RESUMO
The active site of type A or B influenza virus neuraminidase is composed of 11 conserved residues that directly interact with the substrate, sialic acid. An aromatic benzene ring has been used to replace the pyranose of sialic acid in our design of novel neuraminidase inhibitors. A bis(hydroxymethyl)pyrrolidinone ring was constructed in place of the N-acetyl group on the sialic acid. The hydroxymethyl groups replace two active site water molecules, which resulted in the high affinity of the nanomolar inhibitors. However, these inhibitors have greater potency for type A influenza virus than for type B influenza virus. To resolve the differences, we determined the X-ray crystal structure of three benzoic acid substituted inhibitors bound to the active site of B/Lee/40 neuraminidase. The investigation of a hydrophobic aliphatic group and a hydrophilic guanidino group on the aromatic inhibitors shows changes in the interaction with the active site residue Glu275. The results provide an explanation for the difference in efficacy of these inhibitors against types A and B viruses, even though the 11 active site residues of the neuraminidase are conserved.
Assuntos
Sequência Conservada , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Vírus da Influenza B/enzimologia , Neuraminidase/antagonistas & inibidores , Água/metabolismo , Ácido Benzoico/química , Ácido Benzoico/metabolismo , Sítios de Ligação , Cristalização , Cristalografia por Raios X , Desenho de Fármacos , Elétrons , Ligação de Hidrogênio , Vírus da Influenza A/enzimologia , Concentração Inibidora 50 , Modelos Moleculares , Conformação Molecular , Dados de Sequência Molecular , Ácido N-Acetilneuramínico/análogos & derivados , Ácido N-Acetilneuramínico/química , Ácido N-Acetilneuramínico/metabolismo , Neuraminidase/química , Neuraminidase/metabolismo , Relação Estrutura-AtividadeRESUMO
To obtain a better understanding of the behavior of "non-professional" costs in a medical practice, the authors analyzed the expenses of a 19-doctor practice. The analysis revealed that 80 percent of these expenses were fixed costs. Fixed costs, as opposed to variable costs, remain static in total but vary on a per unit basis as volume changes. Organizations with high fixed cost must maximize capacity to achieve profitability. Thus, the relationship among volume, capacity, cost and profit must be understood by medical practices negotiating rates for service units.
Assuntos
Contabilidade/métodos , Alocação de Custos/métodos , Prática de Grupo/economia , Benchmarking/economia , Programas de Assistência Gerenciada/economia , Escalas de Valor Relativo , Salários e Benefícios , Estados UnidosRESUMO
The X-ray crystal structure of the DNA decamer d(GACCGCGGTC), containing half the human papilloma virus E2 binding site, has been solved from two crystals grown at different ionic conditions (50 mM MgCl2and 50 mM spermine or 1.56 mM MgCl2and 1.56 mM spermine). Despite the variation in salt concentration, the two DNA structures are in a very similar, A-type DNA conformation, with helical axes curving towards the major groove. Although the salt concentrations do not effect the helical parameters or hydration to a large degree, there is a change in the overall helical curvature; 18 degrees and 31 degrees for the low and high salt structures, respectively. This curvature appears to be sequence specific and biologically relevant when compared with similar DNA structures, including the E2 binding site of a protein-DNA complex.