The synthesis of cyclic ADP-carbocyclic-ribose (cADPcR, 4) designed as a stable mimic of cyclic ADP-ribose (cADPR, 1), a Ca2+-mobilizing second messenger, was achieved using as the key step a condensation reaction with the phenylthiophosphate-type substrate 14 to form an intramolecular pyrophosphate linkage. The N-1-carbocyclic-ribosyladenosine derivative 16 was prepared via the condensation between the imidazole nucleoside derivative 17, prepared from AICA-riboside (19), and the readily available optically active carbocyclic amine 18. Compound 16 was then converted to the corresponding 5' '-phosphoryl-5'-phenylthiophosphate derivatives 14. Treatment of 14 with AgNO3 in the presence of molecular sieves (3 A) in pyridine at room temperature gave the desired cyclization product 32 in 93% yield, and subsequent acidic treatment provided the target cADPcR (4). This represents a general method for synthesizing biologically important cyclic nucleotides of this type. 1H NMR analysis of cADPcR suggested that its conformation in aqueous medium is similar to that of cADPR. cADPcR, unlike cADPR, was stable under neutral and acidic conditions, where under basic conditions, it formed the Dimroth-rearranged N6-cyclized product 34. cADPcR was also stable in rat brain membrane homogenate which has cADPR degradation activity. Furthermore, cADPcR was resistant to the hydrolysis by CD38 cADPR hydrolase, while cADPR was rapidly hydrolyzed under the same conditions. When cADPcR was injected into sea urchin eggs, it caused a significant release of Ca2+ in the cells, an effect considerably stronger than that of cADPR. Thus, cADPcR was identified as a stable mimic of cADPR.
The variation of velocity and volume transport of the Tsugaru Warm Current (TWC) is investigated based on the data of continuous and cross‐sectional current monitoring in the Tsugaru Strait. During November 1999–March 2000, the structure of the velocity transection across the strait was almost stable in which the TWC occupied the central part of the strait, while the returning flows existed in the northern and southern part of the strait, and the velocity of these flows decreases with time. The volume transport of the TWC varies from 2.1 to 1.1 Sv, with the mean value of 1.5 Sv, following the variation of the TWC. The temporal variation of the volume transport shows a linear correlation with that of the sea level difference between the Japan Sea and the Pacific Ocean. The contribution of the sea level difference to the volume transport is estimated to be roughly 70%.
Evolutional changes in interior structures of mixed population biofilms grown on domestic wastewater were quantitatively analyzed using a cryosectioning technique and an image analysis. Meanwhile, transport of particulates into the biofilms was also experimentally investigated using fluorescent microbeads as tracers to relate the biofilm structure and particulate transport into the biofilm. Microscopic observation of the cryomicrotomy biofilm sections indicated the biofilms were very porous and consisted of interwinded filamentous biomass acting as a framework of the biofilm. A honeycomb structure was often found, which would make the biofilm more resistant to water flow. There were micropores with the diameter of about 10 μm microcolony aggregates attached to filamentous biomass and macropores with the diameter of 20–200 μm in the biomass matrix. These pores did not clog during two months of cultivation. Areal porosity was about 30% in the bottom biofilm and more than 90% in the surface. Significant difference in transport efficiency was not observed for various sizes of microbeads due to the presence of macropores. Therefore, even 10 μm tracer beads could quickly traverse throughout a biofilm 640 μm thick via water channels or macropores and then penetrated into the micropores. Convective transport from the bulk to the bottom biofilm, rather than molecular diffusion, was responsible for this rapid transport. Based on experimental results, it can be concluded that the biofilm structure seems to be well designed to maximize the transport efficiency of substrates and products and the strength of biofilm structure.
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