Система поддержки принятие решений при проведении клинических исследований

Разработка программного обеспечения для проведения клинических исследований. Данное приложение может применяться для обеспечения целостности данных, безопасности субъекта исследования, качества продукта в ходе проведения исследования, что приведет к автоматизации рутинных процессов и будет способствовать повышению скорости и качества проводимых исследований.Development of software for conducting clinical trials. This application can be used to ensure data integrity, security of the research subject, and product quality during the research, which will lead to automation of routine processes and will help to improve the speed and quality of research

The Gene Cluster for Fluorometabolite Biosynthesis in Streptomyces cattleya: A Thioesterase Confers Resistance to Fluoroacetyl-Coenzyme A

SummaryA genomic library of Streptomyces cattleya was screened to isolate a gene cluster encoding enzymes responsible for the production of fluorine-containing metabolites. In addition to the previously described fluorinase FlA which catalyzes the formation of 5′-fluoro-5′-deoxyadenosine from S-adenosylmethionine and fluoride, 11 other putative open reading frames have been identified. Three of the proteins encoded by these genes have been characterized. FlB was determined to be the second enzyme in the pathway, catalyzing the phosphorolytic cleavage of 5′-fluoro-5′-deoxyadenosine to produce 5-fluoro-5-deoxy-D-ribose-1-phosphate. The enzyme FlI was found to be an S-adenosylhomocysteine hydrolase, which may act to relieve S-adenosylhomocysteine inhibition of the fluorinase. Finally, flK encodes a thioesterase which catalyzes the selective breakdown of fluoroacetyl-CoA but not acetyl-CoA, suggesting that it provides the producing strain with a mechanism for resistance to fluoroacetate

Structural basis for the activity and substrate specificity of fluoroacetyl-CoA thioesterase FlK.

The thioesterase FlK from the fluoroacetate-producing Streptomyces cattleya catalyzes the hydrolysis of fluoroacetyl-coenzyme A. This provides an effective self-defense mechanism, preventing any fluoroacetyl-coenzyme A formed from being further metabolized to 4-hydroxy-trans-aconitate, a lethal inhibitor of the tricarboxylic acid cycle. Remarkably, FlK does not accept acetyl-coenzyme A as a substrate. Crystal structure analysis shows that FlK forms a dimer, in which each subunit adopts a hot dog fold as observed for type II thioesterases. Unlike other type II thioesterases, which invariably utilize either an aspartate or a glutamate as catalytic base, we show by site-directed mutagenesis and crystallography that FlK employs a catalytic triad composed of Thr(42), His(76), and a water molecule, analogous to the Ser/Cys-His-acid triad of type I thioesterases. Structural comparison of FlK complexed with various substrate analogues suggests that the interaction between the fluorine of the substrate and the side chain of Arg(120) located opposite to the catalytic triad is essential for correct coordination of the substrate at the active site and therefore accounts for the substrate specificity

Anti-Staphylococcal Calopins from Fruiting Bodies of Caloboletus radicans

Three new and seven known calopins were isolated from Caloboletus radicans. The structures of the new cyclocalopins, 8-deacetylcyclocalopin B (1), cyclocalopin A-15-ol (2), and 12,15-dimethoxycyclocalopin A (3), were mainly elucidated by NMR and MS data analysis. The stereochemistry of 1–3 was assigned based on ROE correlations, coupling constants and by comparison of their CD spectra with those of similar known calopins. While 1–10 were inactive against two cancer cell lines, they displayed antistaphylococcal activity against methicillin-resistant Staphylococcus aureus strains (MRSA) with MIC values of 16−256 μg/mL. Moreover, some calopins were active against the fish pathogen Enterococcus faecalis F1B1

Caenorhabditis elegans N-glycan Core β-galactoside Confers Sensitivity towards Nematotoxic Fungal Galectin CGL2

The physiological role of fungal galectins has remained elusive. Here, we show that feeding of a mushroom galectin, Coprinopsis cinerea CGL2, to Caenorhabditis elegans inhibited development and reproduction and ultimately resulted in killing of this nematode. The lack of toxicity of a carbohydrate-binding defective CGL2 variant and the resistance of a C. elegans mutant defective in GDP-fucose biosynthesis suggested that CGL2-mediated nematotoxicity depends on the interaction between the galectin and a fucose-containing glycoconjugate. A screen for CGL2-resistant worm mutants identified this glycoconjugate as a Galβ1,4Fucα1,6 modification of C. elegans N-glycan cores. Analysis of N-glycan structures in wild type and CGL2-resistant nematodes confirmed this finding and allowed the identification of a novel putative glycosyltransferase required for the biosynthesis of this glycoepitope. The X-ray crystal structure of a complex between CGL2 and the Galβ1,4Fucα1,6GlcNAc trisaccharide at 1.5 Å resolution revealed the biophysical basis for this interaction. Our results suggest that fungal galectins play a role in the defense of fungi against predators by binding to specific glycoconjugates of these organisms

NMR analysis of (1S,1aR,6aR)-2’,3’,6,6a-tetrahydro-spiro[cycloprop[a]indene-1(1aH),1’-[1H]indene]

The aldol condensation product of 1H-indan-1-one, (2E)-2-(2,3-dihydro-1H-inden-1-ylidene)-2,3-dihydro-1H-inden-1-one, subjected to Huang–Minlon reduction conditions was shown, via 1D and 2DNMRanalysis, to be amixture of (1S,1aR,6aR)-2’,3’,6,6a-tetrahydro-spiro[cycloprop[a]indene-1(1aH),1’-[1H]indene] and its 1R,1aS,6aS enantiomerand not 2,3,1’,3’-tetrahydro-[1,2’]-biindenylidene as originally expected. The full NMR assignment, the coupling constants in the proton NMR, and the couplings in the HMBC and NOESY of the title compound are summarized in the Table

NMR analiza 2-(2',3'-dihidro-1'H-inden-1'-il)-1H-indena

1H, 13C and two dimensional NMR analyses were applied to determine the NMR parameters of 2-(2',3'-dihydro-1'H-inden-1'-yl)-1H-indene. The chemical shifts of all the H- and C-atoms, as well as the appropriate coupling constants were determined and the complete NMR resonance assignments of the molecule are given. The predicted patterns of the four different H atoms of the two methylene groups of the indane structural element coincided completely with the complex patterns in the NMR spectra.1H, 13C i dvodimenzionalna NMR analiza su primenjeni da bi se odredili NMR parametri 2-(2', 3'-dihidro-1'H-inden-1'-il)-1H-indena. Određena su hemijska pomeranja svih H- i C-atoma, kao i odgovarajuće konstante kuplovanja i dat je potpun NMR raspored. Predskazane mustre četiri različita vodonikova atoma dve metilenske grupe indenskog strukturnog elementa potpuno su se poklopile sa složenim mustrama dobijenim na NMR spektru. Derivati indena predstavljaju glavne komponente pirolitičkih ulja i NMR podaci o njima mogu biti korisni u istraživanjima okoline

NMR analiza (1S,1aR,6aR)-2’,3’,6,6a-tetrahidro-spiro[cikloprop[a]inden-1(1ah),1’-[ 1h]indena]

The aldol condensation product of 1H-indan-1-one, (2E)-2-(2,3-dihydro-1H-in den-1-ylidene)-2,3-dihydro-1H-inden-1-one, subjected to Huang–Minlon reduction conditions was shown, via 1D and 2D NMR analysis, to be a mixture of (1S,1aR,6aR)-2’,3’,6,6a-tetrahydro-spiro cycloprop a indene-1(1aH),1’ 1H indene and its 1R,1aS,6aS enantiomer and not 2,3,1’,3’-tetrahydro- 1,2’ -biindenylidene as originally expected. The full NMR assignment, the coupling constants in the proton NMR, and the couplings in the HMBC and NOESY of the title compound are summarized in the Table.Pokazano je pomoću 1D i 2D NMR analize da (2E)-2-(2,3-dihidro-1H-inden-1-iliden)-2, 3-dihidro-1H-inden-1-on, dobiven aldolnom kondenzacijom 1H-indan-1-ona, podvrgnut Huang–Minlonovoj redukciji daje smešu (1S,1aR,6aR)-2’,3’,6 6a-tetrahidro-spiro[cikloprop[a]inden-1(1aH),1’-[1H]indena] i njegovog 1R,1aS,6aS enantiomera, a ne 2,3,1’,3’-tetrahidro-[1,2’]-biindeniliden kako se prvobitno očekivalo. Potpuni NMR raspored, konstante kuplovanja protona i HMBC i NOESY kuplovanje jedinjenja u naslovu dati su zbirno u tabeli