Algoriphagus machipongonensis sp. nov., co-isolated with a colonial choanoflagellate

A Gram-negative, non-motile, non-spore-forming bacterial strain, PR1[superscript T], was isolated from a mud core sample containing colonial choanoflagellates near Hog Island, Virginia, USA. Strain PR1[superscript T] grew optimally at 30 °C and with 3 % (w/v) NaCl. Strain PR1[superscript T] contained MK-7 as the major menaquinone as well as carotenoids but lacked pigments of the flexirubin-type. The predominant fatty acids were iso-C15 : 0 (29.4 %), iso-C17 : 1ω9c (18.5 %) and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c; 11.3 %). The major polar lipids detected in strain PR1[superscript T] were phosphatidylethanolamine, an unknown phospholipid, an aminophospholipid, an aminolipid and two lipids of unknown character. The DNA G+C content was 38.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain PR1[superscript T] fell within the cluster comprising the genus Algoriphagus and was most closely related to Algoriphagus halophilus JC 2051[superscript T] (95.4 % sequence similarity) and Algoriphagus lutimaris S1-3[superscript T] (95.3 % sequence similarity). The 16S rRNA gene sequence similarity between strain PR1[superscript T] and the type strains of other species of the genus Algoriphagus were in the range 91–95 %. Differential phenotypic properties and phylogenetic and genetic distinctiveness of strain PR1[superscript T] demonstrated that this strain was distinct from other members of the genus Algoriphagus, including its closest relative, A. halophilus. Based on phenotypic, chemotaxonomic, phylogenetic and genomic data, strain PR1[superscript T] should be placed in the genus Algoriphagus as a representative of a novel species, for which the name Algoriphagus machipongonensis sp. nov. is proposed. The type strain is PR1[superscript T] ( = ATCC BAA-2233[superscript T]  = DSM 24695[superscript T]).Gordon and Betty Moore Foundation (Investigator Award (581))National Institutes of Health (U.S.) (NIH National Research Service Award and Fellowship grant (5F32GM086054))United States. National Aeronautics and Space Administration (NASA Astrobiology Institute (NNA08CN84A

Opportunities for improving animal welfare in rodent models of epilepsy and seizures

Animal models of epilepsy and seizures, mostly involving mice and rats, are used to understand the pathophysiology of the different forms of epilepsy and their comorbidities, to identify biomarkers, and to discover new antiepileptic drugs and treatments for comorbidities. Such models represent an important area for application of the 3Rs (replacement, reduction and refinement of animal use). This report provides background information and recommendations aimed at minimising pain, suffering and distress in rodent models of epilepsy and seizures in order to improve animal welfare and optimise the quality of studies in this area. The report includes practical guidance on principles of choosing a model, induction procedures, in vivo recordings, perioperative care, welfare assessment, humane endpoints, social housing, environmental enrichment, reporting of studies and data sharing. In addition, some model-specific welfare considerations are discussed, and data gaps and areas for further research are identified. The guidance is based upon a systematic review of the scientific literature, survey of the international epilepsy research community, consultation with veterinarians and animal care and welfare officers, and the expert opinion and practical experience of the members of a Working Group convened by the United Kingdom's National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs)

GABAA receptor subtype involvement in addictive behaviour

GABAA receptors form the major class of inhibitory neurotransmitter receptors in the mammalian brain. This review sets out to summarise the evidence that variations in genes encoding GABAA receptor isoforms are associated with aspects of addictive behaviour in humans, while animal models of addictive behaviour also implicate certain subtypes of GABAA receptor. In addition to outlining the evidence for the involvement of specific subtypes in addiction, we summarise the particular contributions of these isoforms in control over the functioning of brain circuits, especially the mesolimbic system, and make a first attempt to bring together evidence from several fields to understanding potential involvement of GABAA Receptor Subtypes in addictive behaviour. While the weight of the published literature is on alcohol dependency, the underlying principles outlined are relevant across a number of different aspects of addictive behaviour

Cooperativity and communication in archaeal Cdc48·20S, an ancient proteolytic machine

Thesis: Ph. D., Massachusetts Institute of Technology, Department of Earth, Atmospheric, and Planetary Sciences, 2015."September 2015." Cataloged from PDF version of thesis.Includes bibliographical references.ATP dependent proteolysis is a process essential for life and is carried out by AAA+ proteases. AAA+ unfoldases use the energy of ATP hydrolysis to power the unfolding and translocation of protein substrates into compartmentalized peptidases for regulated proteolysis. Cdc48 is a highly conserved AAA+ homohexameric unfoldase which is made up of two AAA+ rings. Each ring can, in principle, bind and hydrolyzing ATP, but it is unclear what roles are played by each ring and how they coordinate their activities. A regulatory N domain functions to control the activity of the enzyme and binding to its partner peptidase, the 20S proteasome. In this thesis I present experiments which investigate the role of inter-ring communication in ATP hydrolysis, protein unfolding, and allosteric interactions with the 20S and show how these features affect enzyme function. Experiments also show how the N domain controls D1-D2 interactions that govern ATP hydrolysis and substrate unfolding. Finally, I present experiments that take steps toward developing a system for screening protein substrates of Cdc48-20S and identify several substrates from E. coli lysates.by Jonathan Dean Grabenstatter.Ph. D