bluuebunny/crossref_metadata_2025 huggingface dataset:
Interplay between population firing stability and single neuron dynamics in hippocampal networks
Authors: Slomowitz Edden , Styr Boaz , Vertkin Irena , Milshtein-Parush Hila , Nelken Israel , Slutsky Michael , Slutsky Inna
Published: January 2015

Neuronal circuits' ability to maintain the delicate balance between stability and flexibility in changing environments is critical for normal neuronal functioning. However, to what extent individual neurons and neuronal populations maintain internal firing properties remains largely unknown. In this study, we show that distributions of spontaneous population firing rates and synchrony are subject to accurate homeostatic control following increase of synaptic inhibition in cultured hippocampal networks. Reduction in firing rate triggered synaptic and intrinsic adaptive responses operating as global homeostatic mechanisms to maintain firing macro-stability, without achieving local homeostasis at the single-neuron level. Adaptive mechanisms, while stabilizing population firing properties, reduced short-term facilitation essential for synaptic discrimination of input patterns. Thus, invariant ongoing population dynamics emerge from intrinsically unstable activity patterns of individual neurons and synapses. The observed differences in the precision of homeostatic control at different spatial scales challenge cell-autonomous theory of network homeostasis and suggest the existence of network-wide regulation rules.
Folate content in faba beans (Vicia faba L.)—effects of cultivar, maturity stage, industrial processing, and bioprocessing
Authors: Hefni Mohammed E. , Shalaby Mohamed T. , Witthöft Cornelia M.
Published: January 2015

Faba beans are an important source of folate and commonly consumed in Egypt. This study examined the effects of Egyptian industrial food processing (e.g., canning and freezing), germination, cultivar, and maturity stages on folate content, with the aim to develop a candidate functional canned faba bean food with increased folate content. The folate content in four cultivars of green faba beans ranged from 110 to 130 μg 100 g−1 fresh weight (535–620 μg 100 g−1 dry matter [DM]), which was four‐ to sixfold higher than in dried seeds. Industrial canning of dried seeds resulted in significant folate losses of ~20% (P = 0.004), while industrial freezing had no effect. Germination of faba beans increased the folate content by >40% (P < 0.0001). A novel industrial canning process involving pregermination of dried faba beans resulted in a net folate content of 194 μg 100 g−1 DM, which is 52% more than in conventional canned beans. The consumption of green faba beans should be recommended, providing ~120 μg dietary folate equivalents per 100 g/portion.
Renale Denervation: Schweizer Erfahrungen im Langzeitverlauf
Authors: Sudano Isabella , Battegay Edouard , Erne Paul , Lüscher Thomas F.
Published: January 2015

Trotz grosser Fortschritte der pharmakologischen und nicht-pharmakologischen Behandlung von Patienten mit arterieller Hypertonie erreicht ein beträchtlicher Anteil der Patienten nicht die Zielwerte der Hypertonieleitlinien. Ist es nicht möglich, trotz einer antihypertensiven Therapie mit drei maximal-dosierten Medikamenten, inklusive eines Diuretikums, den Blutdruck adäquat zu senken, handelt es sich um eine therapierefraktäre arterielle Hypertonie. Die renale Sympathikusdenervation ist ein neues und vielversprechendes Verfahren zur Behandlung einer therapierefraktären arteriellen Hypertonie. Es ist essenziell, die Patienten korrekt zu selektionieren und die Prozedur gemäss Richtlinie nur in spezialisierten Zentren durchzuführen.
Das schmerzhafte ISG – diagnostischer Algorithmus und therapeutische Möglichkeiten
Authors: Laux Christoph J. , Osterhoff Georg , Werner Clément M.L.
Published: January 2015

Die Iliosakralgelenke sind in 13–30% der Fälle Ursache für tieflumbale Rückenschmerzen und werden oftmals in differenzialdiagnostischen Überlegungen vernachlässigt. Die komplexe Anatomie und das vielfältige Schmerzbild führen häufig zu einer Fehlinterpretation einer symptomatischen SI-Arthropathie mit im Verlauf frustranen Therapieversuchen. Dieser Artikel richtet das Augenmerk auf das klinische Bild der SI-Arthropathie und deren Diagnose und zeigt die therapeutischen Möglichkeiten auf. Unter den vielen Provokationstests ist der PSIS-Distraktionstest ein zuverlässiges und behandlungsweisendes Diagnostikum im klinischen Alltag. Letztendlich fordert die Diagnose einer mechanischen SI-Arthropathie jedoch stets eine signifikante Beschwerdelinderung nach intraartikulärer Infiltration. Meist bringt eine konservative Therapie ausreichende Beschwerdelinderung. In seltenen Fällen kommen chirurgische Interventionen zum Einsatz.
Drei Patienten mit der gleichen Diagnose aber sehr unterschiedlichen Symptomen
Authors: Knechtle Beat , Rüst Christoph A. , Rosemann Thomas
Published: January 2015

Wir stellen drei Fälle von Patienten mit einer thorakalen Diskushernie vor, die initial alle missinterpretiert wurden. In einem Fall lagen pulmonale Symptome im Vordergrund, in einem weiteren Fall waren es kardiale Symptome und in einem dritten Fall neurologische Symptome. Unergiebige Abklärungen von thorakalen Beschwerden sollten den Verdacht auf eine thorakale Diskushernie lenken.
Imaging local electric fields produced upon synchrotron X-ray exposure
Authors: Dettmar Christopher M. , Newman Justin A. , Toth Scott J. , Becker Michael , Fischetti Robert F. , Simpson Garth J.
Published: January 2015

Significance

Exposure to high-energy X-rays, experienced during X-ray diffraction experiments for determining atomic structures, can potentially create large local electric fields from photoelectron ejection. Under cryogenic conditions, these fields may persist indefinitely. Mapping of electric fields is important for a complete understanding of damage mechanisms involved in X-ray measurements, for potential effects on the quality and interpretation of X-ray data, and for possible direct impact on diffraction resolution through the piezoelectric effect. Mapping of electric fields may also be useful in retaining a record of locations of X-ray exposure. In this paper, electric-field induced second harmonic generation imaging is explored in simulation and experiment. It provides insight into the position and distribution of local electric fields within an X-ray exposed sample.
Giant ankyrin-G stabilizes somatodendritic GABAergic synapses through opposing endocytosis of GABA
A
receptors
Authors: Tseng Wei Chou , Jenkins Paul M. , Tanaka Masashi , Mooney Richard , Bennett Vann
Published: January 2015

Significance

GABA
A
-receptor-based interneuron circuitry is essential for higher order function of the human nervous system and is implicated in schizophrenia, depression, anxiety disorders, and autism. GABAergic synapses are located on neuronal cell bodies and dendritic shafts as well as axon initial segments. This study demonstrates that giant ankyrin-G forms micron-scale domains on neuronal cell bodies and dendritic shafts, and promotes somatodendritic GABAergic synapse stability through interaction with GABARAP and inhibition of GABA
A
receptor endocytosis. This previously undescribed mechanism for regulating cell surface expression of GABA
A
receptors and maintaining GABAergic interneuron synapses offers a rationale for previous association of ankyrin-G genetic variation with neurodevelopmental disorders and psychiatric disease.
Plasmodium falciparum
evades mosquito immunity by disrupting JNK-mediated apoptosis of invaded midgut cells
Authors: Ramphul Urvashi N. , Garver Lindsey S. , Molina-Cruz Alvaro , Canepa Gaspar E. , Barillas-Mury Carolina
Published: January 2015

Significance

The
Anopheles gambiae
mosquito is a very effective vector of human
Plasmodium falciparum
malaria. We recently found that the
Pfs47
gene allows the parasite to survive, by evading the mosquito immune system. In this study, we explored the mechanism of
Pfs47
immune evasion. We found that
Pfs47
inhibits Jun-N-terminal kinase-mediated activation of apoptosis in invaded mosquito midgut cells by preventing activation of several caspases. Furthermore, the lack of caspase-S2 activation prevents the induction of enzymes that potentiate epithelial nitration, a reaction required for parasites to be “visible” to the mosquito complement-like system. These findings shed new light on how a single parasite gene inactivates the mosquito immune system and allows it to be successfully transmitted to a new host.
Association of Uric Acid With Vascular Stiffness in the Framingham Heart Study
Authors: Mehta Tapan , Nuccio Eugene , McFann Kim , Madero Magdalena , Sarnak Mark J. , Jalal Diana
Published: January 2015

BACKGROUND

Uric acid is associated with increased risk of cardiovascular disease and arterial stiffness in patients with hypertension or stroke. It remains unknown if uric acid is associated with arterial stiffness in the general population.

METHODS

We analyzed the association between serum uric acid levels and measures of arterial stiffness such as carotid-femoral pulse wave velocity (CF PWV), carotid-radial pulse wave velocity (CR PWV) and augmentation index (AI) in 4,140 participants from the Generation 3 Framingham cohort using linear regression.

RESULTS

Mean (SD) age was 40.0 (8.8) years and mean (SD) serum uric acid levels were 5.3 (1.5) mg/dl. Mean (SD) CF PWV was 7.0 (1.4) m/s. Individuals in the highest quartile of uric acid were more likely to be male, have a higher prevalence of hypertension, higher BMI, fasting glucose and insulin, and lower estimated glomerular filtration rate (eGFR). Multivariate adjusted means of CF PWV were 6.90, 6.94, 7.06, and 7.15 m/s for uric acid quartile 1, 2, 3, and 4 respectively. In unadjusted analysis each 1mg/dl increase in uric acid was associated with higher CF-PWV (β = 0.27; 95% CI = 0.25, 0.29; P < 0.0001). This was attenuated but remained significant after adjusting for age, sex, smoking, hypertension, BMI, fasting glucose, insulin, animal protein intake, and eGFR (β= 0.06; 95% CI = 0.02, 0.09; P < 0.0007). There was no association between serum uric acid levels and AI upon adjustment for cardiovascular risk factors.

CONCLUSIONS

Serum uric acid levels are significantly associated with CF PWV and CR PWV in a younger Caucasian population.
Separate universe simulations
Authors: Wagner Christian , Schmidt Fabian , Chiang Chi-Ting , Komatsu Eiichiro
Published: January 2015

The large-scale statistics of observables such as the galaxy density are chiefly determined by their dependence on the local coarse-grained matter density. This dependence can be measured directly and efficiently in N-body simulations by using the fact that a uniform density perturbation with respect to some fiducial background cosmology is equivalent to modifying the background and including curvature, i.e. by simulating a ‘separate universe’. We derive this mapping to fully non-linear order, and provide a step-by-step description of how to perform and analyse the separate universe simulations. This technique can be applied to a wide range of observables. As an example, we calculate the response of the non-linear matter power spectrum to long-wavelength density perturbations, which corresponds to the angle-averaged squeezed limit of the matter bispectrum and higher n-point functions. Using only a modest simulation volume, we obtain simulation results for the bispectrum and trispectrum with per cent-level precision over a wide range of scales.
Maximum speed of hypervelocity stars ejected from binaries
Authors: Tauris Thomas M.
Published: January 2015

The recent detection of hypervelocity stars (HVSs) as late-type B-stars and HVS candidate G/K dwarfs raises the important question of their origin. In this Letter, we investigate the maximum possible velocities of such HVSs if they are produced from binaries which are disrupted via an asymmetric supernova explosion. We find that HVSs up to ∼770 and ∼1280 km s−1 are possible in the Galactic rest frame from this scenario for these two subclasses of HVSs, respectively. We conclude that whereas a binary origin cannot easily explain all of the observed velocities of B-type HVSs (in agreement with their proposed central massive black hole origin) it can indeed account for the far majority (if not all) of the recently detected G/K-dwarf HVS candidates.
Astrometry of Galactic star-forming regions G48.99−0.30 and G49.19−0.34 with VERA: Astrometric accuracy verification of VERA
Authors: Nagayama Takumi , Kobayashi Hideyuki , Omodaka Toshihiro , Murata Yasuhiro , Burns Ross A. , Honma Mareki , Kurayama Tomoharu , Nakagawa Akiharu , Oyama Tomoaki , Tamura Yoshiaki , Sato Katsuhisa
Published: January 2015

We conducted astrometry of H2O masers in Galactic star-forming regions G48.99−0.30 and G49.19−0.34 with VLBI Exploration of Radio Astrometry (VERA). The observational results yield the parallaxes of G48.99−0.30 and G49.19−0.34 to be 0.178 ± 0.017 mas ($5.62^{+0.59}_{-0.49}\:$kpc) and 0.211 ± 0.016 mas ($4.74^{+0.39}_{-0.33}\:$kpc), respectively. We discuss in detail the sources of astrometric errors based on the observations of the two sources. The positional systemic errors in RA and Dec of (23, 31) μas for G49.19−0.34 are dominated by a tropospheric contribution from the Earth's atmosphere. Those of (23, 53) μas for G48.99−0.30 are attributed to a tropospheric contribution and a maser structure effect. The formal positional error from thermal noise is significantly smaller than these systemic positional errors. VERA phase-referencing observations could achieve a positional accuracy of a few tens of μas in both RA and Dec directions for point-like and bright sources at high elevation.
Witness of gas infall and outflow in the young starburst dwarf galaxy NGC 5253
Authors: Miura Rie E. , Espada Daniel , Sugai Hajime , Nakanishi Kouichiro , Hirota Akihiko
Published: January 2015

We present 12CO(2–1) observations towards the dwarf galaxy NGC 5253 using the Submillimeter Array. The data shows that a large amount of molecular gas is located in the central ∼ 200 pc starburst region, physically associated with two young super stellar clusters (SSCs). The molecular gas traced by 12CO(2–1) is elongated along the minor axis (dust lane) of the galaxy and its kinematics suggest that there is an inflow of molecular gas to the direction of the central SSCs, as is also observed in H i gas at a larger scale. Due to their correlation in spatial and velocity domains, the central SSCs were likely formed from molecular gas in the nucleus. We compare the 12CO(2–1) with available H2 1–0 S(1) data, and show that while the relatively cold gas traced by 12CO(2–1) is distributed around the central starburst region but also along the dust lane, the warm gas traced by H2 1–0 S(1) is associated with the central H ii region and other star-forming regions. Interestingly, a cavity in the H2 1–0 S(1) emission is found to be spatially correlated with a Hα shell. This Hα shell may trace a bipolar outflow from the central SSCs and the H2 1–0 S(1) gas, the gas shocked by the outflow encountering the surrounding quiescent gas. We calculate a molecular gas inflow rate of ∼ 2 M⊙ yr−1, a star formation rate of 0.3–0.5 M⊙ yr−1, and an ionized gas outflow being emitted from the SSCs with a rate of (5–25)× 10−3 f M⊙ yr−1, where f is a volume filling factor.
Implementing Information Science in Policing: Mapping the Evidence Base
Authors: Bowers Kate , Tompson Lisa , Johnson Shane D.
Published: January 2015

In many disciplines, there is a wealth of primary evaluation research on what works, and systematic reviews that synthesize that evidence. This is, of course, extremely positive. However, the sheer scale of the information and the way in which it is indexed and presented can mean that it is difficult for practitioners to locate the best available evidence. For this reason, in health, education, and other disciplines, using techniques from information science, researchers have systematically assembled databases such as those hosted on healthevidence.org and educationendowmentfoundation.org which bring together the most reliable evidence. Hitherto, no such database has existed for crime and criminal justice interventions. This article sets out some of challenges and early findings of one exercise which aims to produce such a database, being completed as part of the What Works Centre for Crime Reduction initiative in collaboration with the College of Policing.
Thenaked endospermGenes Encode Duplicate INDETERMINATE Domain Transcription Factors Required for Maize Endosperm Cell Patterning and Differentiation
Authors: Yi Gibum , Neelakandan Anjanasree K. , Gontarek Bryan C. , Vollbrecht Erik , Becraft Philip W.
Published: January 2015

The aleurone is the outermost layer of cereal endosperm and functions to digest storage products accumulated in starchy endosperm cells as well as to confer important dietary health benefits. Whereas normal maize (Zea mays [Zm]) has a single aleurone layer, naked endosperm (nkd) mutants produce multiple outer cell layers of partially differentiated cells that show sporadic expression of aleurone identity markers such as a viviparous1 promoter  -β-glucuronidase transgene. The 15:1 F2 segregation ratio suggested that two recessive genes were involved, and map-based cloning identified two homologous genes in duplicated regions of the genome. The nkd1 and nkd2 genes encode the INDETERMINATE1 domain (IDD) containing transcription factors ZmIDDveg9 and ZmIDD9 on chromosomes 2 and 10, respectively. Independent mutant alleles of nkd1 and nkd2, as well as nkd2-RNA interference lines in which both nkd genes were knocked down, also showed the nkd mutant phenotype, confirming the gene identities. In wild-type kernels, the nkd transcripts were most abundant around 11 to 16 d after pollination. The NKD proteins have putative nuclear localization signals, and green fluorescent protein fusion proteins showed nuclear localization. The mutant phenotype and gene identities suggest that NKD controls a gene regulatory network involved in aleurone cell fate specification and cell differentiation.
Simulation‐based Training for Emergency Medicine Residents in Sterile Technique During Central Venous Catheterization: Impact on Performance, Policy, and Outcomes
Authors: Hoskote Sumedh S. , Khouli Hassan , Lanoix Richard , Rose Keith , Aqeel Adnan , Clark Mark , Chalfin Donald , Shapiro Janet , Han Qifa
Published: January 2015

Objectives

Central line–associated bloodstream infection (CLABSI) is a preventable nosocomial infection. Simulation‐based training in sterile technique during central venous catheter (CVC) placement for emergency medicine (EM) residents, and its effect on changing the medical intensive care unit (MICU) practice of routine replacement of CVCs placed under sterile technique in the emergency department (ED), has not been evaluated.

Methods

Emergency medicine residents received simulation‐based sterile technique training during CVC placement between May 2008 and September 2010. Between June 2008 and January 2011, the authors reviewed records of patients who had CVCs placed in the ED under sterile technique by EM residents and were admitted to the MICU (group 1) and CVCs placed in the MICU under sterile technique by internal medicine (IM) residents (group 2). IM residents completed similar simulation‐based training before May 2008. Changes in EM residents' sterile technique performance scores were compared, as well as CLABSI rates in both groups. EM residents' CVC procedural skills were not assessed.

Results

Seventy‐six EM residents completed simulation‐based training with significant improvement in performance (median scores 13 out of 24 before training, 24 out of 24 after training; p < 0.001). CLABSI rates per 1,000 catheter‐days were 1.02 in group 1 and 1.02 in group 2 (p = 0.99). Both groups had similar demographics, acuity, and mortality (p > 0.5).

Conclusions

Routine replacement of CVCs placed in the ED under sterile technique after simulation‐based training would appear to be unnecessary. These findings demonstrate patient‐centered outcomes that are comparable for CVCs in ED‐admitted MICU patients, regardless of whether the CVC was placed in the ED or MICU.
Lysosomal signaling molecules regulate longevity in
Caenorhabditis elegans
Authors: Folick Andrew , Oakley Holly D. , Yu Yong , Armstrong Eric H. , Kumari Manju , Sanor Lucas , Moore David D. , Ortlund Eric A. , Zechner Rudolf , Wang Meng C.
Published: January 2015

Lysosomes signal the nucleus to control aging

Folick
et al.
propose a mechanism by which a lysosomal enzyme influences nuclear events that control longevity in the worm (see the Perspective by Shuo and Brunet). Increased expression of the lysosomal acid lipase LIPL-4 increased longevity, and this effect depended on the presence of the lysosomal lipid-binding protein LBP-8. LBP-8 acts as a chaperone that helps carry lipds to the nucleus. The authors identified the fatty acid oleoylethanolamide (OEA) as a potential signaling molecule whose transport to the nucleus could activate nuclear hormone receptors and transcription factors NHR-49 and NHR-80. The transcriptional targets of NHR-49 and NHR-80 in turn regulate longevity.

Science
, this issue p. 83
Rqc2p and 60
S
ribosomal subunits mediate mRNA-independent elongation of nascent chains
Authors: Shen Peter S. , Park Joseph , Qin Yidan , Li Xueming , Parsawar Krishna , Larson Matthew H. , Cox James , Cheng Yifan , Lambowitz Alan M. , Weissman Jonathan S. , Brandman Onn , Frost Adam
Published: January 2015

Tagging truncated proteins with CAT tails

During the translation of a messenger RNA (mRNA) into protein, ribosomes can sometimes stall. Truncated proteins thus formed can be toxic to the cell and must be destroyed. Shen
et al.
show that the proteins Ltn1p and Rqc2p, subunits of the ribosome quality control complex, bind to the stalled and partially disassembled ribosome. Ltn1p, a ubiquitin ligase, binds near the nascent polypeptide exit tunnel on the ribosome, well placed to tag the truncated protein for destruction. The Rqc2p protein interacts with the transfer RNA binding sites on the partial ribosome and recruits alanine- and threonine-bearing tRNAs. Rqc2p then catalyzes the addition of these amino acids onto the unfinished protein, in the absence of both the fully assembled ribosome and mRNA. These so-called CAT tails may promote the heat shock response, which helps buffer against malformed proteins.

Science
, this issue p.
75
Mechanistic insight from the crystal structure of mitochondrial complex I
Authors: Zickermann Volker , Wirth Christophe , Nasiri Hamid , Siegmund Karin , Schwalbe Harald , Hunte Carola , Brandt Ulrich
Published: January 2015

Energy conversion in complex 1

ATP, the energy source of the cell, is synthesized by a protein residing in the mitochondrial inner membrane. The synthesis is driven by a proton gradient generated by redox reactions that transfer electrons between a series of enzymes in the membrane. The largest complex in this electron transfer chain is the 1-MD complex 1. It couples electron transfer from NADH to ubiquinone to the translocation of four protons. Zickermann
et al.
report the crystal structure of a complex comprising the 14 central subunits and the largest accessory subunit of mitochondrial complex 1 from a yeast-genetic model at 3.6 Å resolution. The structure identifies four potential proton translocation pathways and gives insight into how energy from the redox reactions is transmitted to drive proton pumping.

Science
, this issue p.
44
The type VI secretion system of
Vibrio cholerae
fosters horizontal gene transfer
Authors: Borgeaud Sandrine , Metzger Lisa C. , Scrignari Tiziana , Blokesch Melanie
Published: January 2015

Killing, sex, and gene swaps in bacteria

The bacterial type VI secretion system (T6SS) is used by bacteria to inject toxins into neighboring cells to eliminate competition. This molecular machine is thus considered to be a mechanism by which bacteria can exert social control in complex microbial communities. Borgeaud
et al.
have discovered that in
Vibrio cholerae
, T6SS genes are co-regulated with genes involved in DNA uptake. Hence, T6SS-dependent killing of other bacteria is directed at neighboring cells, which release their DNA to be taken up by the killer, which can then integrate valuable genes and rapidly evolve, leading to antibiotic resistance or virulence.

Science
, this issue p.
63
Extremely short-lived reaction resonances in Cl + HD (
v
= 1) → DCl + H due to chemical bond softening
Authors: Yang Tiangang , Chen Jun , Huang Long , Wang Tao , Xiao Chunlei , Sun Zhigang , Dai Dongxu , Yang Xueming , Zhang Dong H.
Published: January 2015

A few very brief pauses in the action

Chemical reactions proceed by the cumulative effect of trillions upon trillions of collisions between atoms and molecules. Usually, a given collision bounces the participants right back out again, either in their original form or with the atoms shuffled around into distinct products. In certain cases, the reacting partners experience a brief lull, termed a resonance, before they rearrange. Yang
et al.
report the discovery of particularly short-lived resonances in certain reactive collisions of chlorine atoms with vibrationally excited hydrogen deuteride (HD). Their results suggest that similar, as yet overlooked, resonances may lurk in other reactions of vibrationally excited molecules.

Science
, this issue p.
60
Variation in cancer risk among tissues can be explained by the number of stem cell divisions
Authors: Tomasetti Cristian , Vogelstein Bert
Published: January 2015

Crunching the numbers to explain cancer

Why do some tissues give rise to cancer in humans a million times more frequently than others? Tomasetti and Vogelstein conclude that these differences can be explained by the number of stem cell divisions. By plotting the lifetime incidence of various cancers against the estimated number of normal stem cell divisions in the corresponding tissues over a lifetime, they found a strong correlation extending over five orders of magnitude. This suggests that random errors occurring during DNA replication in normal stem cells are a major contributing factor in cancer development. Remarkably, this “bad luck” component explains a far greater number of cancers than do hereditary and environmental factors.

Science
, this issue p.
78
Rh-catalyzed C–C bond cleavage by transfer hydroformylation
Authors: Murphy Stephen K. , Park Jung-Woo , Cruz Faben A. , Dong Vy M.
Published: January 2015

Shifting hydroformylation into reverse

The hydroformylation reaction is applied on large scale in the chemical industry to make aldehydes by adding hydrogen and carbon monoxide to olefins. The reverse process could also prove useful in modifying complex molecules for pharmaceutical research, but methods directed toward that end often strip off the CO without the hydrogen. Murphy
et al.
now show that a rhodium catalyst can achieve selective dehydroformylation of a diverse range of compounds under mild conditions (see the Perspective by Landis). The protocol relies on effective transfer of the CO and H
2
equivalents to a sacrificial strained olefin added to the mix.

Science
, this issue p.
56
; see also p.
29
Structure and inhibition of EV-D68, a virus that causes respiratory illness in children
Authors: Liu Yue , Sheng Ju , Fokine Andrei , Meng Geng , Shin Woong-Hee , Long Feng , Kuhn Richard J. , Kihara Daisuke , Rossmann Michael G.
Published: January 2015

Targeting EV-D68, a respiratory virus

A recent outbreak of respiratory illness in U.S. children was caused by entorovirus D68 (EV-D68). Enteroviruses also include human pathogens such as human rhinovirus, which causes the common cold, and poliovirus. Most of these viruses are stabilized by a factor that binds in a hydrophobic pocket of the capsid protein VP1, and antiviral compounds can act by displacing this factor. Liu
et al.
report the crystal structure of EV-D68 and its complex with the antiviral compound peconaril. In EV-D68, the hydrophobic pocket contained a fatty acid that was displaced by peconaril. Peconaril efficiently inhibited EV-D68 infection of cells, making it a possible drug candidate against EV-D68.

Science
, this issue p.
71
Persisters unmasked
Authors: Holden David W.
Published: January 2015

Intracellular toxins cause bacterial growth arrest and antibiotic tolerance
Geoscientists aim to magnify specialized Web searching
Authors: You Jia
Published: January 2015

National Science Foundation project, GeoLink, could become search hub for the geoscience community.
A science exodus from Ukraine's rebellious East
Authors: Stone Richard
Published: January 2015

Thousands of scientists—along with entire universities—have fled war-torn eastern Ukraine. Others have staked their futures on the breakaway republics.
Ukraine mourns a lost science jewel
Authors: Stone Richard
Published: January 2015

Russia's annexation of Crimea was a big blow to Ukrainian science.
Data meets design
The Social Machine Designs for Living Online
Judith Donath
MIT Press, 2014. 432 pp.
Authors: Golbeck Jennifer
Published: January 2015

Jennifer Golbeck enjoys a manifesto on the beauty of well-designed virtual interfaces
Construction and deconstruction of aldehydes by transfer hydroformylation
Authors: Landis Clark R.
Published: January 2015

A soluble rhodium catalyst converts alkenes to aldehydes without the need for toxic or explosive gas-phase reactants

[Also see Report by

Murphy
et al.

]
Conundrum of jumbled mosquito genomes
Authors: Clark Andrew G. , Messer Philipp W.
Published: January 2015

Multiple
Anopheles
mosquito genome sequences reveal extreme levels of mixing

[Also see Research Articles by

Fontaine
et al.

and

Neafsey
et al.

]
Lysosomal lipid lengthens life span
Authors: Han Shuo , Brunet Anne
Published: January 2015

A fatty acid moves from the lysosome to the nucleus, altering gene expression and extending longevity in the worm

[Also see Report by

Folick
et al.

]
Killer fat
Authors: Alcorn John F. , Kolls Jay K.
Published: January 2015

Adipocytes in the skin release an antimicrobial factor to fight staphylococcus infection

[Also see Report by

Zhang
et al.

]
Data, eternal
Authors: McNutt Marcia
Published: January 2015

During 2014,
Science
worked with members of the research community, other publishers, and representatives of funding agencies on many initiatives to increase transparency and promote reproducibility in the published research literature. Those efforts will continue in 2015. Connected to that progress, and an essential element to its success, an additional focus will be on making data more open, easier to access, more discoverable, and more thoroughly documented. My own commitment to these goals is deeply held, for I learned early in my career that interpretations come and go, but data are forever.
Safety and Immunogenicity of an Intranasal Sendai Virus-Based Human Parainfluenza Virus Type 1 Vaccine in 3- to 6-Year-Old Children
Authors: Adderson Elisabeth , Branum Kristen , Sealy Robert E. , Jones Bart G. , Surman Sherri L. , Penkert Rhiannon , Freiden Pamela , Slobod Karen S. , Gaur Aditya H. , Hayden Randall T. , Allison Kim , Howlett Nanna , Utech Jill , Allay Jim , Knight James , Sleep Susan , Meagher Michael M. , Russell Charles J. , Portner Allen , Hurwitz Julia L.
Published: January 2015

Human parainfluenza virus type 1 (hPIV-1) is the most common cause of laryngotracheobronchitis (croup), resulting in tens of thousands of hospitalizations each year in the United States alone. No licensed vaccine is yet available. We have developed murine PIV-1 (Sendai virus [SeV]) as a live Jennerian vaccine for hPIV-1. Here, we describe vaccine testing in healthy 3- to 6-year-old hPIV-1-seropositive children in a dose escalation study. One dose of the vaccine (5 × 105, 5 × 106, or 5 × 10750% egg infectious doses) was delivered by the intranasal route to each study participant. The vaccine was well tolerated by all the study participants. There was no sign of vaccine virus replication in the airway in any participant. Most children exhibited an increase in antibody binding and neutralizing responses toward hPIV-1 within 4 weeks from the time of vaccination. In several children, antibody responses remained above incoming levels for at least 6 months after vaccination. Data suggest that SeV may provide a benefit to 3- to 6-year-old children, even when vaccine recipients have preexisting cross-reactive antibodies due to previous exposures to hPIV-1. Results encourage the testing of SeV administration in young seronegative children to protect against the serious respiratory tract diseases caused by hPIV-1 infections.
Cost-Effective Pooling of DNA from Nasopharyngeal Swab Samples for Large-Scale Detection of Bacteria by Real-Time PCR
Authors: Edouard Sophie , Prudent Elsa , Gautret Philippe , Memish Ziad A. , Raoult Didier
Published: January 2015

We investigated the potential of pooling DNA from nasopharyngeal specimens to reduce the cost of real-time PCR (RT-PCR) for bacterial detection. Lyophilization is required to reconcentrate DNA. This strategy yields a high specificity (86%) and a high sensitivity (96%). We estimate that compared to individual testing, 37% fewer RT-PCR tests are needed.
Four Carbapenem-Resistant Gram-Negative Species Carrying Distinct Carbapenemases in a Single Patient
Authors: Ding Baixing , Hu Fupin , Yang Yang , Guo Qinglan , Huang Jinwei , Wang Minggui
Published: January 2015

Carbapenem-resistant
Escherichia coli
,
Klebsiella pneumoniae
,
Enterobacter aerogenes
, and
Acinetobacter baumannii
were isolated from a single patient, each producing different carbapenemases (NDM-1, KPC-2, IMP, and OXA-23, respectively). The NDM-1-producing
E. coli
strain was preceded by a clonally related carbapenem-susceptible strain a month earlier, suggesting
in vivo
acquisition of
bla
NDM-1
.
The Herpes Simplex Virus 1 UL51 Protein Interacts with the UL7 Protein and Plays a Role in Its Recruitment into the Virion
Authors: Roller Richard J. , Fetters Rachel
Published: January 2015

The alphaherpesvirus UL51 protein is a tegument component that interacts with the viral glycoprotein E and functions at multiple steps in virus assembly and spread in epithelial cells. We show here that pUL51 forms a complex in infected cells with another conserved tegument protein, pUL7. This complex can form in the absence of other viral proteins and is largely responsible for recruitment of pUL7 to cytoplasmic membranes and into the virion tegument. Incomplete colocalization of pUL51 and pUL7 in infected cells, however, suggests that a significant fraction of the population of each protein is not complexed with the other and that they may accomplish independent functions.

IMPORTANCE
The ability of herpesviruses to spread from cell to cell in the face of an immune response is critical for disease and shedding following reactivation from latency. Cell-to-cell spread is a conserved ability of herpesviruses, and the identification of conserved viral genes that mediate this process will aid in the design of attenuated vaccines and of novel therapeutics. The conserved UL51 gene of herpes simplex virus 1 plays important roles in cell-to-cell spread and in virus assembly in the cytoplasm, both of which likely depend on specific interactions with other viral and cellular proteins. Here we identify one of those interactions with the product of another conserved herpesvirus gene, UL7, and show that formation of this complex mediates recruitment of UL7 to membranes and to the virion.
Identification and Characterization of Nuclear and Nucleolar Localization Signals in the Adeno-Associated Virus Serotype 2 Assembly-Activating Protein
Authors: Earley Lauriel F. , Kawano Yasuhiro , Adachi Kei , Sun Xiao-Xin , Dai Mu-Shui , Nakai Hiroyuki
Published: January 2015

Assembly-activating protein (AAP) of adeno-associated virus serotype 2 (AAV2) is a nucleolar-localizing protein that plays a critical role in transporting the viral capsid VP3 protein to the nucleolus for assembly. Here, we identify and characterize AAV2 AAP (AAP2) nuclear (NLS) and nucleolar (NoLS) localization signals near the carboxy-terminal region of AAP2 (amino acid positions 144 to 184) (AAP2
144–184
). This region contains five basic-amino-acid-rich (BR) clusters, KSKRSRR (AAP2BR1), RRR (AAP2BR2), RFR (AAP2BR3), RSTSSR (AAP2BR4), and RRIK (AAP2BR5), from the amino terminus to the carboxy terminus. We created 30 AAP2BR mutants by arginine/lysine-to-alanine mutagenesis or deletion of AAP2BRs and 8 and 1 green fluorescent protein (GFP)-AAP2BR and β-galactosidase–AAP2BR fusion proteins, respectively, and analyzed their intracellular localization in HeLa cells by immunofluorescence microscopy. The results showed that AAP2
144–184
has redundant multipartite NLSs and that any combinations of 4 AAP2BRs, but not 3 or less, can constitute a functional NLS-NoLS; AAP2BR1 and AAP2BR2 play the most influential role for nuclear localization, but either one of the two AAP2BRs is dispensable if all 4 of the other AAP2BRs are present, resulting in 3 different, overlapping NLS motifs; and the NoLS is shared redundantly among the five AAP2BRs and functions in a context-dependent manner. AAP2BR mutations not only resulted in aberrant intracellular localization, but also attenuated AAP2 protein expression to various degrees, and both of these abnormalities have a significant negative impact on capsid production. Thus, this study reveals the organization of the intermingling NLSs and NoLSs in AAP2 and provides insights into their functional roles in capsid assembly.

IMPORTANCE
Adeno-associated virus (AAV) has become a popular and successful vector for
in vivo
gene therapy; however, its biology has yet to be fully understood. In this regard, the recent discovery of the assembly-activating protein (AAP), a nonstructural, nucleolar-localizing AAV protein essential for viral capsid assembly, has provided us a new opportunity to better understand the fundamental processes required for virion formation. Here, we identify clusters of basic amino acids in the carboxy terminus of AAP from AAV serotype 2 (AAV2) that act as nuclear and nucleolar localization signals. We also demonstrate their importance in maintaining AAP expression levels and efficient production of viral capsids. Insights into the functions of AAP can elucidate the requirements and process for AAV capsid assembly, which may lead to improved vector production for use in gene therapy. This study also contributes to the growing body of work on nuclear and nucleolar localization signals.
Mutations Conferring a Noncytotoxic Phenotype on Chikungunya Virus Replicons Compromise Enzymatic Properties of Nonstructural Protein 2
Authors: Utt Age , Das Pratyush Kumar , Varjak Margus , Lulla Valeria , Lulla Aleksei , Merits Andres
Published: January 2015

Chikungunya virus (CHIKV) (genus
Alphavirus
) has a positive-sense RNA genome. CHIKV nonstructural protein 2 (nsP2) proteolytically processes the viral nonstructural polyprotein, possesses nucleoside triphosphatase (NTPase), RNA triphosphatase, and RNA helicase activities, and induces cytopathic effects in vertebrate cells. Although alphaviral nsP2 mutations can result in a noncytotoxic phenotype, the effects of such mutations on nsP2 enzymatic activities are not well understood. In this study, we introduced a P718G (PG) mutation and selected for additional mutations in CHIKV nsP2 that resulted in a CHIKV replicon with a noncytotoxic phenotype in BHK-21 cells. Combinations of PG and either an E116K (EK) substitution or a GEEGS sequence insertion after residue T648 (5A) markedly reduced RNA synthesis; however, neither PG nor 5A prevented nsP2 nuclear translocation. Introducing PG into recombinant nsP2 inhibited proteolytic cleavage of nsP1/nsP2 and nsP3/nsP4 sites, reduced GTPase and RNA helicase activities, and abolished RNA stimulation of GTPase activity. 5A and EK modulated the effects of PG. However, only the RNA helicase activity of nsP2 was reduced by both of these mutations, suggesting that defects in this activity may be linked to a noncytotoxic phenotype. These results increase our understanding of the molecular basis for the cytotoxicity that accompanies alphaviral replication. Furthermore, adaptation of the CHIKV replicon containing both 5A and PG allowed the selection of a CHIKV replicon with adaptive mutations in nsP1 and nsP3 that enable persistence in human cell line. Such cell lines represent valuable experimental systems for discovering host factors and for screening inhibitors of CHIKV replication at lower biosafety levels.

IMPORTANCE
CHIKV is a medically important pathogen that causes febrile illness and can cause chronic arthritis. No approved vaccines or antivirals are available for CHIKV. The attenuation of CHIKV is critical to the establishment of experimental systems that can be used to conduct virus replication studies at a lower biosafety level. We applied a functional selection approach to develop, for the first time, a noncytotoxic CHIKV replicon capable of persisting in human cell lines. We anticipate that this safe and efficient research tool will be valuable for screening CHIKV replication inhibitors and for identifying and analyzing host factors involved in viral replication. We also analyzed, from virological and protein biochemistry perspectives, the functional defects caused by mutations conferring noncytotoxic phenotypes; we found that all known enzymatic activities of CHIKV nsP2, as well as its RNA-binding capability, were compromised by these mutations, which led to a reduced capacity for replication.
Impact of Natural IgM Concentration on Gene Therapy with Adenovirus Type 5 Vectors
Authors: Qiu Qi , Xu Zhili , Tian Jie , Moitra Rituparna , Gunti Sreenivasulu , Notkins Abner L. , Byrnes Andrew P.
Published: January 2015

Natural IgM inhibits gene transfer by adenovirus type 5 (Ad5) vectors. We show that polyreactive natural IgM antibodies bind to Ad5 and that inhibition of liver transduction by IgM depends on Kupffer cells. By manipulating IgM concentrationin vivo, we demonstrate that IgM inhibits liver transduction in a concentration-dependent manner. We further show that differences in natural IgM between BALB/c and C57BL/6 mice contribute to lower efficiency of Ad5 gene transfer in BALB/c mice.
Differential Effects of the G118R, H51Y, and E138K Resistance Substitutions in Different Subtypes of HIV Integrase
Authors: Quashie Peter K. , Oliviera Maureen , Veres Tamar , Osman Nathan , Han Ying-Shan , Hassounah Said , Lie Yolanda , Huang Wei , Mesplède Thibault , Wainberg Mark A.
Published: January 2015

Dolutegravir (DTG) is the latest antiretroviral (ARV) approved for the treatment of human immunodeficiency virus (HIV) infection. The G118R substitution, previously identified with MK-2048 and raltegravir, may represent the initial substitution in a dolutegravir resistance pathway. We have found that subtype C integrase proteins have a low enzymatic cost associated with the G118R substitution, mostly at the strand transfer step of integration, compared to either subtype B or recombinant CRF02_AG proteins. Subtype B and circulating recombinant form AG (CRF02_AG) clonal viruses encoding G118R-bearing integrases were severely restricted in their viral replication capacity, and G118R/E138K-bearing viruses had various levels of resistance to dolutegravir, raltegravir, and elvitegravir. In cell-free experiments, the impacts of the H51Y and E138K substitutions on resistance and enzyme efficiency, when present with G118R, were highly dependent on viral subtype. Sequence alignment and homology modeling showed that the subtype-specific effects of these mutations were likely due to differential amino acid residue networks in the different integrase proteins, caused by polymorphic residues, which significantly affect native protein activity, structure, or function and are important for drug-mediated inhibition of enzyme activity. This preemptive study will aid in the interpretation of resistance patterns in dolutegravir-treated patients.

IMPORTANCE
Recognized drug resistance mutations have never been reported for naive patients treated with dolutegravir. Additionally, in integrase inhibitor-experienced patients, only R263K and other previously known integrase resistance substitutions have been reported. Here we suggest that alternate resistance pathways may develop in non-B HIV-1 subtypes and explain how “minor” polymorphisms and substitutions in HIV integrase that are associated with these subtypes can influence resistance against dolutegravir. This work also highlights the importance of phenotyping versus genotyping when a strong inhibitor such as dolutegravir is being used. By characterizing the G118R substitution, this work also preemptively defines parameters for a potentially important pathway in some non-B HIV subtype viruses treated with dolutegravir and will aid in the inhibition of such a virus, if detected. The general inability of strand transfer-related substitutions to diminish 3′ processing indicates the importance of the 3′ processing step and highlights a therapeutic angle that needs to be better exploited.
Ebola Virus and Severe Acute Respiratory Syndrome Coronavirus Display Late Cell Entry Kinetics: Evidence that Transport to NPC1
+
Endolysosomes Is a Rate-Defining Step
Authors: Mingo Rebecca M. , Simmons James A. , Shoemaker Charles J. , Nelson Elizabeth A. , Schornberg Kathryn L. , D'Souza Ryan S. , Casanova James E. , White Judith M.
Published: January 2015

Ebola virus (EBOV) causes hemorrhagic fevers with high mortality rates. During cellular entry, the virus is internalized by macropinocytosis and trafficked through endosomes until fusion between the viral and an endosomal membrane is triggered, releasing the RNA genome into the cytoplasm. We found that while macropinocytotic uptake of filamentous EBOV viruslike particles (VLPs) expressing the EBOV glycoprotein (GP) occurs relatively quickly, VLPs only begin to enter the cytoplasm after a 30-min lag, considerably later than particles bearing the influenza hemagglutinin or GP from lymphocytic choriomeningitis virus, which enter through late endosomes (LE). For EBOV, the long lag is not due to the large size or unusual shape of EBOV filaments, the need to prime EBOV GP to the 19-kDa receptor-binding species, or a need for unusually low endosomal pH. In contrast, since we observed that EBOV entry occurs upon arrival in Niemann-Pick C1 (NPC1)-positive endolysosomes (LE/Lys), we propose that trafficking to LE/Lys is a key rate-defining step. Additional experiments revealed, unexpectedly, that severe acute respiratory syndrome (SARS) S-mediated entry also begins only after a 30-min lag. Furthermore, although SARS does not require NPC1 for entry, SARS entry also begins after colocalization with NPC1. Since the only endosomal requirement for SARS entry is cathepsin L activity, we tested and provide evidence that NPC1
+
LE/Lys have higher cathepsin L activity than LE, with no detectable activity in earlier endosomes. Our findings suggest that both EBOV and SARS traffic deep into the endocytic pathway for entry and that they do so to access higher cathepsin activity.

IMPORTANCE
Ebola virus is a hemorrhagic fever virus that causes high fatality rates when it spreads from zoonotic vectors into the human population. Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) causes severe respiratory distress in infected patients. A devastating outbreak of EBOV occurred in West Africa in 2014, and there was a significant outbreak of SARS in 2003. No effective vaccine or treatment has yet been approved for either virus. We present evidence that both viruses traffic late into the endocytic pathway, to NPC1
+
LE/Lys, in order to enter host cells, and that they do so to access high levels of cathepsin activity, which both viruses use in their fusion-triggering mechanisms. This unexpected similarity suggests an unexplored vulnerability, trafficking to NPC1
+
LE/Lys, as a therapeutic target for SARS and EBOV.