CYCLIC NUCLEOTIDE PHOSPHODIESTERASE OF HUMAN CEREBROSPINAL FLUID

Abstract— Cyclic 3',5'-AMP (cAMP) and cyclic 3',5'–GMP (cGMP) phosphodiesterase activities were found in human cerebrospinal fluid (CSF) using low substrate concentration (0.4μM). More rapid hydrolysis of cGMP than that of cAMP was observed in human CSF. However, cGMP hydrolytic activity of CSF was very much lower (0.3 pmol/min/ml CSF) than that of human cerebral cortex (33.7 nmol/min/g wet cortex). The pH optimum was found to be 8.0 (cGMP phosphodiesterase) and 7.5 (cAMP phosphodiesterase). The maximum stimulation of both cAMP and cGMP phosphodiesterase was achieved at 4 mM-MgCl₂. Cyclic AMP had relatively little effect on the hydrolysis of cGMP in CSF and the cortex, while cGMP inhibited hydrolysis of cAMP in both tissues. Snake venom was found to stimulate cAMP and cGMP phosphodiesterase activity of CSF, by 60% and 110% respectively. This stimulation by snake venom was also observed in the cortex phosphodiesterase, but was not observed in human plasma or thyroid phosphodiesterase. When CSF was applied to Sepharose 6B column, cGMP phosphodiesterase was separated into three different molecular forms. A plot of activity against substrate concentration using peak I (largest molecular size) revealed a high affinity ( K _m= 2.6μM) and a low affinity ( K _m= 100μM) for cAMP suggesting the existence of at least two molecular forms of the enzyme. On the other hand, using a cGMP as substrate the only one K _m value (1.90 μm) was obtained. These K _m values of CSF enzymes described above were close to those obtained from human cerebral cortex preparations. The enzyme under peak I corresponded to the cortex enzyme when judged from its molecular size and stimulation by snake venom. It seems likely from our results that at least a part of CSF phosphodiesterase originates from the central nervous system.     

Studies of pesticides effects on Chlorella metabolism II. Effect of DCMU on galactolipid metabolism

DCMU (N'-(3,4-dichlorophenyl)-N, N-dimethylurea) was testedfor effects on the metabolism of galactolipids in Chlorellaand chloroplasts isolated from higher plants. In Chlorella,DCMU affected galactolipid synthesis in the light more thanthat of other lipids, but it showed no effect on lipid synthesisin the dark. DCMU did not affect the turnover of galactolipidsin the light. In vitro studies using ¹⁴C-acetate or ¹⁴C-UDP-galactoseas a precursor showed that DCMU had no effect on the synthesisof gross lipid or galactolipids in chloroplasts isolated fromhigher plants. The significance of these observations are discussed. (Received September 21, 1974; )     

Studies on the Foaming Property of the Chicken Egg White

The egg white was treated under various whipping conditions, and its foaminess measured. At the same time, the amounts of the coagulated proteins formed from each egg white and their constituent hexose were measured. From these results, discussions were made about the relation between the foaminess of the egg white and the amount of the coagulated proteins under various whipping conditions.     

Changes in the Shape and Surface Hydrophobicity of Ovalbumin during Its Transformation to S-Ovalbumin

Intrinsic viscosity, Stokes radius and the hydrophobic coefficient of Keshavarz and Nakai [Biochim. Biophys. Acta, 576, 269 (1979)] were measured to compare the shape and surface hydrophobicity of ovalbumin and s-ovalbumin. Both the intrinsic viscosity and Stokes radius of s- ovalbumin were smaller than those of ovalbumin, which suggests that the configuration of s- ovalbumin became more compact during the ovalbumin-s-ovalbumin transformation. The hydrophobic coefficient of s-ovalbumin was larger than that of ovalbumin, which suggests that the surface hydrophobicity of s-ovalbumin was larger than that of ovalbumin. Further, these properties were measured for ovalbumin samples obtained at various stages of ovalbumin-s-ovalbumin transformation. Changes in the shape and surface hydrophobicity of ovalbumin were not found in the first stage of ovalbumin-s-ovalbumin transformation. They changed rapidly in the last stage of the ovalbumin-s-ovalbumin transformation.     

Lagrangian tracking measurements revealed the temporal dynamics of nitrogen and phosphorus spiralling in a large Japanese river

Limnology - The in-stream processing of nutrients plays an important role in the fluvial nutrient transport from lands to the ocean, but few empirical studies have addressed the temporal dynamics...     

A Geometry-Based Multiple Testing Correction for Contingency Tables by Truncated Normal Distribution

Statistics in Biosciences - Inference procedure is a critical step of experimental researches to draw scientific conclusions especially in multiple testing. The false positive rate increases unless...     

Basic pH-induced modification of excitation-energy dynamics in fucoxanthin chlorophyll a/c-binding proteins isolated from a pinguiophyte,Glossomastix chrysoplasta

Photosynthetic organisms have diversified light-harvesting complexes (LHCs) to collect solar energy efficiently, leading to an acquisition of their ecological niches. Herein we report on biochemical and spectroscopic characterizations of fucoxanthin chlorophyll a/c-binding protein (FCP) complexes isolated from a marine pinguiophyte Glossomastix chrysoplasta. The pinguiophyte FCP showed one subunit band in SDS-PAGE and one protein-complex band with a molecular weight at around 66 kDa in clear-native PAGE. By HPLC analysis, the FCP possesses chlorophylls a and c, fucoxanthin, and violaxanthin. To clarify excitation-energy-relaxation processes in the FCP, we measured time-resolved fluorescence spectra at 77 K of the FCP adapted to pH 5.0, 6.5, and 8.0. Fluorescence curves measured at pH 5.0 and 8.0 showed shorter lifetime components compared with those at pH 6.5. The rapid decay components at pH 5.0 and 8.0 are unveiled by fluorescence decay-associated (FDA) spectra; fluorescence decays occur in the 270 and 160-ps FDA spectra only at pH 5.0 and 8.0, respectively. In addition, energy-transfer pathways with time constants of tens of picoseconds are altered under the basic pH condition but not the acidic pH condition. These findings provide novel insights into pH-dependent energy-transfer and energy-quenching machinery in not only FCP family but also photosynthetic LHCs.