Spectroscopic and Mechanistic Studies of Heterodimetallic Forms of Metallo-β-lactamase NDM-1

In an effort to characterize the roles of each metal ion in metallo-β-lactamase NDM-1, heterodimetallic analogues (CoCo-, ZnCo-, and CoCd-) of the enzyme were generated and characterized. UV–vis, 1H NMR, EPR, and EXAFS spectroscopies were used to confirm the fidelity of the metal substitutions, including the presence of a homogeneous, heterodimetallic cluster, with a single-atom bridge. This marks the first preparation of a metallo-β-lactamase selectively substituted with a paramagnetic metal ion, Co(II), either in the Zn1 (CoCd-NDM-1) or in the Zn2 site (ZnCo-NDM-1), as well as both (CoCo-NDM-1). We then used these metal-substituted forms of the enzyme to probe the reaction mechanism, using steady-state and stopped-flow kinetics, stopped-flow fluorescence, and rapid-freeze-quench EPR. Both metal sites show significant effects on the kinetic constants, and both paramagnetic variants (CoCd- and ZnCo-NDM-1) showed significant structural changes on reaction with substrate. These changes are discussed in terms of a minimal kinetic mechanism that incorporates all of the data

Lesões traumáticas do parênquima pulmonar: aspectos na tomografia computadorizada Traumatic lung lesions: computed tomography findings

As lesões pulmonares são achados freqüentes no trauma torácico, sendo cada vez mais diagnosticadas pela tomografia computadorizada, em especial pelo rápido tempo de aquisição decorrente da técnica helicoidal, que permite a avaliação de pacientes em estado grave, possibilitando a adoção de conduta terapêutica eficiente. Os autores estudaram 150 pacientes vítimas de trauma torácico submetidos a tomografia computadorizada, que apresentaram lesões pulmonares, representadas por contusões, atelectasias, lacerações e hematomas pulmonares. As contusões pulmonares se caracterizaram por consolidações e atenuação em vidro fosco, sendo as lesões pulmonares mais comuns. As atelectasias foram observadas com os padrões subsegmentar e compressiva, e foram a segunda lesão mais comum. As lacerações se apresentaram como consolidações com ar ou nível líquido no interior. Os hematomas pulmonares representaram a lesão pulmonar mais rara, presentes em apenas cinco casos, caracterizados por opacidades arredondadas. Neste trabalho o trauma torácico fechado predominou, com 120 casos, enquanto o trauma aberto ocorreu em 30 casos. As causas de trauma fechado, em ordem decrescente de freqüência, foram: colisão automobilística, atropelamento, queda de altura, acidente de motocicleta e espancamento. A forma penetrante de traumatismo torácico decorreu de duas causas de agressão: lesão por arma de fogo e lesão por arma branca.<br>Traumatic lesions of the lung are common findings in patients with thoracic trauma. These lesions are increasingly diagnosed using computed tomography, mostly due to the fast acquisition time helical techniques that allow evaluation of critically ill patients and an efficient therapeutic management. The authors studied 150 patients with thoracic trauma submitted to computed tomography that demonstrated lung contusions, atelectasies, lacerations and hematomas. Lung contusions were the most frequent lesions appearing as consolidation or ground-glass attenuation areas. Atelectasies, in subsegmentar and compressive patterns, were the second most common lesions observed. Lacerations appeared as consolidations with air or liquid level. Lung hematomas, characterized by round opacities, were the most rare lung lesions seen in only five cases. In this study, blunt thoracic trauma accounted for the majority of 120 cases whereas penetrating trauma occurred in 30 cases. The causes of blunt trauma in decrescent order of frequency were motor vehicle accidents, pedestrian hit by car, falls, motorcycle accidents and trashing. Penetrating traumas were caused by bullets or knives