A metallo-beta-lactamase enzyme in action: crystal structures of the monozinc carbapenemase CphA and its complex with biapenem

One strategy developed by bacteria to resist the action of beta-lactam antibiotics is the expression of metallo-beta-lactamases. CphA from Aeromonas hydrophila is a member of a clinically important subclass of metallo-beta-lactamases that have only one zinc ion in their active site and for which no structure is available. The crystal structures of wild-type CphA and its N220G mutant show the structural features of the active site of this enzyme, which is modeled specifically for carbapenem hydrolysis. The structure of CphA after reaction with a carbapenem substrate, biapenem, reveals that the enzyme traps a reaction intermediate in the active site. These three X-ray structures have allowed us to propose how the enzyme recognizes carbapenems and suggest a mechanistic pathway for hydrolysis of the beta-lactam. This will be relevant for the design of metallo-beta-lactamase inhibitors as well as of antibiotics that escape their hydrolytic activity.     

Differences between measured and resultant joint moments during isometric contractions at the ankle joint

The purpose of this study was to examine two hypotheses: (a) during voluntary and electrically induced isometric contractions the moments measured at the dynamometer are different from the resultant moments in the same plane around the ankle joint and (b) at a given resultant moment during electrically induced isometric contractions the ankle angle while loading is different from the ankle angle while unloading. Twenty-seven long distance runners participated in the study. All subjects performed isometric maximal voluntary contractions (MVC) and contractions induced by electrostimulation at four different ankle-knee angle combinations on a Biodex-dynamometer. The kinematics of the leg were recorded using the vicon 624 system with eight cameras operating at 120 Hz. The main findings were: (a) the resultant moment at the ankle joint and the moment measured by the Biodex-dynamometer during isometric contractions are different, (b) during a plantar flexion effort the ankle angle changes significantly, whereas the knee angle shows only small and in most cases not significant changes, and (c) at identical resultant ankle joint moments the ankle angles are different between the loading and the unloading phases. The observed differences may lead to erroneous conclusions concerning the following: (a) diagnostic of muscle architecture, (b) estimation of the moment-ankle angle relationship and (c) estimation of the strain and hysteresis of tendons and aponeuroses.     

Determination of amphetamine-derived designer drugs in human urine by SPE extraction and capillary electrophoresis with mass spectrometry detection

In recent years, a number of newer designer drugs have entered the illicit drug market. The methylenedioxy-derivates of amphetamine represent the largest group of designer drugs. This paper describes a method for screening for and quantification of ten 2,5-methylenedioxy-derivates of amphetamine and phenylethylamine in human urine, using capillary electrophoresis coupled to electrospray ionisation-mass spectrometry (CE-ESI-MS). Prior to CE-MS analysis, a simple solid-phase extraction (SPE) was used for sample cleanup. The method was validates according to international guidelines.     

First record of the genus Jenynsia from marine water on the coast of Punta del Este,Maldonado, Uruguay (Cyprinodontiformes: Anablepidae)

The first report of the genus Jenynsia occurring in marine waters is presented here. The evolution of high salinity tolerance within Anablepidae is discussed and a hypothesis is proposed that this characteristic is a plesiomorphic trait in Jenynsia which was probably present in the common ancestor of the Anablepidae.     

Fluorine nuclear magnetic resonance-based assay in living mammalian cells

Nuclear magnetic resonance (NMR)-based screening has been recognized as a powerful approach for the identification and characterization of molecules interacting with pharmaceutical targets. Indeed, several NMR methods have been developed and successfully applied to many drug discovery projects. Whereas most of these approaches have targeted isolated biomolecular receptors, very few cases are reported with the screening performed in intact cells and cell extracts. Here we report the first successful application of the fluorine NMR-based assay n-FABS (n-fluorine atoms for biochemical screening) in living mammalian cells expressing the membrane protein fatty acid amide hydrolase (FAAH). This method allows the identification of both weak and potent inhibitors and the measurement of their potency in a physiological environment.     

Different cell cycle modifications repress apoptosis at different steps independent of developmental signaling in Drosophila

Apoptotic cell death is important for the normal development of a variety of organisms. Apoptosis is also a response to DNA damage and an important barrier to oncogenesis. The apoptotic response to DNA damage is dampened in specific cell types during development. Developmental signaling pathways can repress apoptosis, and reduced cell proliferation also correlates with a lower apoptotic response. However, because developmental signaling regulates both cell proliferation and apoptosis, the relative contribution of cell division to the apoptotic response has been hard to discern in vivo. Here we use Drosophila oogenesis as an in vivo model system to determine the extent to which cell proliferation influences the apoptotic response to DNA damage. We find that different types of cell cycle modifications are sufficient to repress the apoptotic response to ionizing radiation independent of developmental signaling. The step(s) at which the apoptosis pathway was repressed depended on the type of cell cycle modification—either upstream or downstream of expression of the p53-regulated proapoptotic genes. Our findings have important implications for understanding the coordination of cell proliferation with the apoptotic response in development and disease, including cancer and the tissue-specific responses to radiation therapy.