Autoradiographic studies of human chromosomes

Chromosomal sites of DNA synthesis during the final 30 minutes or less of the S-phase of the cell division cycle of fibroblasts were delinated autoradiographically. Very light labeling was found, indicating that a recognizable but very minor portion of the cell's DNA is synthesized during a few minutes at the extreme end of S. This interval immediately follows those periods near the end of S when prominent synthetic asynchrony exists in different chromosomal regions. A non-random distribution of label, but one different from the more familiar end-of-S pattern, was detected during this final interval. The late-replicating X was less heavily labeled than some autosomes during the final minutes of S, while sites in chromosome No. 3 were somewhat more heavily labeled than those in other chromosomes. The biological significance of these minute, last-to-replicate chromosomal regions is unknown.     

Kinetics of the Hydrolysis of 8-Bromo-Cyclic GMP by the Light-Activated Phosphodiesterase of Toad Rods

The hypothesis that cyclic GMP is the internal transmitter of retinal rod phototransduction, when combined with the observations that 8-bromo-cyclic GMP opens the cyclic GMP-dependent outer segment conductance and that rods into which 8-bromo-cyclic GMP has been injected still respond to light, predicts that the light-activated phosphodiesterase (EC 3.1.4.17) must catalyze the hydrolysis of 8-bromo-cyclic GMP. This hypothesis was tested by measuring light-activated toad rod disk membrane phosphodiesterase with a pH assay technique. Phosphodiesterase-catalyzed hydrolysis of 8-bromo-cyclic GMP was confirmed: at pH 8.0, total proton production after flash activation was identical to total amount of 8-bromo-cyclic GMP added as substrate. Photoactivated phosphodiesterase was remarkably less efficient in catalyzing the hydrolysis of 8-bromo-cyclic GMP than of cyclic GMP: Vmax for 8-bromo-cyclic GMP was 0.063 M/M rhodopsin/s, whereas that for cyclic GMP was 11 M/M rhodopsin/s--170 times greater. The Km for 8-bromo-cyclic GMP was 160 microM, and for cyclic GMP, 590 microM. 8-bromo-cyclic GMP competitively inhibited phosphodiesterase-catalyzed hydrolysis of cyclic GMP with a Ki of 1.2 mM. Complete reaction progress curves were analyzed for obedience to Michaelis-Menten kinetics: cyclic GMP hydrolysis, 8-bromo-cyclic GMP hydrolysis, and cyclic GMP hydrolysis in the presence of 8-bromo-cyclic GMP as competitive inhibitor were found to follow the integrated form of the Michaelis-Menten equation over the time course of the reactions, assuming phosphodiesterase was activated as a step. The kinetic parameters extracted from reaction progress curves were consistent with those derived from analysis of the initial velocity.(ABSTRACT TRUNCATED AT 250 WORDS)     

The Role of Snowmelt and Spring Rainfall in Inorganic Nutrient Fluxes from a Large Temperate Watershed, the Androscoggin River Basin (Maine and New Hampshire)

The importance of snowmelt and spring rainfall to water and nutrient exports from macro-scale watersheds (>1000 km²) is not well established. Data collected from the Androscoggin River watershed (Maine and New Hampshire) between February 1999 and March 2002 show that the 90-day spring melt period accounted for 39–57% of total annual discharge and is likely driven both by snowpack melting and spring rainfall. While large loads of dissolved inorganic nitrogen (DIN) are delivered to the watershed from snowmelt and rain (from 1.16× 10⁶ to 1.61× 10⁶ kg N over the study years), only one third of this N load is exported from the basin during the snowmelt period (0.40× 10⁶–0.48 × 10⁶ kg N). Despite reduced residence time and temperature limitations on biological N retention, there is a poor mass balance between DIN input to the watershed and the nitrogen exported from mouth of the river. Inferences from a geochemical hydrograph separation suggests that approximately 51–63% of the water leaving the mouth of the Androscoggin river is from these ‘new’ water sources (rain and snowmelt) while 37–49% is from DIN depleted soil and groundwater. Mixing of water from different sources, as well as nutrient retention by dams in the upper watershed, may account for the large discrepancy between DIN inputs and exports from this watershed.