Efficient and inexpensive method for purification of heparin binding proteins.

Heparin binding (HB) proteins mediate a wide range of important cellular processes, which makes this class of proteins biopharmaceutically important. Engineering HB proteins may bring many advantages, but it necessitates cost effective and efficient purification methodologies compared to currently available methods. One of the most important classes of HB proteins are fibroblast growth factors (FGFs) and their receptors (FGFRs). In this study, we report an efficient off-column purification of FGF-1 from soluble fractions and purification of the D2 domain of FGFR from insoluble inclusion bodies, using a weak Amberlite cation (IRC) exchanger. FGF-1 and the D2 domain have been expressed in Escherichia coli and purified to homogeneity using IRC resin. This approach is an alternative to conventional affinity column chromatography, which exhibits several disadvantages, including time-consuming experimental procedures for purification and regeneration and results in the expensive production of recombinant proteins. Results of the heparin binding chromatography and steady state fluorescence experiments show that the FGF-1 and the D2 are in a native conformation. The findings of this study will not only aid an in-depth investigation of this class of proteins but will also provide avenues for inexpensive and efficient purification of other important biological macromolecules.

Continuous arterial pressure waveform monitoring in pediatric cardiac transplant, cardiomyopathy and pulmonary hypertension patients.

PURPOSE: A continuous cardiac output monitor based on arterial pressure waveform (FloTrac/Vigileo; Edwards Lifesciences, Irvine, CA) is now approved for use in adults but not in children. This device is minimally invasive, calculates cardiac output continuously and in real time, and is easy to use. Our study sought to validate the FloTrac with the pulmonary artery catheter (PAC) intermittent thermodilution technique in pediatric cardiac patients. METHODS: This was a prospective pilot study comparing cardiac output measurements obtained via the FloTrac and arterial pressure waveform analysis with intermittent thermodilution. Subjects carried the diagnosis of pulmonary hypertension or cardiomyopathy, or were in the postoperative course after orthotopic heart transplantation. RESULTS: Enrolled in the study were 31 subjects, and 136 data points were obtained. The age range was 8 months to 16 years. The mean body surface area (BSA) was 1.1 m(2). Bland-Altman plots for the mean cardiac outputs of all subjects with a BSA ≥ 1 m(2) showed limits of agreement of -2.7 to 8.0 l/min (± 5.4 l/min). Patients with a BSA ≤ 1 m(2) demonstrated even wider limits of agreement (± 8.5 l/min). The intraclass correlation for the PAC was 0.929 and 0.992 for the FloTrac. CONCLUSION: There was poor agreement between the PAC and FloTrac in measuring cardiac output in a population of children with pulmonary hypertension or cardiomyopathy, or after cardiac transplantation. This is in contrast to adult studies published thus far. This suggests that the utility of the FloTrac and measurements obtained from arterial pulse wave analysis in children is uncertain at this time.

X-ray dose from pediatric cardiac catheterization: a comparison of materials and methods for measurement or calculation.

Pediatric cardiac catheterization procedures have the potential to transmit high X-ray doses, which may lead to acute effects or latent skin reactions. Direct measurement of radiation dose was facilitated using nanodot dosimeters and radiochromic film. Direct measurement results were compared with vendor-listed dosimetry and calculation using phantom data. Vendor-listed data demonstrated a wide discrepancy with measured data, whereas the calculation reproducibly overestimated the actual dose. A simple formula was derived to calculate the dose using fluoroscopy time, cine frame quantity and average cine mA in a biplane environment.