Crystal structure of a mammalian voltage-dependent Shaker family K+ channel

Voltage-dependent potassium ion (K+) channels (Kv channels) conduct K+ ions across the cell membrane in response to changes in the membrane voltage, thereby regulating neuronal excitability by modulating the shape and frequency of action potentials. Here we report the crystal structure, at a resolution of 2.9 angstroms, of a mammalian Kv channel, Kv1.2, which is a member of the Shaker K+ channel family. This structure is in complex with an oxido-reductase beta subunit of the kind that can regulate mammalian Kv channels in their native cell environment. The activation gate of the pore is open. Large side portals communicate between the pore and the cytoplasm. Electrostatic properties of the side portals and positions of the T1 domain and beta subunit are consistent with electrophysiological studies of inactivation gating and with the possibility of K+ channel regulation by the beta subunit.     

Diversity,Bioactivity Profiling and Untargeted Metabolomics of the Cultivable Gut Microbiota of Ciona intestinalis

It is widely accepted that the commensal gut microbiota contributes to the health and well-being of its host. The solitary tunicate Ciona intestinalis emerges as a model organism for studying host–microbe interactions taking place in the gut, however, the potential of its gut-associated microbiota for marine biodiscovery remains unexploited. In this study, we set out to investigate the diversity, chemical space, and pharmacological potential of the gut-associated microbiota of C. intestinalis collected from the Baltic and North Seas. In a culture-based approach, we isolated 61 bacterial and 40 fungal strains affiliated to 33 different microbial genera, indicating a rich and diverse gut microbiota dominated by Gammaproteobacteria. In vitro screening of the crude microbial extracts indicated their antibacterial (64% of extracts), anticancer (22%), and/or antifungal (11%) potential. Nine microbial crude extracts were prioritized for in-depth metabolome mining by a bioactivity- and chemical diversity-based selection procedure. UPLC-MS/MS-based metabolomics combining automated (feature-based molecular networking and in silico dereplication) and manual approaches significantly improved the annotation rates. A high chemical diversity was detected where peptides and polyketides were the predominant classes. Many compounds remained unknown, including two putatively novel lipopeptides produced by a Trichoderma sp. strain. This is the first study assessing the chemical and pharmacological profile of the cultivable gut microbiota of C. intestinalis.     

Omega-3 Fatty Acid Supplementation Reduces Basal TNFα but Not Toll-Like Receptor-Stimulated TNFα in Full-Sized and Miniature Mares

It has been well documented that omega-3 long-chain polyunsaturated fatty acids (n-3 PUFA) confer a wide variety of health benefits to humans and animals. The current study was designed to evaluate the ability of n-3 PUFA to modulate the innate immune response in two diverse breeds of horses. Ten Quarter Horse and 10 American Miniature Horse mares were assigned to either an n-3-PUFA-supplemented or a control diet (five full-sized and five miniature mares/treatment) for 56 days. The treatment diet was designed to deliver 64.4 mg/kg body weight of combined eicosapentaenoic acid and docosahexaenoic acid daily. Blood was collected through jugular venipuncture into heparinized tubes on days 0, 28, and 56. Serum PUFA analysis was conducted by gas chromatography. Peripheral blood mononuclear cell production of tumor necrosis factor-α (TNFα) in response to toll-like receptor (TLR) ligands lipopolysaccharide, flagellin, and lipoteichoic acid was estimated using an equine-specific enyme-linked immunosorbent assay. Peripheral blood samples from day 56 were also analyzed for total and differential leukocyte counts and subjected to flow cytometric analysis. Body type did not affect basal or TLR-stimulated TNFα production. Serum PUFA analysis revealed a decrease in α-linolenic acid and substantial increases in arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, and docosapentaenoic acid at both day 28 and day 56 in horses fed n-3 PUFA (P < .0001 for all). Dietary n-3 PUFA supplementation reduced (P < .05) unstimulated basal, but not TLR-stimulated, TNFα production by peripheral blood mononuclear cells. Supplementation with n-3 PUFA neither affected total or differential leukocyte counts nor selected cell surface markers. These results suggest that n-3 PUFA supplementation in the horse can modify circulating PUFA and alter the inflammatory response by reducing basal TNFα production. Furthermore, under conditions of the current study and considering the end points evaluated, the American Miniature Horse could potentially be used as a model for full-sized horse breeds.     

Omega-3 Fatty Acid Supplementation Affects Selected Phospholipids in Peripheral White Blood Cells and in Plasma of Full-Sized and Miniature Mares

Dietary omega-3 fatty acids (n-3 PUFA), notably eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), impart health benefits in humans and animals. In horses, dietary n-3 PUFAs elevate EPA and DHA and may promote anti-inflammatory effects. No reports document effects of dietary n-3 PUFA on fatty acyl components of circulating and cellular phospholipids in horses nor whether responses to dietary n-3 PUFA are similar among horse breeds. Ten Quarter Horse and 10 American Miniature Horse mares were assigned to n-3 PUFA (64.4 mg· kg body weight [BW]⁻¹·d⁻¹) or control diet for 56 days. Blood was sampled at 0, 28, and 56 days. Apparent phospholipid molecular species from several classes (phosphatidylcholine [PC]; “ether-linked” phosphatidylcholine [i.e., alk(en)yl, acyl glycerophosphocholine] [ePC]; phosphatidylethanolamine [PE]; phosphatidylinositol [PI]; and phosphatidylserine [PS]) were determined in plasma and peripheral blood mononuclear cells (PBMC) by mass spectrometry. Statistical analysis showed that six phospholipid species had diet × day interactions (P < .05) for both plasma and PBMC. Further evaluation of these species demonstrated that the mole percentage of PC(38:6), PC(40:7), PC(42:10), PE(38:5), PE(40:6), and PE(40:7) (where x:y represents total acyl carbon:total carbon-carbon double bonds) in both plasma and PBMC phospholipids was elevated in horses fed n-3 PUFA (P < .001 for all). Analysis of the acyl product ions revealed that these contained an acyl chain of mass consistent with an n-3 PUFA. Thus, supplementation increased n-3 PUFA in selected plasma and PBMC phospholipids. The absence of breed effects suggests that miniature and full-size horses responded similarly to dietary treatment.     

The evolution of maximum body size of terrestrial mammals

The extinction of dinosaurs at the Cretaceous/Paleogene (K/Pg) boundary was the seminal event that opened the door for the subsequent diversification of terrestrial mammals. Our compilation of maximum body size at the ordinal level by sub-epoch shows a near-exponential increase after the K/Pg. On each continent, the maximum size of mammals leveled off after 40 million years ago and thereafter remained approximately constant. There was remarkable congruence in the rate, trajectory, and upper limit across continents, orders, and trophic guilds, despite differences in geological and climatic history, turnover of lineages, and ecological variation. Our analysis suggests that although the primary driver for the evolution of giant mammals was diversification to fill ecological niches, environmental temperature and land area may have ultimately constrained the maximum size achieved.     

Metabolic interactions of agrochemicals in humans

Agrochemicals and other xenobiotics are metabolized by xenobiotic-metabolizing enzymes (XMEs) to products that may be more or less toxic than the parent chemical. In this regard, phase-I XMEs such as cytochrome P450s (CYPs) are of primary importance. Interactions at the level of metabolism may take place via either inhibition or induction of XMEs. Such interactions have often been investigated, in vitro, in experimental animals, using subcellular fractions such as liver microsomes, but seldom in humans or at the level of individual XME isoforms. The authors have been investigating the metabolism of a number of agrochemicals by human liver microsomes and recombinant CYP isoforms and have recently embarked on studies of the induction of XMEs in human hepatocytes. The insecticides chlorpyrifos, carbaryl, carbofuran and fipronil, as well as the repellant DEET, are all extensively metabolized by human liver microsomes and, although a number of CYP isoforms may be involved, CYP2B6 and CYP3A4 are usually the most important. Permethrin is hydrolyzed by esterase(s) present in both human liver microsomes and cytosol. A number of metabolic interactions have been observed. Chlorpyrifos and other phosphorothioates are potent inhibitors of the CYP-dependent metabolism of both endogenous substrates, such as testosterone and estradiol, and exogenous substrates, such as carbaryl, presumably as a result of the interaction of highly reactive sulfur, released during the oxidative desulfuration reaction, with the heme iron of CYP. The hydrolysis of permethrin in human liver can be inhibited by chlorpyrifos oxon and by carbaryl. Fipronil can inhibit testosterone metabolism by CYP3A4 and is an effective inducer of CYP isoforms in human hepatocytes.     

Are <Emphasis Type="Italic">Melilotus albus</Emphasis> and <Emphasis Type="Italic">M. officinalis</Emphasis> conspecific?

The principal economic species of the genus Melilotus are white sweet-clover (Melilotus albus) and the extremely similar yellow sweet-clover (M. officinalis). Although they are widely recognized as distinct species, some influential references in North America reduce the former to a subspecific rank or even merely a conspecific synonym of the latter. Given their importance and the large numbers of germplasm collections, the doubt needs to be resolved. This review of relevant published evidence finds that in addition to the difference in floral colour, the traditional segregation of the two as distinct species is best supported by very strong reproductive barriers as well as divergent DNA sequences in three barcoding genes. Additional but weaker confirmation of separateness is provided by studies reporting differences in external morphology, biochemistry, seed protein profiles, karyotype and DNA microsatellites.