Important variations in parvalbumin content in common fish species: a factor possibly contributing to variable allergenicity.

BACKGROUND: Although 95% of fish-allergic patients are sensitized to the major fish allergen parvalbumin, clinical reactions to different fish species vary considerably in symptoms, intensity and frequency in allergic subjects. This study aimed at the quantification of parvalbumin levels in salmon, trout, cod, carp, mackerel, herring, redfish and tuna. METHODS: Fish muscle extracts were separated by SDS-PAGE and parvalbumin content was estimated by densitometric band quantification. Individual antisera were raised in BALB/c mice against parvalbumins purified from seven fish species. Parvalbumin content was quantified in fish (raw/processed) and skin prick test (SPT) solutions by ELISA using the corresponding anti-serum for detection and the purified parvalbumins as standards. RESULTS: Using SDS-PAGE scanning, parvalbumin contents were <0.5 mg per gram tissue for mackerel, 0.5-2 mg for salmon and trout, and >2 mg for cod, carp, redfish and herring. Using ELISA, parvalbumin content ranged from <0.05 mg for tuna, 0.3-0.7 mg for mackerel, 1-2.5 mg for salmon, trout and cod to >2.5 mg per gram raw muscle for carp, herring and redfish. The parvalbumin content of processed samples (cooked/commercial) was 20-60% lower. Allergen content in SPT samples ranged from 20 to 70 µg parvalbumin/ml of extract. No parvalbumin was found in tuna SPT solution. CONCLUSION: The parvalbumin content of most commonly consumed fish species varies considerably. Differences range from severalfold to one hundredfold. This has to be taken into account when designing food challenge tests and advising fish-allergic patients.

In-vitro evaluation of coronary stents and 64-detector-row computed tomography using a newly developed model of coronary artery stenosis.

BACKGROUND: Stent implantation is the predominant therapy for non-surgical myocardial revascularization in patients with coronary artery disease. However, despite substantial advances in multidetector computed tomography (MDCT) coronary imaging, a reliable detection of coronary in-stent restenosis is currently not possible. PURPOSE: To examine the ability of 64-detector-row CT to detect and to grade in-stent stenosis in coronary stents using a newly developed ex-vivo vessel phantom with a realistic CT density pattern, artificial stenosis, and a thorax phantom. MATERIAL AND METHODS: Four different stents (Liberté and Lunar ROX, Boston Scientific; Driver, Medtronic; Multi-Link Vision, Guidant) were examined. The stents were placed on a polymer tube with a diameter of 2.5, 3.0, 3.5, or 4.0 mm. Different degrees of stenosis (0%, 30%, 50%, 70-80%) were created inside the tube. For quantitative analysis, attenuation values were measured in the non-stenotic vessel outside the stent, in the non-stenotic vessel inside the stent, and in the stenotic area inside the stent. The grade of stenosis was visually assessed by two observers. RESULTS: All stents led to artificial reduction of attenuation, the least degree of which was found in the Liberté stent (11.3+/-10.2 HU) and the Multi-Link Vision stent (17.6+/-17.9 HU; P = 0.25). Overall, the non-stenotic vessel was correctly diagnosed in 55.5%, the low-grade stenosis in 58.3%, the intermediate stenosis in 63.8%, and the high-grade stenosis in 80.5%. In the 3.0-, 3.5-, and 4.0-mm vessels, in none of the cases was a non-stenotic or low-grade stenotic vessel misdiagnosed as intermediate or high-grade stenosis. The average deviation from the real grade of stenosis was 0.40 for the Liberté stent, 0.46 for the Lunar ROX stent, 0.45 for the Driver stent, and 0.58 for the Multi-Link Vision stent. CONCLUSION: Our ex-vivo data show that non-stenotic stents and low-grade in-stent stenosis can be reliably differentiated from intermediate and high-grade in-stent stenosis in vessels with a diameter of 3 to 4 mm. With regard to artifacts and the grading of stenoses, the Liberté stent was best suited for CT coronary angiography.

In vitro evaluation of coronary stents and in-stent stenosis using a dynamic cardiac phantom and a 64-detector row CT scanner.

INTRODUCTION: The aim of the study was to examine the ability of a 64-slice MDCT to detect in-stent stenoses in an ex vivo model of coronary stents. METHODS: Five different stents (Liberté, Boston Scientific; Driver, Medtronic; Multi-Link Vision, Guidant; Taxus Express, Boston Scientific; Cypher, Cordis) were examined using a dynamic cardiac phantom. The stents were pulled over a vessel model that consists of a polymer tube with diameters of 3.0, 3.5, and 4.0 mm and four different degrees of stenoses (0%; 30%; 50%; 70-80%). This model was moved with a rate of 60 bpm to mimic cardiac motion. To assess the degree of artificial signal reduction (artificial reduction of attenuation (ARA)) by the different stents, attenuation values were measured in the vessel outside the stent, and in the non-stenotic vessel inside the stent. Furthermore the grade of stenosis was assessed by two clinical observers. RESULTS: Highest ARA was found for the Cypher Stent (35 HU), whereas the Liberté Stent presented the lowest ARA (16 HU). Depending on the stent and the vessel diameter, up to 87.5% of the stenoses were correctly diagnosed. In the 3.0 and 3.5 mm vessels, a nonstenotic or low-grade stenotic vessel was diagnosed as intermediate or high-grade stenosis in 22.5%, whereas in the 4.0 mm vessels, this kind of overestimation did not occur. A 50% stenosis was diagnosed as a 30% stenosis in 30%. On the other hand, high-grade stenoses were underestimated in only 10%. On a four-point scale, the average deviation from the real grade of stenosis was 0.21 for the Liberté stent, 0.54 for the Taxus Express stent, 0.29 for Driver stent, 0.62 for the Multi-Link Vision stent, and 0.37 for the Cypher stent. CONCLUSIONS: In a dynamic cardiac phantom model, high grade stenoses in vessels with a diameter of 4 mm could be reliably detected irrespective of the stent type used in this study. Vice versa, high grade stenoses (> or = 50%) could only be ruled out with certainty in vessels with a diameter of 4 mm. In smaller vessels, the ability to correctly diagnose high-grade stenoses was dependent on the type of stent and the imaging artifacts associated with it.

Intramyocardial gene therapy with naked DNA encoding vascular endothelial growth factor improves collateral flow to ischemic myocardium.

Both VEGF protein and VEGF DNA in combination with an adenoviral vector have been shown to enhance collateral formation in a porcine model of chronic myocardial ischemia. We sought to determine whether direct intramyocardial injection of naked DNA encoding for VEGF could similarly improve myocardial perfusion. Initially, 23 nonischemic pigs received either 200 microg of plasmid DNA encoding beta-galactosidase (pCMVbeta, n = 11) or 500 microg of phVEGF165 (n = 12) into four separate sites in the myocardium via a small anterolateral thoracotomy incision in the fourth intercostal space. Two additional groups of pigs received an intramyocardial injection of either phVEGF165 (n = 6) or pCMVbeta (n = 7) 3 to 4 weeks after implantation of an ameroid constrictor around the left circumflex coronary artery. The injections caused no change in heart rate or blood pressure, and no ventricular arrhythmias or histologic evidence of inflammation. VEGF protein was detected by Western blot in VEGF-treated animals, with the strongest bands closest to the injection site. Plasma VEGF concentration (ELISA) increased from 3+/-2 to 27+/-13 pg/ml (p = 0.035) by day 4 after treatment. No increase in VEGF protein was noted in pCMVbeta-treated animals whereas these did stain positive for beta-Gal. Resting myocardial blood flow (colored microspheres) was significantly reduced in the ischemic versus nonischemic territory in control animals (1.07+/-0.05 versus 1.32+/-0.05; p < 0.05) but not VEGF-treated pigs (1.32+/-0.24 versus 1.13+/-0.12; p = NS). Maximal vasodilatation with adenosine significantly increased flow to the ischemic region in VEGF-treated pigs (2.16+/-0.57 versus 1.32+/-0.24; p < 0.05) but not controls (1.31+/-0.05 versus 1.17+/-0.06;p = NS). Collateral filling of the occluded circumflex artery improved in five of six VEGF-treated pigs (mean change in Rentrop score, +1.5). We conclude that direct intramyocardial transfection phVEGF165 is safe and capable of producing sufficient VEGF protein to enhance collateral formation and myocardial perfusion. This approach may offer an alternative therapy for patients with intractable myocardial ischemia not amenable to PTCA or CABG.