Pediatric grand rounds: a tribute to Norman J. Siegel, M.D

At ceremonies held in Baltimore Maryland on May, 4, 2002, Dr. Howard A Pearson, Professor of Pediatrics, was awarded the John A. Howland award of the American Pediatric Society, probably the most prestigious award of American Pediatrics. Dr. Pearson had been nominated for the award by Dr. Norman Siegel. The Department of Pediatrics Grand Rounds on Wednesday noon, October 26, 2002 was originally scheduled as a repetition of the presentation by Dr. Siegel and the acceptance by Dr. Pearson for those who could not be in Baltimore. However; in a number of meetings, unknown to Dr. Siegel, it was unanimously decided that it would be very appropriate to instead honor him as he stepped down from his position as Vice- and Interim Chairman of Pediatrics, and to formally thank him for his long and faithful service to the Department of Pediatrics, the Yale University School of Medicine, and the Yale New Haven Hospital.     

The cardiac mechanical stretch sensor machinery involves a Z disc complex that is defective in a subset of human dilated cardiomyopathy

Muscle cells respond to mechanical stretch stimuli by triggering downstream signals for myocyte growth and survival. The molecular components of the muscle stretch sensor are unknown, and their role in muscle disease is unclear. Here, we present biophysical/biochemical studies in muscle LIM protein (MLP) deficient cardiac muscle that support a selective role for this Z disc protein in mechanical stretch sensing. MLP interacts with and colocalizes with telethonin (T-cap), a titin interacting protein. Further, a human MLP mutation (W4R) associated with dilated cardiomyopathy (DCM) results in a marked defect in T-cap interaction/localization. We propose that a Z disc MLP/T-cap complex is a key component of the in vivo cardiomyocyte stretch sensor machinery, and that defects in the complex can lead to human DCM and associated heart failure.     

Prickle mediates feedback amplification to generate asymmetric planar cell polarity signaling

Myosin-1c (also known as myosin-Ibeta) has been proposed to mediate the slow component of adaptation by hair cells, the sensory cells of the inner ear. To test this hypothesis, we mutated tyrosine-61 of myosin-1c to glycine, conferring susceptibility to inhibition by N(6)-modified ADP analogs. We expressed the mutant myosin-1c in utricular hair cells of transgenic mice, delivered an ADP analog through a whole-cell recording pipette, and found that the analog rapidly blocked adaptation to positive and negative deflections in transgenic cells but not in wild-type cells. The speed and specificity of inhibition suggests that myosin-1c participates in adaptation in hair cells.     

Morphology and morphometry of in vivo- and in vitro-produced bovine concepti from early pregnancy to term and association with high birth weights

This study was designed to characterize conceptus development based on pre- and postnatal measurements of in vivo- and in vitro-derived bovine pregnancies. In vivo-produced embryos were obtained after superovulation, whereas in vitro-produced embryos were derived from established procedures for bovine IVM, IVF and IVC. Blastocysts were transferred to recipients to obtain pregnancies of single (in vivo/singleton or in vitro/singleton groups) or twin fetuses (in vitro/twins group). Ultrasonographic examinations were performed weekly, from Day 30 of gestation through term. Videotaped images were digitized, and still-frames were used for the measurement of conceptus traits. Calves and fetal membranes (FM) were examined and measured upon delivery. In vitro-produced fetuses were smaller than in vivo controls (P < 0.05) during early pregnancy (Day 37 to Day 58), but in vitro/singletons presented significantly higher weights at birth than in vivo/control and in vitro/twin calves (P < 0.05). From late first trimester of pregnancy (Day 72 to Day 93), placentomes surrounding in vitro-derived singleton fetuses were longer and thinner than controls (P < 0.05). At term, the presence of giant cotyledons in the fetal membranes in the in vitro group was associated with a larger cotyledonary surface area in the fetal horn (P < 0.05). The biphasic growth pattern seen in in vitro-produced pregnancies was characterized by conceptus growth retardation during early pregnancy, followed by changes in the development of the placental tissue. Resulting high birth weights may be a consequence of aberrant placental development due to the disruption of the placental restraint on fetal growth toward the end of pregnancy.     

Development of a coupled enzyme assay for the measurement of alternanase activity**, ***

Alternanase, an endoglucanase that hydrolyzes the bacterial exopolysaccharide alternan, will also hydrolyze the trisaccharide, panose, to produce glucose and a disaccharide that can be formed into a novel, cyclic tetrasaccharide. The glucose can then be selectively and quantitatively measured by enzyme-based reaction which forms the basis of a coupled enzyme assay to quantitate alternanase activity. By this method a preparation of alternanase purified by affinity chromatography on immobilized isomaltose had a maximum reaction rate (V _max) of 0.75 mol glucose min^–1 and a K _m of 34 mM for panose. Two competitive inhibitors of alternanase activity were also evaluated using this coupled enzyme assay: isomaltose had a K _i of 94 mM while the cyclic tetrasaccharide had a K _i of 66 mM.