Effects of troponin I phosphorylation on conformational exchange in the regulatory domain of cardiac troponin C.

Conformational exchange has been demonstrated within the regulatory domain of calcium-saturated cardiac troponin C when bound to the NH2-terminal domain of cardiac troponin I-(1-80), and cardiac troponin I-(1-80)DD, having serine residues 23 and 24 mutated to aspartate to mimic the phosphorylated form of the protein. Binding of cardiac troponin I-(1-80) decreases conformational exchange for residues 29, 32, and 34. Comparison of average transverse cross correlation rates show that both the NH2- and COOH-terminal domains of cardiac troponin C tumble with similar correlation times when bound to cardiac troponin I-(1-80). In contrast, the NH2- and COOH-terminal domains in free cardiac troponin C and cardiac troponin C bound cardiac troponin I-(1-80)DD tumble independently. These results suggest that the nonphosphorylated cardiac specific NH2 terminus of cardiac troponin I interacts with the NH2-terminal domain of cardiac troponin C.     

Organochlorine Pesticide Residues in Human Fat in the United Kingdom 1969-71

During the period 1969-71 samples of human body fat were taken during routine necropsies on 201 subjects over 5 years old and 20 stillborn babies and infants under 5 years old, and analysis was carried out for organochlorine pesticide residues. By comparing the results from this period with those from similar studies undertaken in 1963-4 and 1965-7, it is clear that a downward trend continues.     

Misincorporation during DNA synthesis, analyzed by gel electrophoresis.

A method has been developed for simultaneous comparison of the propensity of a DNA polymerase to misincorporate at different points on a natural template-primer. In this method elongation of a [5'-32P] primer, annealed to a bacteriophage template strand, is carried out in the presence of only three dNTPs (highly purified by HPLC). Under these conditions the rate of primer elongation (monitored by gel electrophoresis/autoradiography) is limited by the rate of misincorporation at template positions complementary to the missing dNTP. Variations in the rate of elongation (revealed by autoradiographic banding patterns) reflect variations in the propensity for misincorporation at different positions along the template. The effect on primer elongation produced by addition of a chemically modified dNTP to 'minus' reactions reveals the mispairing potential of the modified nucleotide during DNA synthesis. By use of this electrophoretic assay of misincorporation we have demonstrated that the fidelity of E. coli DNA polymerase I varies greatly at different positions along a natural template, and that BrdUTP and IodUTP can be incorporated in place of dCTP during chain elongation catalyzed by this enzyme.     

The perfusion of isolated whole organs

The long term preservation of organs is a highly desired goal to provide adequate numbers of appropriate organs for transplantation, and the lack of methods to accomplish this adequately is one of the outstanding totally unsolved problems in the field. Although other methods are useful for short periods, only perfusion preservation currently holds immediate potential for longer and even indefinite extracorporeal storage. Perfusion is the method which theoretically creates a physiological environment rather than increasing resistance to an abnormal one. This review considers in detail the physiological environment, how it might be created, and the current and future problems in attaining such a goal.