The isolation, partial purification and preliminary characterization of a growth factor from chick embryo brains

Chicl( brain growth factor (CBGF) is a mitogen isolated from embryonic chick brains thought to have a potential role as a trophic factor involved in nerve dependent amphibian limb regeneration. In addition, CBGF stimulates 3H-thymidine incorporation in chick embryo brain astrocytes in vitro. In this study, cultured chick embryo brain non-neuronal cells were employed in a bioassay to monitor CBGF activity throughout various stages of its pllrification. Cell culture and assay conditions were optimized. Nonneuronal cells grew best on collagen-coated culture dishes in complete medium, were most responsive to a growth stimulus [10% fetal bovine serum (FBS)] at the second and third subcultures, and were healthiest when rendered "quiescent" in medium supplemented with 1% FBS. The most effective bioassay conditions consisted of a minimum 14.5 hour "quiescence" time (24 hours was used), a 6 hour "prestimulation" time, and a 24 hour 3H-thymidine labeling time. Four-day subconfluent primary non-neuronal cells consisted of 6.63% GFAP positive cells; as a result cultures were thought to be mainly composed of astroblasts. CBGF was purified from 18-day chick embryo brains by ultrafiltration through Amicon PM-30 and YM-2 membranes, size exclusion chromatography through a Biogel P6 column, and analytical reverse-phase high-performance liquid chromatography (rp-HPLC). The greatest activity resided in rp-HPLC fraction #7 (10 ng/ml) which was as effective as 10% FBS at stimulating 3H-thymidine incorporation in chick embryo brain nonneuronal cells. Although other researchers report the isolation of a mitogenic fraction consisting of 5'-GMP from the embryonic chick brain, UV absorbance spectra, rp-HPLC elution profiles, and fast atom bombardment (FAB) mass spectra indicated that CBGF is neither 5'-GMP nor 51-AMP. 2 Moreover, commercially available 5t-GMP was inhibitory to 3H-thymidine incorporation in the chick non-neuronal cells, while Sf-AMP had no effect. Upon treatment with pronase, the biological activity of fraction P6-3 increased; this increase was nearly 30% greater than what would be expected from a simple additive effect of any mitogenic activity of pronase alone together with P6-3 alone. This may suggest the presence of an inhibitor protein. The bioactive component may be a protein protected by a nucleoside/nucleotide or simply a nucleoside/nucleotide acting alone. While the FAB mass spectrum of rp-HPLC fraction #7 did not reveal molecular weight or sequence information, the ion of highest molecular weight was observed at m/z 1610; this is consistent with previous estimations of CBGF's size. 3

The Effectiveness of Learning Portfolios: A Study of Quality Assurance Programs of Selected Health Regulatory Colleges in Ontario

Health regulatory colleges promote quality practice and continued competence through Quality Assurance (QA) programs. For many colleges, a QA program includes the use of portfolios that incorporate self-directed learning. The purpose of this study was to determine some of the issues surrounding the effectiveness of QA portfolio programs. The literature review revealed that portfolios are valuable tools, but gaps in knowledge include a comparative analysis of QA programs and the perspective of regulatory college administrators. Data were collected through interviews with 6 administrators and a review of 14 portfolio models described on college websites. The results from the two data sources were applied to Robert Stake's responsive evaluation framework to identify issues related to the portfolio's effectiveness (Stake, 1967). The learning components of portfolios were analyzed through the humanist and constructivist lenses. All 14 portfolio models were found to have 3 main components: self-diagnosis, learning plan and activities, and self-evaluation. However, differences were uncovered in learners' autonomy in selecting learning activities, methods of portfolio evaluation, and the relationship between the portfolio and other QA components. The results revealed a dual philosophy of learning in portfolio models and an apparent contradiction between the needs of the individual learner and the organization. Paths for future research include the tenuous relationship between competence and learning, and the impact of technical approaches on selfdirected learning initiatives. A key recommendation is to acknowledge the unique identity of each profession so that health regulatory colleges can address legislative demands and learner needs.