Інформаційні технології у сфері освіти - нові можливості, нові завдання

AbstractIn Limulus photoreceptors, light induces a depolarization. The gating mechanism of the cation channels involved is not understood but evidence exists that cyclic nucleotides may act as ligands. Using an antiserum against a cGMP-protein conjugate, we found strong immunoreactivity localized to the light-sensitive rhabdoms of lateral eye photoreceptors. In homogenated tissue, the cGMP concentration was on average 0.2 pmol/mg retinal tissue protein both in light- and dark-adapted eyes. Phosphodiesterase inhibitors caused a 2–3-fold increase in the cGMP level. Despite our failure to detect a light dependence of the cGMP concentration, these results support the proposed role of cGMP in Limulus phototransduction

Cyclic GMP in the pig vitreous and retina after experimental retinal detachment

Purpose: Earlier studies have revealed a decreased level of cGMP in vitreous fluid obtained from patients with a retinal detachment. To further investigate this phenomenon, we developed an experimental retinal detachment model in pigs. Methods: Experimental unilateral retinal detachments were induced in pig eyes by subretinal injection of 0.25% sodium hyaluronate. Fourteen days later the vitreous and retinas were analyzed for cGMP expression. Following enucleation, the retinas were incubated in the presence of a nonselective phosphodiesterase inhibitor (IBMX), and the particulate guanylyl cyclase stimulator atrial natriuretic peptide (ANP) or the soluble guanylyl cyclase stimulator sodium nitroprusside (SNP). cGMP was visualized in retinal wholemounts by immunochemistry combined with a computer based stereology system. cGMP levels in vitreous were determined by ELISA. Results: The mean vitreous cGMP level in pig eyes with a retinal detachment (1.45 pmol/ml) was significantly lower compared to the mean level of cGMP in healthy pig eyes (4.61 pmol/ml; p= 0.028 was considered significant). In the inner retina, ANP as well as SNP induced cGMP immunoreactivity in both detached and healthy retinas. After incubation with ANP, cGMP could also be detected in the outer nuclear layer of the detached retina, whereas this was not the case in the normal retina. Conclusions: Experimental retinal detachment in the pig eye leads to a decrease of cGMP levels in vitreous similar to that observed in clinical studies. This model may be helpful to analyze the mechanisms involved in cGMP dynamics following retinal detachment

Structural and time-resolved mechanistic investigations of protein hydrolysis by the acidic proline-specific endoprotease from Aspergillus niger

Proline-specific endoproteases have been successfully used in, for example, the in-situ degradation of gluten, the hydrolysis of bitter peptides, the reduction of haze during beer production, and the generation of peptides for mass spectroscopy and proteomics applications. Here we present the crystal structure of the extracellular proline-specific endoprotease from Aspergillus niger (AnPEP), a member of the S28 peptidase family with rarely observed true proline-specific endoprotease activity. Family S28 proteases have a conventional Ser-Asp-His catalytic triad, but their oxyanion-stabilizing hole shows a glutamic acid, an amino acid not previously observed in this role. Since these enzymes have an acidic pH optimum, the presence of a glutamic acid in the oxyanion hole may confine their activity to an acidic pH. Yet, considering the presence of the conventional catalytic triad, it is remarkable that the Aspergillus niger enzyme remains active down to pH 1.5. The determination of the primary cleavage site of cytochrome c along with molecular dynamics-assisted docking studies indicate that the active site pocket of AnPEP can accommodate a reverse turn of approximately 12 amino acids with proline at the S1 specificity pocket. Comparison with the structures of two S28-proline-specific exopeptidases reveals not only a more spacious active site cavity but also the absence of any putative binding sites for amino- and carboxyl-terminal residues as observed in the exopeptidases, explaining AnPEP's observed endoprotease activity. This article is protected by copyright. All rights reserved.</p

Time-integrated 3D approach of late Quaternary sediment-depocenter migration in the Tagus depositional system: From river valley to abyssal plain

Quantification of sediment volumes in continental to deep ocean basins is key to understanding processes of sediment distribution in source-to-sink depositional systems. Using our own and published data we present the first quantification of sediment-volume changes in basins along the course of a major southwest European river during the deglaciation. The salient points of this quantitative record in the Tagus and equivalent North Atlantic basins show crucial roles for sea level, climate and land-use in the distribution of sediments. The bypass of sediments starved the Tagus basins, and subsequently sedimentation mainly occurred on the Tagus Abyssal Plain during the sea-level lowstand of the Last Glacial Maximum. The main sediment depocenter rapidly shifted via the continental shelf to the Lower Tagus Valley during sea-level rise in the deglaciation period. Finally, the main sediment depocenter shifted further landward into the Lower Tagus Valley during sea-level high stand in the Holocene. During the high-stand phase (last 7 ky), sediment flux increased up to 2.5 times, due to climate and land-use changes. The average catchment denudation rate during the last 12 ky (0.04–0.1 mm/y) is in agreement with those of other European catchments. Our study clearly demonstrates the added value of detailed knowledge of 3D depocenter distribution, size and chronology. This allowed us to identify an increased sediment flux during the last 7 ky, which was not identified using local observations from boreholes alone. The uniqueness of the Tagus depositional system lies in the combination of a large accommodation space in the bedrock-confined Lower Tagus Valley, the steep lowstand-surface gradient and the narrow continental shelf with canyons indenting the shelf break

Presence of soluble and particulate guanylyl cyclase in the same hippocampal astrocytes

The localisation of particulate and soluble guanylyl cyclase was studied in hippocampal astrocytes. Counting the colocalisation of cGMP immunoreactivity with the astrocytic marker glial fibrillary acidic protein after stimulation of brain slices with sodium nitroprusside (0.1 mM) or atrial natriuretic peptide (100 nM), we were able to show that at least 67% of the hippocampal astrocytes contained both guanylyl cyclase isoforms. In addition, it was shown that a large number of atrial natriuretic peptide, brain-derived natriuretic peptide or sodium nitroprusside responsive cells contain the β1-subunit of the soluble guanylyl cyclase. The results show that, in at least a subset of hippocampal astrocytes, soluble and particulate guanylyl cyclases are simultaneously present in the same cells

Geometric considerations of nitric oxide-cyclic GMP signalling in the glomerular neuropil of the locust antennal lobe

Using NADPH-diaphorase staining as a marker for nitric oxide (NO) synthase and an antiserum against cyclic GMP, we recently reported the anatomical distribution of nitric oxide donor and target cells in the antennal lobe, the principal olfactory neuropile of the locust. The most striking NADPH-diaphorase activity in the olfactory pathway is concentrated in a cluster of intensely stained local interneurons innervating the glomeruli. After incubation of tissue in a nitric oxide donor and inhibition of phospodiesterase activity, neurons of this cluster expressed cyclic GMP-immunoreactivity in the cell body and neurites. Here we examine the importance of the arrangement of NO donor and target cells for information processing in the glomeruli. The cellular organization of the NO-cyclic GMP system in olfactory interneurons, and the dendritic branching pattern, suggest that nitric oxide may not only act as intercellular, but also as intracellular messenger molecule in the glomerular neuropile of the antennal lobe. <br