New Aminocoumarin Antibiotics Formed by a Combined Mutational and Chemoenzymatic Approach Utilizing the Carbamoyltransferase NovN

AbstractFive new aminocoumarin antibiotics were produced by a combined mutational and chemoenzymatic approach. For this purpose, the 3″-carbamoyltransferase NovN from the novobiocin producer Streptomyces spheroides was overexpressed in the heterologous host S. lividans as an N-terminal His6 fusion protein and purified by nickel affinity chromatography. Five different 3″-unsubstituted aminocoumarin derivatives were isolated from mutants of the clorobiocin producer S. roseochromogenes, carrying single or multiple gene defects. All five compounds were readily accepted as substrates by NovN, and the 3″-carbamoylated products were isolated on a preparative scale. Their structures were elucidated by 1H-NMR and mass spectroscopy, and their inhibitory activity on gyrase in vitro as well as their antibacterial activity was determined. The results give further insight into the structure-activity relationships of aminocoumarin antibiotics

Evolutionary Relationships of Microbial Aromatic Prenyltransferases

The linkage of isoprenoid and aromatic moieties, catalyzed by aromatic prenyltransferases (PTases), leads to an impressive diversity of primary and secondary metabolites, including important pharmaceuticals and toxins. A few years ago, a hydroxynaphthalene PTase, NphB, featuring a novel ten-stranded β-barrel fold was identified in Streptomyces sp. strain CL190. This fold, termed the PT-barrel, is formed of five tandem ααββ structural repeats and remained exclusive to the NphB family until its recent discovery in the DMATS family of indole PTases. Members of these two families exist only in fungi and bacteria, and all of them appear to catalyze the prenylation of aromatic substrates involved in secondary metabolism. Sequence comparisons using PSI-BLAST do not yield matches between these two families, suggesting that they may have converged upon the same fold independently. However, we now provide evidence for a common ancestry for the NphB and DMATS families of PTases. We also identify sequence repeats that coincide with the structural repeats in proteins belonging to these two families. Therefore we propose that the PT-barrel arose by amplification of an ancestral ααββ module. In view of their homology and their similarities in structure and function, we propose to group the NphB and DMATS families together into a single superfamily, the PT-barrel superfamily

Out of the boxes, out of the silos: The need of interdisciplinary collaboration to reduce poor-quality medical products in the supply chain

In this paper, we argue that understanding and addressing the problem of poor-quality medical products requires a more interdisciplinary approach than has been evident to date. While prospective studies based on rigorous standardized methodologies are the gold standard for measuring the prevalence of poor-quality medical products and understanding their distribution nationally and internationally, they should be complemented by social science research to unpack the complex set of social, economic, and governance factors that underlie these patterns. In the following sections, we discuss specific examples of prospective quality surveys and of social science studies, highlighting the value of cross-sector partnerships in driving high-quality, policy-relevant research in this area

Lost Highway Not Forgotten: Satellite Tracking of a Bowhead Whale (Balaena mysticetus) from the Critically Endangered Spitsbergen Stock

The Spitsbergen bowhead whale stock is critically endangered. It is believed to number in the tens. Here we report results from the first satellite transmitter ever deployed on an individual from this stock. A female whale was tagged on 3 April 2010 (at 79˚54' N, 01˚03' E), but no locations were transmitted by the tag until 30 April 2010, after which data were received continuously for 86 days. Additionally, three small clusters of locations were transmitted later in the year; the latest was received 20 December 2010 (262 days after deployment). During the 86 days of continuous tracking, the whale initially remained in the middle of the Fram Strait, between 77˚45' N, 5˚ W and 80˚10' N, 5˚ E. For a two-week period starting around 10 June 2010, the whale traveled southwest down to 73˚40' N (at least 950 km). Subsequently it remained at southern latitudes between ~70˚ and 73˚ N until the tag stopped continuous transmissions on 24 July. Movement patterns analyzed using first-passage times (FTP), fitted as functions of various environmental variables using Cox Proportional Hazards models, showed that the whale spent most of its time in waters close to the ice edge with modest ice coverage, over areas where the bottom slope was relatively steep. Winter positions (27 November – 20 December 2010) revealed that the whale was back in the North at about 80˚ N. This information, in combination with recent data from passive acoustic listening devices, suggests that the Spitsbergen bowhead stock overwinters at high-latitude locations. The north-south movements of this whale during summer are consistent with the patterns that early whalers described for bowhead whales in this region in the 16th and 17th centuries.La population de baleines boréales de Spitzberg est en danger critique d’extinction. L’on croit qu’elle se chiffrerait dans la dizaine. Ici, nous faisons état des résultats obtenus à l’aide du premier émetteur satellite à n’avoir jamais été installé sur un individu de cette population. Une baleine femelle a été marquée le 3 avril 2010 (à 79˚54' N, 01˚03' E), mais aucun signal n’a été transmis par ce marquage avant le 30 avril 2010, après quoi nous avons reçu des données continuelles pendant 86 jours. Plus tard dans le courant de l’année, nous avons également reçu trois petits blocs d’information, dont le dernier a été transmis le 20 décembre 2010 (262 jours après la date du marquage). Au cours des 86 jours d’information continuelle, la baleine restait d’abord au milieu du détroit de Fram, entre 77˚45' N, 5˚ O et 80˚10' N, 5˚ E. Pendant une période de deux semaines commençant vers le 10 juin 2010, la baleine s’est déplacée vers le sud-ouest jusqu’à 73˚40' N (au moins 950 km). Par la suite, elle est restée dans les latitudes du sud entre ~70˚ et 73˚ N jusqu’à ce que le marquage cesse les transmissions continuelles le 24 juillet. Les habitudes de déplacement analysées en recourant aux temps du premier passage (FTP), ajustées à titre de fonctions de diverses variables environnementales s’appuyant sur les modèles des hasards proportionnels de Cox, ont laissé entrevoir que la baleine passait la plus grande partie de son temps dans les eaux à proximité des lisières de glace dont la couverture était modeste par rapport aux endroits où la pente du fond était relativement abrupte. Les positions enregistrées en hiver (du 27 novembre au20 décembre 2010) ont révélé que la baleine était retournée dans le nord à environ 80˚ N. Cette information, alliée aux récentes données provenant d’appareils d’écoute acoustique, suggère que la population de baleines boréales de Spitzberg passe l’hiver à de hautes latitudes. Pendant l’été, les mouvements nord-sud de cette baleine sont conformes aux habitudes de déplacement de la baleine boréale, telles que décrites par les anciens baleiniers dans cette région au cours des XVIe et XVIIe siècles

Mitogenomics and the genetic differentiation of contemporary <i>Balaena mysticetus</i> (Cetacea) from Svalbard

Full mitochondrial genomes were assembled for 12 recently sampled animals from the Svalbard bowhead whale (Balaena mysticetus) stock via high-throughput sequencing data, facilitating analysis of the demographic history of the population for the first time. The Svalbard population has retained noticeable amounts of mitochondrial genome diversity despite extreme historical harvest levels. Haplotype and nucleotide diversities were similar to those estimated earlier for other bowhead whale populations. The reconstructed demographic history was in accordance with a boom–bust scenario, combining a slight Pleistocene population growth 25 000–35 000 years ago and a Holocene decline. Employing a mutation rate of 3.418 × 10–8 substitutions per site per year, the time to the most recent common ancestor for the mitochondrial genomes of the contemporary Svalbard bowhead whales was estimated to be 68 782 (54 353–83 216) years before the present. Based on 370 bp fragments of the D-loop region, significant genetic differentiation was detected between all extant bowhead whale populations across the circumpolar Arctic. Thus, the Svalbard bowhead whales can be regarded as a population with its own genetic legacy

Active Cytomegalovirus Infection in Patients with Septic Shock

Cytomegalovirus reactivation occurred in one third of patients and was associated with prolonged ventilation and stay in an intensive care unit

Prophylactic Palmitoylethanolamide Prolongs Survival and Decreases Detrimental Inflammation in Aged Mice With Bacterial Meningitis

Easy-to-achieve interventions to promote healthy longevity are desired to diminish the incidence and severity of infections, as well as associated disability upon recovery. The dietary supplement palmitoylethanolamide (PEA) exerts anti-inflammatory and neuroprotective properties. Here, we investigated the effect of prophylactic PEA on the early immune response, clinical course, and survival of old mice after intracerebral E. coli K1 infection. Nineteen-month-old wild type mice were treated intraperitoneally with two doses of either 0.1 mg PEA/kg in 250 μl vehicle solution (n = 19) or with 250 μl vehicle solution only as controls (n = 19), 12 h and 30 min prior to intracerebral E. coli K1 infection. The intraperitoneal route was chosen to reduce distress in mice and to ensure exact dosing. Survival time, bacterial loads in cerebellum, blood, spleen, liver, and microglia counts and activation scores in the brain were evaluated. We measured the levels of IL-1β, IL-6, MIP-1α, and CXCL1 in cerebellum and spleen, as well as of bioactive lipids in serum in PEA- and vehicle-treated animals 24 h after infection. In the absence of antibiotic therapy, the median survival time of PEA-pre-treated infected mice was prolonged by 18 h compared to mice of the vehicle-pre-treated infected group (P = 0.031). PEA prophylaxis delayed the onset of clinical symptoms (P = 0.037). This protective effect was associated with lower bacterial loads in the spleen, liver, and blood compared to those of vehicle-injected animals (P ≤ 0.037). PEA-pre-treated animals showed diminished levels of pro-inflammatory cytokines and chemokines in spleen 24 h after infection, as well as reduced serum concentrations of arachidonic acid and of one of its metabolites, 20-hydroxyeicosatetraenoic acid. In the brain, prophylactic PEA tended to reduce bacterial titers and attenuated microglial activation in aged infected animals (P = 0.042). Our findings suggest that prophylactic PEA can counteract infection associated detrimental responses in old animals. Accordingly, PEA treatment slowed the onset of infection symptoms and prolonged the survival of old infected mice. In a clinical setting, prophylactic administration of PEA might extend the potential therapeutic window where antibiotic therapy can be initiated to rescue elderly patients

Universal baleen whale microsatellite panel for individual identification and power to detect parentage

Highly polymorphic single tandem repeat loci (STR, also known as microsatellite loci) remain a familiar, cost efficient class of markers for genetic analyses in ecology, behavior and conservation. We characterize a new universal set of ten STR loci (from 28 potential candidate loci) in seven baleen whale species, which are optimized for PCR amplification in two multiplex reactions along with a Y chromosome marker for sex determination. The optimized, universal set of STR loci provides an ideal starting point for new studies in baleen whales aimed at individual-based and population genetic studies, and facilitates data sharing among research groups. Data from the new STR loci were combined with genotypes from other published STR loci to assess the power to assign parentage (paternity) using exclusion in four species: fin whales, humpback whales, blue whales and bowhead whales. We argue that parentage studies should present a power analysis to demonstrate that the specific data are sufficiently informative to assign parentage with statistical rigor