive for influenza A (H1N1) among her family. She had not received the H1N1 influenza A vaccine. A chest x-ray showed extensive consolidation of the left lung and concomitant pleural effusion. Nasal and throat swabs were positive for WHO-approved Real Time Polymerase Chain Reaction (RT-PCR) H1N1 pandemic influenza A assay (8). Antibiotic treatment with intravenous cefotaxime and teichoplanin in addition to oral oseltamivir was started. On day three, the patient was referred to our hospital because of progressive deterioration of clinical conditions. On admission, her temperature was 38.5°C and she had moderate respiratory distress (respiratory rate 30 breaths/min, oxygen saturation 94% in 4 L/min O2 via ventimask). Chest examination revealed dullness on percussion and absent breath sound on the left

LETTER TO THE EDITORSEVERE NECROTIZING PNEUMONIA COMPLICATING INFLUENZA

A (H1N1): THE ROLE OF IMMUNOLOGIC INTERACTION

R. GUIDI1, P. OSIMANI1, C. AZZARI2, M. RESTI2 and F.M. DE BENEDICTIS1

1Department of Pediatrics, Azienda Ospedaliero-Universitaria, Salesi Children’s Hospital, Ancona; 2Department of Pediatrics, University of Florence and Anna Meyer Children’s Hospital, Florence, Italy


This report describes the successful management of a documented necrotizing pneumonia due to Streptococcus pneumoniae in a child with pandemic influenza A (H1N1). The importance of early recognition of bacterial superinfection in patients with influenza and the immunologic interactive mechanisms between viruses and bacteria in determining respiratory diseases are highlighted. The role of modern molecular techniques in improving diagnostic microbiology sensitivity and informing consequent clinical care is emphasized.

Key words: necrotizing pneumonia, influenza A H1N1, Streptococcus pneumoniae

Mailing address: Fernando Maria de Benedictis, MDDivision of Pediatrics,Salesi Children’s Hospital,11, via Corridoni,I-60123 Ancona, ItalyTel: ++39 071 5962351 Fax: ++39 071 5962234e-mail: [email protected]

Vol. 24, no. 4, 1093-1097 (2011)






Aromatase inhibitors are a valuable and well-established adjuvant therapy in the treatment of estrogen-receptor-positive breast cancer in menopausal women. Among the most frequent side effects, secondary to the use of these drugs, are osteoarthralgia and muscle pain (1). These symptoms are bearable and usually disappear after the discontinuation of therapy. On the contrary, in more rare cases the joint involvement acquires the characteristics of typical arthritis, with severe and persistent pain, swelling and prolonged stiffness. It is still not entirely clear whether these drugs can induce a true rheumatoid arthritis (RA) and render it more apparent or aggravate an existing condition with low disease activity. Recently, although with limited observations, it seems that the association of RA and therapy with aromatase inhibitors can not be considered a mere coincidence (2-3). The case of a patient who came under our observation confirms this hypothesis.

Case report The patient is a 56-year-old woman, diagnosed

with breast cancer (histologically classified as lymphoepithelioma-like carcinoma, pT2,N2,Mx, Stage III A) in 2005 and treated with quadrantectomy, chemotherapy, radiation therapy, and aromatase inhibitors (Anastrozole). About a year later, the patient began to experience widespread osteoarthralgia. In March 2007, a bone scan was performed which resulted negative. The joint pain symptomatology lasted with the same characteristics until August 2009, when the pain became more severe, with swelling and prolonged morning stiffness (>1 hour), symmetrically involving the hands, wrists, knees, ankles and feet. On that occasion another bone scan was performed which showed, unlike the previous one, hyperfixation of the tracer (99mTechnetium methylenediphosphonate) in multiple joints (wrists, ankles, feet with a symmetric distribution, as well as left knee, left femoral neck, and vertebral somites).

LETTER TO THE EDITORRHEUMATOID ARTHRITIS: A COMPLICATION

OF AROMATASE INHIBITOR THERAPY?

V. BRUZZESE, C. HASSAN, A. ZULLO and G. ZAMPA

CHU of Internal Medicine, Section of Rheumatology, Nuovo Regina Margherita Hospital, Rome, Italy

Received July 6, 2011 – Accepted September 29, 2011

We report the case of a 56-year-old woman treated with aromatase inhibitors for a breast cancer. Following one year of such therapy, the patient presented with widespread osteoarthrealgia. The clinical picture worsened 3 years later when the pain became more severe with swelling and stiffness involving several joints in a symmetric fashion. Biochemical analysis showed an increase of ESR, CRP and rheumatoid factor, as well as of anti-CCP antibodies. The x-ray was compatible with a diagnosis of rheumatoid arthritis (RA). Therapy with methotrexate, prednisol one, bisphosphonates and vitamin D was started, achieving a quick clinical remission. Aromatase inhibitors have been shown to alter the distribution of Th1/Th2 lymphocytes and increase the level of RANKL. A possible role of aromatase inhibitors in RA development has been further addressed.

Mailing address: Dr.Vincenzo BruzzeseVia Bosco degli Arvali 2400148 Rome, ItalyFax: ++39 0658446675Tel: ++39 0658446504e-mail:[email protected]

Key words: breast cancer, aromatase inhibitors, rheumatoid arthritis, RANKL

Vol. 24, no. 4, 1099-1101 (2011)






mainly topical corticosteroids, are frequently used for treating allergic rhinitis. However, topical drugs may be scarcely effective if nasal secretions are not sufficiently removed by nasal lavage. Recent studies have shown that nasal irrigation with hypertonic solution was able to determine a significant improvement in rhinitis symptoms in children

The burden of allergic diseases is continuously increasing in all industrialized countries and allergic rhinitis can lead to a substantial deterioration in children’s quality of life with relevant effects on learning and friendship (1). Nasal obstruction is a main symptom and is closely dependent on allergic inflammation (2). Thus, anti-inflammatory drugs,

LETTER TO THE EDITOREFFECTIVENESS OF ISCHIA THERMAL WATER NASAL AEROSOL IN CHILDREN

WITH SEASONAL ALLERGIC RHINITIS:A RANDOMIZED AND CONTROLLED STUDY

M. MIRAGLIA DEL GIUDICE1, F. DECIMO1, N. MAIELLO1, S. LEONARDI2, G. PARISI3,M. GOLLUCCIO2, M. CAPASSO1, U. BALESTRIERI1, A. ROCCO1, L. PERRONE1

and G. CIPRANDI4

1Pediatric Department “F.Fede”, II University of Naples, Naples; 2Pediatric Department, University of Catania; 3UOC Pediatrics and Neonatology, ASL Na-2, PO “Anna Rizzoli”, Lacco

Ameno; 4Azienda Ospedaliera Universitaria San Martino, Genova, Italy


Allergic rhinitis is characterized by local inflammation. Nasal lavage may be a useful treatment, however, there are few studies on this topic. This study aims to evaluate the effects of Ischia thermal water nasal irrigation on allergic rhinitis symptoms and airway inflammation during the period of natural exposure to Parietaria pollen in children with allergic rhinitis and intermittent asthma. Forty allergic children were randomly divided into two groups: the first group (Group 1) practiced crenotherapy with thermal water aerosol for 15 days per month, for three consecutive months, the control group (Group 2) was treated with 0.9% NaCl (isotonic) solution. In addition, all children were treated with cetirizine (0.5 gtt./kg/day once daily). Nasal symptom assessment, including Total Symptom Score (TSS), spirometry, and exhaled nitric oxide (FeNO) were considered before the treatment (T0), at the end of the treatment (T1) and again 2 weeks after the end of the treatment (T2). The study was registered in the Clinical Trials.gov (NCT01326247). Thermal water significantly reduced both TSS and FeNO levels and there was a significant relationship between reduction of nasal symptoms and FeNO values at the end of treatment with thermal water. In conclusion, this study shows that nasal crenotherapy with the hyper-mineral chloride-sodium water of Ischia was effective in children with seasonal allergic rhinitis based on the sensitivity to Parietaria. These results demonstrate that this natural treatment may be effective in a common and debilitating disease such as the allergic rhinitis.

Mailing address: Giorgio Ciprandi, M.D.Semeiotica e Metodologia Medica I,Viale Benedetto XV 6, 16132 Genoa, ItalyTel: ++39 10 35331820Fax: ++39 10 3538664e-mail: [email protected]

Key words: thermal water, nasal lavage, Ischia, allergic rhinitis, children

Vol. 24, no. 4, 1103-1109 (2011)






The term tattoo derives from Polynesian “tatau”. The art of tattooing has ancient origins; the first description of tattooed men dates back to 6,000 years ago and were discovered by archaeologists in Egypt, China, Siberia, Austria, and South America. The Bronze Age mummified body discovered in Austria, known as “the Similaun Mummy”, had about fifty tattoos all over his body. In addition, some tools from the upper Paleolithic era (about 35,000 to 10,000 years BC) probably used to tattoo, such as needles and pigments, were also discovered

Today tattoos are becoming part of mainstream culture. From recent statistics, only in Western countries, about eighty million people have at least one tattoo on their bodies. (1-5). It is well known that tattoos can cause many complications, including mainly local inflammation, infection and hypersensitivity. Allergic sensitivity to one of the pigments is the most frequent cause of these reactions.

We describe the case of young woman with contact allergic dermatitis to the gold contained in her tattoo.

Case reportA 29-years-old woman presented to the

Dermatology Unit, S. Andrea Hospital, Rome because of a 2-week history of allergic dermatitis presenting with heat, pruritus, erythema and scaling, in the area of a coloured tattoo on her left arm. These lesions developed 2 months after tattooing (Fig. 1-2).

General physical examination was normal, and in particular there was no evidence of respiratory, cardiac, ocular or joint disease. Cutaneous examination revealed erythematous lesions localized to the gold pigmented areas.

The patch test was performed using the standard North American series test [containing DMDM hydantoin bacitracin, mixed dialkyl thiourea, formaldehyde-releasing preservative, rubber antioxidant, glutaraldehyde 0.5%, bronopol, formaldehyde 1%, fragrance mix, propylene glycol 30%, 4-chloro-3,5-xylenol, diazolidinylurea 1%, ethyleneurea/melamine formaldehyde mix 5%, disperse blue 106/124, mix ethyl acrylate, glyceryl monothioglycolate (GMTG), goldsodiumthiosulfate,

LETTER TO THE EDITORCONTACT ALLERGIC DERMATITIS TO GOLD IN A TATTOO: A CASE REPORT

A. TAMMARO1, P. TUCHINDA2, S. PERSECHINO1 and A. GASPARI3

1Dermatology Unit, S. Andrea Hospital, II School of Medicine, University of Rome ‘Sapienza’, Italy; 2Department of Dermatology Siriraj Hospital, Mahidol University, Bangkok, Thailand;

3Department of Dermatology, University of Maryland, Baltimore, Maryland, USA


The art of tattooing has increasing in recent decades. Allergic sensitivity to one of the pigments is the most frequent cause of dermatological reactions at the site of the tatoo. Gold is a new pigment used in tatooing, because of its bright yellow color and luster. Allergy to this metal is uncommon. To our knowledge, this is the first reported case of allergic contact dermatitis to gold in a tattoo.

Mailing address: Antonella Tammaro, MDU.O.C. Dermatology, II School of Medicine, University of Rome ‘Sapienza’Via di Grottarossa, 1035 00189 Rome, ItalyTel: ++39 06 33775907 Fax: ++39 06 33775378e-mail: [email protected]

Key words: tattoo, gold, allergic contact dermatitis

Vol. 24, no. 4, 1111-1114 (2011)






LETTER TO THE EDITORACQUIRED ANGIOEDEMA WITH C1 INHIBITOR DEFICIENCY ASSOCIATED WITH

ANTICARDIOLIPIN ANTIBODIES

E. DI LEO1, E. NETTIS1, V. MONTINARO2, G. CALOGIURI3, P. DELLE DONNE1,A. FERRANNINI1 and A. VACCA1,4

1Section of Allergology and Clinical Immunology, Department of Internal Medicine and Infectious Diseases, University of Bari Medical School, Bari; 2Division of Nephrology, Department of

Emergency and Organ Transplantation, University of Bari, Azienda Ospedaliero-Universitaria “Consorziale Policlinico”, Bari; 3Fourth Pneumology Department, Pneumologic Hospital

A. Galateo, San Cesario di Lecce; 4Department of Internal Medicine and Clinical Oncology, University of Bari Medical School, Bari, Italy


Acquired angioedema (AAE) with C1 inhibitor deficiency is often associated to B cell lymphoproliferative disorders or autoimmune diseases. We report a case of AAE associated with IgM anti-cardiolipin antibodies, with frequent edematous attacks, that disappeared completely after a slight immunosuppression and danazol therapy.

Mailing address: Eustachio Nettis, M.D.Cattedra di Allergologia ed Immunologia Clinica,Università degli Studi di Bari,Policlinico, Piazza Giulio Cesare, 11 70124 Bari, ItalyTel: ++39 080 5592821Fax: ++39 080 5593576e-mail: [email protected]

Key words: acquired angioedema, anticardiolipin antibody, immunosuppressive therapy,C1-INH deficiency

Angioedema not associated to urticaria is a recurrent, non-pruritic and self-limiting swelling of the skin and mucosae due to a transient increase of endothelial capillary permeability of subcutaneous or submucosal tissue. Angioedema attacks may also occur as cutaneous abdominal, genital and upper airway involvement. While cutaneous and abdominal localizations are very disabling, the laryngeal involvement can be fatal if not promptly treated. Non-allergic angioedema occurs as hereditary (HAE), acquired (AAE), Angiotensin converting enzyme (ACE) inhibitor-induced (RAE), pseudoallergic angioedema (PAE) and idiopathic angioedema (IAE) (1). HAE and AAE are due to a genetic or acquired deficit of C1 inhibitor, respectively (2), although forms of AAE with levels of C1 inhibitor quantitatively normal have been described.

The common final pathogenetic event is an increased production of bradykinin, which, in turn, binds to specific and constitutively expressed B2 receptors and triggers increase of endothelial permeability. As a consequence, plasma flows out of the intravascular to the extravascular compartment forming the interstitial oedema.

AAE is a rare disease (about 160 cases described in the literature) (3) characterized by an increased catabolism of C1 inhibitor, classically associated to: a) B cell lymphoproliferative disorders or monoclonal gammopathies of undetermined significance (type I); b) autoimmune diseases associated or not to the presence of autoantibodies against C1 inhibitor (Type II) (4-6). However, we find in literature some cases of type I AAE in presence of antibodies against C1-INH; therefore this classification must be revisited (7).

Vol. 24, no. 4, 1115-1118 (2011)






Crohn’s disease (CD) is an inflammatory bowel disease (IBD) of unknown aetiology characterized by intestinal and extra-intestinal manifestations suggesting the presence of a systemic inflammatory disorder (1-2). The lungs may be also involved, which will worsen the overall prognosis (1, 3). Several reports have demonstrated that, nearly half of IBD patients show some degree of lung involvement (4-5), including small airway (6), parenchymal and pulmonary vascular disease as well as serositis (7-8). A number of studies have been focusing on pulmonary function tests (PFT) which in turn have revealed abnormalities such as

a decrease in lung diffusing capacity in nearly 57% of CD patients showing no radiological alterations of the chest (9). These abnormalities, which have also been described during clinical remission, are the likely consequence of a subclinical inflammatory reaction. This hypothesis draws additional support from the increased airway hyper-responsiveness that has been documented in asymptomatic IBD patients (10). Most of these patients are usually asymptomatic; in addition, the procedures necessary to prove pulmonary involvement are quite costly and complicated. Exhaled Nitric Oxide (eNO) can be detected non-invasively by using a

LETTER TO THE EDITOREXHALED NITRIC OXIDE AS A MARKER OF LUNG INVOLVEMENT

IN CROHN’S DISEASE

M. MALERBA1, B. RAGNOLI2, L. BUFFOLI1, A. RADAELI2, C. RICCI3, F. LANZAROTTO3 and A. LANZINI3

1Department of Internal Medicine, University of Brescia; 2Department of Emergency, Spedali Civili of Brescia; 3Gastroenterology Unit, University of Brescia and Spedali Civili of Brescia, Italy


Crohn’s Disease is an inflammatory bowel disease associated with a variety of systemic manifestations, including large and small airway involvement. The latter is most often a subclinical one, and requires expensive and invasive diagnostic approaches. Nitric Oxide (NO) can be detected non-invasively in the exhaled air (eNO) and be considered as a surrogate marker of airway inflammation. eNO tested at multiple expiratory flows can be used to distinguish the alveolar concentration of NO (CalvNO) from the total amount of fractional eNO (FeNO). The aim of our study is to compare FeNO and concentration of alveolar nitric oxide (CalvNO) levels and to assess their relationship with pulmonary involvement in Crohn’s patients differing in clinical stage and therapeutic regimens versus a group of healthy subjects. Thirty Crohn’s patients not showing clinical evidence of pulmonary diseases and 21 non-smoking, non-atopic healthy controls were enrolled. FeNO (14.9±10.2 ppb vs 10.1±6.3 ppb, p=0.049) and CalvNO (4.4±2.2 ppb vs 2.6±1.9; p=0.006) values were found to be significantly higher in Crohn’s patients than in healthy controls. Both FeNO and CalvNO correlated positively with the Crohn’s Disease Activity Index. In conclusion, our results for FeNO and CalvNO confirm the presence of subclinical pulmonary involvement in Crohn’s disease. eNO measurement may be of clinical value in the follow-up of Crohn’s patients.

Mailing address: Dr Mario Malerba, Department of Internal Medicine, University of Brescia, Spedali Civili di Brescia, P.zzale Spedali Civili 1, 25123 Brescia, Italy Tel: ++39 030 3995250. Fax: ++39 030 396011 e-mail: [email protected].

Key words: airway inflammation, inflammatory bowel disease, markers of inflammation

Vol. 24, no. 4, 1119-1124 (2011)






Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by involvement of multiple organs and systems and by detection of autoantibodies towards several nuclear autoantigens. The natural history of SLE is highly heterogeneous and virtually all tissues and organs can be involved. However, even if the disease occurs more frequently in women, with a female to male ratio ranging from 8:1 to 13:1, breast involvement is a rare event. Breast pathology in SLE patients frequently presents as lupus panniculitis with skin erythema, tenderness, pain, and parenchymal nodules (1-2). In some cases, lupus mastitis can herald SLE (3).

Diagnostic difficulties arise when breast masses are detected without significant cutaneous inflammation indicative of lupus panniculitis; in these cases, it is mandatory to rule out breast carcinoma (4-6). Furthermore, since in SLE patients surgical trauma can exacerbate breast inflammation, minimally invasive procedures should be carried out for diagnostic purposes (7).

Here, we report the diagnostic procedures selected for a 47-year-old patient with a long history of SLE, who presented with episodes of recurrent acute mastitis and a suspicious breast nodule, suggestive of cancer.

LETTER TO THE EDITORLUPUS MASTITIS IN SYSTEMIC LUPUS ERYTHEMATOSUS:

A RARE CONDITION REQUIRING A MINIMALLY INVASIVE DIAGNOSTIC APPROACH

G. LUCIVERO1,4, C. ROMANO1, F. FERRARACCIO3, A. SELLITTO1, U. DE FANIS1, R. GIUNTA1, A. GUARINO1, P.P. AURIEMMA2, M. BENINCASA1

and F. IOVINO2

1Department of Gerontology, Geriatrics and Metabolic Diseases, 2Department of Anaesthesiological, Surgical and Emergency Sciences, and 3Department of Pathology, Second

University of Naples, Italy

Received March 16, 2011 – Accepted October 18, 2011

Breast involvement is a rare event in SLE patients. The most frequent presentation is lupus panniculitis with skin erithema, tenderness, and parenchymal nodules. However, when breast masses are detected in SLE patients without significant superficial inflammation, it is mandatory to rule out breast carcinoma. Here, we report the case of a 47-year-old woman with an 18-year-long history of SLE, who presented with a suspicious breast mass. Since surgical trauma has been reported to be able to exacerbate breast inflammation in lupus mastitis, an ultrasound-guided minimally invasive Mammotome biopsy was performed to obtain tissue samples for histological and immunohistochemical examinations. Histology was consistent with lupus mastitis. The patient was already on mycophenolate mofetil and hydroxychloroquine. At the latest follow-up visit 6 years later, no progression of the breast lesion was observed.

Mailing address: Prof. Giacomo Lucivero, M.D., Ph.D., Division of Internal Medicine, Allergy and Clinical Immunology, Polyclinic Hospital, Second University of Naples, Piazza L. Miraglia, 80138 Naples, ItalyTel: ++39 081 5665081 Fax: ++39 081 5665080e-mail: [email protected]

Key words: lupus mastitis, systemic lupus erythematosus, breast disease, Mammotome biopsy

Vol. 24, no. 4, 1125-1129 (2011)

atherosclerosis: a classic inflammatory atherosclerosis: a classic inflammatory disease a....

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