Simultaneous determination of gemcitabine and gemcitabine-squalene by liquid chromatography-tandem mass spectrometry in human plasma

Gemcitabine-squalene is a new prodrug that self-organizes in water forming nanoassemblies. It exhibits better anti-cancer properties in vitro and in vivo than gemcitabine. A liquid chromatography/tandem mass spectrometry assay of gemcitabine-squalene and gemcitabine was developed in human plasma in order to quantitate gemcitabine and its squalene conjugate. After protein precipitation with acetonitrile/methanol (90/10, v/v), the compounds were analyzed by reversed-phase high performance liquid chromatography and detected by tandem mass spectrometry using multiple reaction monitoring. The method was linear over the concentration range of 10-10,000 ng/ml of human plasma for both compounds with an accuracy lower than 10.4% and a precision below 14.8%. The method showed a lower limit of quantitation of 10 ng/ml of human plasma for dFdC and dFdC-SQ. A preliminary in vivo study in mice was shown as application of the method as no significant difference between human and mice plasma for the analysis of dFdC and dFdC-SQ was demonstrated.     

High resolution crystal structures of Mycobacterium tuberculosis adenosine kinase: insights into the mechanism and specificity of this novel prokaryotic enzyme

Adenosine kinase (ADK) catalyzes the phosphorylation of adenosine (Ado) to adenosine monophosphate (AMP). It is part of the purine salvage pathway that has been identified only in eukaryotes, with the single exception of Mycobacterium spp. Whereas it is not clear if Mycobacterium tuberculosis (Mtb) ADK is essential, it has been shown that the enzyme can selectively phosphorylate nucleoside analogs to produce products toxic to the cell. We have determined the crystal structure of Mtb ADK unliganded as well as ligand (Ado) bound at 1.5- and 1.9-A resolution, respectively. The structure of the binary complexes with the inhibitor 2-fluoroadenosine (F-Ado) bound and with the adenosine 5'-(beta,gamma-methylene)triphosphate (AMP-PCP) (non-hydrolyzable ATP analog) bound were also solved at 1.9-A resolution. These four structures indicate that Mtb ADK is a dimer formed by an extended beta sheet. The active site of the unliganded ADK is in an open conformation, and upon Ado binding a lid domain of the protein undergoes a large conformation change to close the active site. In the closed conformation, the lid forms direct interactions with the substrate and residues of the active site. Interestingly, AMP-PCP binding alone was not sufficient to produce the closed state of the enzyme. The binding mode of F-Ado was characterized to illustrate the role of additional non-bonding interactions in Mtb ADK compared with human ADK.     

Researching little-known species: the African bat Otomops martiensseni (Chiroptera: Molossidae)

Listed as 'vulnerable' by the International Union for theConservation of Nature, the molossid bat Otomopsmartiensseni occurs widely in Africa and, according to someauthorities, in Madagascar. Apart from a few known cave roosts, there are fewrecords of O. martiensseni, although around Durban, SouthAfrica, the species is common and roosts in buildings. Originally described asthree species, populations of O. martiensseni differsignificantly in size (length of forearm) between East Africa and Durban orMadagascar, but not between Durban and Madagascar. Seventeen buildings used asroosts by O. martiensseni averaged 34.5 ± 15.8years old. In the Durban area, bats entered roosts by landing andcrawling. Roost populations ranged from 7 to 29 individuals, typicallyconsisting of one adult male, several adult females and young (bats withunossified epiphyses), suggesting a harem social structure. The ratio of adultfemales to young was virtually 1:1, and among young the ratio of males:femalesaveraged 2:1. Radio-tracking showed that individuals used several day and nightroosts, and foraged widely in a landscape dominated by sugarcane and urbandevelopment. The echolocation and many social calls of O.martiensseni are readily audible to human observers, allowing anon-contact, low technology method for monitoring the local distribution andactivity of these bats. Although listed as a species of special concern inKwaZulu Natal, there these bats appear to be candidates for inclusion on a'blue' list of species, ones showing stabilized or increasedabundance. We recommend that O. martiensseni be recognizedas a 'flagship' species in the Durban area, r epresenting theresilience of nature.