bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunogold characteristics of VGLUT3‐positive GABAergic nerve terminals suggest corelease of glutamate
Authors: Stensrud Mats Julius , Sogn Carl Johan , Gundersen Vidar
Published: May 2015

There is compelling evidence that glutamate can act as a cotransmitter in the mammalian brain. Interestingly, the third vesicular glutamate transporter (VGLUT3) is primarily found in neurons that were anticipated to be nonglutamatergic. Whereas the function of VGLUT3 in acetylcholinergic and serotoninergic neurons has been elucidated, the role of VGLUT3 in neurons releasing gamma‐aminobutyric acid (GABA) is not settled. We have previously shown that VGLUT3 is found together with the vesicular GABA transporter (VIAAT) on synaptic vesicle membranes in the hippocampus. Now we provide novel electron microscopic data from the rat hippocampus suggesting that glutamate is enriched in inhibitory nerve terminals containing VGLUT3 compared to those lacking VGLUT3. The opposite was found for GABA; VGLUT3‐positive inhibitory terminals contained lower density of GABA labeling compared to VGLUT3‐negative inhibitory terminals. In addition, semiquantitative confocal immunofluorescence showed that N‐methyl‐D‐aspartate (NMDA)‐receptor labeling was present more frequently in VGLUT3‐positive/VIAAT‐positive synapses versus in VGLUT3‐negative/VIAAT‐positive synapses. Electron microscopic immunogold data further suggest that NMDA receptors are enriched in VGLUT3 containing inhibitory terminals. Our data reveal new chemical characteristics of a subset of GABAergic interneurons in the hippocampus. The analyses suggest that glutamate is coreleased with GABA from hippocampal basket cell‐synapses to act on NMDA receptors. J. Comp. Neurol. 523:2698–2713, 2015. © 2015 Wiley Periodicals, Inc.
Projections from the subparaventricular zone define four channels of output from the circadian timing system
Authors: Vujovic Nina , Gooley Joshua J. , Jhou Thomas C. , Saper Clifford B.
Published: May 2015

The subparaventricular zone of the hypothalamus (SPZ) is the main efferent target of neural projections from the suprachiasmatic nucleus (SCN) and an important relay for the circadian timing system. Although the SPZ is fairly homogeneous cytoarchitecturally and neurochemically, it has been divided into distinct functional and connectional subdivisions. The dorsal subdivision of the SPZ (dSPZ) plays an important role in relaying signals from the SCN controlling body temperature rhythms, while the ventral subdivision (vSPZ) is critical for rhythms of sleep and locomotor activity (Lu et al. [] J Neurosci 21:4864–4874). On the other hand, the medial part of the SPZ receives input mainly from the dorsomedial SCN, whereas the lateral SPZ receives input from the ventrolateral SCN and the retinohypothalamic tract (Leak and Moore [] J Comp Neurol 433:312–334). We therefore investigated whether there are corresponding differences in efferent outputs from these four quadrants of the SPZ (dorsolateral, ventrolateral, dorsomedial, and ventromedial) by a combination of anterograde and retrograde tracing. We found that, while all four subdivisions of the SPZ share a similar backbone of major projection pathways to the septal region, thalamus, hypothalamus, and brainstem, each segment of the SPZ has a specific set of targets where its projections dominate. Furthermore, we observed intra‐SPZ projections of varying densities between the four subdivisions. Taken together, this pattern of organization suggests that the circadian timing system may have several parallel neural outflow pathways that provide a road map for understanding how they subserve different functions. J. Comp. Neurol. 523:2714–2737, 2015. © 2015 Wiley Periodicals, Inc.
Evidence for involvement of a limbic paraventricular hypothalamic inhibitory network in hypothalamic‐pituitary‐adrenal axis adaptations to repeated stress
Authors: Radley Jason J. , Sawchenko Paul E.
Published: May 2015

Emotional stressors activate a stereotyped set of limbic forebrain cell groups implicated in constraining stress‐induced hypothalamic‐pituitary‐adrenal (HPA) axis activation by inhibiting hypophysiotropic neurons in the paraventricular hypothalamic nucleus (PVH). We previously identified a circumscribed, anterior part of the bed nuclei of the stria terminalis (aBST) that houses stress‐sensitive, PVH‐projecting, γ‐aminobutyric acid (GABA)‐ergic neurons as representing a site of convergence of stress‐inhibitory influences originating from medial prefrontal and hippocampal cortices. Here we investigate whether exaggerated HPA axis responses associated with chronic variable stress (CVS; daily exposure to different stressors at unpredictable times over 14 days, followed by restraint stress on day 15) and diminished HPA output seen following repeated (14 days) restraint‐stress exposure are associated with differential engagement of the limbic modulatory network. Relative to acutely restrained rats, animals subjected to CVS showed the expected increase (sensitization) in HPA responses and diminished levels of activation (Fos) of GABAergic neurons and glutamic acid decarboxylase (GAD) mRNA expression in the aBST. By contrast, repeated restraint stress produced habituation in HPA responses, maintained levels of activation of GABAergic neurons, and increased GAD expression in the aBST. aBST‐projecting neurons in limbic sites implicated in HPA axis inhibition tended to show diminished activational responses in both repeated‐stress paradigms, with the exception of the paraventricular thalamic nucleus, in which responsiveness was maintained in repeatedly restrained animals. The results are consistent with the view that differential engagement of HPA inhibitory mechanisms in the aBST may contribute to alterations in HPA axis responses to emotional stress in sensitization and habituation paradigms. J. Comp. Neurol. 523:2769–2787, 2015. © 2015 Wiley Periodicals, Inc.
The mummified brain of a pleistocene woolly mammoth (Mammuthus primigenius) compared with the brain of the extant African elephant (Loxodonta africana)
Authors: Kharlamova Anastasia S. , Saveliev Sergei V. , Protopopov Albert V. , Maseko Busisiwe C. , Bhagwandin Adhil , Manger Paul R.
Published: May 2015

This study presents the results of an examination of the mummified brain of a pleistocene woolly mammoth (Mammuthus primigenius) recovered from the Yakutian permafrost in Siberia, Russia. This unique specimen (from 39,440–38,850 years BP) provides the rare opportunity to compare the brain morphology of this extinct species with a related extant species, the African elephant (Loxodonta africana). An anatomical description of the preserved brain of the woolly mammoth is provided, along with a series of quantitative analyses of various brain structures. These descriptions are based on visual inspection of the actual specimen as well as qualitative and quantitative comparison of computed tomography imaging data obtained for the woolly mammoth in comparison with magnetic resonance imaging data from three African elephant brains. In general, the brain of the woolly mammoth specimen examined, estimated to weigh between 4,230 and 4,340 g, showed the typical shape, size, and gross structures observed in extant elephants. Quantitative comparative analyses of various features of the brain, such as the amygdala, corpus callosum, cerebellum, and gyrnecephalic index, all indicate that the brain of the woolly mammoth specimen examined has many similarities with that of modern African elephants. The analysis provided here indicates that a specific brain type representative of the Elephantidae is likely to be a feature of this mammalian family. In addition, the extensive similarities between the woolly mammoth brain and the African elephant brain indicate that the specializations observed in the extant elephant brain are likely to have been present in the woolly mammoth. J. Comp. Neurol. 523:2326–2343, 2015. © 2015 Wiley Periodicals, Inc.
Effects of the jimpy mutation on mouse retinal structure and function
Authors: Hovhannisyan Anahit , Benkner Boris , Biesemeier Antje , Schraermeyer Ulrich , Kukley Maria , Münch Thomas A.
Published: May 2015

The Jimpy mutant mouse has a point mutation in the proteolipid protein gene (plp1). The resulting misfolding of the protein leads to oligodendrocyte death, myelin destruction, and failure to produce adequately myelinated axons in the central nervous system (CNS). It is not known how the absence of normal myelination during development influences neural function. We characterized the Jimpy mouse retina to find out whether lack of myelination in the optic nerve during development has an effect on normal functioning and morphology of the retina. Optokinetic reflex measurements showed that Jimpy mice had, in general, a functional visual system. Both PLP1 antibody staining and reverse transcriptase–polymerase chain reaction for plp1 mRNA showed that plp1 is not expressed in the wild‐type retina. However, in the optic nerve, plp1 is normally expressed, and consequently, in Jimpy mutant mice, myelination of axons in the optic nerve was mostly absent. Nevertheless, neither axon count nor axon ultrastructure in the optic nerve was affected. Physiological recordings of ganglion cell activity using microelectrode arrays revealed a decrease of stimulus‐evoked activity at mesopic light levels. Morphological analysis of the retina did not show any significant differences in the gross morphology, such as thickness of retinal layers or cell number in the inner and outer nuclear layer. The cell bodies in the inner nuclear layer, however, were larger in the peripheral retina of Jimpy mutant mice. Antibody labeling against cell type–specific markers showed that the number of rod bipolar and horizontal cells was increased in Jimpy mice. In conclusion, whereas the Jimpy mutation has dramatic effects on the myelination of retinal ganglion cell axons, it has moderate effects on retinal morphology and function. J. Comp. Neurol. 523:2788–2806, 2015. © 2015 Wiley Periodicals, Inc.
UV‐Initiated Synthesis of Electroactive High Surface Area Ta and Ti Mesoporous Oxides Composites with Polypyrrole Nanowires within the Pores
Authors: Smith Luke A. C. , Romer Frederik , Trudeau Michel L. , Smith Mark E. , Hanna John V. , Antonelli David M.
Published: May 2015

This paper describes the synthesis and characterization of high surface area mesoporous Ti and Ta oxides with polypyrrole nanowires in the pores. The incorporation of polymer was used to improve the electron conductivity into the channels inside these high surface area (400–1000 m2 g−1) materials in order to exploit surface redox sites for possible pseudocapacitive Li storage. Synthesis was achieved using catalyst‐free UV‐initiated polymerization of vapor‐loaded pyrrole monomer. The best materials showed improved conductivity for both the Ti and Ta oxides as well as improved Li capacity (190 mA h g−1) relative to the pristine material (128 mA h g−1) and superior capacity retention (49 % as compared to 22 %) for the Ti composites. The retention in surface area was also 87 % compared to 49 % reported previously for analogous materials synthesized by catalyst‐initiated methods, which only yielded Li capacities of 170 mA h g−1, further highlighting the superiority of this new photochemical approach.
Enzyme‐Mimicking Peptides to Catalytically Grow ZnO Nanocrystals in Non‐Aqueous Environments
Authors: Maeda Yoshiaki , Wei Zengyan , Ikezoe Yasuhiro , Matsui Hiroshi
Published: May 2015

The application of enzymes for biomimetic material growth is a potential path to breakthroughs in catalytic inorganic nanocrystal synthesis because the biocatalytic functions of proteins can lead to highly efficient growth of materials with accurate structures at low temperature. However, many metal nanocrystals are grown in organic solvents, which is not an ideal environment for biomineralization with enzymes due to denaturing of catalytic pocket structures. Here, we applied a catalytic CP4 peptide, optimized for protease reactions such as amide and ester hydrolysis, to test the catalytic ZnO nanocrystal growth in non‐aqueous environments and the performance was compared with the enzyme subtilisin. We demonstrated that the CP4 catalytic peptide, evolved by the hydrogel‐based combinatory phage display library, could grow ZnO nanoparticles from zinc acetate precursors in higher crystallinity and yield, as compared to the protease enzyme, in an organic solvent by catalyzing ester‐elimination of precursors at room temperature. This result indicates that catalytic oligopeptides, which possess no rigid 3D structures but are rather flexible, could fold into the catalytically active conformation more frequently in harsh environments such as organic solvents.
Xerogel Coatings Produced by the Sol–Gel Process as Anti‐Fouling, Fouling‐Release Surfaces: From Lab Bench to Commercial Reality
Authors: Detty Michael R. , Ciriminna Rosaria , Bright Frank V. , Pagliaro Mario
Published: May 2015

We show the large environmental and practical impact of nanochemistry innovation by telling the story of how the sol–gel hybrid coating technology was subtly adapted to meet the urgent demand for clean antifouling marine coatings for vessels of widely different size, composition, and operation requirements.
Fear conditioned responses and PTSD symptoms in children: Sex differences in fear‐related symptoms
Authors: Gamwell Kaitlyn , Nylocks Maria , Cross Dorthie , Bradley Bekh , Norrholm Seth D. , Jovanovic Tanja
Published: May 2015

Fear conditioning studies in adults have found that posttraumatic stress disorder (PTSD) is associated with heightened fear responses and impaired discrimination. The objective of the current study was to examine the association between PTSD symptoms and fear conditioned responses in children from a highly traumatized urban population. Children between 8 and 13 years old participated in a fear conditioning study in addition to providing information about their trauma history and PTSD symptoms. Results showed that females showed less discrimination between danger and safety signals during conditioning compared to age‐matched males. In boys, intrusive symptoms were predictive of fear responses, even after controlling for trauma exposure. However, in girls, conditioned fear to the danger cue was predictive of self‐blame and fear of repeated trauma. This study suggests there are early sex differences in the patterns of fear conditioning and that these sex differences may translate to differential risk for trauma‐related psychopathology. © 2015 Wiley Periodicals, Inc. Dev Psychobiol 57: 799–808, 2015.
Exposure to synthetic oxytocin during delivery and its effect on psychomotor development
Authors: González‐Valenzuela María‐José , López‐Montiel Dolores , González‐Mesa Ernesto Santiago
Published: May 2015

The main objective is to examine the influence of oxytocin administration during delivery on psychomotor development at age five years. This was a retrospective cohort study involving two groups: children of mothers exposed vs. not exposed to oxytocin during labor. Of the 7,465 newborns registered in our maternity service during 2006 we randomly selected an initial sample of 400 children. Of these, 146 children were assessed using the motor scale of the Battelle Developmental Inventory. Other predictor variables that could potentially act as confounders and/or interact with the main relationship were also examined. The data were subjected to bivariate analysis, estimates of measures of strength of association, stratified analysis and multivariate binary logistic regression. The results indicate that exposure to synthetic oxytocin during delivery is an independent risk factor for a delay in gross and fine motor development. This was the case after controlling for the variables duration of labor and sex of the newborn, none of which modified the effect of oxytocin on gross and fine motor development. However, sex of the newborn were shown to be confounding gross motor development. In light of these results, and with the aim of preventing possible psychomotor alterations, further studies are now needed to analyze the effect that the oxytocin dose and the duration of perfusion may have on children's subsequent development. © 2015 Wiley Periodicals, Inc. Dev Psychobiol 57: 908–920, 2015.
Plant‐soil feedbacks from 30‐year family‐specific soil cultures: phylogeny, soil chemistry and plant life stage
Authors: Mehrabi Zia , Bell Thomas , Lewis Owen T.
Published: May 2015

Intraspecific negative feedback effects, where performance is reduced on soils conditioned by conspecifics, are widely documented in plant communities. However, interspecific feedbacks are less well studied, and their direction, strength, causes, and consequences are poorly understood. If more closely related species share pathogens, or have similar soil resource requirements, plants may perform better on soils conditioned by more distant phylogenetic relatives. There have been few empirical tests of this prediction across plant life stages, and none of which attempt to account for soil chemistry. Here, we test the utility of phylogeny for predicting soil feedback effects on plant survival and performance (germination, seedling survival, growth rate, biomass). We implement a full factorial experiment growing species representing five families on five plant family‐specific soil sources. Our experiments exploit soils that have been cultured for over 30 years in plant family‐specific beds at Oxford University Botanic Gardens. Plant responses to soil source were idiosyncratic, and species did not perform better on soils cultured by phylogenetically more distant relatives. The magnitude and sign of feedback effects could, however, be explained by differences in the chemical properties of “home” and “away” soils. Furthermore, the direction of soil chemistry‐related plant‐soil feedbacks was dependent on plant life stage, with the effects of soil chemistry on germination success and accumulation of biomass inversely related. Our results (1) suggest that the phylogenetic distance between plant families cannot predict plant–soil feedbacks across multiple life stages, and (2) highlight the need to consider changes in soil chemistry as an important driver of population responses. The contrasting responses at plant life stages suggest that studies focusing on brief phases in plant demography (e.g., germination success) may not give a full picture of plant–soil feedback effects.
Sex‐specific foraging behavior in response to fishing activities in a threatened seabird
Authors: García‐Tarrasón Manuel , Bécares Juan , Bateman Santiago , Arcos José Manuel , Jover Lluís , Sanpera Carolina
Published: May 2015

Some seabird species have learnt to efficiently exploit fishing discards from trawling activities. However, a discard ban has been proposed as necessary in Europe to ensure the sustainability of the seas. It is of crucial importance for the management and conservation purposes to study the potential consequences of a discard ban on the foraging ecology of threatened seabirds. We assessed the influence of fishing activities on the feeding habits of 22 male and 15 female Audouin's gulls (Larus audouinii) from the Ebro Delta (Mediterranean Sea) during the breeding period using GPS loggers together with Stable Isotope Analysis (SIA), which provided new insights into their foraging behavior and trophic ecology, respectively. GPS data revealed different sex‐specific foraging patterns between workdays and weekends. Females were highly consistent in that they foraged at sea throughout the week even though discarding stops at weekends. In contrast, males switched from foraging at sea during the week (when discards are produced) to an increased use of rice field habitats at weekends (when fishermen do not work). This sex‐specific foraging behavior could be related to specific nutritional requirements associated with previous egg production, an energetically demanding period for females. However, on a broader time scale integrated by the SIA, both sexes showed a high degree of individual specialization in their trophic ecology. The need to obtain detailed information on the dependence and response of seabirds to fishing activities is crucial in conservation sciences. In this regard, sex‐specific foraging behavior in relation to fisheries has been overlooked, despite the ecological and conservation implications. For instance, this situation may lead to sex differentiation in bycatch mortality in longlines when trawlers do not operate. Moreover, any new fisheries policy will need to be implemented gradually to facilitate the adaptation of a specialized species to a discard ban scenario.
Long‐term livestock exclusion facilitates native woody plant encroachment in a sandy semiarid rangeland
Authors: Su Hua , Liu Wei , Xu Hong , Wang Zongshuai , Zhang Huifang , Hu Haixiao , Li Yonggeng
Published: May 2015

The role of livestock grazing in regulating woody cover and biomass in grass‐dominant systems is well recognized. However, the way in which woody plant populations in respond when livestock are removed from grazing in the absence of other disturbances, such as fire, remains unclear.

We conducted a 10‐year, replicated fencing experiment in a sandy semiarid rangeland in northern China (which has a mean annual rainfall of 365 mm), where fires have been actively suppressed for decades.

Fencing dramatically influenced the growth and age structure of the native tree species, Ulmus pumila, which is the sole dominant tree in the area. After a decade, the density of the U. pumila tree population in the fencing plots increased doubly and canopy cover increased triply. The proportion of both saplings (U2) and young trees (U3) increased in fencing plots but decreased in grazing plots after the 10‐year treatment period. The effects of fencing on U. pumila trees varied by age class, with potential implications for the future structure of the U. pumila tree community. Decadal fencing led to approximately 80‐fold increase in recruitment and a nearly 2.5‐fold decrease in the mortality of both U2 and U3. Further, livestock grazing generated a “browsing trap” to the recruitment of both U2 and U3, and had a small impact on the mortality of old trees. A long‐term, fencing‐driven shift in woody species composition was mediated via its effects on both recruitment and mortality rates.

Synthesis and applications. Our results demonstrate that in the long‐term absence of both fire and livestock, native woody plant encroachment tends to occur in sandy rangelands, transforming the woody plant demography in the process. The feasibility of full livestock exclusion in sandy rangelands requires further discussion. A balanced amount of livestock grazing may provide critical ecosystem services by regulating woody cover and mediating woody plant encroachment.
A simple validated method for predicting the risk of hospitalization for worsening of heart failure in ambulatory patients: the Redin‐SCORE
Authors: Álvarez‐García Jesús , Ferrero‐Gregori Andreu , Puig Teresa , Vázquez Rafael , Delgado Juan , Pascual‐Figal Domingo , Alonso‐Pulpón Luis , González‐Juanatey José R. , Rivera Miguel , Worner Fernando , Bardají Alfredo , Cinca Juan ,   on behalf of the investigators of the Spanish Heart Failure Network (REDINSCOR)
Published: May 2015

Aims

Prevention of hospital readmissions is one of the main objectives in the management of patients with heart failure (HF). Most of the models predicting readmissions are based on data extracted from hospitalized patients rather than from outpatients. Our objective was to develop a validated score predicting 1‐month and 1‐year risk of readmission for worsening of HF in ambulatory patients.

Methods and results

A cohort of 2507 ambulatory patients with chronic HF was prospectively followed for a median of 3.3 years. Clinical, echocardiographic, ECG, and biochemical variables were used in a competing risk regression analysis to construct a risk score for readmissions due to worsening of HF. Thereafter, the score was externally validated using a different cohort of 992 patients with chronic HF (MUSIC registry). Predictors of 1‐month readmission were the presence of elevated natriuretic peptides, left ventricular (LV) HF signs, and estimated glomerular filtration rate (eGFR) <60 mL/min/m2. Predictors of 1‐year readmission were elevated natriuretic peptides, anaemia, left atrial size >26 mm/m2, heart rate >70 b.p.m., LV HF signs, and eGFR <60 mL/min/m2. The C‐statistics for the models were 0.72 and 0.66, respectively. The cumulative incidence function distinguished low‐risk (<1% event rate) and high‐risk groups (>5% event rate) for 1‐month HF readmission. Likewise, low‐risk (7.8%), intermediate‐risk (15.6%) and high‐risk groups (26.1%) were identified for 1‐year HF readmission risk. The C‐statistics remained consistent after the external validation (<5% loss of discrimination).

Conclusion

The Redin‐SCORE predicts early and late readmission for worsening of HF using proven prognostic variables that are routinely collected in outpatient management of chronic HF.
Maintenance of serum potassium with sodium zirconium cyclosilicate (ZS‐9) in heart failure patients: results from a phase 3 randomized, double‐blind, placebo‐controlled trial
Authors: Anker Stefan D. , Kosiborod Mikhail , Zannad Faiez , Piña Ileana L. , McCullough Peter A. , Filippatos Gerasimos , van der Meer Peter , Ponikowski Piotr , Rasmussen Henrik S. , Lavin Philip T. , Singh Bhupinder , Yang Alex , Deedwania Prakash
Published: May 2015

Aims

Hyperkalaemia in heart failure patients limits use of cardioprotective renin–angiotensin–aldosterone system inhibitors (RAASi). Sodium zirconium cyclosilicate (ZS‐9) is a selective potassium ion trap, whose mechanism of action may allow for potassium binding in the upper gastrointestinal tract as early as the duodenum following oral administration. ZS‐9 previously demonstrated the ability to reduce elevated potassium levels into the normal range, with a median time of normalization of 2.2 h and sustain normal potassium levels for 28 days in HARMONIZE—a Phase 3, double‐blind, randomized, placebo‐controlled trial. In the present study we evaluated management of serum potassium with daily ZS‐9 over 28 days in heart failure patients from HARMONIZE, including those receiving RAASi therapies.

Methods and results

Heart failure patients with evidence of hyperkalaemia (serum potassium ≥5.1 mmol/L, n = 94) were treated with open‐label ZS‐9 for 48 h. Patients (n = 87; 60 receiving RAASi) who achieved normokalaemia (potassium 3.5–5.0 mmol/L) were randomized to daily ZS‐9 (5, 10, or 15 g) or placebo for 28 days. Mean potassium and proportion of patients maintaining normokalaemia during days 8–29 post‐randomization were evaluated. Despite RAASi doses being kept constant, patients on 5 g, 10 g, and 15 g ZS‐9 maintained a lower potassium level (4.7 mmol/L, 4.5 mmol/L, and 4.4 mmol/L, respectively) than the placebo group (5.2 mmol/L; P<0.01 vs. each ZS‐9 group); greater proportions of ZS‐9 patients (83%, 89%, and 92%, respectively) maintained normokalaemia than placebo (40%; P < 0.01 vs. each ZS‐9 group). The safety profile was consistent with previously reported overall study population.

Conclusion

Compared with placebo, all three ZS‐9 doses lowered potassium and effectively maintained normokalaemia for 28 days in heart failure patients without adjusting concomitant RAASi, while maintaining a safety profile consistent with the overall study population.
Dielectrophoretic immobilization of proteins: Quantification by atomic force microscopy
Authors: Laux Eva‐Maria , Knigge Xenia , Bier Frank F. , Wenger Christian , Hölzel Ralph
Published: May 2015

The combination of alternating electric fields with nanometer‐sized electrodes allows the permanent immobilization of proteins by dielectrophoretic force. Here, atomic force microscopy is introduced as a quantification method, and results are compared with fluorescence microscopy. Experimental parameters, for example the applied voltage and duration of field application, are varied systematically, and the influence on the amount of immobilized proteins is investigated. A linear correlation to the duration of field application was found by atomic force microscopy, and both microscopical methods yield a square dependence of the amount of immobilized proteins on the applied voltage. While fluorescence microscopy allows real‐time imaging, atomic force microscopy reveals immobilized proteins obscured in fluorescence images due to low S/N. Furthermore, the higher spatial resolution of the atomic force microscope enables the visualization of the protein distribution on single nanoelectrodes. The electric field distribution is calculated and compared to experimental results with very good agreement to atomic force microscopy measurements.
Exosome enrichment of human serum using multiple cycles of centrifugation
Authors: Kim Jeongkwon , Tan Zhijing , Lubman David M.
Published: May 2015

In this work, we compared the use of repeated cycles of centrifugation at conventional speeds for enrichment of exosomes from human serum compared to the use of ultracentrifugation (UC). After removal of cells and cell debris, a speed of 110 000 × g or 40 000 × g was used for the UC or centrifugation enrichment process, respectively. The enriched exosomes were analyzed using the bicinchoninic acid assay, 1D gel separation, transmission electron microscopy, Western blotting, and high‐resolution LC‐MS/MS analysis. It was found that a five‐cycle repetition of UC or centrifugation is necessary for successful removal of nonexosomal proteins in the enrichment of exosomes from human serum. More significantly, 5× centrifugation enrichment was found to provide similar or better performance than 5× UC enrichment in terms of enriched exosome protein amount, Western blot band intensity for detection of CD‐63, and numbers of identified exosome‐related proteins and cluster of differentiation (CD) proteins. A total of 478 proteins were identified in the LC‐MS/MS analyses of exosome proteins obtained from 5× UCs and 5× centrifugations including many important CD membrane proteins. The presence of previously reported exosome‐related proteins including key exosome protein markers demonstrates the utility of this method for analysis of proteins in human serum.
Evaluation of DNA methylation markers and their potential to predict human aging
Authors: Soares Bispo Santos Silva Deborah , Antunes Joana , Balamurugan Kuppareddi , Duncan George , Sampaio Alho Clarice , McCord Bruce
Published: May 2015

We present epigenetic methylation data for two genetic loci, GRIA2, and NPTX2, which were tested for prediction of age from different donors of biofluids. We analyzed 44 saliva samples and 23 blood samples from volunteers with ages ranging from 5 to 72 years. DNA was extracted and bisulfite modified using commercial kits. Specific primers were used for amplification and methylation profiles were determined by pyrosequencing. Methylation data from both markers and their relationship with age were determined using linear regression analysis, which indicates a positive correlation between methylation and age. Older individuals tend to have increased methylation in both markers compared to younger individuals and this trend was more pronounced in the GRIA2 locus when compared to NPTX2. The epigenetic predicted age, calculated using a GRIA2 regression analysis model, was strongly correlated to chronological age (R2 = 0.801), with an average difference of 6.9 years between estimated and observed ages. When using a NPTX2 regression model, we observed a lower correlation between predicted and chronological age (R2 = 0.654), with an average difference of 9.2 years. These data indicate these loci can be used as a novel tool for age prediction with potential applications in many areas, including clinical and forensic investigations.
Hydro‐meteorological drivers and sources of suspended sediment flux in the pro‐glacial zone of the retreating Castle Creek Glacier, Cariboo Mountains, British Columbia, Canada
Authors: Leggat Michael S. , Owens Philip N. , Stott Tim A. , Forrester Barry J. , Déry Stephen J. , Menounos Brian
Published: May 2015

Glaciers are major agents of erosion that increase sediment load to the downstream fluvial system. The Castle Creek Glacier, British Columbia, Canada, has retreated ~1.0 km in the past 70 years. Suspended sediment concentration (SSC) and streamflow (Q) were monitored independently at five sites within its pro‐glacial zone over a 60 day period from July to September 2011, representing part of the ablation season. Meteorological data were collected from two automatic weather stations proximal to the glacier. The time‐series were divided into hydrologic days and the shape and magnitude of the SSC response to hydro‐meteorological conditions (‘cold and wet’, ‘hot and dry’, ‘warm and damp’, and ‘storm’) were categorized using principal component analysis (PCA) and cluster analysis (CA). Suspended sediment load (SSL) was computed and summarized for the categories. The distribution of monitoring sites and results of the multivariate statistical analyses describe the temporal and spatial variability of suspended sediment flux and the relative importance of glacial and para‐glacial sediment sources in the pro‐glacial zone. During the 2011 study period, ~ 60% of the total SSL was derived from the glacial stream and sediment deposits proximal to the terminus of the glacier; during ‘storm’ events, that contribution dropped to ~40% as the contribution from diffuse and point sources of sediment throughout the pro‐glacial zone and within the meltwater channels increased. While ‘storm’ events accounted for just 3% of the study period, SSL was ~600% higher than the average over the monitoring period, and ~20% of the total SSL was generated in that time. Determining how hydro‐meteorological conditions and sediment sources control sediment fluxes will assist attempts to predict how pro‐glacial zones respond to future climate changes. Copyright © 2015 John Wiley & Sons, Ltd.
Use of terrestrial photogrammetry based on structure‐from‐motion for mass balance estimation of a small glacier in the Italian alps
Authors: Piermattei Livia , Carturan Luca , Guarnieri Alberto
Published: May 2015

Different high‐resolution techniques can be employed to obtain information about the three‐dimensional (3D) surface of glaciers. This is typically carried out using efficient, but also expensive and logistically demanding, light detection and ranging (LiDAR) technologies, such as airborne scanners and terrestrial laser scanners. Recent technological improvements in the field of image analysis and computer vision have prompted the development of a low‐cost photogrammetric approach, which is referred to as ‘structure‐from‐motion’ (SfM). Combined with dense image‐matching algorithms, this method has become competitive for the production of high‐quality 3D models. However, several issues typical of this approach should be considered for application in glacial environments. In particular, the surface morphology, the different substrata, the occurrence of sharp contrast from solar shadows and the variable distance from the camera positions can negatively affect the image texture, and reduce the possibility of obtaining a reliable point cloud from the images. The objective of this study is to test the structure‐from‐motion multi view stereo (SfM‐MVS) approach in a small debris‐covered glacier located in the eastern Italian Alps, using a consumer‐grade reflex camera and the computer vision‐based software PhotoScan. The quality of the 3D models produced by the SfM‐MVS process was assessed via the comparison with digital terrain models obtained from terrestrial laser scanning (TLS) surveys that were performed at the same epochs. The effect of different terrain gradients and different substrata (debris, snow and firn) was also evaluated in terms of the accuracy of the reconstruction by SfM‐MVS versus TLS. Our results show that the quality of this new photogrammetric approach is similar to the quality of TLS and that point cloud densities are comparable or even higher compared with TLS. However, special care should be taken while planning the SfM survey geometry, to optimize the 3D model quality and spatial coverage. Copyright © 2015 John Wiley & Sons, Ltd.
Dynamic riverine landscapes: the role of ecosystem engineers
Authors: Harvey Gemma L. , Bertoldi Walter
Published: May 2015

An important and highly active research agenda has developed at the interface of fluvial geomorphology and ecology that addresses the capacity for vegetation and animals to act as ecosystem engineers within fluvial systems. This paper briefly introduces this research domain and describes the 15 papers that contribute to the special issue on 'Dynamic riverine landscapes: the role of ecosystem engineers'. The papers illustrate the breadth of research activity at this interface, investigating the influence of a range of ecosystem engineering organisms through a combination of field study, laboratory experiments, numerical simulation and analysis of remotely sensed data. Together, the papers address a series of key themes: conceptual frameworks for feedbacks between aquatic biota, hydraulics, sediment dynamics and nutrient dynamics and their quantification through experimental and field research; the potential contribution of ecosystem engineering species to assist river recovery and restoration; and the contribution of riparian vegetation to bank stability and morphodynamics across a range of spatio‐temporal scales. Copyright © 2015 John Wiley & Sons, Ltd.
Soil carbon losses by sheet erosion: a potentially critical contribution to the global carbon cycle
Authors: Müller‐Nedebock Daniel , Chaplot Vincent
Published: May 2015

Despite soil erosion through water being a ubiquitous process and its environmental consequences being well understood, its effects upon the global carbon cycle still remain largely uncertain. How much soil organic carbon (SOC) is removed each year from soils by sheet wash, an important if not the most efficient mechanism of detachment and transport of surficial soil material? What are the main environnemental controls worldwide? These are important questions which largely remain unanswered. Empirical data from 240 runoff plots studied over entire rainy seasons from different regions of the world were analysed to estimate particulate organic carbon (POC) losses (POCL), and POC enrichment in the sediments compared to the bulk soil (ER), which can be used as a proxy of the fate of the eroded POC. The median POCL was 9.9 g C m‐2 y‐1 with highest values observed for semi‐arid soils (POCL = 10.8 g C m‐2 y‐1), followed by tropical soils (POCL = 6.4 g C m‐2 y‐1) and temperate soils (POCL = 1.7 g C m‐2 y‐1). Considering the mean POCL of 27.2 g C m‐2 y‐1, the total amount of SOC displaced annually by sheet erosion from its source would be 1.32 ± 0.20 Gt C, i.e. 14.6% of the net annual fossil fuel induced C emissions of 9 Gt C. Because of low sediment enrichment in POC, erosion‐induced CO2 emissions are likely to be limited in clayey environments while POC burial within hillslopes is likely to constitute an important carbon sink. In contrast, most of the POC displaced from sandy soils is likely to be emitted to the atmosphere. These results underpin the major role sheet wash plays in the displacement of SOC from its source and in the fate of the eroded SOC, with large variations across the different pedo‐climatic regions of the world. Copyright © 2015 John Wiley & Sons, Ltd.
Superheated liquid and supercritical denatured ethanol extraction of antioxidants from Crimson red grape stems
Authors: Wenzel Jonathan , Samaniego Cheryl Storer , Wang Lihua , Nelson La'Shyla , Ketchum Korrine , Ammerman Michelle , Zand Ali
Published: May 2015

Grapes are widely known for health benefits due to their antioxidant content. In wine production, grape stems are often discarded, though they has a higher content of antioxidants than the juice. The effectiveness of using an environmentally friendly solvent, ethanol, as a superheated liquid and supercritical fluid to extract antioxidant compounds from grape stems of organically grown Crimson Seedless grapes was evaluated. The Ferric Reducing Ability of Plasma (FRAP) assay and the Total Phenolic Content (TPC), or Folin‐Ciocalteu assay, were used to quantify the antioxidant power of grape stem extracts. The extractions were performed at temperatures between 160°C and 300°C at constant density. It was found that the optimal extraction temperature was 204°C, at superheated liquid conditions, with a FRAP value of 0.670 mmol Trolox Equivalent/g of dry grape stem. The FRAP values were higher than other studies that extracted antioxidants from grape stems using single‐pass batch extraction.
The Impact of Waiting Time on Health Gains from Surgery: Evidence from a National Patient‐reported Outcome Dataset
Authors: Nikolova Silviya , Harrison Mark , Sutton Matt
Published: May 2015

Reducing waiting times has been a major focus of the English National Health Service for many years, but little is known about the impact on health outcomes. The collection of data on patient‐reported outcome measures for all patients undergoing four large‐volume procedures facilitates analysis of the impact of waiting times on patient outcomes. The availability of patient‐reported outcome measures before and after surgery allows us to estimate the impact of waiting times on the effectiveness of treatment, controlling for pre‐surgery health and the endogeneity of waiting times caused by prioritisation with respect to pre‐intervention health. We find that waiting time has a negative and statistically significant impact on the health gain from hip and knee replacement surgery and no impact on the effectiveness of varicose vein and hernia surgery. The magnitude of this effect at patient level is small, 0.1% of the outcome measure range for each additional week of waiting. However, the value of this effect is substantially larger than existing estimates of the disutility experienced during the waiting period. The health losses associated with an additional week of waiting for annual populations of hip and knee replacement patients are worth £11.1m and £11.5m, respectively. Copyright © 2015 John Wiley & Sons, Ltd.
Does Commuting Affect Health?
Authors: Künn‐Nelen Annemarie
Published: May 2015

This paper analyzes the relation between commuting time and health in the UK. I focus on four different types of health outcomes: subjective health measures, objective health measures, health behavior, and healthcare utilization. Fixed effect models are estimated with British Household Panel Survey data. I find that whereas objective health and health behavior are barely affected by commuting time, subjective health measures are clearly lower for people who commute longer. A longer commuting time is, moreover, related to more visits to the general practitioner. Effects turn out to be more pronounced for women and for commuters driving a car. For women, commuting time is also negatively related to regular exercise and positively to calling in sick. Copyright © 2015 John Wiley & Sons, Ltd.
Acute hepatitis C virus infection induces anti‐host cell receptor antibodies with virus‐neutralizing properties
Authors: Tawar Rajiv G. , Colpitts Che C. , Timm Jörg , Fehm Tanja , Roggendorf Michael , Meisel Helga , Meyer Nicolas , Habersetzer François , Cosset François‐Loïc , Berg Thomas , Zeisel Mirjam B. , Baumert Thomas F.
Published: May 2015

Hepatitis C virus (HCV) causes persistent infection in the majority of infected individuals. The mechanisms of persistence and clearance are only partially understood. Antibodies (Abs) against host cell entry receptors have been shown to inhibit HCV infection in cell culture and animal models. In this study, we aimed to investigate whether anti‐receptor Abs are induced during infection in humans in vivo and whether their presence is associated with outcome of infection. We established an enzyme‐linked immunosorbant assay using a recombinant CD81‐claudin‐1 (CLDN1) fusion protein to detect and quantify Abs directed against extracellular epitopes of the HCV CD81‐CLDN1 coreceptor complex. The presence of anti‐receptor Abs was studied in serum of patients from a well‐defined cohort of a single‐source HCV outbreak of pregnant women and several control groups, including uninfected pregnant women, patients with chronic hepatitis B and D virus (HBV/HDV) infection, and healthy individuals. Virus‐neutralizing activity of Abs was determined using recombinant cell culture–derived HCV (HCVcc). Our results demonstrate that HCV‐infected patients have statistically significantly higher anti‐CD81/CLDN1 Ab titers during the early phase of infection than controls. The titers were significantly higher in resolvers compared to persisters. Functional studies using immunoadsorption and HCV cell culture models demonstrate that HCV‐neutralizing anti‐receptor Abs are induced in the early phase of HCV infection, but not in control groups. Conclusion: The virus‐neutralizing properties of these Abs suggest a role for control of viral infection in conjunction with antiviral responses. Characterization of these anti‐receptor Abs opens new avenues to prevent and treat HCV infection. (Hepatology 2015;62:726–736)
Age effects on the water‐use efficiency and water‐use dynamics of temperate pine plantation forests
Authors: Skubel Rachel , Arain M. Altaf , Peichl Matthias , Brodeur Jason J. , Khomik Myroslava , Thorne Robin , Trant Janelle , Kula Michelle
Published: May 2015

This study analysed age‐related water‐use dynamics across three temperate conifer forest plantations (aged 11, 39 and 74 years, as of 2013, henceforth referred to as TP02, TP74 and TP39, respectively, where the last two digits represent the year of planting) in southern Ontario, Canada, from 2008 to 2013. Eddy covariance‐measured mean evapotranspiration over the growing season (April–October) was 438 ± 19, 392 ± 19 and 323 ± 25 mm at TP39, TP74 and TP02, respectively. Daytime bulk surface conductance was highest and most variable at the TP39 site (8.5 ± 4.0 mm s−1), followed by the TP74 (7.0 ± 2.8 mm s−1) and TP02 (5.4 ± 2.5 mm s−1) sites. Evapotranspiration at all the forests was sensitive to air temperature and tended to decrease with increasing atmospheric dryness. The youngest forest's evapotranspiration was most conservative, which led to an increase in water‐use efficiency throughout the study period, in particular during drought events. The oldest forest was the least restrictive in its water use during drought, which led to lower water‐use efficiency during such events as compared with the younger forests. The oldest forest was thinned in early 2012, when about one third of trees were commercially harvested. No significant change in evapotranspiration or water‐use efficiency was observed at this site following thinning; however, daytime bulk surface conductance declined. Our results suggested that changes in stand structure with forest ageing, such as reduction in stem density and increase in sapwood area, were responsible for differences in soil water demand during drought and non‐drought periods, leading to differences in forest water use. Hence, forest age, because of its structural implications, is an important control on stand‐level water‐use efficiency and forests' response to drought events. Our study suggested that younger forests may be best suited to maximize growth and carbon uptake efficiency under rising air temperatures and increasing precipitation variability as predicted by climate models for eastern North America. Copyright © 2015 John Wiley & Sons, Ltd.