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Over the last decade, nanotechnology has become more relevant in medicine. Among magnetic nanomaterials the future of iron oxide nanoparticles (IONPs) for clinical applications relies on their biocompatibility in moderate doses as well as their ability to be produced in a wide range of sizes and shapes with biofunctionalization potential. Additionally, they show great promise to serve as a cell tracking system in cell-based therapies, and to generate local temperature increases in the magnetic thermotherapy of solid tumours. Thus, the study and development of novel magnetic nanoparticles for biomedical applications is one of the key topics in the field of nanotechnology. Extremely small-sized Fe3O4 superparamagnetic nanoparticles were prepared by coprecipitation, thinly coated with silica and conjugated with FITC, as molecular model specimen. Nanoparticles were characterized by dynamic light scattering (DSL), transmission electron microscopy (TEM) and X-ray diffraction analysis (XRD) and surface functional groups and composition were analysed by infrared spectroscopy (FTIR). We assessed the biocompatibility of the magnetic nanoparticles carriers with biofunctional coating (FITC-conjugated) using a colon carcinoma cell line (CaCo-2) as human cellular model. Phase contrast, fluorescence and confocal microscopy analyses were performed to study nanoparticles up-take and internalization

Objective: There are no comprehensive surveys relating the reported high prevalence of asthma and allergic diseases in athletes to comorbidities and immune changes associated with intense chronic exercise. This 12-year survey aims to evaluate several clinical, functional and immunological parameters in order to assess features, trend and burden of asthma, allergy, infections and autoimmune diseases, in a large homogeneous population of Olympic athletes. Methods: Six hundred and fifty-nine Italian Olympic athletes were studied through four cross-sectional surveys performed between 2000 and 2012 before the Summer and Winter Olympics. Clinical diagnosis of allergic, autoimmune and infectious diseases was complemented by: skin-prick tests (n = 569); pulmonary function tests (n = 415); total (n = 158) and specific (n = 72) serum IgE; serum autoantibodies (n = 30), cytokines and growth factors (n = 92); flow cytometry (n = 135). Results: The prevalence of asthma and/or exercise-induced bronchoconstriction was 14.7%, with a significant increase (P = 0.04) from 2000 (11.3%) to 2008 (17.2%). The prevalence of rhinitis, conjunctivitis, skin allergic diseases and anaphylaxis was 26.2%, 20.0%, 14.8% and 1.1%, respectively. Sensitization to inhalant allergens was documented in 49.0% of athletes, being 32.7% in 2000 and 56.5% in 2008 (P < 0.0001). Food, drug and venom allergy was present in 7.1%, 5.0% and 2.1% of athletes, respectively. The high prevalence of asthma and allergy was associated with recurrent upper respiratory tract (10.3%) and herpes (18.2%) infections, an abnormal T cell subset profile and a general down-regulation of serum cytokines with a significantly lower IFN-[gamma]/IL-4 ratio. Conclusion: A chronic and intense physical exercise may cause a transient immunodepression with a preferential shift to a Th2 response, associated with abnormalities of the respiratory tract.

I sieri di 72 daini sono stati saggiati, mediante IFI, per la ricerca di IgG anti A. phagocytophilum fissando il titolo soglia a 1/80; campioni di sangue in EDTA sono stati sottoposti ad estrazione del DNA ed a NESTED-PCR per l'amplificazione di un frammento del 16S rRNA. Parallelamente sono stati raccolti dal corpo degli animali n. 2 zecche identificate su chiave morfologica e testate per la ricerca del patogeno mediante PCR. I prodotti di amplificazione sono stati sequenziati e le sequenze confrontate con quelle presenti in Genbank. N. 20 animali (27,77%) sono risultati sieropositivi per A. phagocytophilum, con titoli compresi tra 1/80 ed 1/640 ed 11 (15,27%) hanno mostrato positività ematica alla PCR. L'analisi dei prodotti di sequenziamento ha mostrato omologia nucleotidica completa tra gli isolati ed una identità del 99% con isolati di A. phagocytophilum di origine europea. I risultati delle indagini ixodologiche hanno evidenziato una elevata presenza della specie I. ricinus (36/42 esemplari raccolti) ed hanno confermato la suscettibilità di tale specie all'infezione (3/36 positivi). Il sequenziamento degli isolati provenienti dalle zecche è ancora in fase di attuazione. I dati ottenuti supportano l'esistenza e la circolazione del patogeno nel Parco della Mesola. La definizione del ruolo del daino quale serbatoio d'infezione necessita, tuttavia, di ulteriori studi atti a rilevarne la persistente infettività. I risultati rivestono un certo rilievo in quanto rappresentano il terzo contributo scientifico finalizzato allo studio dell'infezione in questa specie animale e concorrono a definire il ruolo delle popolazioni dei selvatici quali sentinelle epidemiologiche dell'Ehrlichiosi Granulocitica nel territorio.

Optimally designed cancer vaccines should combine the best tumor antigens with the most effective immunotherapy agents and delivery strategies to achieve positive clinical results. The unique immunoglobulin (Ig) idiotype on the surface of each B-cell lymphoma represents an ideal tumor-specific antigen for use as a cancer vaccine. It has been theorized that effective cancer vaccines can be developed using the minimum essential subset of T cell and B cell epitopes that comprise the ‘immunome’, the universe of neoplasm-derived peptides that interface with B and T cells of the host immune system. Idiotypic antigenic determinants of a B-cell lymphoma lie within the hypervariable regions and mainly within the complementarity-determining regions (CDR)s 3. Thus, the CDR3s are considered a “hot spot” of particular interest for construction of subunit vaccines. DNA vaccines, whose safety and tolerability are substantiated in completed and ongoing clinical trials, have emerged as a novel lymphoma vaccine formulation for antigen-specific immunotherapy. The molecular precision tools offered by gene-based vaccines allow to explore the use of CDR3 sequence as an anti-lymphoma vaccine.

DNA vaccine platform has produced exciting results in a wide array of applications, from prophylactic vaccine strategies to poten- tial therapeutics targeting cancers and infectious disease (Fioretti et al., 2010; Rinaldi et al., 2009; Rinaldi et al., 2006; Fazio et al., 2004). Many tumor antigens are excellent targets for active immunotherapy and can be presented by DNA vaccines in a molecular form suitable to engage both humoral and cellular immune responses. Our group is involved in the development of DNA vaccination strategies as active immunotherapy for the treatment of B-cell lym- phomas. The idiotype region in the immunoglobulin expressed on the surface of B-cell lymphomas is a tumour-specific as well a patient-specific antigen and an ideal target for personalised immunotherapy. We focused on the idiotypic determinants lying within the CDR3 hypervariable regions and demonstrated that immunisation with CDR3-based DNA vaccines elicited anti- idiotypic humoral response able to recognize native antigens on tumour cells (Rinaldi et al., 2001). In a highly aggressive preclinical murine B-cell lymphoma model, we evaluated the antitumour response recruited by CDR3- based DNA vaccines. We showed that a combination of CDR3-based DNA vaccines with electroporation in a homologous prime-boost approach was protective against a lethal tumour challenge (Rinaldi et al., 2008; Iurescia et al., 2010). Since a goal of our studies is the development of tai- lored immunisation strategies against previously established tumours, efforts are ongoing to optimize the DNA vaccine tech- nology platform. Strategies to improve antigen expression and immunogenicity involve epitope prediction, MHC I presentation enhancement, inclusion of pathogen-derived sequences aimed to stimulate CD4+ T cell help and immunomodulating cytokines. Preliminary results encourage the use of therapeutic protocols based on DNA vaccines as tailored drugs for the treatment of lymphoma. Optimal integration of active immunization approaches into standard therapies suggests DNA vaccination as an effective per- sonalized medicine to eradicate minimal residual diseases during clinical remission following standard chemotherapy in lymphoma patients.

Atherosclerosis, one of the most prevalent and severe age-related diseases, is the leading cause of death in industrialized countries and it is becoming an increasingly worldwide risk to health. As the size of elderly population grows, the incidence of atherosclerosis will increase, necessitating the exploration of radical, but potentially powerful, therapeutic strategies. One important goal is the availability of therapeutic molecules effective against the atherosclerotic plaque progression and able to stabilize advanced lesions. This generally requires robust measures to improve plasma lipoprotein profiles. Pharmaceutical agents currently available do not support significant changes in plasma lipoprotein levels. So that, strong new agents and therapies are needed in order to treat hyperlipidemia and atherosclerosis. Apolipoprotein E (ApoE), a key regulator in cholesterol-rich lipoprotein metabolism, is considered a strong candidate for treating hypercholesterolemia and cardiovascular disease. ApoE gene therapy could be proposed as a new valid approach for the correction of hyperlipidemic conditions and atherosclerosis by using viral and non-viral vectors. In this review we discuss the major developments in both viral and non-viral gene transfer systems aimed at achieving efficient protocols for treating hypercholesterolemia and atherosclerosis by using ApoE as therapeutic gene. Key scientific advances on the ability of ApoE gene transfer to markedly improve outcome after cerebral ischemia, suggesting a potential strategy in human stroke therapy, are also described in this review. The characterization of ApoE and its many functions has provided insight into the ultimate potential of this protein as a possible therapeutic agent for the treatment of several diseases. Possible therapeutic strategies that encompass the use of ApoE in gene therapy for Alzheimer's disease and other central nervous system (CNS) diseases will be also discussed.

The morphometric characteristics and the ultramicroscopic findings of Cryptosporidium spp. at various stages of their life cycle in the intestinal and bursal epithelial cells of naturally infected 30-day-old commercial turkeys are reported. Small, sporulated oocysts, observed in the small intestinal content after flotation, were identified as Cryptosporidium meleagridis on the basis of morphometric characteristics (round in shape and 4.5-5.0 microm in size) and the small intestinal localization. Light section examinations revealed the presence of the protozoon in multiple organs, but its prevalence was highest in the intestinal and bursal epithelial cells. Ultramicroscopic studies on ileum and bursal samples showed the presence of all the life cycle stages in the microvillar brush epithelial cells in both the organs examined. On the basis of the comparison of the morphology and the sizes of the microorganisms parasitizing the ileum and the bursa, hypotheses are considered on the possible species involved.

We developed an animal model to evaluate the 125-I-metaiodobenzylguanidine (125-I-mIBG) biodistribution in tumor bearing mice. Six weeks old nude-atimic mice were subcutaneously injected with 30 x 10(6) cells of the human neuroblastoma (NB) cell line SH-SY5Y. TE-671, a rhabdomyosarcoma cell line, was used as a control tumor without a specific mIBG uptake mechanism. In order to prevent possible tumor rejection mediated by NK activity the anti asialo GM1 antiserum was administered intraperitoneally once a week for 4 weeks. The maximum anti asialo mediated effect was obtained by administering the first dose the same day as the cell implant. In this group of animals by 9 weeks 98% of mice had a measurable tumor. We have utilized this model to evaluate the biodistribution of 125-I-mIBG given as two different formulations: standard preparation with a specific activity of 84 mCi/mg and the no carrier added (n.c.a) formulation with a specific activity of approximately 8,000 mCi/mg. Our preliminary results indicate that the biodistribution of the two different formulations in the various organs are similar. Therefore it appears that n.c.a. mIBG should not cause an increased toxicity in possible normal target organs such as heart or adrenals. Additional experiments will be performed in this model to ascertain if there is a potential advantage of the clinical use of n.c.a. mIBG over the standard preparation.