bluuebunny/crossref_metadata_2025 huggingface dataset:
Habitat and plant diversity of Trifolium tumens (Steven ex M. Bieb.) collected in Azerbaijan and its characterisation and field evaluation in Tasmania, Australia
Authors: Hall E. J. , Hughes S. J. , Humphries A. W. , Corkrey R.
Published: August 2013

Trifolium tumens (Talish clover) is a perennial legume species not previously domesticated for use in world agriculture. It is native to areas of the eastern Mediterranean, Caucasus and mountainous areas of the Middle East with a warm temperate or cool Mediterranean environment. The species has been identified by pasture researchers in Australia as a having sufficient drought and grazing tolerance to fill a gap created by the lack of a well adapted perennial legume for dryland pasture systems across low rainfall (450 to 750 mm annual average rainfall), temperate regions of Australia. The need to expand available genetic diversity of this species resulted in a germplasm collecting mission to Azerbaijan in 2004. The successful mission collected seed from 35 populations of T. tumens across 7 agro ecological zones, including 5 climatic zones. This collection increased the number of accessions held in ex situ local and international collections from 21 to 56. The range of adaptation and frequency of the collection of T. tumens was far greater than expected with accessions found at altitudes ranging from 12 to 1700 m above sea level, in soils with pH (1:5 H20) ranging between 5.0 and 9.0 and rainfall varying from 300 to 1300 mm. T. tumens was found growing alongside a diverse range of companion species in heavily grazed lowland pastures up to lightly grazed alpine meadows. This material has been characterised by the Tasmanian Institute of Agriculture (TIA), with promising accessions contributing to a breeding program to develop a commercial cultivar adapted to temperate environments (<750 mm rainfall).
Trial by fire: Community Wildfire Protection Plans put to the test
Authors: Jakes Pamela J. , Sturtevant Victoria
Published: August 2013

Research has found that community wildfire protection planning can make significant contributions to wildfire mitigation and preparedness, but can the planning process and resulting Community Wildfire Protection Plans make a difference to wildfire response and recovery? In case studies conducted in four USA communities with Community Wildfire Protection Plans in place when wildfires occurred, we saw a range of Community Wildfire Protection Plan projects designed to change the path and intensity of the wildfires. In most of our communities, the Community Wildfire Protection Plan and planning process improved relationships among firefighting agencies, clarified responsibilities and improved communication systems, contributing to fire response efficiency and effectiveness. We found that social learning resulting from the wildfire experience motivated communities to revisit and implement their Community Wildfire Protection Plans, changing the planning frame and scale and increasing the plan’s relevance for response and recovery. We conclude that Community Wildfire Protection Plans and experience with wildfire can also result in greater community capacity that builds resilience and increases adaptive capacity for future environmental changes and disasters.
Behaviour of free-ranging short-beaked common dolphins (Delphinus delphis) in Gulf St Vincent, South Australia
Authors: Filby Nicole E. , Bossley Mike , Stockin Karen A.
Published: August 2013

Common dolphins are subject to large-scale fishing activity and tourism operations in South Australia; however, there is a paucity of data on this species. Understanding the behaviour of a population can contribute greatly to our knowledge of a species and how to manage potential population-level threats. This paper describes the behaviour of short-beaked common dolphins (Delphinus delphis) in Australian waters for the first time. Data were collected from 109 independent dolphin groups during boat-based surveys conducted in Gulf St Vincent, South Australia, between September 2005 and May 2008. Activity budgets were used to assess behaviour of common dolphins in relation to diel patterns, season, water depth, sea surface temperature (SST), group size and composition. Foraging (33.9%) and resting (2.8%) were the most and least frequently observed behaviours, respectively. Travelling (33.0%), socialising (20.2%) and milling (10.1%) accounted for the remainder of the activity budget. Diurnal differences were detected, with foraging (59.5%) and socialising (31.8%) groups most frequently observed from 10.00 to 11.59 hours. Behaviour did not vary seasonally or with water depth, SST, group size or composition. Behaviour varied significantly between single- and multispecies aggregations. Foraging was more frequent in multispecies aggregations, as 78.4% of all foraging behaviour observed for common dolphins occurred in the presence of other species. Multispecies aggregations were most frequently observed with flesh-footed shearwaters (Puffinus carneipes), which were present during 29.4% of common dolphin encounters. Behaviour varied significantly during aggregations with shearwaters, as 62.2% of foraging groups occurred in the presence of shearwaters. Resting, milling or socialising was rarely observed in the presence of any other species, indicating that the primary mechanism for aggregations is likely prey-related.
Enhanced basal lubrication and the contribution of the Greenland ice sheet to future sea-level rise
Authors: Shannon Sarah R. , Payne Antony J. , Bartholomew Ian D. , van den Broeke Michiel R. , Edwards Tamsin L. , Fettweis Xavier , Gagliardini Olivier , Gillet-Chaulet Fabien , Goelzer Heiko , Hoffman Matthew J. , Huybrechts Philippe , Mair Douglas W. F. , Nienow Peter W. , Perego Mauro , Price Stephen F. , Smeets C. J. P. Paul , Sole Andrew J. , van de Wal Roderik S. W. , Zwinger Thomas
Published: August 2013

We assess the effect of enhanced basal sliding on the flow and mass budget of the Greenland ice sheet, using a newly developed parameterization of the relation between meltwater runoff and ice flow. A wide range of observations suggest that water generated by melt at the surface of the ice sheet reaches its bed by both fracture and drainage through moulins. Once at the bed, this water is likely to affect lubrication, although current observations are insufficient to determine whether changes in subglacial hydraulics will limit the potential for the speedup of flow. An uncertainty analysis based on our best-fit parameterization admits both possibilities: continuously increasing or bounded lubrication. We apply the parameterization to four higher-order ice-sheet models in a series of experiments forced by changes in both lubrication and surface mass budget and determine the additional mass loss brought about by lubrication in comparison with experiments forced only by changes in surface mass balance. We use forcing from a regional climate model, itself forced by output from the European Centre Hamburg Model (ECHAM5) global climate model run under scenario A1B. Although changes in lubrication generate widespread effects on the flow and form of the ice sheet, they do not affect substantial net mass loss; increase in the ice sheet’s contribution to sea-level rise from basal lubrication is projected by all models to be no more than 5% of the contribution from surface mass budget forcing alone.
What do we mean? On the importance of not abandoning scientific rigor when talking about science education
Authors: Klahr David
Published: August 2013

Although the “science of science communication” usually refers to the flow of scientific knowledge from scientists to the public, scientists direct most of their communications not to the public, but instead to other scientists in their field. This paper presents a case study on this understudied type of communication: within a discipline, among its practitioners. I argue that many of the contentious disagreements that exist today in the field in which I conduct my research—early science education—derive from a lack of operational definitions, such that when competing claims are made for the efficacy of one type of science instruction vs. another, the arguments are hopelessly disjointed. The aim of the paper is not to resolve the current claims and counterclaims about the most effective pedagogies in science education, but rather to note that the assessment of one approach vs. the other is all too often defended on the basis of strongly held beliefs, rather than on the results of replicable experiments, designed around operational definitions of the teaching methods being investigated. A detailed example of operational definitions from my own research on elementary school science instruction is provided. In addition, the paper addresses the issue of how casual use of labels—both within the discipline and when communicating with the public—may inadvertently “undo” the benefits of operational definitions.
Bridging the gap between science and decision making
Authors: von Winterfeldt Detlof
Published: August 2013

All decisions, whether they are personal, public, or business-related, are based on the decision maker’s beliefs and values. Science can and should help decision makers by shaping their beliefs. Unfortunately, science is not easily accessible to decision makers, and scientists often do not understand decision makers’ information needs. This article presents a framework for bridging the gap between science and decision making and illustrates it with two examples. The first example is a personal health decision. It shows how a formal representation of the beliefs and values can reflect scientific inputs by a physician to combine with the values held by the decision maker to inform a medical choice. The second example is a public policy decision about managing a potential environmental hazard. It illustrates how controversial beliefs can be reflected as uncertainties and informed by science to make better decisions. Both examples use decision analysis to bridge science and decisions. The conclusions suggest that this can be a helpful process that requires skills in both science and decision making.
Neutral and weakly nonneutral sequence variants may define individuality
Authors: Bromberg Yana , Kahn Peter C. , Rost Burkhard
Published: August 2013

Large-scale computational analyses of the growing wealth of genome-variation data consistently tell two distinct stories. The first is expected: coding variants reported in disease-related databases significantly alter the function of affected proteins. The second is surprising: the genomes of healthy individuals appear to carry many variants that are predicted to have some effect on function. As long as the complete experimental analysis of all human genome variants remains impossible, computational methods, such as PolyPhen, SNAP, and SIFT, might provide important insights. These methods capture the effects of particular variants very well and can highlight trends in populations of variants. Diseases are, arguably, extreme phenotypic variations and are often attributable to one or a few severely functionally disruptive variants. Our findings suggest a genomic basis of the different nondisease phenotypes. Prediction methods indicate that variants in seemingly healthy individuals tend to be neutral or weakly disruptive for protein molecular function. These variant effects are predicted to be largely either experimentally undetectable or are not deemed significant enough to be published. This may suggest that nondisease phenotypes arise through combinations of many variants whose effects are weakly nonneutral (damaging or enhancing) to the molecular protein function but fall within the wild-type range of overall physiological function.
Competing molecular interactions of aPKC isoforms regulate neuronal polarity
Authors: Parker Sara S. , Mandell Edward K. , Hapak Sophie M. , Maskaykina Irina Y. , Kusne Yael , Kim Ji-Young , Moy Jamie K. , St. John Paul A. , Wilson Jean M. , Gothard Katalin M. , Price Theodore J. , Ghosh Sourav
Published: August 2013

Atypical protein kinase C (aPKC) isoforms ζ and λ interact with polarity complex protein Par3 and are evolutionarily conserved regulators of cell polarity.
Prkcz
encodes aPKC-ζ and PKM-ζ, a truncated, neuron-specific alternative transcript, and
Prkcl
encodes aPKC-λ. Here we show that, in embryonic hippocampal neurons, two aPKC isoforms, aPKC-λ and PKM-ζ, are expressed. The localization of these isoforms is spatially distinct in a polarized neuron. aPKC-λ, as well as Par3, localizes at the presumptive axon, whereas PKM-ζ and Par3 are distributed at non-axon-forming neurites. PKM-ζ competes with aPKC-λ for binding to Par3 and disrupts the aPKC-λ–Par3 complex. Silencing of PKM-ζ or overexpression of aPKC-λ in hippocampal neurons alters neuronal polarity, resulting in neurons with supernumerary axons. In contrast, the overexpression of PKM-ζ prevents axon specification. Our studies suggest a molecular model wherein mutually antagonistic intermolecular competition between aPKC isoforms directs the establishment of neuronal polarity.
RasGRF2 Rac-GEF activity couples NMDA receptor calcium flux to enhanced synaptic transmission
Authors: Schwechter Brandon , Rosenmund Christian , Tolias Kimberley F.
Published: August 2013

Dendritic spines are the primary sites of excitatory synaptic transmission in the vertebrate brain, and the morphology of these actin-rich structures correlates with synaptic function. Here we demonstrate a unique method for inducing spine enlargement and synaptic potentiation in dispersed hippocampal neurons, and use this technique to identify a coordinator of these processes; Ras-specific guanine nucleotide releasing factor 2 (RasGRF2). RasGRF2 is a dual Ras/Rac guanine nucleotide exchange factor (GEF) that is known to be necessary for long-term potentiation in situ. Contrary to the prevailing assumption, we find RasGRF2’s Rac-GEF activity to be essential for synaptic potentiation by using a molecular replacement strategy designed to dissociate Rac- from Ras-GEF activities. Furthermore, we demonstrate that Rac1 activity itself is sufficient to rapidly modulate postsynaptic strength by using a photoactivatable derivative of this small GTPase. Because Rac1 is a major actin regulator, our results support a model where the initial phase of long-term potentiation is driven by the cytoskeleton.
Reconciling migration models to the Americas with the variation of North American native mitogenomes
Authors: Achilli Alessandro , Perego Ugo A. , Lancioni Hovirag , Olivieri Anna , Gandini Francesca , Hooshiar Kashani Baharak , Battaglia Vincenza , Grugni Viola , Angerhofer Norman , Rogers Mary P. , Herrera Rene J. , Woodward Scott R. , Labuda Damian , Smith David Glenn , Cybulski Jerome S. , Semino Ornella , Malhi Ripan S. , Torroni Antonio
Published: August 2013

In this study we evaluated migration models to the Americas by using the information contained in native mitochondrial genomes (mitogenomes) from North America. Molecular and phylogeographic analyses of B2a mitogenomes, which are absent in Eskimo–Aleut and northern Na-Dene speakers, revealed that this haplogroup arose in North America ∼11–13 ka from one of the founder Paleo-Indian B2 mitogenomes. In contrast, haplogroup A2a, which is typical of Eskimo–Aleuts and Na-Dene, but also present in the easternmost Siberian groups, originated only 4–7 ka in Alaska, led to the first Paleo-Eskimo settlement of northern Canada and Greenland, and contributed to the formation of the Na-Dene gene pool. However, mitogenomes also show that Amerindians from northern North America, without any distinction between Na-Dene and non–Na-Dene, were heavily affected by an additional and distinctive Beringian genetic input. In conclusion, most mtDNA variation (along the double-continent) stems from the first wave from Beringia, which followed the Pacific coastal route. This was accompanied or followed by a second inland migratory event, marked by haplogroups X2a and C4c, which affected all Amerindian groups of Northern North America. Much later, the ancestral A2a carriers spread from Alaska, undertaking both a westward migration to Asia and an eastward expansion into the circumpolar regions of Canada. Thus, the first American founders left the greatest genetic mark but the original maternal makeup of North American Natives was subsequently reshaped by additional streams of gene flow and local population dynamics, making a three-wave view too simplistic.
High-resolution crystal structure reveals molecular details of target recognition by bacitracin
Authors: Economou Nicoleta J. , Cocklin Simon , Loll Patrick J.
Published: August 2013

Bacitracin is a metalloantibiotic agent that is widely used as a medicine and feed additive. It interferes with bacterial cell-wall biosynthesis by binding undecaprenyl-pyrophosphate, a lipid carrier that serves as a critical intermediate in cell wall production. Despite bacitracin’s broad use, the molecular details of its target recognition have not been elucidated. Here we report a crystal structure for the ternary complex of bacitracin A, zinc, and a geranyl-pyrophosphate ligand at a resolution of 1.1 Å. The antibiotic forms a compact structure that completely envelopes the ligand’s pyrophosphate group, together with flanking zinc and sodium ions. The complex adopts a highly amphipathic conformation that offers clues to antibiotic function in the context of bacterial membranes. Bacitracin’s efficient sequestration of its target represents a previously unseen mode for the recognition of lipid pyrophosphates, and suggests new directions for the design of next-generation antimicrobial agents.
Surge of neurophysiological coherence and connectivity in the dying brain
Authors: Borjigin Jimo , Lee UnCheol , Liu Tiecheng , Pal Dinesh , Huff Sean , Klarr Daniel , Sloboda Jennifer , Hernandez Jason , Wang Michael M. , Mashour George A.
Published: August 2013

The brain is assumed to be hypoactive during cardiac arrest. However, the neurophysiological state of the brain immediately following cardiac arrest has not been systematically investigated. In this study, we performed continuous electroencephalography in rats undergoing experimental cardiac arrest and analyzed changes in power density, coherence, directed connectivity, and cross-frequency coupling. We identified a transient surge of synchronous gamma oscillations that occurred within the first 30 s after cardiac arrest and preceded isoelectric electroencephalogram. Gamma oscillations during cardiac arrest were global and highly coherent; moreover, this frequency band exhibited a striking increase in anterior–posterior-directed connectivity and tight phase-coupling to both theta and alpha waves. High-frequency neurophysiological activity in the near-death state exceeded levels found during the conscious waking state. These data demonstrate that the mammalian brain can, albeit paradoxically, generate neural correlates of heightened conscious processing at near-death.
Near-term climate mitigation by short-lived forcers
Authors: Smith Steven J. , Mizrahi Andrew
Published: August 2013

Emissions reductions focused on anthropogenic climate-forcing agents with relatively short atmospheric lifetimes, such as methane (CH4) and black carbon, have been suggested as a strategy to reduce the rate of climate change over the next several decades. We find that reductions of methane and black carbon would likely have only a modest impact on near-term global climate warming. Even with maximally feasible reductions phased in from 2015 to 2035, global mean temperatures in 2050 would be reduced by 0.16 °C, with a range of 0.04–0.35 °C because of uncertainties in carbonaceous aerosol emissions and aerosol forcing per unit of emissions. The high end of this range is only possible if total historical aerosol forcing is relatively small. More realistic emission reductions would likely provide an even smaller climate benefit. We find that the climate benefit from reductions in short-lived forcing agents are smaller than previously estimated. These near-term climate benefits of targeted reductions in short-lived forcers are not substantially different in magnitude from the benefits from a comprehensive climate policy.
Biochemical analysis of a multifunctional cytochrome P450 (CYP51) enzyme required for synthesis of antimicrobial triterpenes in plants
Authors: Geisler Katrin , Hughes Richard K. , Sainsbury Frank , Lomonossoff George P. , Rejzek Martin , Fairhurst Shirley , Olsen Carl-Erik , Motawia Mohammed Saddik , Melton Rachel E. , Hemmings Andrew M. , Bak Søren , Osbourn Anne
Published: August 2013

Significance

We carried out functional analysis of the oat enzyme AsCYP51H10, which is a divergent member of the CYP51 cytochrome P450 family and showed that this enzyme is able to catalyze both hydroxylation and epoxidation of the simple triterpene β-amyrin to give 12,13β-epoxy-3β,16β-dihydroxy-oleanane (12,13β-epoxy-16β-hydroxy-β-amyrin). In contrast, the canonical CYP51 enzymes are highly conserved and catalyze only sterol demethylation. We further show that the C12,13 epoxy group is critical for antifungal activity, a discovery that has important implications for triterpene metabolic engineering for food, health, and industrial biotechnology applications.
AMPA receptor pHluorin-GluA2 reports NMDA receptor-induced intracellular acidification in hippocampal neurons
Authors: Rathje Mette , Fang Huaqiang , Bachman Julia L. , Anggono Victor , Gether Ulrik , Huganir Richard L. , Madsen Kenneth L.
Published: August 2013

NMDA receptor activation promotes endocytosis of AMPA receptors, which is an important mechanism underlying long-term synaptic depression. The pH-sensitive GFP variant pHluorin fused to the N terminus of GluA2 (pH-GluA2) has been used to assay NMDA-mediated AMPA receptor endocytosis and recycling. Here, we demonstrate that in somatic and dendritic regions of hippocampal neurons a large fraction of the fluorescent signal originates from intracellular pH-GluA2, and that the decline in fluorescence in response to NMDA and AMPA primarily describes an intracellular acidification, which quenches the pHluorin signal from intracellular receptor pools. Neurons expressing an endoplasmic reticulum-retained mutant of GluA2 (pH-GluA2 ΔC49) displayed a larger response to NMDA than neurons expressing wild-type pH-GluA2. A similar NMDA-elicited decline in pHluorin signal was observed by expressing cytosolic pHluorin alone without fusion to GluA2 (cyto-pHluorin). Intracellular acidification in response to NMDA was further confirmed by using the ratiometric pH indicator carboxy-SNARF-1. The NMDA-induced decline was followed by rapid recovery of the fluorescent signal from both cyto-pHluorin and pH-GluA2. The recovery was sodium-dependent and sensitive to Na
+
/H
+
-exchanger (NHE) inhibitors. Moreover, recovery was more rapid after shRNA-mediated knockdown of the GluA2 binding PDZ domain-containing protein interacting with C kinase 1 (PICK1). Interestingly, the accelerating effect of PICK1 knockdown on the fluorescence recovery was eliminated in the presence of the NHE1 inhibitor zoniporide. Our results indicate that the pH-GluA2 recycling assay is an unreliable assay for studying AMPA receptor trafficking and also suggest a role for PICK1 in regulating intracellular pH via modulation of NHE activity.
Informative Correlations among Metrics of Yellowstone Lake Cutthroat Trout Caught by Two Quantitative Methods across Three Recent Decades
Authors: Kaeding Lynn R.
Published: August 2013

The Yellowstone Cutthroat Trout Oncorhynchus clarkii bouvieri (YCT) of Yellowstone Lake, in Yellowstone National Park, Wyoming, is an iteroparous fish and obligate stream spawner. The size and several other attributes of the annual YCT spawning run in one lake tributary, Clear Creek, have been periodically estimated for several decades. The trends in that run's metrics have been assumed to depict the trends in the lake's YCT population as a whole, although such associations had not been substantiated by statistical analyses. The present study revealed strong correlations between metrics of YCT in the run (years 1977–2007) and of “prespawner” YCT (i.e., mature fish whose excised gonads indicated that the fish would have spawned the next year) caught in gill nets set in various lake locations the preceding fall. Data for both capture methods also revealed a negative effect of spawning population density on YCT somatic growth, which is known to be positively associated with fecundity. This study showed that most metrics of YCT in the Clear Creek spawning run were indicative of those of prespawner YCT in Yellowstone Lake and aided the development of population models that are needed to determine the causal factors in the recent, three-decade population decline of YCT.

Received April 10, 2013; accepted June 12, 2013
Two Cases of Intranasal Naloxone Self-Administration in Opioid Overdose
Authors: Green Traci C. , Ray Madeline , Bowman Sarah E. , McKenzie Michelle , Rich Josiah D.
Published: August 2013

Background

Overdose is a leading cause of death for former prisoners, exacting its greatest toll during the first 2 weeks post release. Protective effects have been observed with training individuals at high risk of overdose and prescribing them naloxone, an opioid antagonist that reverses the effects of the opioid-induced respiratory depression that causes death.

Cases

The authors report 2 people with opiate use histories who self-administered intranasal naloxone to treat their own heroin overdoses following release from prison. Patient A is a 34-year-old male, who reported having experienced an overdose on heroin the day after he was released from incarceration. Patient B is a 29-year-old female, who reported an overdose on her first injection of heroin, 17 days post release from incarceration. Both patients self-administered the medication but were assisted at some point during the injury by a witness whom they had personally instructed in how to prepare and administer the medication. Neither patient experienced withdrawal symptoms following exposure to naloxone.

Discussion

Self-administration of naloxone should not be a goal of overdose death prevention training. A safer, more reliable approach is to prescribe naloxone to at-risk patients and train and also equip members of their household and social or drug-using networks in overdose prevention and response.
Suprasegmental Lexical Stress Cues in Visual Speech can Guide Spoken-Word Recognition
Authors: Jesse Alexandra , McQueen James M.
Published: August 2013

Visual cues to the individual segments of speech and to sentence prosody guide speech recognition. The present study tested whether visual suprasegmental cues to the stress patterns of words can also constrain recognition. Dutch listeners use acoustic suprasegmental cues to lexical stress (changes in duration, amplitude, and pitch) in spoken-word recognition. We asked here whether they can also use visual suprasegmental cues. In two categorization experiments, Dutch participants saw a speaker say fragments of word pairs that were segmentally identical but differed in their stress realization (e.g., ˈ ca-vi from cavia “guinea pig” vs. ˌka-vi from kaviaar “caviar”). Participants were able to distinguish between these pairs from seeing a speaker alone. Only the presence of primary stress in the fragment, not its absence, was informative. Participants were able to distinguish visually primary from secondary stress on first syllables, but only when the fragment-bearing target word carried phrase-level emphasis. Furthermore, participants distinguished fragments with primary stress on their second syllable from those with secondary stress on their first syllable (e.g., pro-ˈjec from projector “projector” vs. ˌpro-jec from projectiel “projectile”), independently of phrase-level emphasis. Seeing a speaker thus contributes to spoken-word recognition by providing suprasegmental information about the presence of primary lexical stress.
The GEF Bcr activates RhoA/MAL signaling to promote keratinocyte differentiation via desmoglein-1
Authors: Dubash Adi D. , Koetsier Jennifer L. , Amargo Evangeline V. , Najor Nicole A. , Harmon Robert M. , Green Kathleen J.
Published: August 2013

Although much is known about signaling factors downstream of Rho GTPases that contribute to epidermal differentiation, little is known about which upstream regulatory proteins (guanine nucleotide exchange factors [GEFs] or GTPase-activating proteins [GAPs]) are involved in coordinating Rho signaling in keratinocytes. Here we identify the GEF breakpoint cluster region (Bcr) as a major upstream regulator of RhoA activity, stress fibers, and focal adhesion formation in keratinocytes. Loss of Bcr reduced expression of multiple markers of differentiation (such as desmoglein-1 [Dsg1], keratin-1, and loricrin) and abrogated MAL/SRF signaling in differentiating keratinocytes. We further demonstrated that loss of Bcr or MAL reduced levels of Dsg1 mRNA in keratinocytes, and ectopic expression of Dsg1 rescued defects in differentiation seen upon loss of Bcr or MAL signaling. Taken together, these data identify the GEF Bcr as a regulator of RhoA/MAL signaling in keratinocytes, which in turn promotes differentiation through the desmosomal cadherin Dsg1.
Tristetraprolin regulation of interleukin 23 mRNA stability prevents a spontaneous inflammatory disease
Authors: Molle Céline , Zhang Tong , Ysebrant de Lendonck Laure , Gueydan Cyril , Andrianne Mathieu , Sherer Félicie , Van Simaeys Gaetan , Blackshear Perry J. , Leo Oberdan , Goriely Stanislas
Published: August 2013

Interleukin (IL) 12 and IL23 are two related heterodimeric cytokines produced by antigen-presenting cells. The balance between these two cytokines plays a crucial role in the control of Th1/Th17 responses and autoimmune inflammation. Most studies focused on their transcriptional regulation. Herein, we explored the role of the adenine and uridine–rich element (ARE)–binding protein tristetraprolin (TTP) in influencing mRNA stability of IL12p35, IL12/23p40, and IL23p19 subunits. LPS-stimulated bone marrow–derived dendritic cells (BMDCs) from TTP−/− mice produced normal levels of IL12/23p40. Production of IL12p70 was modestly increased in these conditions. In contrast, we observed a strong impact of TTP on IL23 production and IL23p19 mRNA stability through several AREs in the 3′ untranslated region. TTP−/− mice spontaneously develop an inflammatory syndrome characterized by cachexia, myeloid hyperplasia, dermatitis, and erosive arthritis. We observed IL23p19 expression within skin lesions associated with exacerbated IL17A and IL22 production by infiltrating γδ T cells and draining lymph node CD4 T cells. We demonstrate that the clinical and immunological parameters associated with TTP deficiency were completely dependent on the IL23–IL17A axis. We conclude that tight control of IL23 mRNA stability by TTP is critical to avoid severe inflammation.
Human L-ferritin deficiency is characterized by idiopathic generalized seizures and atypical restless leg syndrome
Authors: Cozzi Anna , Santambrogio Paolo , Privitera Daniela , Broccoli Vania , Rotundo Luisa Ida , Garavaglia Barbara , Benz Rudolf , Altamura Sandro , Goede Jeroen S. , Muckenthaler Martina U. , Levi Sonia
Published: August 2013

The ubiquitously expressed iron storage protein ferritin plays a central role in maintaining cellular iron homeostasis. Cytosolic ferritins are composed of heavy (H) and light (L) subunits that co-assemble into a hollow spherical shell with an internal cavity where iron is stored. The ferroxidase activity of the ferritin H chain is critical to store iron in its Fe3+ oxidation state, while the L chain shows iron nucleation properties. We describe a unique case of a 23-yr-old female patient affected by a homozygous loss of function mutation in the L-ferritin gene, idiopathic generalized seizures, and atypical restless leg syndrome (RLS). We show that L chain ferritin is undetectable in primary fibroblasts from the patient, and thus ferritin consists only of H chains. Increased iron incorporation into the FtH homopolymer leads to reduced cellular iron availability, diminished levels of cytosolic catalase, SOD1 protein levels, enhanced ROS production and higher levels of oxidized proteins. Importantly, key phenotypic features observed in fibroblasts are also mirrored in reprogrammed neurons from the patient’s fibroblasts. Our results demonstrate for the first time the pathophysiological consequences of L-ferritin deficiency in a human and help to define the concept for a new disease entity hallmarked by idiopathic generalized seizure and atypical RLS.
Eigenvalues of weighted 𝑝-Laplacian
Authors: Wang Lihan
Published: August 2013

In a paper by Z. Lu and J. Rowlett, it is shown that the eigenvalues of the weighted Laplacian can be approximated by eigenvalues of a naturally associated family of narrow graphs. In this paper, we generalize this result to the

p
p

-Laplacian. Our approach features overcoming the nonlinearity of the

p
p

-Laplacian when

p
≠
2

p\neq 2

, which is different from the Laplacian case.
Prostaglandin E specifically upregulates the expression of the mannose-receptor on mouse bone marrow-derived macrophages.
Authors: Schreiber S , Blum J S , Chappel J C , Stenson W F , Stahl P D , Teitelbaum S L , Perkins S L
Published: August 2013

The macrophage mannose receptor (MMR) facilitates the binding and internalization of microorganisms and glycoproteins with terminal mannose residues. The receptor is progressively upregulated as bone marrow precursor cells mature into macrophages and thus may serve as a marker of differentiation. Prostaglandins of the E series (PGE) are known inhibitors of monocyte and macrophage precursor proliferation, an effect often associated with cellular maturation. MMR expression was therefore assessed after exposure of bone marrow macrophage precursor (BMMP) cells to these prostanoids. Receptor expression was determined by ligand binding and via immunoprecipitation of newly synthesized receptor molecules. PGE1 and PGE2 at 10(-9)-10(-6) M upregulated MMR surface expression and biosynthesis four- to sixfold in a dose-dependent manner. BMMPs responsive to prostaglandins were characterized by plastic adherence, F4/80 antigen expression, and nonspecific esterase activity. Prostaglandins accelerated the expression of the MMR in cells by 48-72h, with maximal levels of receptor expression being identical in control or treated cells. Thus, prostaglandins enhanced mannose receptor expression in adherent but not fully differentiated macrophage precursors. This effect is specific for PGE and is mimicked by dibutyrl cyclic AMP. These results indicate that prostaglandins accelerate MMR expression and hence the differentiation of macrophage precursor cells. Cells resident in the bone marrow secrete abundant prostaglandins, suggesting that a paracrine mechanism may exist to regulate MMR expression and function.
Thrombin immobilized to extracellular matrix is a potent mitogen for vascular smooth muscle cells: nonenzymatic mode of action.
Authors: Bar-Shavit R , Benezra M , Eldor A , Hy-Am E , Fenton J W , Wilner G D , Vlodavsky I
Published: August 2013

Esterolytically inactive diisopropyl fluorophosphate-conjugated thrombin (DIP-alpha-thrombin) stimulated 3H-thymidine incorporation and proliferation of growth-arrested vascular smooth muscle cells (SMCs), similar to native alpha-thrombin. Half-maximal mitogenic response of SMCs was obtained at 1 nM thrombin and was specifically blocked by the leech-derived, high-affinity thrombin inhibitor, hirudin. Native thrombin and a variety of thrombin species that were chemically modified to alter thrombin procoagulant or esterolytic functions were found to induce 3H-thymidine incorporation to a similar extent. Exposure of SMCs to DIP-alpha-thrombin caused a rapid and transient expression of the c-fos protooncogene, determined by Northern blot analysis. These results indicate that thrombin is a potent mitogen for SMCs through a distinct non-enzymatic domain. Binding of 125I-alpha-thrombin to SMC cultures revealed an apparent dissociation constant of 6 nM and an estimated 5.4 x 10(5) binding sites per cell. This binding was inhibited to the same extent by native thrombin and by its nonenzymatic form, DIP-alpha-thrombin. Moreover, the chemotactic fragment of thrombin (CB67-129), which failed to elicit a mitogenic response, competed for 125I-alpha-thrombin binding to SMCs. Cross-linking analysis of 125I-alpha-thrombin to SMCs revealed a specific cell-surface binding site 55 kDa in size. Finally, thrombin immobilized to a naturally produced extracellular matrix retained potent mitogenic activity toward SMCs. These observations lend support to the possibility that in vivo, subendothelial basement membranes sequester thrombin (as well as other bioactive molecules), which may stimulate localized and persistent growth of arterial SMCs. Thrombin may thus be involved directly in progression of atherosclerotic plaque formation.
Structural requirements for neural cell adhesion molecule-heparin interaction.
Authors: Reyes A A , Akeson R , Brezina L , Cole G J
Published: August 2013

Two biological domains have been identified in the amino terminal region of the neural cell adhesion molecule (NCAM): a homophilic-binding domain, responsible for NCAM-NCAM interactions, and a heparin-binding domain (HBD). It is not known whether these two domains exist as distinct structural entities in the NCAM molecule. To approach this question, we have further defined the relationship between NCAM-heparin binding and cell adhesion. A putative HBD consisting of two clusters of basic amino acid residues located close to each other in the linear amino acid sequence of NCAM has previously been identified. Synthetic peptides corresponding to this domain were shown to bind both heparin and retinal cells. Here we report the construction of NCAM cDNAs with targeted mutations in the HBD. Mouse fibroblast cells transfected with the mutant cDNAs express NCAM polypeptides with altered HBD (NCAM-102 and NCAM-104) or deleted HBD (HBD-) at levels similar to those of wild-type NCAM. Mutant NCAM polypeptides purified from transfected cell lines have substantially reduced binding to heparin and fail to promote chick retinal cell attachment. Furthermore, whereas a synthetic peptide that contains both basic amino acid clusters inhibits retinal-cell adhesion to NCAM-coated dishes, synthetic peptides in which either one of the two basic regions is altered to contain only neutral amino acids do not inhibit this adhesion. These results confirm that this region of the NCAM polypeptide does indeed mediate not only the large majority of NCAM's affinity for heparin but also a significant portion of the cell-adhesion-mediating capability of NCAM.
A protein kinase C isozyme is translocated to cytoskeletal elements on activation.
Authors: Mochly-Rosen D , Henrich C J , Cheever L , Khaner H , Simpson P C
Published: August 2013

Protein kinase C (PKC)1 isozymes comprise a family of related cytosolic kinases that translocate to the cell particulate fraction on stimulation. The activated enzyme is thought to be on the plasma membrane. However, phosphorylation of protein substrates occurs throughout the cell and is inconsistent with plasma membrane localization. Using an isozyme-specific monoclonal antibody we found that, on activation, this PKC isozyme translocates to myofibrils in cardiac myocytes and to microfilaments in fibroblasts. Translocation of this activated PKC isozyme to cytoskeletal elements may explain some of the effects of PKC on cell contractility and morphology. In addition, differences in the translocation site of individual isozymes--and, therefore, phosphorylation of different substrates localized at these sites--may explain the diverse biological effects of PKC.
Phorbol ester-induced actin cytoskeletal reorganization requires a heavy metal ion.
Authors: Hedberg K K , Birrell G B , Griffith O H
Published: August 2013

The cell-permeant heavy metal chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine(TPEN) was found to counteract phorbol ester-induced actin reorganization in PTK2 and Swiss 3T3 cells. By using fluorescence and the higher resolution technique of photoelectron microscopy to monitor actin patterns, 15-min pretreatment with 25-50 microM TPEN was found to dramatically reduce actin alterations resulting from subsequent phorbol ester treatment in PTK2 cells. Similar results were obtained with Swiss 3T3 cells using 50 microM TPEN for 1.5 h. Phorbol ester-induced actin alterations are thought to depend on activation of protein kinase C (PKC). In contrast to the phorbol ester effect, the PKC-independent actin cytoskeletal disruption caused by staurosporine and cytochalasin B was unaffected by TPEN pretreatment. TPEN did not block phorbol ester-induced activation of PKC in Swiss 3T3 cells, as observed by the phosphorylation of the 80K PKC substrate protein (MARCKS protein). TPEN also did not inhibit partially purified PKC from Swiss 3T3 cells in an in vitro PKC-specific commercial assay. To establish that the effect of TPEN is the removal of metal ions and not some other nonspecific effect of TPEN, a series of transition metal ions was added at the end of the TPEN pretreatment. The results indicate that the transient but dramatic phorbol ester-induced reorganization of the actin cytoskeleton in cultured cells depends on an interaction of PKC with a heavy metal, probably zinc.
A growth factor for cardiac myocytes is produced by cardiac nonmyocytes.
Authors: Long C S , Henrich C J , Simpson P C
Published: August 2013

Cardiac nonmyocytes, primarily fibroblasts, surround cardiac myocytes in vivo. We examined whether nonmyocytes could modulate myocyte growth by production of one or more growth factors. Cardiac myocyte hypertrophic growth was stimulated in cultures with increasing numbers of cardiac nonmyocytes. This effect of nonmyocytes on myocyte size was reproduced by serum-free medium conditioned by the cardiac nonmyocytes. The majority of the nonmyocyte-derived myocyte growth-promoting activity bound to heparin-Sepharose and was eluted with 0.75 M NaCl. Several known polypeptide growth factors found recently in cardiac tissue, namely acidic fibroblast growth factor (aFGF), basic FGF (bFGF), platelet-derived growth factor (PDGF), tumor necrosis factor alpha (TNF alpha), and transforming growth factor beta 1 (TGF beta 1), also caused hypertrophy of cardiac myocytes in a dose-dependent manner. However, the nonmyocyte-derived growth factor (tentatively named NMDGF) could be distinguished from these other growth factors by different heparin-Sepharose binding profiles (TNF alpha, aFGF, bFGF, and TGF beta 1) by neutralizing growth factor-specific antisera (PDGF, TNF alpha, aFGF, bFGF, and TGF beta 1), by the failure of NMDGF to stimulate phosphatidylinositol hydrolysis (PDGF and TGF beta 1), and, finally, by the apparent molecular weight of NMDGF (45-50 kDa). This nonmyocyte-derived heparin-binding growth factor may represent a novel paracrine growth mechanism in myocardium.
Subcellular distribution of the alpha subunit(s) of Gi: visualization by immunofluorescent and immunogold labeling.
Authors: Lewis J M , Woolkalis M J , Gerton G L , Smith R M , Jarett L , Manning D R
Published: August 2013

The subcellular distribution of the alpha subunit(s) of Gi has an obvious bearing on the ability of this protein to interact with receptors and targets and on its potential to serve in still unexplored capacities. In this study, we have examined the distribution of Gi alpha by means of light and electron microscopy. The cells employed were mouse 3T3 fibroblasts, normal rat kidney fibroblasts, rat C6 glioma cells, human umbilical vein endothelial cells, and human 293 kidney fibroblasts. By indirect immunofluorescence, two patterns of Gi alpha were evident. The more prominent was that associated with phase-dense, cytoplasmic structures exhibiting a tubule-like morphology. A similar distribution was noted for mitochondria, indicating attachment to a subset of microtubules. The second pattern appeared as a diffuse, particulate fluorescence associated with the plasma membrane. By immunogold labeling and electron microscopy, two populations of Gi alpha were again evident. In this instance, labeling of the plasma membrane was the more prominent. Gold particles were most often evenly distributed along the plasma membrane and were concentrated along microspikes. The second, less abundant population of Gi alpha represented the subunit (or fragments) within lysosomes. Specificity in immunolabeling was confirmed in all instances by immunotransfer blotting, the use of antibodies differing in specificities for epitopes within Gi alpha, the absence of labeling with preimmune sera, and the decrease in labeling after preincubation of antisera with appropriate peptides. These results support the proposal that several populations of Gi alpha exist: those evident within the cytoplasm by immunofluorescence, those present at the plasma membrane, and those evident within lysosomes by immunogold labeling.
Association of the crosslinked IgE receptor with the membrane skeleton is independent of the known signaling mechanisms in rat basophilic leukemia cells.
Authors: Apgar J R
Published: August 2013

Crosslinking of the IgE receptor on the surface of rat basophilic leukemia (RBL) cells by multivalent antigen induces an association of these receptors with the detergent-insoluble membrane skeleton. Detergent insolubility of the receptor can also be induced on purified plasma membranes isolated from RBL cells by the use of either IgE oligomers or IgE monomers plus multivalent antigen. The critical event in initiating this interaction between the receptor and the membrane skeleton is cross-linking of the receptor. This association is rapid, and, when triggered by multivalent antigen, it is quickly reversed by the addition of excess monovalent antigen. The fact that this association occurs with the use of purified plasma membranes indicates that all of the components necessary for this interaction are present in the plasma membrane and that intracellular components are not required. Although crosslinking of the receptor activates phospholipase C and phospholipase A2 leading to the generation of several second messengers, none of these signaling mechanisms appears to be involved in IgE receptor interaction with the membrane skeleton. This interaction cannot be induced by phorbol 12-myristate 13-acetate (PMA), ionomycin, or a combination of these two reagents, although this will result in degranulation. Furthermore, receptor detergent insolubility is temperature independent when triggered by multivalent antigen, thus indicating that enzyme-catalyzed reactions are not important. This was verified by the fact that a variety of inhibitors that block phosphatidylinositol metabolism, arachidonic acid metabolism, Ca2+ influx, and protein kinase C (PKC) activation had no effect on antigen-induced association of the receptor with the membrane skeleton. These results indicate that the signaling mechanisms leading to the degranulation response are not involved in the association of the crosslinked receptor with the membrane skeleton.