Nature contents: 13 March 2014

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Volume 507 Number 7491      nature •Visit Nature homepage •Subscribe to Nature •View Table of Contents The science that matters. Every week.               Sources of Error in Next-Gen Sequencing (NGS)  Learn how to identify and mitigate error in NGS projects. This article discusses the sources of experimental error in NGS and presents a simple analysis for characterizing and abating error. The author, Kimberly Robasky, formerly a student of George Church, is currently a Bioinformatics Director at EA|Quintiles. Read More...       Jump to the content that matters to you View Table of Contents ▶ ▼ News & Comment   ▼ Biological Sciences   ▼ Health Sciences   ▼ Physical Sciences   ▼ Earth & Environmental Sciences   ▼ Careers & Jobs       This week's highlights     Physical Sciences More Physical sciences ▼   Reflection from the strong gravity regime in a lensed quasar at redshift z=0.658   Measurements of the spin of a supermassive black hole more than 6 billion light years away - the most distant black hole for which a spin has been measured - offer new clues as to how these objects grow. The reflection-dominated spectrum of a 'lensed' quasar at z=0.658, together with archival X-ray data, show that it is rotating rapidly and that much of its radiation comes from a compact region close to the black hole. The data suggest that the black hole has grown by coherent accretion rather than in a chaotic manner.       Earth & Environmental Sciences More Earth & Environmental sciences ▼   Hydrous mantle transition zone indicated by ringwoodite included within diamond   It is not clear just how much water there is within the solid Earth, or where it is to be found. Graham Pearson and co-authors present evidence from a diamond inclusion from Juína, Brazil, for the first known terrestrial occurrence of ringwoodite - a high-pressure form of olivine mineral first identified in meteorites and thought to be a major constituent of the Earth's mantle transition zone. The water-rich nature of this inclusion provides direct evidence that, at least locally, the transition zone is hydrous, to about one weight per cent.       Biological Sciences More Biological sciences ▼   Derived immune and ancestral pigmentation alleles in a 7,000-year-old Mesolithic European   The emergence of agriculture may have caused many of the changes in human physiology evident in the fossil record. Which changes it is hard to say in the absence of a baseline - a record of human physiology just before the advent of farming. We may now have that in the form of genome of a Mesolithic hunter-gatherer from Spain. The genes of this male, alive around 7,000 years ago, had more in common with ancient genomes from Siberia than with other Europeans, suggesting a wide genetic continuity across Eurasia. He was probably lactose intolerant and digested starchy food less effectively than did Neolithic farmers, supporting the idea that these changes came in with agriculture. And he had the unusual combination of dark skin and blue eyes, suggesting that in the Mesolithic, the transition to a lighter 'modern European' skin tone was incomplete and that eye-colour changes came first.                 Podcast & Video     In this week's podcast: This week, how a thirteenth century cosmologist predicted the Big Bang, the ocean of water hidden deep in the Earth's mantle, and what the wolves of Yellowstone tell us about ecosystems.         News & Comment Read daily news coverage▶ ▲top               THIS WEEK           Editorials                   Don’t hide the decline ▶     US scientists should not be placated by the ‘flat budget’ myth. Funds are decreasing, and the situation will get worse.             An elegant chaos ▶     Universal theories are few and far between in ecology, but that is what makes it fascinating.             Share alike ▶     Research communities need to agree on standard etiquette for data-sharing.             World View                   Practical science has a global reach and appeal ▶     As English schools consider downgrading practical science, John Baruch points out that other nations are rushing to include more.             Seven Days                   7 days: 7–13 March 2014 ▶     The week in science: Infant HIV cure; Craig Venter launches genomics company; and asteroid caught falling apart.             NEWS IN FOCUS           Obama’s budget request falls flat ▶     Hopes dim for a science-funding increase in 2015.             China goes back to basics on research funding ▶     Core science gets budget boost in a bid to change research culture and increase innovation.             Storm brewing over WHO sugar proposal ▶     Industry backlash expected over suggested cut in intake.             Global seismic network takes to the seas ▶     Two systems could plug the ocean-sized gap in earthquake detection.             Cellulosic ethanol fights for life ▶     Pioneering biofuel producers hope that US government largesse will ease their way into a tough market.             Features                   Energy: Islands of light ▶     More than a billion people lack electricity, but now microgrids are powering up rural areas.             Rethinking predators: Legend of the wolf ▶     Predators are supposed to exert strong control over ecosystems, but nature doesn’t always play by the rules.             COMMENT           History: A medieval multiverse ▶     Ideas in a thirteenth-century treatise on the nature of matter still resonate today, say Tom C. B. McLeish and colleagues.             Policy: The art of science advice to government ▶     Peter Gluckman, New Zealand's chief science adviser, offers his ten principles for building trust, influence, engagement and independence.             Books and Arts                   In retrospect: Sylva ▶     Gabriel Hemery celebrates the 350th anniversary of John Evelyn's treatise on the science and practice of forestry.             Particle physics: Higgs on the big screen ▶     Alexandra Witze savours a behind-the-scenes look at physics's most famous arrival.             Correspondence                   Climate change: China must publicize its emissions reports Yuan-Feng Wang, Yu-Rong Zhang | Statistics: Biomedicine must look beyond P values Anne-Louise Ponsonby, Terence Dwyer | Neutron sources: Road maps of no use to some physicists Aleksandar Matic, Peter Böni, Adrian Goldman | Politics: End education meddling in Nepal Kosh P. Neupane | Technology: Fight floods on a global scale Guy J.-P. Schumann, Paul D. Bates, Jeffrey C. Neal et al.                 The New York Stem Cell Foundation - Robertson Investigator Awards in Translational Stem Cell Research.    The New York Stem Cell Foundation is accepting proposals for high-risk, high-reward projects exploring the therapeutic potential of stem cells. The awards provide $1.5M USD over 5 years. Application deadline: March 17, 2014. View the full RFA at www.nyscf.org/stemcell         Biological Sciences ▲top             RESEARCH           Latest Online                   Bone biology: Vessels of rejuvenation ▶     Ferdinand le Noble, Jos le Noble             Genetics: Closing the distance on obesity culprits ▶     David U. Gorkin, Bing Ren             Cell biology: The disassembly of death ▶     Christopher D. Gregory             Haematopoietic stem cells require a highly regulated protein synthesis rate ▶     Robert A. J. Signer, Jeffrey A. Magee, Adrian Salic et al.     Haematopoietic stem cells (HSCs) have a lower rate of protein synthesis in vivo than most other haematopoietic cells, and both increases and decreases in the rate of protein synthesis impair HSC function, demonstrating that HSC maintenance—and hence, cellular homeostasis—requires the rate of protein synthesis to be highly regulated.             Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone ▶     Anjali P. Kusumbe, Saravana K. Ramasamy, Ralf H. Adams     Bone homeostasis and repair declines with ageing and the mechanisms regulating the relationship between bone growth and blood vessel formation have remained unknown; this mouse study identifies the endothelial cells that promote the formation of new bone, a small microvessel subtype that can be identified by high CD31 and high Emcn expression.             Unexpected link between an antibiotic, pannexin channels and apoptosis ▶     Ivan K. H. Poon, Yu-Hsin Chiu, Allison J. Armstrong et al.     The pannexin 1 channel on the plasma membrane of apoptotic cells mediates the release of find-me molecular signals to attract phagocytic cells for clearance of the apoptotic cells; here the quinolone antibiotic trovafloxacin is identified as a direct inhibitor of pannexin 1, which results in dysregulated fragmentation of apoptotic cells and may partly explain quinolone toxicity.             Inhibition of miR-25 improves cardiac contractility in the failing heart ▶     Christine Wahlquist, Dongtak Jeong, Agustin Rojas-Muñoz et al.     Reduced activity of the calcium-transporting ATPase SERCA2a is a hallmark of heart failure; here, microRNAs that downregulate SERCA2a function are identified, and antagonism of one, miR-25, is shown to halt heart failure in mice.             Cell-cycle-regulated activation of Akt kinase by phosphorylation at its carboxyl terminus ▶     Pengda Liu, Michael Begley, Wojciech Michowski et al.     Phosphorylation of Akt at its carboxy-terminal tail is an essential layer of Akt activation to regulate its physiological functions.             Structure of a type IV secretion system ▶     Harry H. Low, Francesca Gubellini, Angel Rivera-Calzada et al.     The three-dimensional structure of the type IV secretion system encoded by the Escherichia coli R388 conjugative plasmid.             A new fossil species supports an early origin for toothed whale echolocation ▶     Jonathan H. Geisler, Matthew W. Colbert, James L. Carew     Phylogenetic analysis of a new species of fossil toothed whale, Cotylocara macei, from the Oligocene epoch places it in a basal clade of odontocetes, and its features suggest that rudimentary echolocation evolved in the early Oligocene and was followed by convergent evolution in their skulls.             Obesity-associated variants within FTO form long-range functional connections with IRX3 ▶     Scott Smemo, Juan J. Tena, Kyoung-Han Kim et al.     Obesity-associated noncoding sequences within FTO are functionally connected with IRX3, and long-range enhancers in this region recapitulate aspects of IRX3 expression, suggesting that the obesity-associated interval is part of IRX3 regulation; Irx3-deficient mice have lower body weight and are resistant to diet-induced obesity, suggesting IRX3 as a novel determinant of body mass and composition.             Herbivores and nutrients control grassland plant diversity via light limitation ▶     Elizabeth T. Borer, Eric W. Seabloom, Daniel S. Gruner et al.     Experimental data collected from 40 grasslands on 6 continents show that nutrients and herbivores can serve as counteracting forces to control local plant diversity; nutrient addition reduces local diversity through light limitation, and herbivory rescues diversity at sites where it alleviates light limitation.             Endothelial Notch activity promotes angiogenesis and osteogenesis in bone ▶     Saravana K. Ramasamy, Anjali P. Kusumbe, Lin Wang et al.     Blood vessel growth in bone is revealed to require Notch signalling and involve a specialized form of angiogenesis that does not involve endothelial sprouts.             Articles and Letters                   Alveolar progenitor and stem cells in lung development, renewal and cancer ▶     Tushar J. Desai, Douglas G. Brownfield, Mark A. Krasnow     Lung alveoli are lined by two types of alveolar epithelial cells, squamous alveolar type (AT) 1 cells that mediate gas exchange and cuboidal AT2 cells that secrete surfactant to prevent alveolar collapse during breathing; here alveolar markers, genetic lineage tracing and clonal analysis are used in mice to identify alveolar progenitor and stem cells in vivo, and to map their locations and potential during lung development, maintenance and cancer.             C9orf72 nucleotide repeat structures initiate molecular cascades of disease ▶     Aaron R. Haeusler, Christopher J. Donnelly, Goran Periz et al.     Structurally polymorphic C9orf72 hexanucleotide repeats cause an impairment in transcriptional processivity and lead to accumulation of truncated repeat-containing transcripts that bind to specific ribonucleoproteins, such as nucleolin, in a conformation-dependent manner resulting in nucleolar stress and C9orf72-linked pathology in amyotrophic lateral sclerosis and frontotemporal dementia.             Proof of principle for epitope-focused vaccine design ▶     Bruno E. Correia, John T. Bates, Rebecca J. Loomis et al.     Computational protein design methods are used to generate new candidates for a human respiratory syncytial virus (RSV) vaccine; artificial protein scaffolds that mimic the structure of a RSV epitope are shown to induce RSV-specific neutralizing antibodies in macaques.             Derived immune and ancestral pigmentation alleles in a 7,000-year-old Mesolithic European ▶     Iñigo Olalde, Morten E. Allentoft, Federico Sánchez-Quinto et al.     A complete pre-agricultural European human genome from a ∼7,000-year-old Mesolithic skeleton suggests the existence of a common genomic signature across western and central Eurasia from the Upper Paleolithic to the Mesolithic, and ancestral alleles in several skin pigmentation genes suggest that the light skin of modern Europeans was not yet ubiquitous in Mesolithic times.             doublesex is a mimicry supergene ▶     K. Kunte, W. Zhang, A. Tenger-Trolander et al.     The phenomenon of sex-limited mimicry is phylogenetically widespread in the swallowtail butterfly genus Papilio — now, a single gene, doublesex, is shown to control supergene mimicry, a finding that is in contrast to the long-held view that supergenes are likely to be controlled by a tightly linked cluster of loci.             Dynamic sensory cues shape song structure in Drosophila  ▶     Philip Coen, Jan Clemens, Andrew J. Weinstein et al.     Drosophila male courtship songs were thought to have a fixed structure with song repetition variations introduced unintentionally because of neural noise; this behavioural assay and computational modelling study instead reveals that males use fast changes in sensory information to actively pattern individual song sequences.             An excitatory paraventricular nucleus to AgRP neuron circuit that drives hunger ▶     Michael J. Krashes, Bhavik P. Shah, Joseph C. Madara et al.     The AgRP-expressing neurons in the arcuate nucleus drive food-seeking behaviours during caloric restriction; a mouse study of monosynaptic retrograde rabies spread and optogenetic circuit mapping reveals that these neurons are activated by input from hypothalamic paraventricular nucleus cells and their activation or inhibition can modulate feeding behaviour.             L-Myc expression by dendritic cells is required for optimal T-cell priming ▶     Wumesh KC, Ansuman T. Satpathy, Aaron S. Rapaport et al.     L-Myc, a paralogue of the proto-oncogene c-Myc, is shown to regulate dendritic cell homeostasis and functionality; unlike c-Myc, L-Myc is not repressed by interferons and its expression allows for optimal dendritic cell proliferation and T-cell priming in the presence of inflammation.             A transcriptional switch underlies commitment to sexual development in malaria parasites ▶     Björn F. C. Kafsack, Núria Rovira-Graells, Taane G. Clark et al.     The DNA-binding protein PfAP2-G is found to be a master regulator of sexual development in the malaria parasite; this protein appears to regulate early gametocytogenesis and is epigenetically silenced in the majority of blood-stage parasites.             A cascade of DNA-binding proteins for sexual commitment and development in Plasmodium  ▶     Abhinav Sinha, Katie R. Hughes, Katarzyna K. Modrzynska et al.     Malaria parasites must produce gametocytes for transmission to the mosquito vector, although the molecular mechanisms underlying commitment to gametocyte production remain unclear; here this process is found to be controlled by PbAP2-G, a member of the ApiAP2 family of DNA-binding proteins, in the rodent-infecting Plasmodium berghei parasite.             DNA-guided DNA interference by a prokaryotic Argonaute ▶     Daan C. Swarts, Matthijs M. Jore, Edze R. Westra et al.     Here, Argonaute from the prokaryote Thermus thermophilus is shown to use small DNA guides to interfere directly with invading foreign DNA, rather than being involved in RNA-guided RNA interference, as observed in eukaryotes.             Reviews and Perspectives                   The present and future role of microfluidics in biomedical research ▶     Eric K. Sackmann, Anna L. Fulton, David J. Beebe     Recent progress in the various lab-on-a-chip microtechnologies is reviewed and the clinical and research areas in which they have made the greatest impact are discussed.             News & Views                   Evolutionary biology: Sex, lies and butterflies ▶     David W. Loehlin, Sean B. Carroll             Neurodegenerative diseases: G-quadruplex poses quadruple threat ▶     J. Paul Taylor             Neuroscience: Ordered randomness in fly love songs ▶     Bence P. Ölveczky             50 & 100 Years Ago ▶             Evolutionary biology: Speciation undone ▶     Peter R. Grant, B. Rosemary Grant             Bone biology: Vessels of rejuvenation ▶     Ferdinand le Noble, Jos le Noble             Genetics: Closing the distance on obesity culprits ▶     David U. Gorkin, Bing Ren             Cell biology: The disassembly of death ▶     Christopher D. Gregory             Corrigenda                   Corrigendum: Crystal structure of a nitrate/nitrite exchanger ▶     Hongjin Zheng, Goragot Wisedchaisri, Tamir Gonen             Corrigendum: An environmental bacterial taxon with a large and distinct metabolic repertoire ▶     Micheal C. Wilson, Tetsushi Mori, Christian Rückert et al.             Corrigendum: Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs ▶     Maurilio Sampaolesi, Stephane Blot, Giuseppe D'Antona et al.             Research Highlights                   Evolutionary biology: How the fish got its fins | Archaeology: Cats tamed early in Egypt | Conservation biology: Extinction looms for many mammals | Ecology: Warmer climate disturbs food web | Cancer: High cholesterol in prostate tumours | Neuroscience: Why warm caresses feel so good         NEWS & COMMENT           In retrospect: Sylva | Statistics: Biomedicine must look beyond P values | An elegant chaos | Storm brewing over WHO sugar proposal | Rethinking predators: Legend of the wolf             More Biological Sciences ▶                 The Structural Genomics Consortium (SGC), Natureconferences, and the São Paulo Research Foundation are pleased to present: Chemical Probe-based Open Science: Uncovering New Human and Plant Biology April 28-29, 2014 Faculdade de Ciências Médicas, State University of Campinas (UNICAMP), Brazil Click here for more information or to register for this conference today!         Health Sciences ▲top             RESEARCH           Latest Online                   Bone biology: Vessels of rejuvenation ▶     Ferdinand le Noble, Jos le Noble             Genetics: Closing the distance on obesity culprits ▶     David U. Gorkin, Bing Ren             Inhibition of miR-25 improves cardiac contractility in the failing heart ▶     Christine Wahlquist, Dongtak Jeong, Agustin Rojas-Muñoz et al.     Reduced activity of the calcium-transporting ATPase SERCA2a is a hallmark of heart failure; here, microRNAs that downregulate SERCA2a function are identified, and antagonism of one, miR-25, is shown to halt heart failure in mice.             Cell-cycle-regulated activation of Akt kinase by phosphorylation at its carboxyl terminus ▶     Pengda Liu, Michael Begley, Wojciech Michowski et al.     Phosphorylation of Akt at its carboxy-terminal tail is an essential layer of Akt activation to regulate its physiological functions.             Obesity-associated variants within FTO form long-range functional connections with IRX3 ▶     Scott Smemo, Juan J. Tena, Kyoung-Han Kim et al.     Obesity-associated noncoding sequences within FTO are functionally connected with IRX3, and long-range enhancers in this region recapitulate aspects of IRX3 expression, suggesting that the obesity-associated interval is part of IRX3 regulation; Irx3-deficient mice have lower body weight and are resistant to diet-induced obesity, suggesting IRX3 as a novel determinant of body mass and composition.             Articles and Letters                   C9orf72 nucleotide repeat structures initiate molecular cascades of disease ▶     Aaron R. Haeusler, Christopher J. Donnelly, Goran Periz et al.     Structurally polymorphic C9orf72 hexanucleotide repeats cause an impairment in transcriptional processivity and lead to accumulation of truncated repeat-containing transcripts that bind to specific ribonucleoproteins, such as nucleolin, in a conformation-dependent manner resulting in nucleolar stress and C9orf72-linked pathology in amyotrophic lateral sclerosis and frontotemporal dementia.             News & Views                   Neurodegenerative diseases: G-quadruplex poses quadruple threat ▶     J. Paul Taylor             50 & 100 Years Ago ▶             Bone biology: Vessels of rejuvenation ▶     Ferdinand le Noble, Jos le Noble             Genetics: Closing the distance on obesity culprits ▶     David U. Gorkin, Bing Ren             Research Highlights                   Cancer: High cholesterol in prostate tumours | Neuroscience: Why warm caresses feel so good         NEWS & COMMENT           Statistics: Biomedicine must look beyond P values | Storm brewing over WHO sugar proposal             More Health Sciences ▶               Physical Sciences ▲top             RESEARCH           Latest Online                   Low-temperature physics: Chaos in the cold ▶     Paul S. Julienne             Efficient rotational cooling of Coulomb-crystallized molecular ions by a helium buffer gas ▶     A. K. Hansen, O. O. Versolato, Ł. Kłosowski et al.     In combination with sympathetic cooling of translational degrees of freedom (leading to Coulomb crystallization), cooling of the rotational degrees of freedom of magnesium hydride ions using a helium buffer gas leads to temperatures in a tunable range from 60 kelvin down to about 7 kelvin for a single ion, the lowest such temperature so far recorded.             Quantum chaos in ultracold collisions of gas-phase erbium atoms ▶     Albert Frisch, Michael Mark, Kiyotaka Aikawa et al.     An ultracold gas of erbium atoms is shown to have many scattering resonances whose quantum fluctuations exhibit chaotic behaviour resulting from the anisotropy of the atoms’ interactions.             Articles and Letters                   Reflection from the strong gravity regime in a lensed quasar at redshift z=0.658 ▶     R. C. Reis, M. T. Reynolds, J. M. Miller et al.     Observations of a gravitationally lensed quasar at redshift 0.658 imply a high spin parameter, which indicates that this supermassive black hole grew by coherent accretion rather than in a chaotic manner.             Interrogating selectivity in catalysis using molecular vibrations ▶     Anat Milo, Elizabeth N. Bess, Matthew S. Sigman     A set of parameters based on the response of a molecule’s properties to infrared vibrations can be used to model and predict selectivity trends for molecular reactions with interlinked steric and electronic effects at positions of interest             Conformation-induced remote meta-C–H activation of amines ▶     Ri-Yuan Tang, Gang Li, Jin-Quan Yu     In anilines and benzylic amines, a recyclable chemical template can direct the olefination and acetoxylation of meta-C–H bonds as far as 11 bonds away from a functional group; in particular, it is able to direct the meta-selective C–H functionalization of bicyclic heterocycles via a highly strained, tricyclic-cyclophane-like palladated intermediate.             Reviews and Perspectives                   The present and future role of microfluidics in biomedical research ▶     Eric K. Sackmann, Anna L. Fulton, David J. Beebe     Recent progress in the various lab-on-a-chip microtechnologies is reviewed and the clinical and research areas in which they have made the greatest impact are discussed.             News & Views                   Astrophysics: Cosmic lens reveals spinning black hole ▶     Guido Risaliti             Low-temperature physics: Chaos in the cold ▶     Paul S. Julienne             Research Highlights                   Physics: Fast imaging captures falling droplets | Organic chemistry: Catalyst eases fuel production | Optics: Hot air guides laser beams | Engineering: Shake to make power         NEWS & COMMENT           Practical science has a global reach and appeal | History: A medieval multiverse | Particle physics: Higgs on the big screen | Statistics: Biomedicine must look beyond P values | Neutron sources: Road maps of no use to some physicists | Cellulosic ethanol fights for life | Energy: Islands of light             More Physical Sciences ▶               Earth & Environmental Sciences ▲top             RESEARCH           Articles and Letters                   Hydrous mantle transition zone indicated by ringwoodite included within diamond ▶     D. G. Pearson, F. E. Brenker, F. Nestola et al.     X-ray diffraction, Raman and infrared spectroscopic evidence for the inclusion of water-rich ringwoodite in diamond from Juína, Brazil, indicates that, at least locally, the Earth’s transition zone is hydrous to about 1 weight per cent.             News & Views                   Geology: Earth's deep water reservoir ▶     Hans Keppler             Brief Communications Arising                   Air pollution and forest water use ▶     Christopher D. Holmes             Keenan et al. reply ▶     Trevor F. Keenan, David Y. Hollinger, Gil Bohrer et al.             Research Highlights                   Ecology: Warmer climate disturbs food web         NEWS & COMMENT           Global seismic network takes to the seas | Climate change: China must publicize its emissions reports | Technology: Fight floods on a global scale             More Earth & Environmental Sciences ▶                 FREE POSTER: Dyslipidaemia and its treatment  Disorders of plasma lipid and lipoprotein metabolism can cause atherosclerotic cardiovascular disease. This poster displays the main lipid-metabolism pathways in the body, the points at which monogenetic protein mutations lead to dyslipidaemia, and the mechanisms of action of both established and novel lipid-modifying drugs.   Download now!    This poster was produced with support from: Regeneron Pharmaceuticals, Inc. and Sanofi         Careers & Jobs ▲top               Geoscience: Fracking fundamentals ▶     Scientists in the United States who are looking to ride the gas-exploration boom can find a variety of options for employment, from chemical research to environmental monitoring.                   Turning point: Johan Bollen ▶     Behavioural researcher aims to transform current funding models.                   UK science workforce ▶     UK report seeks to quantify diversity in the country's scientific workforce.                   Skewed rankings ▶     Female researchers avoid collaborating with lower-ranked female colleagues, finds study.                   Costs of childcare ▶     Breaks due to childcare are associated with lower wages for female physicians, says study.               naturejobs.com Science jobs of the week       Research Assistant     Rutgers New Jersey Medical School            PhD-Student     ETH Zurich            PhD Studentship     The Pirbright Institute            Postdoctoral Research Felllow     Women and Infants Hospital of Rhode Island          No matter what your career stage, student, postdoc or senior scientist, you will find articles on naturejobs.com to help guide you in your science career. 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