Development of a hypoallergenic recombinant parvalbumin for first-in-man subcutaneous immunotherapy of fish allergy.

BACKGROUND: The FAST (food allergy-specific immunotherapy) project aims at developing safe and effective subcutaneous immunotherapy for fish allergy, using recombinant hypoallergenic carp parvalbumin, Cyp c 1. OBJECTIVES: Preclinical characterization and good manufacturing practice (GMP) production of mutant Cyp (mCyp) c 1. METHODS: Escherichia coli-produced mCyp c 1 was purified using standard chromatographic techniques. Physicochemical properties were investigated by gel electrophoresis, size exclusion chromatography, circular dichroism spectroscopy, reverse-phase high-performance liquid chromatography and mass spectrometry. Allergenicity was assessed by ImmunoCAP inhibition and basophil histamine release assay, immunogenicity by immunization of laboratory animals and stimulation of patients' peripheral blood mononuclear cells (PBMCs). Reference molecules were purified wild-type Cyp c 1 (natural and/or recombinant). GMP-compliant alum-adsorbed mCyp c 1 was tested for acute toxicity in mice and rabbits and for repeated-dose toxicity in mice. Accelerated and real-time protocols were used to evaluate stability of mCyp c 1 as drug substance and drug product. RESULTS: Purified mCyp c 1 behaves as a folded and stable molecule. Using sera of 26 double-blind placebo-controlled food-challenge-proven fish-allergic patients, reduction in allergenic activity ranged from 10- to 5,000-fold (1,000-fold on average), but with retained immunogenicity (immunization in mice/rabbits) and potency to stimulate human PBMCs. Toxicity studies revealed no toxic effects and real-time stability studies on the Al(OH)3-adsorbed drug product demonstrated at least 20 months of stability. CONCLUSION: The GMP drug product developed for treatment of fish allergy has the characteristics targeted for in FAST: i.e. hypoallergenicity with retained immunogenicity. These results have warranted first-in-man immunotherapy studies to evaluate the safety of this innovative vaccine.

Performance assessment of four 64-slice computed tomographic devices for a typical clinical protocol.

OBJECTIVE: To compare the performances of four 64-slice CT devices, as regards radiation dose and image quality. METHODS: Effective dose was measured with thermoluminescent dosimeters in an Alderson Rando phantom (Alderson Research Laboratories, New York, NY). Quantitative image quality was evaluated in a Catphan 600 phantom (The Phantom Laboratory, New York, NY) using 3 parameters (modulation transfer function, contrast-to-noise ratio, and figure of merit). Effective and main organ doses were measured in 40 patients (10 for each CT device), who underwent an abdominal study with the same standard protocol; moreover, in these patients, clinical image quality, using a 5-grade quality score, was assessed. RESULTS: In-phantom measured doses and quantitative image quality showed some differences among the 4 devices. On the contrary, effective and organ doses provided to the patients were similar; no statistically significant differences were found also for clinical image quality. CONCLUSIONS: Some differences were found among the 4 devices from a physical point of view; on the other hand, the patient data were similar.

FAST: towards safe and effective subcutaneous immunotherapy of persistent life-threatening food allergies.

The FAST project (Food Allergy Specific Immunotherapy) aims at the development of safe and effective treatment of food allergies, targeting prevalent, persistent and severe allergy to fish and peach. Classical allergen-specific immunotherapy (SIT), using subcutaneous injections with aqueous food extracts may be effective but has proven to be accompanied by too many anaphylactic side-effects. FAST aims to develop a safe alternative by replacing food extracts with hypoallergenic recombinant major allergens as the active ingredients of SIT. Both severe fish and peach allergy are caused by a single major allergen, parvalbumin (Cyp c 1) and lipid transfer protein (Pru p 3), respectively. Two approaches are being evaluated for achieving hypoallergenicity, i.e. site-directed mutagenesis and chemical modification. The most promising hypoallergens will be produced under GMP conditions. After pre-clinical testing (toxicology testing and efficacy in mouse models), SCIT with alum-absorbed hypoallergens will be evaluated in phase I/IIa and IIb randomized double-blind placebo-controlled (DBPC) clinical trials, with the DBPC food challenge as primary read-out. To understand the underlying immune mechanisms in depth serological and cellular immune analyses will be performed, allowing identification of novel biomarkers for monitoring treatment efficacy. FAST aims at improving the quality of life of food allergic patients by providing a safe and effective treatment that will significantly lower their threshold for fish or peach intake, thereby decreasing their anxiety and dependence on rescue medication.

Non-traumatic abdominal emergencies: imaging and intervention in acute pancreatic conditions.

Pancreatic emergency, unrelated to traumatic events, can occur as a consequence of the more significant pancreatic pathologies (acute and chronic pancreatitis, tumors) or of the interventional or surgical treatment carried out as therapy for the above-mentioned lesions. Acute pancreatic conditions are represented by pancreatic infections, the involvement of organs, structures, and adjacent spaces within the pancreatic disease, and, lastly, vascular complications. Acute pancreatic conditions are common in pancreatic diseases and can be catastrophic; even if there is a gamut in the severity of clinical presentation, each can be potentially life threatening. Immediate radiological detection of the lesions together with a correct therapeutic percutaneous radiological approach whenever an interventional procedure is preferable to surgery or, when performed before surgery, whenever it can optimize its results, is of fundamental importance in the management of these patients. This article focuses on the essential role of radiology and the integration of imaging and intervention in acute pancreatic conditions.