ALBERT

Description: Bone remodeling is a natural process that enables growth and maintenance of the skeleton. It involves the deposition of mineralized matrix by osteoblasts and resorption by osteoclasts. Several cancers that metastasize to bone negatively perturb the remodeling process through a series of interactions with osteoclasts, and osteoblasts. These interactions have been described as the “vicious cycle” of cancer metastasis in bone. Due to the inaccessibility of the skeletal tissue it is difficult to study this system in vivo . In contrast, standard tissue culture lacks sufficient complexity. We have developed a specialized three-dimensional culture system that permits growth of a non-vascularized, multiple-cell-layer of mineralized osteoblastic tissue from pre-osteoblasts. In this study, the essential properties of bone remodeling were created in vitro by co-culturing the mineralized collagenous osteoblastic tissue with actively resorbing osteoclasts followed by reinfusion with proliferating pre-osteoblasts. Cell-cell and cell-matrix interactions were determined by confocal microscopy as well as by assays for cell specific cytokines and growth factors. Osteoclasts, differentiated in the presence of osteoblasts, led to degradation of the collagen-rich extracellular matrix. Further addition of metastatic breast cancer cells to the co-culture mimicked the vicious cycle; i.e. there was a further reduction in osteoblastic tissue thickness, an increase in osteoclastogenesis, chemotaxis of cancer cells to osteoclasts and formation of cancer cells into large colonies. The resulting model system permits detailed study of fundamental osteobiological and osteopathological processes in a manner that will enhance development of therapeutic interventions to skeletal diseases. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Patterns of migratory connectivity are a vital yet poorly understood component of the ecology and evolution of migratory birds. Our ability to accurately characterize patterns of migratory connectivity is often limited by the spatial resolution of the data but recent advances in probabilistic assignment approaches have begun pairing stable isotopes with other sources of data (e.g., genetic and mark-recapture) to improve the accuracy and precision of inferences based on a single marker. Here, we combine stable isotopes and geographic variation in morphology (wing length) to probabilistically assign Wood thrush (Hylocichla mustilena) captured on the wintering grounds to breeding locations. In addition, we use known origin samples to validate our model and assess potentially important impacts of covariates of isotopic and morphological data (age, sex and breeding location). Our results show that despite relatively high levels of mixing across their breeding and non-breeding ranges, moderate levels of migratory connectivity along an east-west gradient exist. In addition, combining stable isotopes with geographic variation in wing improved the precision of breeding assignments by 10% and 37% compared to assignments based on isotopes alone or wing length alone, respectively. These results demonstrate that geographical variation in morphological traits can greatly improve estimates of migratory connectivity when combined with other intrinsic markers (e.g., stable isotopes or genetic data). The wealth of morphological data available from museum specimens across the world represents a tremendously valuable, but largely untapped, resource that is widely applicable for quantifying patterns of migratory connectivity.

Description: Cervical carcinoma represents the paradigm of virus-induced cancers, where virtually all cervical cancers come from previous “high-risk” HPV infection. The persistent expression of the HPV viral oncoproteins E6 and E7 is responsible for the reprogramming of fundamental cellular functions in the host cell, thus generating a noticeable, yet only partially explored, imbalance in protein molecular networks and cell signaling pathways. Eighty-eight cellular factors, identified as HPV direct or surrogate targets, were chosen and monitored in a retrospective analysis for their mRNA expression in HPV-induced cervical lesions, from dysplasia to cancer. Real-time quantitative PCR (qPCR) was performed by using formalin-fixed, paraffin embedded archival samples. Gene expression analysis identified 40 genes significantly modulated in LSIL, HSIL and squamous cervical carcinoma. Interestingly, among these, the expression level of a panel of four genes, TOP2A, CTNNB1, PFKM and GSN, was able to distinguish between normal tissues and cervical carcinomas. Immunohistochemistry was also done to assess protein expression of two genes among those up-regulated during the transition between dysplasia and carcinoma, namely E2F1 and CDC25A, and their correlation with clinical parameters. Besides the possibility of significantly enhancing the use of some of these factors in diagnostic or prognostic procedures, these data clearly outline specific pathways, and thus key biological processes, altered in cervical dysplasia and carcinoma. Deeper insight on how these molecular mechanisms work may help widen the spectrum of novel innovative approaches to these virus-induced cell pathologies. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Cellular pathways are numerous and are highly integrated in function in the control of cellular systems. They collectively regulate cell division, proliferation, survival and apoptosis of cells and mutagenesis of key genes that control these pathways can initiate neoplastic transformations. Understanding these pathways is crucial to future therapeutic and preventive strategies of the disease. Ovarian cancers are of three major types; epithelial, germ-cell and stromal. However, ovarian cancers of epithelial origin, arising from the mesothelium, are the predominant form. Of the subtypes of ovarian cancer, the high-grade serous tumors are fatal, with low survival rate due to late detection and poor response to treatments. Close examination of preserved ovarian tissues and in vitro studies have provided insights into the mechanistic changes occurring in cells mediated by a few key genes. This review will focus on pathways and key genes of the pathways that are mutated or have aberrant functions in the pathology of ovarian cancer. Non-genetic mechanisms that are gaining prominence in the pathology of ovarian cancer, miRNAs and epigenetics, will also be discussed in the review. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Normal pregnancy is associated with systemic vasodilation and decreased vascular contraction, partly due to increased release of endothelium-derived vasodilator substances. Endothelin-1 (ET-1) is an endothelium-derived vasoconstrictor acting via endothelin receptor type A (ET A R) and possibly type B (ET B R) in vascular smooth muscle cells (VSMCs), with additional vasodilator effects via endothelial ET B R. However, the role of ET-1 receptor subtypes in the regulation of vascular function during pregnancy is unclear. We investigated whether the decreased vascular contraction during pregnancy reflects changes in the expression/activity of ET A R and ET B R. Contraction was measured in single aortic VSMCs isolated from virgin, mid-pregnant (mid-Preg, day 12) and late-Preg (day 19) Sprague-Dawley rats, and the mRNA expression, protein amount, tissue and cellular distribution of ET A R and ET B R were examined using RT-PCR, Western blots, immunohistochemistry and immunofluorescence. Phenylephrine (Phe, 10 −5  M), KCl (51 mM) and ET-1 (10 −6  M) caused VSMC contraction that was in late-Preg 〈 mid-Preg and virgin rats. In VSMCs treated with ET B R antagonist BQ788, ET-1 caused significant contraction that was still in late-Preg 〈 mid-Preg and virgin rats. In VSMCs treated with the ET A R antagonist BQ123, ET-1 caused a small contraction; and the ET B R agonists IRL-1620 and sarafotoxin 6c (S6c) caused similar contraction that was in late-Preg 〈 mid-Preg and virgin rats. RT-PCR revealed similar ET A R, but greater ET B R mRNA expression in pregnant vs. virgin rats. Western blots revealed similar ET A R, and greater protein amount of ET B R in endothelium-intact vessels, but reduced ET B R in endothelium-denuded vessels of pregnant vs. virgin rats. Immunohistochemistry revealed prominent ET B R staining in the intima, but reduced ET A R and ET B R in the aortic media of pregnant rats. Immunofluorescence signal for ET A R and ET B R was less in VSMCs of pregnant vs. virgin rats. The pregnancy-associated decrease in ET A R- and ET B R-mediated VSMC contraction appears to involve downregulation of ET A R and ET B R expression/activity in VSM, and may play a role in the adaptive vasodilation during pregnancy. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Neuroregenerative medicine is an ever-growing field in which regeneration of lost cells/tissues due to a neurodegenerative disease is the ultimate goal. With the scarcity of available replacement alternatives, stem cells provide an attractive source for regenerating neural tissue. While many stem cell sources exist, including: mesenchymal stem cells (MSCs), embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs), the limited cellular potency, technical difficulties, and ethical considerations associated with these make finding alternate sources a desirable goal. Periodontal ligament stem cells (PDLSCs) derived from the neural crest were induced into neural-like cells using a combination of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Morphological changes were evident in our treated group, seen under both light microscopy and scanning electron microscopy (SEM). A statistically significant increase in the expression of neuron-specific β-tubulin III and the neural stem/progenitor cell marker nestin, along with positive immunohistochemical staining for glial fibrillary acidic protein (GFAP), demonstrated the success of our treatment in inducing both neuronal and glial phenotypes. Positive staining for synaptophysin demonstrated neural connections and electrophysiological recordings indicated that when subjected to whole cell patch clamping, our treated cells displayed inward currents conducted through voltage-gated sodium (Na + ) channels. Taken together, our results indicate the success of our treatment in inducing PDLSCs to neural-like cells. The ease of sourcing and expansion, their embryologic neural crest origin, and the lack of ethical implications in their use make PDLSCs an attractive source for use in neuroregenerative medicine. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Urotensin II (UII), a vasoactive peptide modulates renal hemodynamics. However, the physiological functions of UII in glomerular cells are unclear. In particular, whether UII alters mesangial tone remains largely unknown. The present study investigates the physiological effects of UII on intracellular Ca 2+ ([Ca 2+ ] i ) and contraction in glomerular mesangial cells (GMCs). This study also tested the hypothesis that the regulator of G-protein signaling (RGS) controls UII receptor (UTR) activity in GMCs. RT-PCR, Western immunoblotting, and immunofluorescence revealed UTR expression and localization in cultured murine GMCs. Mouse UII (mUII) stimulated [Ca 2+ ] i elevation in GMCs in the absence and presence of extracellular Ca 2+ . mUII also caused a reduction in planar GMC surface area. mUII-induced [Ca 2+ ] i elevation and contraction in GMCs were attenuated by SB 657510, a UTR antagonist, araguspongin B, an inositol 1,4,5-trisphosphate receptor antagonist, thapsigargin, a sarco/endoplasmic reticulum Ca 2+ -ATPase inhibitor, and La 3+ , a store-operated Ca 2+ channel blocker, but not nimodipine, an L-type Ca 2+ channel blocker. In situ proximity ligation assay indicated molecular proximity between endogenous RGS2 and UTR in the cells. Treatment of GMCs with mUII increased plasma membrane association of RGS2 by ∼ 2-fold. mUII also increased the interaction between RGS2 and UTR in the cells. siRNA-mediated knockdown of RGS2 in murine GMCs increased mUII-induced [Ca 2+ ] i elevation and contraction by ∼ 35 and 31%, respectively. These findings indicate that mUII induces [Ca 2+ ] i elevation and contraction in murine GMCs. Data also suggest that UTR activation stimulates RGS2 recruitment to GMC plasma membrane as a negative feedback mechanism to regulate UTR signaling. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3) n . The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of 8 known β3GnT genes in mice. In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. N-glycan branching and core composition, however, can also modulate the extent of PLN modification. Here we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. Postnatally, GCNT2 is maintained in mature olfactory neurons that coexpress β3GnT2 and PLN. This highly specific coexpression suggests that GCNT2 and β3GnT2 function cooperatively in PLN synthesis. In support of this, β3GnT2 and GCNT2 cotransfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. These data clearly suggest that GCNT2 functions in vivo together with β3GnT2 to determine PLN levels in olfactory neurons by regulating β1,6-branches that promote PLN extension. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Predicting how climate change is likely to interact with myriad other stressors that threaten species of conservation concern is an essential challenge in aquatic ecosystems. This study provides a framework to accomplish this task in salmon-bearing streams of the northwestern United States, where land-use related reductions in riparian shading have caused changes in stream thermal regimes, and additional warming from projected climate change may result in significant losses of coldwater fish habitat over the next century. Predatory non-native smallmouth bass have also been introduced into many northwestern streams and their range is likely to expand as streams warm, presenting an additional challenge to the persistence of threatened Pacific salmon. The goal of this work was to forecast the interactive effects of climate change, riparian management, and non-native species on stream-rearing salmon, and to evaluate the capacity of restoration to mitigate these effects. We intersected downscaled global climate forecasts with a local-scale water temperature model to predict mid- and end-of-century temperatures in streams in the Columbia River basin; we compared one stream that is thermally impaired due to the loss of riparian vegetation and another that is cooler and has a largely intact riparian corridor. Using the forecasted stream temperatures in conjunction with fish-habitat models, we predicted how stream-rearing Chinook salmon and bass distributions would change as each stream warmed. In the highly modified stream, end-of-century warming may cause near total loss of Chinook salmon rearing habitat and a complete invasion of the upper watershed by bass. In the less modified stream, bass were thermally restricted from the upstream-most areas. In both systems, temperature increases resulted in higher predicted spatial overlap between stream-rearing Chinook salmon and potentially predatory bass in the early summer (2-4-fold increase) and greater abundance of bass. We found that riparian restoration could prevent the extirpation of Chinook salmon from the more altered stream, and could also restrict bass from occupying the upper 31 km of salmon rearing habitat. The proposed methodology and model predictions are critical for prioritizing climate-change adaptation strategies before salmonids are exposed to both warmer water and greater predation risk by non-native species.

Description: Spermatogenesis is a special process by which spermatogonial stem cells (SSCs) divide and differentiate to male gametes called mature spermatozoa. SSCs are the unique cells because they are adult stem cells that transmit genetic information to subsequent generations. Accumulating evidence has demonstrated that SSCs can be reprogrammed to acquire pluripotency to become embryonic stem-like cells that differentiate into all cell lineages of the three germ layers, highlighting potential important applications of SSCs for regenerative medicine. Recent studies from peers and us have made great achievements on the characterization, isolation and culture of mouse and human SSCs, which could lead to better understanding the biology of SSCs and the applications of SSCs in both reproductive and regenerative medicine. In this review, we first compared the cell identity and biochemical phenotypes between mouse SSCs and human SSCs. Notably, the cell types of mouse and human SSCs are distinct, and human SSCs share some but not all phenotypes with mouse SSCs. The approaches for isolating SSCs as well as short- and long- term culture of mouse SSCs and short-period culture of human SSCs were also discussed. We further addressed the new advances on the self-renewal of SSCs with an aim to establish the long-term culture of human SSCs which has not yet been achieved. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Efforts to test and improve terrestrial biosphere models (TBMs) using a variety of data sources have become increasingly common. However, geographically extensive forest inventories have been under-exploited in previous model-data fusion efforts. Inventory observations of forest growth, mortality, and biomass integrate processes across a range of time scales, including slow time-scale processes such as species turnover, that are likely to have important effects on ecosystem responses to environmental variation. However, the large number (thousands) of inventory plots precludes detailed measurements at each location, so that uncertainty in climate, soil properties, and other environmental drivers may be large. Errors in driver variables, if ignored, introduce bias into model-data fusion. We estimated errors in climate and soil drivers at U.S. Forest Inventory and Analysis (FIA) plots, and we explored the effects of these errors on model-data fusion with the Geophysical Fluid Dynamics Laboratory LM3V dynamic global vegetation model. When driver errors were ignored or assumed small at FIA plots, responses of biomass production in LM3V to precipitation and soil available water capacity appeared steeper than the corresponding responses estimated from FIA data. These differences became non-significant if driver errors at FIA plots were assumed large. Ignoring driver errors when optimizing LM3V parameter values yielded estimates for fine-root allocation that were larger than biometric estimates, which is consistent with the expected direction of bias. To explore if complications posed by driver errors could be circumvented by relying on intensive study sites where driver errors are small, we performed a power analysis. To accurately quantify the response of biomass production to spatial variation in mean annual precipitation within the eastern U.S. would require at least 40 intensive study sites, which is larger than the number of sites typically available for individual biomes in existing plot networks. Driver errors may be accommodated by several existing model-data fusion approaches, including hierarchical Bayesian methods and ensemble filtering methods; however, these methods are computationally expensive. We propose a new approach, in which the TBM functional response is fit directly to the driver-error-corrected functional response estimated from data, rather than to the raw observations.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Much of our understanding of natural forest dynamics in the temperate region of Europe is based on observational studies in old-growth remnants that have emphasized small-scale gap dynamics and equilibrium stand structure and composition. Relatively little attention has been given to the role of infrequent disturbance events in forest dynamics. In this study, we analyzed dendroecological data from four stands and three windthrow patches in an old-growth landscape in the Dinaric Mountains of Bosnia and Herzegovina to examine disturbance history, tree life history traits, and compositional dynamics. Over all stands, most decades during the past 340 years experienced less than 10% canopy loss, yet each stand showed evidence of periodic intermediate severity disturbances that removed 〉 40% of the canopy, some of which were synchronized over the study area landscape. Analysis of radial growth patterns indicated several life history differences among the dominant canopy trees; beech was markedly older than fir, while growth patterns of dead and dying trees suggested fir was able to tolerate longer periods of suppressed growth in shade. Maple had the fastest radial growth and accessed the canopy primarily through rapid early growth in canopy gaps, whereas most beech and fir experienced a period of suppressed growth prior to canopy accession. Peaks in disturbance were roughly linked to increased recruitment, but mainly of shade tolerant beech and fir; less tolerant species (i.e. maple, ash, and elm) recruited successfully on some of the windthown sites where advance regeneration of beech and fir was less abundant. The results challenge the traditional notions of stability in temperate old-growth of Europe and highlight the non-equilibrial nature of canopy composition due to unique histories of disturbance and tree life history differences. These findings provide valuable information for developing natural disturbance-based silvicultural systems, as well as insight into maintaining less shade tolerant, but valuable broadleaved trees in temperate forests of Europe.

Description: Angiogenin (ANG) undergoes nuclear translocation and promotes ribosomal RNA (rRNA) transcription thereby enhancing cell growth and proliferation. However, the mode of action of ANG in stimulating rRNA transcription is unclear. Here, we show that ANG enhances the formation of RNA polymerase I (Pol I) pre-initiation complex at the ribosomal DNA (rDNA) promoter. ANG binds at the upstream control element (UCE) of the promoter and enhances promoter occupancy of RNA Pol I as well as the selectivity factor SL1 components TAF I 48 and TAF I 110. We also show that ANG increases the number of actively transcribing rDNA by epigenetic activation through promoter methylation and histone modification. ANG binds to histone H3, inhibits H3K9 methylation, and activates H3K4 methylation as well as H4 acetylation at the rDNA promoter. These data suggest that one of the mechanisms by which ANG stimulates rRNA transcription is through an epigenetic activation of rDNA promoter. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 23, Issue 6, Page 1322-1330, September 2013. Cities produce considerable ecological light pollution (ELP), yet the effects of artificial night lighting on biological communities and ecosystem function have not been fully explored. From June 2010 to June 2011, we surveyed aquatic emergent insects, riparian arthropods entering the water, and riparian spiders of the family Tetragnathidae at nine stream reaches representing common ambient ELP levels of Columbus, Ohio, USA, streams (low, 0.1–0.5 lux; moderate, 0.6–2.0 lux; high, 2.1–4.0 lux). In August 2011, we experimentally increased light levels at the low- and moderate-treatment reaches to 10–12 lux to represent urban streams exposed to extremely high levels of ELP. Although season exerted the dominant influence on invertebrate fluxes over the course of the year, when analyzed by season, we found that light strongly influenced multiple invertebrate responses. The experimental light addition resulted in a 44% decrease in tetragnathid spider density (P = 0.035), decreases of 16% in family richness (P = 0.040) and 76% in mean body size (P = 0.022) of aquatic emergent insects, and a 309% increase in mean body size of terrestrial arthropods (P = 0.015). Our results provide evidence that artificial light sources can alter community structure and ecosystem function in streams via changes in reciprocal aquatic–terrestrial fluxes of invertebrates.

Description: Ecological Applications, Volume 23, Issue 6, Page 1357-1366, September 2013. Examining the causes of interspecific differences in susceptibility to bidirectional land-use changes (land abandonment and use-intensification) is important for understanding the mechanisms of global biodiversity loss in agricultural landscapes. We tested the hypothesis that rare (endangered) plant species prefer wet and oligotrophic areas within topography- and management-mediated resource (soil water content, nutrient, and aboveground biomass) gradients, making them more susceptible to both abandonment and use-intensification of agricultural lands. We demonstrated that topography and management practices generated resource gradients in seminatural grasslands around traditional paddy terraces. Terraced topography and management practices produced a soil moisture gradient within levees and a nutrient gradient within paddy terraces. Both total and rare species diversity increased with soil water content. Total species diversity increased in more eutrophied areas with low aboveground biomass, whereas rare species diversity was high under oligotrophic conditions. Rare and common species were differentially distributed along the human-induced nutrient gradient, with rare species preferring wet, nutrient-poor environments in the agricultural landscapes studied. We suggest that conservation efforts should concentrate on wet, nutrient-poor areas within such landscapes, which can be located easily using land-use and topography maps. This strategy would reduce the costs of finding and conserving rare grassland species in a given agricultural landscape.

Description: Ecological Applications, Volume 23, Issue 6, Page 1429-1442, September 2013. Carryover effects occur when experiences early in life affect an individual's performance at a later stage. Many studies have shown carryover effects to be important for future performance. However, it is currently unclear whether variation in later environments could overwhelm factors from an earlier life stage. We were interested in whether similar patterns would emerge under the same experimental design with similar taxa. To examine this, we implemented a four-way factorial experimental design with different forestry practices on three species of anurans (each examined in different years) in the aquatic larval environment and terrestrial juvenile environment in outdoor mesocosms in central Missouri, USA. Using Cormack-Jolly-Seber mark–recapture models implemented in program MARK, we investigated whether one environment or both environments best predicted terrestrial juvenile survival. We found only limited evidence of carryover effects for one of three species in one time period. These were the effects of time to metamorphosis and body condition at metamorphosis predicting leopard frog (Lithobates sphenocephalus) survival. However, both effects were counterintuitive and/or very weak. For wood frogs (L. sylvaticus), all of the variables predicting survival had confidence intervals that included zero, but very low survival may have limited our ability to estimate parameters. The terrestrial environment was important for predicting survival in both American toads (Anaxyrus americanus) and southern leopard frogs. The partial harvest forest tended to increase survival relative to control forest and early-successional forest in American toads. Alternately, early-successional forest with downed wood removed increased survival for leopard frogs, but this treatment was no different from control forest for American toads. Previous studies have shown negative effects of recent clearcuts on terrestrial amphibians. It appears that vegetative regrowth after just a few years can mitigate these initial negative effects. Our study shows that variation in later environments probably can overwhelm variation from earlier environments. However, previous evidence of carryover effects suggests that more research is needed to predict when carryover effects are likely to occur.

Description: Ecological Applications, Volume 23, Issue 6, Page 1475-1487, September 2013. Statistical indicators such as rising variance and rising skewness in key system parameters may provide early warning of “regime shifts” in communities and populations. However, the utility of these indicators has rarely been tested in the large, complex ecosystems that are of most interest to managers. Crustacean fisheries in the Gulf of Alaska and Bering Sea experienced a series of collapses beginning in the 1970s, and we used spatially resolved catch data from these fisheries to test the predictions that increasing variability and skewness would precede stock collapse. Our data set consisted of catch data from 14 fisheries (12 collapsing and two non-collapsing), spanning 278 cumulative years. Our sampling unit for analysis was the Alaska Department of Fish and Game statistical reporting area (mean n for individual fisheries = 42 areas, range 7–81). We found that spatial variability in catches increased prior to stock collapse: a random-effects model estimating trend in variability across all 12 collapsing fisheries showed strong evidence of increasing variability prior to collapse. Individual trends in variability were statistically significant for only four of the 12 collapsing fisheries, suggesting that rising variability might be most effective as an indicator when information from multiple populations is available. Analyzing data across multiple fisheries allowed us to detect increasing variability 1–4 years prior to collapse, and trends in variability were significantly different for collapsing and non-collapsing fisheries. In spite of theoretical expectations, we found no evidence of pre-collapse increases in catch skewness. Further, while models generally predict that rising variability should be a transient phenomenon around collapse points, increased variability was a persistent feature of collapsed fisheries in our study. We conclude that this result is more consistent with fishing effects as the cause of increased catch variability, rather than the critical slowing down that is the driver of increased variability in regime shift models. While our results support the use of rising spatial variability as a leading indicator of regime shifts, the failure of our data to support other model-derived predictions underscores the need for empirical validation before these indicators can be used with confidence by ecosystem managers.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Determining the range of a species and exploring species-habitat associations are central questions in ecology and can be answered by analyzing presence-absence data. Often, both the sampling of sites and the desired area of inference involve neighboring sites; thus positive spatial autocorrelation between these sites is expected. Using survey data for the Southern ground hornbill (Bucorvus leadbeateri) from the Southern African Bird Atlas Project, we compared advantages and disadvantages of three increasingly complex models for species occupancy: an occupancy model that accounted for nondetection but assumed all sites were independent, and two spatial occupancy models that accounted for both nondetection and spatial autocorrelation. We modeled the spatial autocorrelation with an intrinsic conditional autoregressive (ICAR) model and with a restricted spatial regression (RSR) model. Both spatial models can readily be applied to any other gridded, presence-absence data set using a newly introduced R package. The RSR model provided the best inference and was able to capture small-scale variation that the other models did not. It showed that ground hornbills are strongly dependent on protected areas in the north of their South African range but less so further south. The ICAR models did not capture any spatial autocorrelation in the data, and they took an order of magnitude longer than the RSR models to run. Thus, the RSR occupancy model appears to be an attractive choice for modeling occurrences at large spatial domains, while accounting for imperfect detection and spatial autocorrelation.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Our increasing dependence on a small number of agricultural crops such as corn is leading to reductions in agricultural biodiversity. Reductions in the number of crops in rotation or the replacement of rotations by monocultures are responsible for this loss of biodiversity. The belowground implications of simplifying agricultural plant communities remain unresolved; however, agroecosystem sustainability will be severely compromised if reductions in biodiversity reduce soil C and N concentrations, alter microbial communities, and degrade soil ecosystem functions as reported in natural communities. We conducted a meta-analysis of 122 studies to examine crop rotation effects on total soil C and N concentrations, and the faster cycling microbial biomass C and N pools that play key roles in soil nutrient cycling and physical processes such as aggregate formation. We specifically examined how rotation crop type and management practices influence C and N dynamics in different climates and soil types. We found that adding one or more crops in rotation to a monoculture increased total soil C by 3.6% and total N by 5.3%, but when rotations included a cover crop (i.e. crops that are not harvested but produced to enrich the soil and capture inorganic N), total C increased by 8.5% and total N 12.8%. Rotations substantially increased the soil microbial biomass C (20.7%) and N (26.1%) pools, and these overwhelming effects on microbial biomass were not moderated by crop type or management practices. Crop rotations, especially those that include cover crops, sustain soil quality and productivity by enhancing soil C, N and microbial biomass, making them a cornerstone for sustainable agroecosystems.

Description: Constitutive androstane receptor (CAR) was originally identified as xenobiotic sensor that regulates the expression of cytochrome P450 genes. However, recent studies suggest that this nuclear receptor is also involved in the regulation of energy metabolism including glucose and lipid homeostasis. This study investigated the role of CAR in the regulation of bone mass in vivo using CAR -/- mice. Endogenous mRNA expression of CAR was observed in both primary osteoblasts and osteoclast precursors. CAR -/- mice have exhibited significant increase in whole body bone mineral density (BMD) by 9.5% ( p  〈 0.01) and 5.5% (p 〈 0.05) at 10 and 15 weeks of age, respectively, compared with WT mice in males. Microcomputed tomography analysis of proximal tibia demonstrated a significant increase in trabecular bone volume (62.7%), trabecular number (54.1%) in male CAR -/- mice compared with WT mice. However, primary culture of calvarial cells exhibited no significant changes in osteogenic differentiation potential between CAR -/- and WT. In addition, the number of tartrate-resistant acid-phosphatase positive osteoclasts in the femur and serum level of CTx was not different between CAR -/- and WT mice. The higher BMD and microstructural parameters were not observed in female mice. Interestingly, serum level of testosterone in male CAR -/- mice was 2.5-fold higher compared with WT mice and the mRNA expressions of Cyp2b9 and 2b10 in the liver, which regulate testosterone metabolism, were significantly down-regulated in male CAR -/- mice. Furthermore, the difference in BMD between CAR -/- and WT mice disappeared at 8 weeks after performing orchiectomy. CAR -/- mice also exhibited significant increase in serum1,25(OH) 2 D 3 levels but Cyp 27B1 which converts 25(OH)D 3 to 1,25(OH) 2 D 3 was significantly down-regulated compared to WT mice. These results suggest that in vivo deletion of CAR resulted in higher bone mass, which appears to be a result from reduced metabolism of testosterone due to down-regulation of Cyp2b. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Tropical forests play a vital role in the global carbon cycle, but the amount of carbon they contain and its spatial distribution remain uncertain. Recent studies suggest that once tree height is accounted for in biomass calculations, in addition to diameter and wood density, carbon stock estimates are reduced in many areas. However, it is possible that larger crown sizes might offset the reduction in biomass estimates in some forests where tree heights are lower because even comparatively short trees develop large, well-lit crowns in or above the forest canopy. While current allometric models and theory focus on diameter, wood density, and height, the influence of crown size and structure has not been well studied. To test the extent to which accounting for crown parameters can improve biomass estimates, we harvested and weighed 51 trees (11-169 cm diameter) in southwestern Amazonia where no direct biomass measurements have been made. The trees in our study had nearly half of total aboveground biomass in the branches (44 ± 2 %), demonstrating the importance of accounting for tree crowns. Consistent with our predictions, key pantropical equations that include height, but do not account for crown dimensions, underestimated the sum total biomass of all 51 trees by 11 to 14 %, primarily due to severe underestimates of many of the largest trees. In our models, including crown radius greatly improves performance and reduces error, especially for the largest trees. In addition, over the full dataset, crown radius marginally explained more variation in aboveground biomass (10.5 %) than height (6.0 %). Crown form is also important: trees with a monopodial architectural type are estimated to have 21-44 % less mass than trees with other growth patterns. Our analysis suggests that accounting for crown allometry would substantially improve the accuracy of tropical estimates of tree biomass and its distribution in primary and degraded forests.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Tropical forests are important storehouses of carbon and biodiversity. In isolated island ecosystems such as the Hawaiian Islands, relative dominance of native and non-native tree species may influence patterns of forest carbon stocks and biodiversity. We determined aboveground carbon density (ACD) across a matrix of lava flows differing in age, texture and vegetation composition (i.e., native or non-native dominated) in wet lowland forests of Hawaii Island. To do this at the large scales necessary to accurately capture the inherent heterogeneity of these forests, we collected LiDAR data across areas of interest and developed relationships between LiDAR metrics and field-based estimates of forest ACD. This approach enabled us to inventory, rather than merely sample, the entire populations (i.e., forests) of interest. Native Hawaiian wet lowland forests exhibited ACD values similar to those of intact tropical forests elsewhere. In general, ACD of these forests increased with increasing lava flow age, but patterns differed between native and non-native forest stands. On the youngest lavas, native-dominated forest ACD averaged 〈 60 Mg ha-1 compared to ca. 100 Mg C ha-1 for non-native dominated forests. This difference was due to the presence of the non-native, N2-fixing trees, F. moluccana and C. equisetifolia in the non-native dominated forest stands, as well as the corresponding absence of N2-fixing trees in native-dominated forest stands. Following approximately 500 years of primary succession and thereafter, however, both forest types exhibited ACD values averaging ca. 130 Mg C ha-1, although it took non-native forests only 75 to 80 years of post-establishment succession to reach those values. Given the large areas of early successional M. polymorpha-dominated forest on young lava flows, further spread of F. moluccana and C. equisetifolia populations would likely increase ACD stocks but would constitute a significant erosion of the invaluable contribution of Hawaii's native ecosystems to global biodiversity.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Despite the importance of fire and herbivory in structuring savanna systems, few replicated experiments have examined the interactive effects of herbivory and fire on plant dynamics. In addition, the effects of fire on associated ant-tree mutualisms have been largely unexplored. We carried out small controlled burns in each of 18 herbivore treatment plots of the Kenya Long-term Exclosure Experiment (KLEE), where experimentally excluding elephants has resulting in 42% greater tree densities. The KLEE design includes six different herbivore treatments that allowed us to examine how different combinations of megaherbivore wildlife, mesoherbivore wildlife, and cattle affect fire temperatures and subsequent loss of ant symbionts from Acacia trees. Before burning, we quantified herbaceous fuel loads and plant community composition. We tagged all trees, measured their height and basal diameter, and identified the resident ant species on each. We recorded weather conditions during the burns and used ceramic tiles painted with fire-sensitive paints to estimate fire temperatures at different heights and in different microsites (under versus between trees). Across all treatments, fire temperatures were highest at 0-50cm off the ground and hotter in the grass under trees than in the grassy areas between trees. Plots with more trees burned hotter than plots with fewer trees, perhaps because of greater fine woody debris. Plots grazed by wildlife and by cattle prior to burning had lower herbaceous fuel loads and experienced lower burn temperatures than ungrazed plots. Many trees lost their ant colonies during the burns. Ant survivorship differed by ant species, and at the plot level was positively associated with previous herbivory (and lower fire temperatures). Across all treatments, ant colonies on taller trees were more likely to survive, but even some of the tallest trees lost their ant colonies. Our study marks a significant step in understanding the mechanisms that underlie the interactions between fire and herbivory in savanna ecosystems.

Description: Ecological Applications, Volume 23, Issue 7, Page 1531-1543, October 2013. While the area of organic crop production increases at a global scale, the potential interactions between pest management in organic and conventionally managed systems have so far received little attention. Here, we evaluate the landscape-level co-dependence of insecticide-based and natural enemy-based pest management using a simulation model for parasitoid–host interactions in landscapes consisting of conventionally and organically managed fields. In our simulations conventional management consists of broad-spectrum or selective insecticide application, while organic management involves no insecticides. Simulations indicate that insecticide use can easily result in lose–lose scenarios whereby both organically and conventionally managed fields suffer from increased pest loads as compared to a scenario where no insecticides are used, but that under some conditions insecticide use can be compatible with biocontrol. Simulations also suggest that the pathway to achieve the insecticide reduction without triggering additional pest pressure is not straightforward, because increasing the proportion of organically managed fields or reducing the spray frequency in conventional fields can potentially give rise to dramatic increases in pest load. The disruptive effect of insecticide use, however, can be mitigated by spatially clustering organic fields and using selective insecticides, although the effectiveness of this mitigation depends on the behavioral traits of the biocontrol agents. Poorly dispersing parasitoids and parasitoids with high attack rates required a lower amount of organically managed fields for effective pest suppression. Our findings show that the transition from a landscape dominated by conventionally managed crops to organic management has potential pitfalls; intermediate levels of organic management may lead to higher pest burdens than either low or high adoption of organic management.

Description: Ecological Applications, Volume 23, Issue 7, Page 1603-1618, October 2013. The regional spatial scale is a vital linkage for the informed extrapolation of results from local to continental scales to address broad-scale environmental problems. Among-region variation in ecosystem state is commonly accounted for by using a regionalization framework, such as an ecoregion classification. Rarely have alternative regionalization frameworks been tested for variables measuring ecosystem state, nor have the underlying relationships with the variables that are used to define them been assessed. In this study, we asked two questions: (1) How much among-region variation is there for ecosystems and does it differ by regionalization framework? (2) What are the likely causes of this among-region variation? We present a case study using a large data set of lake water chemistry, uni- and multi-scaled hydrogeomorphic and anthropogenic driver variables that likely influence lake chemistry at the subcontinental scale, and seven existing regionalization frameworks. We used multilevel models to quantify and explain within- and among-region variation in lake water chemistry. Our models account for local driver variables of ecosystem variation within regions, differences in regional mean ecosystem state (i.e., random intercepts in multilevel models), and differences in relationships between local drivers and ecosystem state by region (i.e., random slopes in multilevel models). Using one of the best performing regionalization frameworks (Ecological Drainage Units), we found that for lake phosphorus and alkalinity: (1) a majority of all the variation in water chemistry among the studied lakes occurred among regions, (2) very few regional-scale landscape driver variables were required to explain among-region variation in lake water chemistry, (3) a much higher proportion of the total variation among lakes was explained at the regional scale than at the local scale, and (4) some relationships between local-scale driver variables and lake water chemistry varied by region. Our results demonstrate the importance of considering the regional spatial scale for broad-scale research and ecosystem management and conservation. Our quantitative approach can be easily applied to other response variables, ecosystem types, geographic areas, and spatial extents to inform ecosystem responses to global environmental stressors.

Description: Ecological Applications, Volume 23, Issue 7, Page 1544-1553, October 2013. Invasive species are costly and difficult to control. In order to gain a mechanistic understanding of potential control measures, individual-based models uniquely parameterized to reflect the salient life-history characteristics of invasive species are useful. Using invasive Australian Rhinella marina as a case study, we constructed a cohort- and individual-based population simulation that incorporates growth and body size of terrestrial stages. We used this allometric approach to examine the efficacy of nontraditional control methods (i.e., tadpole alarm chemicals and native meat ants) that may have indirect effects on population dynamics mediated by effects on body size. We compared population estimates resulting from these control methods with traditional hand removal. We also conducted a sensitivity analysis to investigate the effect that model parameters, specifically those associated with growth and body size, had on adult population estimates. Incremental increases in hand removal of adults and juveniles caused nonlinear decreases in adult population estimates, suggesting less return with increased investment in hand-removal efforts. Applying tadpole alarm chemicals or meat ants decreased adult population estimates on the same level as removing 15–25% of adults and juveniles by hand. The combined application of tadpole alarm chemicals and meat ants resulted in ∼80% decrease in adult abundance, the largest of any applied control method. In further support of the nontraditional control methods, which greatly affected the metamorph stage, our model was most sensitive to changes in metamorph survival, juvenile survival, metamorph growth rate, and adult survival. Our results highlight the use and insights that can be gained from individual-based models that incorporate growth and body size and the potential success that nontraditional control methods could have in controlling established, invasive Rhinella marina populations.

Description: Ecological Applications, Volume 23, Issue 7, Page 1659-1676, October 2013. This paper proposes a hierarchical Bayesian framework for modeling the life cycle of marine exploited fish with a spatial perspective. The application was developed for a nursery-dependent fish species, the common sole (Solea solea), on the Eastern Channel population (Western Europe). The approach combined processes of different natures and various sources of observations within an integrated framework for life-cycle modeling: (1) outputs of an individual-based model for larval drift and survival that provided yearly estimates of the dispersion and mortality of eggs and larvae, from spawning grounds to settlement in several coastal nurseries; (2) a habitat suitability model, based on juvenile trawl surveys coupled with a geographic information system, to estimate juvenile densities and surface areas of suitable juvenile habitat in each nursery sector; (3) a statistical catch-at-age model for the estimation of the numbers-at-age and the fishing mortality on subadults and adults. The approach provided estimates of hidden variables and parameters of key biological significance. A simulation approach provided insight to the robustness of the approach when only weak data are available. Estimates of spawning biomass, fishing mortality, and recruitment were close to the estimations derived from stock-assessment working groups. In addition, the model quantified mortality along the life cycle, and estimated site-specific density-dependent mortalities between settled larvae and age-0 juveniles in each nursery ground. This provided a better understanding of the productivity and the specific contribution of each nursery ground toward recruitment and population renewal. Perspectives include further development of the modeling framework on the common sole and applications to other fish species to disentangle the effects of multiple interacting stress factors (e.g., estuarine and coastal nursery habitat degradation, fishing pressure) on population renewal and to develop risk analysis in the context of marine spatial planning for sustainable management of fish resources.

Description: Testosterone exerts important effects in the heart. Cardiomyocytes are target cells for androgens, and testosterone induces rapid effects via Ca 2+ release and protein kinase activation and long-term effects via cardiomyocyte differentiation and hypertrophy. Furthermore, it stimulates metabolic effects such as increasing glucose uptake in different tissues. Cardiomyocytes preferentially consume fatty acids for ATP production, but under particular circumstances, glucose uptake is increased to optimize energy production. We studied the effects of testosterone on glucose uptake in cardiomyocytes. We found that testosterone increased uptake of the fluorescent glucose analog 2-(N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)-2-deoxyglucose and [ 3 H]2-deoxyglucose, which was blocked by the glucose transporter 4 (GLUT4) inhibitor indinavir. Testosterone stimulation in the presence of cyproterone or albumin-bound testosterone induced glucose uptake, which suggests an effect that is independent of the intracellular androgen receptor. To determine the degree of GLUT4 cell surface exposure, cardiomyocytes were transfected with the plasmid GLUT4 myc -eGFP. Subsequently, testosterone increased GLUT4 myc- GFP exposure at the plasma membrane. Inhibition of Akt by the Akt-inhibitor-VIII had no effect. However, inhibition of Ca 2+ /calmodulin protein kinase (CaMKII) (KN-93 and autocamtide-2 related inhibitory peptide II) and AMP-activated protein kinase (AMPK) (compound C and siRNA for AMPK) prevented glucose uptake induced by testosterone. Moreover, GLUT4 myc- eGFP exposure at the cell surface caused by testosterone was also abolished after CaMKII and AMPK inhibition. These results suggest that testosterone increases GLUT4-dependent glucose uptake, which is mediated by CaMKII and AMPK in cultured cardiomyocytes. Glucose uptake could represent a mechanism by which testosterone increases energy production and protein synthesis in cardiomyocytes. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Although the role of Cbl-family proteins as key cellular regulators has been established, phenomena regulated in a Cbl-dependent fashion are multiple and the mechanisms mediating the effects of Cbl proteins are diverse. This finding makes it important to consider different phenomena affected by functions of Cbl proteins individually. Among effects of Cbl on various biological functions there are many cases of regulation of cellular phenomena related to cytoskeletal rearrangements, such as cell adhesion, motility and invasion. Some of these regulatory functions are mediated by adaptor-type interactions of Cbl, especially by Cbl-dependent modulation of phosphatidyl-inositol-3' kinase (PI3K), while others are caused by Cbl-dependent ubiquitylation of various cytoskeletal and regulatory proteins, identified and unidentified. The role of Cbl in regulation of cytoskeleton-dependent cellular functions is discussed in this review. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. The process of disease transmission is determined by the interaction of host susceptibility and exposure to parasite infectious stages. Host behavior is an important determinant of the likelihood of exposure to infectious stages, but is difficult to measure and often assumed to be homogenous in models of disease spread. We evaluated the importance of precisely defining host contact when using networks that estimate exposure and predict infection prevalence in a replicated, empirical system. In particular, we hypothesized that infection patterns would be predicted only by a contact network that is defined according to host behavior and parasite life-cycle. Two competing host contact criteria were used to construct networks defined by parasite life-cycle and social contacts. First, parasite defined contacts were based on shared space with a time-delay corresponding to the environmental development time of nematode parasites with a direct fecal-oral life-cycle. Second, social contacts were defined by shared space in the same time period. To quantify the competing networks of exposure and infection, we sampled natural populations of the eastern chipmunk and infection of their gastro-intestinal helminth community using replicated longitudinal capture-mark-recapture techniques. We predicted that 1) infection with parasites with direct fecal-oral life-cycles would be explained by the time-delay contact network but not the social contact network; 2) infection with parasites with trophic life-cycles (via a mobile intermediate host, thus, spatially decoupling transmission from host contact) would not be explained by either contact network. The prevalence of fecal-oral life-cycle nematode parasites was strongly correlated to the number and strength of network connections from the parasite-defined network (including the time-delay), while the prevalence of trophic life-cycle parasites was not correlated with any network metrics. We concluded that incorporating the parasite life-cycle, relative to the way that exposure is measured, is key to inferring transmission and can be empirically quantified using network techniques. In addition, appropriately defining and measuring contacts according the life-history of the parasite and relevant behaviors of the host is a crucial step in applying network analyses to empirical systems.

Description: Ecological Applications, Volume 23, Issue 4, Page 766-776, June 2013. Autogenic ecosystem engineers are critically important parts of many marine and estuarine systems because of their substantial effect on ecosystem services. Oysters are of particular importance because of their capacity to modify coastal and estuarine habitats and the highly degraded status of their habitats worldwide. However, models to predict dynamics of ecosystem engineers have not previously included the effects of exploitation. We developed a linked population and habitat model for autogenic ecosystem engineers undergoing exploitation. We parameterized the model to represent eastern oyster (Crassostrea virginica) in upper Chesapeake Bay by selecting sets of parameter values that matched observed rates of change in abundance and habitat. We used the model to evaluate the effects of a range of management and restoration options including sustainability of historical fishing pressure, effectiveness of a newly enacted sanctuary program, and relative performance of two restoration approaches. In general, autogenic ecosystem engineers are expected to be substantially less resilient to fishing than an equivalent species that does not rely on itself for habitat. Historical fishing mortality rates in upper Chesapeake Bay for oysters were above the levels that would lead to extirpation. Reductions in fishing or closure of the fishery were projected to lead to long-term increases in abundance and habitat. For fisheries to become sustainable outside of sanctuaries, a substantial larval subsidy would be required from oysters within sanctuaries. Restoration efforts using high-relief reefs were predicted to allow recovery within a shorter period of time than low-relief reefs. Models such as ours, that allow for feedbacks between population and habitat dynamics, can be effective tools for guiding management and restoration of autogenic ecosystem engineers.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. A simple population model is developed to evaluate the role of plastic and evolutionary life history changes on sustainable exploitation rates. Plastic changes are embodied in density-dependent compensatory adjustments to somatic growth rate and larval/juvenile survival, which can compensate for the reductions in reproductive lifetime and mean population fecundity that accompany the higher adult mortality imposed by exploitation. Evolutionary changes are embodied in the selective pressures that higher adult mortality imposes on age at maturity, length at maturity, and reproductive investment. Analytical development, based on a biphasic growth model, leads to simple equations that show explicitly how sustainable exploitation rates are bounded by each of these effects. We show that density-dependent growth combined with a fixed length at maturity and fixed reproductive investment can support exploitation-driven mortality that is 80% of the level supported by evolutionary changes in maturation and reproductive investment. Sustainable fishing mortality is proportional to natural mortality (M) times the degree of density-dependent growth, as modified by both the degree of density-dependent early survival and the minimum harvestable length. We apply this model to estimate sustainable exploitation rates for North American walleye populations (Sander vitreus). Our analysis of demographic data from walleye populations spread across a broad latitudinal range indicates that density-dependent variation in growth rate can vary by a factor of two. Implications of this growth response are generally consistent with empirical studies suggesting that optimal fishing mortality is approximately 0.75M for teleosts. This approach can be adapted to the management of other species, particularly when significant exploitation is imposed on many, widely distributed but geographically isolated populations.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Long-term fire exclusion has altered ecological function in many forested ecosystems in North America. The invasion of fire-sensitive tree species into formerly pyrogenic upland forests in the southeastern USA has resulted in dramatic shifts in surface fuels that have been hypothesized to cause reductions in plant community flammability. The mechanism for the reduced flammability or "mesophication" has lacked empirical study. Here we evaluate a potential mechanism of reduced flammability by quantifying moisture retention (response time and initial moisture capacity) of foliar litter beds from 17 southeastern tree species spanning a wide range of fire tolerance. K-means cluster analysis resulted in four species groups: a rapidly drying cluster of eight species; a five species group that absorbed little water, but desorbed slowly; a two species group that absorbed substantial moisture but desorbed rapidly; and a two species cluster that absorbed substantial moisture and dried slowly. Fire-sensitive species were segregated into the slow moisture loss clusters while fire-tolerant species tended to cluster in the rapid drying groups. Principal Components Analysis indicated that several leaf characteristics correlated with absorption capacity and drying rates. Thin-leaved species with high surface area:volume absorbed the greatest moisture content, while those with large, curling leaves had the fastest drying rates. The dramatic shifts in litter fuels as a result of invasion by fire-sensitive species generate a positive feedback that reduce the windows of ignition, thereby facilitating the survival, persistence, and continued invasion of fire-sensitive species in the uplands of the southeastern USA.

Description: Ecological Applications, Volume 23, Issue 4, Page 936-943, June 2013. A typical way to quantify aboveground carbon in forests is to measure tree diameters and use species-specific allometric equations to estimate biomass and carbon stocks. Using “citizen scientists” to collect data that are usually time-consuming and labor-intensive can play a valuable role in ecological research. However, data validation, such as establishing the sampling error in volunteer measurements, is a crucial, but little studied, part of utilizing citizen science data. The aims of this study were to (1) evaluate the quality of tree diameter and height measurements carried out by volunteers compared to expert scientists and (2) estimate how sensitive carbon stock estimates are to these measurement sampling errors. Using all diameter data measured with a diameter tape, the volunteer mean sampling error (difference between repeated measurements of the same stem) was 9.9 mm, and the expert sampling error was 1.8 mm. Excluding those sampling errors 〉1 cm, the mean sampling errors were 2.3 mm (volunteers) and 1.4 mm (experts) (this excluded 14% [volunteer] and 3% [expert] of the data). The sampling error in diameter measurements had a small effect on the biomass estimates of the plots: a volunteer (expert) diameter sampling error of 2.3 mm (1.4 mm) translated into 1.7% (0.9%) change in the biomass estimates calculated from species-specific allometric equations based upon diameter. Height sampling error had a dependent relationship with tree height. Including height measurements in biomass calculations compounded the sampling error markedly; the impact of volunteer sampling error on biomass estimates was ±15%, and the expert range was ±9%. Using dendrometer bands, used to measure growth rates, we calculated that the volunteer (vs. expert) sampling error was 0.6 mm (vs. 0.3 mm), which is equivalent to a difference in carbon storage of ±0.011 kg C/yr (vs. ±0.002 kg C/yr) per stem. Using a citizen science model for monitoring carbon stocks not only has benefits in educating and engaging the public in science, but as demonstrated here, can also provide accurate estimates of biomass or forest carbon stocks.

Description: Ecological Applications, Volume 23, Issue 4, Page 710-725, June 2013. Decision-makers charged with implementing ecosystem-based management (EBM) rely on scientists to predict the consequences of decisions relating to multiple, potentially conflicting, objectives. Such predictions are inherently uncertain, and this can be a barrier to decision-making. The Convention on the Conservation of Antarctic Marine Living Resources requires managers of Southern Ocean fisheries to sustain the productivity of target stocks, the health and resilience of the ecosystem, and the performance of the fisheries themselves. The managers of the Antarctic krill fishery in the Scotia Sea and southern Drake Passage have requested advice on candidate management measures consisting of a regional catch limit and options for subdividing this among smaller areas. We developed a spatially resolved model that simulates krill–predator–fishery interactions and reproduces a plausible representation of past dynamics. We worked with experts and stakeholders to identify (1) key uncertainties affecting our ability to predict ecosystem state; (2) illustrative reference points that represent the management objectives; and (3) a clear and simple way of conveying our results to decision-makers. We developed four scenarios that bracket the key uncertainties and evaluated candidate management measures in each of these scenarios using multiple stochastic simulations. The model emphasizes uncertainty and simulates multiple ecosystem components relating to diverse objectives. We summarize the potentially complex results as estimates of the risk that each illustrative objective will not be achieved (i.e., of the state being outside the range specified by the reference point). This approach allows direct comparisons between objectives. It also demonstrates that a candid appraisal of uncertainty, in the form of risk estimates, can be an aid, rather than a barrier, to understanding and using ecosystem model predictions. Management measures that reduce coastal fishing, relative to oceanic fishing, apparently reduce risks to both the fishery and the ecosystem. However, alternative reference points could alter the perceived risks, so further stakeholder involvement is needed to identify risk metrics that appropriately represent their objectives.

Description: Widespread changes in gene expression underlie B cell development and activation, yet our knowledge of which chromatin-remodeling factors are essential is limited. Here, we demonstrate that the BRG1 catalytic subunit of SWI/SNF complexes was dispensable for murine B cell development but played an important, albeit selective, role during activation. Although BRG1 was dispensable for CD69 induction and differentiation into plasma cells based on the ability of mutant B cells to undergo hypertrophy and secrete IgM antibodies, it was required for robust cell proliferation in response to activation. Accordingly, BRG1 was required for only ∼100 genes to be expressed at normal levels in naïve B cells but 〉1,000 genes during their activation. BRG1 upregulated 5-fold more genes than it downregulated, and the toll-like receptor pathway and JAK/STAT cytokine-signaling pathways were particularly dependent on BRG1. The importance of BRG1 in B cell activation was underscored by the occurrence of opportunistic Pasteurella infections in conditionally mutant mice. B cell activation has long served as a model of inducible gene expression, and the results presented here identify BRG1 as a chromatin-remodeling factor that upregulates the transcriptome of B cells during their activation to promote rapid cell proliferation and to mount an effective immune response. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Understanding tree growth as a function of tree size is important for a multitude of ecological and management applications. Determining what limits growth is of central interest, and forest inventory permanent plots are an abundant source of long-term information but are highly complex. Observation error and multiple sources of shared variation (spatial plot effects, temporal repeated measures, and a mosaic of sampling intervals) make these data challenging to use for growth estimation. We account for these complexities and incorporate potential limiting factors (tree size, competition, and resource supply) into a hierarchical state-space model. We estimate the diameter growth of white fir Abies concolor in the Sierra Nevada of California from forest inventory data, showing that estimating such a model is feasible in a Bayesian framework using readily available modeling tools. In this forest, white fir growth depends strongly on tree size, total plot basal area, and unexplained variation between individual trees. Plot-level resource supply variables (reflecting light, water, and nutrient availability) do not have a strong impact on inventory-size trees. This approach can be applied to many permanent forest plots, leading to greater ecological insights on tree growth.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Acoustic sensors can be used to estimate species richness for vocal species such as birds. They can continuously and passively record large volumes of data over extended periods. This data must subsequently be analysed to detect the presence of vocal species. Automated analysis of acoustic data for large numbers of species is complex and can be subject to high levels of false positive and false negative results. Manual analysis by experienced surveyors can produce accurate results, however the time and effort required to process even small volumes of data can make manual analysis prohibitive. This study examined the use of sampling methods to reduce the cost of analysing large volumes of acoustic sensor data, while retaining high levels of species detection accuracy. Utilising five days of manually analysed acoustic sensor data from four sites, we examined a range of sampling frequencies and methods including random, stratified and biologically informed. We found that randomly selecting 120 one-minute samples from the three hours immediately following dawn over five days of recordings, detected the highest number of species. On average, this method detected 62% of total species from 120 one-minute samples, compared to 34% of total species detected from traditional area search methods. Our results demonstrate that targeted sampling methods can provide an effective means for analysing large volumes of acoustic sensor data efficiently and accurately. Development of automated and semi-automated techniques are required to assist in analysing large volumes of acoustic sensor data.

Description: Optical tweezers were used to scan individual Chronic Myelogenous Leukemia cells to determine if the cell death depends on the scanning conditions. Although increasing the scanning frequency or amplitude means greater force applied to the cells, their effects on cell death are not a simple increasing trend, as observed in the optical microscopy. Indeed, cell death sharply increased at particular screening frequencies and amplitudes, whereas other frequencies or amplitudes were less detrimental. These results suggest that cell damage was more sensitive to certain scanning conditions, rather than simply high applied forces. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Odontoblasts, which derive from dental papilla, are a type of terminally differentiated matrix-secreting cells. Previous studies have identified various transcription factors involved in the differentiation process of odontoblasts. We have recently found that Krüppel-like factor 4 ( Klf4 ) was expressed in the polarizing and elongating odontoblasts, but the function of Klf4 in the differentiation of odontoblasts is still unclear. We hypothesized Klf4 promoted the differentiation of odontoblasts by up-regulating some odontoblast-related genes. In this study, we found that the expression of Klf4 increased significantly during the odontoblastic differentiation of primary mouse dental papilla cells and the mouse dental papilla cell line-mDPC6T. Overexpression of Klf4 significantly up-regulated odontoblast-related genes, such as Dmp1 , Dspp , and Alp , and promoted the accumulation of mineral nodules. Knock-down of Klf4 down-regulated expression of Dmp1 , Dspp , and Alp , and inhibited mineral deposition. We applied in silico analysis and identified one target gene of Klf4 — Dmp1 . Based on further analysis of ChIP data, EMSA and dual luciferase activity assays, we confirmed that Klf4 was able to specifically bind to the Dmp1 promoter and transactivate its expression. Furthermore, forced expression of Dmp1 in the Klf4 knock-down mDPC6T cell line significantly recovered its odontoblastic differentiation ability. Our data confirmed our hypothesis that Klf4 promotes the differentiation of odontoblasts via the up-regulation of Dmp1 . © 2013 Wiley Periodicals, Inc.

Description: Nociception is the sensory mechanism used to detect cues that can harm an organism. The understanding of the neural networks and molecular controls of the reception of pain remains an ongoing challenge for biologists. While we have made significant progress in identifying a number of molecules and pathways that are involved in transduction of noxious stimuli, from the skin through the sensory receptor cell and from this to the spinal cord on into the central nervous system, we still lack a clear understanding of the perceptual processes, the responses to pain and the regulation of pain perception. Mice and rat animal models have been extensively used for nociception studies. However, the study of pain and noiception in these organisms can be rather laborious, costly and time consuming. Conversely the use of Drosophila and C. elegans may be affected by the large evolutionary distance between these animals and humans. We outline here the reasons why zebrafish presents a new and attractive model for studying pain reception and responses and the most interesting findings in the study of nociception that have been obtained using the zebrafish model. © 2013 Wiley Periodicals, Inc.

Description: Transforming growth factor (TGF)-β is a multifunctional cytokine acting during development, tissue homeostasis, regeneration processes and disease progression. Due to its pleiotropic effects, tight regulation of the induced signaling cascades is mandatory. Caveolin proteins regulate a specific endocytic pathway and modulate diverse signaling pathways and thus have been related to severe disorders, e.g. cancer and fibrosis. Caveolin affects TGF-β/-Smad and non-Smad signaling in many ways and thus can determine the cellular outcome upon TGF-β challenge. Reciprocal regulation of caveolin and TGF-β is also evident, ranging from gene expression to miRNA regulation. Finally, there is in vivo evidence that this crosstalk influences disease development and progression. This review gives an overview about the multifaceted relations of caveolin and TGF-β. © 2013 Wiley Periodicals, Inc.

Description: Transforming growth factor (TGF)-β1 regulates diverse cellular functions. Particularly, TGF-β1 induces monocyte migration to sites of injury or inflammation in early period, whereas TGF-β1 inhibits cell migration in late phase. In this study, we attempted to understand how TGF-β1 suppresses cell migration in late phase. We found that TGF-β1 of short exposure induces the production of chemokines, such as macrophage inflammatory protein (MIP)-1α, by Raw 264.7 cells. However, knock-down of small GTPase RhoA by sh-RhoA inhibited the production of MIP-1α and macrophage migration, suggesting that RhoA is essential for expression of this chemokine. An activator of Epac ( e xchange p roteins directly a ctivated by c AMP; a guanine nucleotide exchange factor of Rap1), 8CPT-2Me-cAMP which leads to Rap1 activation abrogated MIP-1α expression and macrophage migration. Indeed, GTP-RhoA and GTP-Rap1 levels were reciprocally regulated in a time-dependent manner following TGF-β1 stimulation. 8CPT-2Me-cAMP suppressed GTP-RhoA levels, whereas si-Rap1 augmented GTP-RhoA levels and cell migration. TGF-β1 produced cAMP in late period and si-RNAs of Epac1 (exchange protein directly activated by cAMP 1) and Epac2 reduced GTP-Rap1 levels leading to promotion of GTP-RhoA levels. Furthermore, si-RNA of ARAP3 (Rap-dependent RhoGAP) increased GTP-RhoA level and cell migration. Therefore, we propose the mechanism that prolonged TGF-β1 treatment produce cAMP, which activates sequentially Epac, Rap1 and ARAP3, resulting in suppression of RhoA, chemokine expression, and macrophage migration. Contrary to the general concept that Rap1 stimulates cell migration, we demonstrated in this study that Rap1 inhibits cell migration by suppression of RhoA activity in response to TGF-β1. © 2013 Wiley Periodicals, Inc.

Description: Polycyclic Aromatic Hydrocarbons (PAHs) are the products of incomplete combustion of organic materials, which are present in cigarette smoke, deep-fried food, and in natural crude oil. Since PAH-metabolites form DNA adducts and cause oxidative DNA damage, we asked if these environmental carcinogens could affect transforming potential of the human Polyomavirus JC oncoprotein, T-antigen (JCV T-antigen). We extracted DMSO soluble PAHs from Deepwater Horizon oil spill in the Gulf of Mexico (oil-PAHs), and detected several carcinogenic PAHs. The oil-PAHs were tested in exponentially growing cultures on normal mouse fibroblasts (R508), and in R508 stably expressing JCV T-antigen (R508/T). The oil-PAHs were cytotoxic only at relatively high doses (1:50–1:100 dilution), and at 1:500 dilution the growth and cell survival rates were practically unaffected. This non-toxic dose triggered however, a significant accumulation of reactive oxygen species (ROS), caused oxidative DNA damage and the formation of DNA double strand breaks (DSBs). Although oil-PAHs induced similar levels of DNA damage in R508 and R508/T cells, only T-antigen expressing cells demonstrated inhibition of high fidelity DNA repair by homologous recombination (HRR). In contrast, low-fidelity repair by non-homologous end joining (NHEJ) was unaffected. This potential mutagenic shift between DNA repair mechanisms was accompanied by a significant increase in clonal growth of R508/T cells chronically exposed to low doses of the oil-PAHs. Our results indicate for the first time carcinogenic synergy in which oil-PAHs trigger oxidative DNA damage and JCV T-antigen compromises DNA repair fidelity. © 2013 Wiley Periodicals, Inc.

Description: The intracellular pH is regulated by a delicate balance of ion distribution across the plasma membrane and the physico-chemical properties of intra- and extracellular components. We analysed the effects of glycosaminoglycans on the intracellular pH of fibroblasts by using the fluorescent pH indicator BCECF-AM. Addition of hyaluronan, hyaluronan oligosaccharides, chondroitin sulfate or heparin to the culture medium of fibroblasts caused intracellular acidification from pH 7.2 to pH 6.7 in a concentration dependent manner. High molecular weight hyaluronan acidified more than hyaluronan oligosaccharides at the same concentrations. Hyaluronidase treatment or inhibition of hyaluronan export with xanthohumol led to intracellular alkalization. These observations indicated that extracellular glycosaminoglycans participate in intracellular pH regulation. The mechanism was explained by Donnan effects and molecular crowding. © 2013 Wiley Periodicals, Inc.

Description: Purpose To test whether the use of a striatum weighted image may improve registration accuracy and diagnostic outcome in patients with parkinsonian syndromes (PS). Methods Weighted images were generated by increasing signal intensity of striatal voxels and used as intermediate dataset for co-registering the brain image onto template. Experimental validation was performed using an anthropomorphic striatal phantom. 123 I-FP-CIT SPECT binding ratios were manually determined in 67 PS subjects an and compared to those obtained using unsupervised standard (UWR) and weighted registered (WR) approach. Normalized cost function was used to evaluate the accuracy of phantom and subjects registered images to the template. Reproducibility between unsupervised and manual ratios was assessed by using intra-class correlation coefficient (ICC) and Bland and Altman analysis. Correlation coefficient was used to assess the dependence of semi-quantitative ratios on clinical findings. Results Weighted method improves accuracy of brain registration onto template as determined by cost function in phantom (0.86 ± 0.06vs.0.98 ± 0.02; Student's t test, P = 0.04) and in subject scans (0.69 ± 0.06vs.0.53 ± 0.06; Student's t test, P 〈 0.0001). Agreement between manual and unsupervised derived binding ratios as measured by ICC was significantly higher on WR as compared to UWR images (0.91vs.0.76). Motor UPDRS score was significantly correlated with manual and unsupervised derived binding potential. In phantom as well as in subjects studies, correlations were more significant using the WR method (BPm: R 2  = 0.36,p = 0.0001; BPwr: R 2  = 0.368, P  = 0.0001; BPuwr: R 2  = 0.300, P  = 0.0008). Conclusion Weighted registration improves accuracy of binding potential estimates and may be a promising approach to enhance the diagnostic outcome of SPECT imaging, correlation with disease severity, and for monitoring disease progression in parkinsonian syndromes. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Ecological systems often exhibit resilient states that are maintained through negative feedbacks. In ponderosa pine forests, fire historically represented the negative feedback mechanism that maintained ecosystem resilience; fire exclusion reduced that resilience, predisposing transition to an alternative ecosystem state upon reintroduction of fire. We evaluated the effects of reintroduced frequent wildfire in unlogged, fire-excluded ponderosa pine forest in the Bob Marshall Wilderness, Montana, USA. Initial reintroduction of fire in 2003 reduced tree density and consumed surface fuels, but also stimulated establishment of a dense cohort of lodgepole pine, maintaining a trajectory towards an alternative state. Resumption of a frequent fire regime by a second fire in 2011 restored a low-density forest dominated by large-diameter ponderosa pine by eliminating many regenerating lodgepole pines and by continuing to remove surface fuels and small-diameter lodgepole pine and Douglas-fir that established during the fire suppression era. Our data demonstrate that some unlogged, fire-excluded ponderosa pine forests possess latent resilience to reintroduced fire. A passive model of simply allowing lightning-ignited fires to burn appears to be a viable approach to restoration of such forests.

Description: Ecological Applications, Volume 23, Issue 6, Page 1396-1409, September 2013. Hydrologic connectivity is critical to the structure, function, and dynamic process of river ecosystems. Dams, road crossings, and water diversions impact connectivity by altering flow regimes, behavioral cues, local geomorphology, and nutrient cycling. This longitudinal fragmentation of river ecosystems also increases genetic and reproductive isolation of aquatic biota such as migratory fishes. The cumulative effects on fish passage of many structures along a river are often substantial, even when individual barriers have negligible impact. Habitat connectivity can be improved through dam removal or other means of fish passage improvement (e.g., ladders, bypasses, culvert improvement). Environmental managers require techniques for comparing alternative fish passage restoration actions at alternative or multiple locations. Herein, we examined a graph-theoretic algorithm for assessing upstream habitat connectivity to investigate both basic and applied fish passage connectivity problems. First, we used hypothetical watershed configurations to assess general alterations to upstream fish passage connectivity with changes in watershed network topology (e.g., linear vs. highly dendritic) and the quantity, location, and passability of each barrier. Our hypothetical network modeling indicates that locations of dams with limited passage efficiency near the watershed outlet create a strong fragmentation signal but are not individually sufficient to disconnect the system. Furthermore, there exists a threshold in the number of dams beyond which connectivity declines precipitously, regardless of watershed topology and dam configuration. Watersheds with highly branched configurations are shown to be less susceptible to disconnection as measured by this metric. Second, we applied the model to prioritize barrier improvement in the mainstem of the Truckee River, Nevada, USA. The Truckee River application demonstrates the ability of the algorithm to address conditions common in fish passage projects including incomplete data, parameter uncertainty, and rapid application. This study demonstrates the utility of a graph-theoretic approach for assessing fish passage connectivity in dendritic river networks assuming full basin utilization for a given species, guild, or community of concern.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. A unique high temporal frequency dataset from an irrigated cotton-wheat rotation was used to test the agroecosystem model DayCent to simulate daily N2O emissions from sub-tropical vertisols under different irrigation intensities. DayCent was able to simulate the effect of different irrigation intensities on N2O fluxes and yield, although it tended to overestimate seasonal fluxes during the cotton season. DayCent accurately predicted soil moisture dynamics and the timing and magnitude of high fluxes associated with fertilizer additions and irrigation events. At the daily scale we found a good correlation of predicted vs. measured N2O fluxes (r2 = 0.52), confirming that DayCent can be used to test agricultural practices for mitigating N2O emission from irrigated cropping systems. A 25 year scenario analysis indicated that N2O losses from irrigated cotton-wheat rotations on black vertisols in Australia can be substantially reduced by an optimized fertilizer and irrigation management system (i.e. frequent irrigation, avoidance of excessive fertiliser application), while sustaining maximum yield potentials.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Mangroves are recognized to possess a variety of ecosystem services including high rates of carbon sequestration and storage. Deforestation and conversion of these ecosystems continue to be high and have been predicted to result in significant carbon emissions to the atmosphere. Yet few studies have quantified the carbon stocks or losses associated with conversion of these ecosystems. In this study we quantified the ecosystem carbon stocks of three common mangrove types of the Caribbean as well as that of abandoned shrimp ponds in areas formerly occupied by mangrove; a common land use conversion of mangroves in the world. In the mangroves of the Montecristi Province in Northwest Dominican Republic, we found C stocks ranged from 706 to 1131 Mg/ha. The medium statured mangroves (3-10m ht) had the highest C stocks while the tall (〉10 m ht)) mangroves had the lowest ecosystem carbon storage. Carbon stocks of the low mangrove (shrub) type (

Description: G-protein coupled receptors (GPCRs) are a large family of proteins that coordinate extracellular signals to produce physiologic outcomes. Adenosine receptors (AR) are one class of GPCRs that have been shown to regulate functions as diverse as inflammation, blood flow, and cellular differentiation. Adenosine signals through four GPCRs that either inhibit (A1AR and A3AR) or activate (A2aAR and A2bAR) adenylyl cyclase. This review will focus on the role of GPCRs, and in particular, adenosine receptors, in adipogenesis. Preadipocytes differentiate to mature adipocytes as the adipose tissue expands to compensate for the consumption of excess nutrients. These newly generated adipocytes contribute to maintaining metabolic homeostasis. Understanding the key drivers of this differentiation process can aid the development of therapeutics to combat the growing obesity epidemic and associated metabolic consequences. Although much literature has covered the transcriptional events that culminate in the formation of an adipocyte, less focus has been on receptor-mediated extracellular signals that direct this process. This review will highlight GPCRs and their downstream messengers as significant players controlling adipocyte differentiation. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: The Toll and Toll-Like Receptor signaling pathways are evolutionarily conserved pathways that regulate innate immunity in insects and mammals. While efforts have been made to clarify the signal transduction events that occur during infection, much less is known about the components that maintain immune quiescence. Here we show that retromer, an intracellular protein complex known for regulating vesicle trafficking, functions in modulating the Toll pathway in Drosophila melanogaster . In mutant animals lacking retromer function, the Toll pathway but not JAK-STAT or IMD pathway is activated, triggering both cellular and humoral responses. Genetic epistasis and clonal analysis suggest that retromer regulates a component that acts upstream of Toll. Our data further show that in the mutant the Toll ligand Spätzle has a processing pattern similar to that of after infection. Together, the results suggest a novel function of retromer in regulating Toll pathway and innate immunity at a step that modulates ligand processing or activity. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Recent changes in sanitary policies by European Union (EU) concerning disposal of carcasses of domestic animals and the increase of non-natural mortality factors as illegal poisoning are threatening European vultures. However, the effects of anthropogenic actions in demographic parameters are poorly studied. Using a long-term study (1994-2011) of the threatened Pyrenean bearded vulture Gypaetus barbatus population, we assess the variation in the proportion of breeding pairs, egg-laying dates, clutch size, productivity, breeding success and survival following a sharp reduction in food availability in 2005. To test such variations we used Cusum tests (from the field of statistical process control) and multi-event capture-recapture modeling to estimate changes in survival probabilities. We found a delay in laying dates, and a regressive trend in clutch size, productivity, flight rate and survival. The maintenance of specific supplementary feeding stations for bearded vultures likely reduced the negative effects of illegal poisoning and food shortages, which mainly affected sub-adult survival. According to the Cusum test, demographic parameters suffered a significant shift after 2005 coinciding with a reduction in the food supply for the avian scavenger guild. Changes in food availability may have produced changes in demographic parameters and an increase in mortality due to an increased exposure to contaminated food. As a result supplementary feeding used as a precautionary measure can be a useful tool to reduce illegal poisoning and shifts in demographic parameters until precedent food availability scenarios are achieved. This study shows how anthropogenic actions through human health regulations affecting habitat quality can suddenly modify demographic parameters in long-lived raptors. Because in natural processes the variation of biological parameters is usually slow and gradual, and because randomness and environmental stochasticity makes it difficult to detect changes in the absence of a large number of records, the use of Cusum test constitutes a useful tool for studies in the field of ecology and evolution.

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print. Under a rapidly warming climate, a critical management issue in semi-arid forests of western North America is how to increase forest resilience to wildfire. We evaluated relationships between fuel reduction treatments and burn severity in the 2006 Tripod Complex fires which burned over 70,000 ha of mixed conifer forests in the North Cascades range of Washington State and involved 387 past harvest and fuel treatment units. A secondary objective was to investigate other drivers of burn severity including landform, weather, vegetation characteristics, and a recent mountain pine beetle outbreak. We used sequential autoregression (SAR) to evaluate drivers of burn severity, represented by the Relative differenced Normalized Burn Ratio index, in two study areas that are centered on early progressions of the wildfire complex. Significant predictor variables include treatment type, landform (elevation), fire weather (minimum relative humidity and maximum temperature), and vegetation characteristics including canopy closure, cover type, and mountain pine beetle attack. Recent mountain pine beetle damage was a statistically significant predictor variable with red and mixed classes of beetle attack associated with higher burn severity. Treatment age and size were only weakly correlated with burn severity and may be partly explained by the lack of treatments older than 30 years and low rates of fuel succession in these semi-arid forests. Even during extreme weather, fuel conditions and landform strongly influenced patterns of burn severity. Fuel treatments that included recent prescribed burning of surface fuels were particularly effective at mitigating burn severity. Although surface and canopy fuel treatments are unlikely to substantially reduce the area burned in regional fire years, recent research including this study suggests that they can be an effective management strategy for increasing forest landscape resilience to wildfires.

Description: Similar to phosphorylation, transient conjugation of ubiquitin to target proteins (ubiquitination) mediated by the concerted action of ubiquitin ligases and de-ubiquitinating enzymes (DUBs) can affect substrate function. As obligate intracellular parasites, viruses rely on different cellular pathways for their own replication and the well conserved ubiquitin conjugating/de-conjugating system is not an exception. Viruses not only usurp the host proteins involved in the ubiquitination/de-ubiquitination process, but they also encode their own ubiquitin ligases and DUBs. Here we report that an N-terminal variant of the herpes simplex virus (HSV) type-1 large tegument protein VP1/2 (VP1/2 1–767 ), encompassing an active DUB domain (herpesvirus tegument ubiquitin specific protease, htUSP), and TSG101, a component of the endosomal sorting complex required for transport (ESCRT)-I, functionally interact. In particular, VP1/2 1–767 modulates TSG101 ubiquitination and influences its intracellular distribution. Given the role played by the ESCRT machinery in crucial steps of both cellular pathways and viral life cycle, the identification of TSG101 as a cellular target for the HSV-1 specific de-ubiquitinating enzyme contributes to the clarification of the still under debate function of viral encoded DUBs highly conserved throughout the Herpesviridae family. This article is protected by copyright. All rights reserved

Description: The transition to a pathological phenotype such as Barrett's esophagus occurs via induction of resistance upon repeated contact with gastric refluxate in esophagus. This study examined the molecular changes within normal esophageal epithelial cells (EECs) under short-term acid loading and the role of these changes in defensive resistance against acidic cytotoxicity. After primary cultured EECs were exposed to pH 4-acidified medium (AM4), cell viability was determined by the MTT assay. Reactive oxygen species (ROS) and NAD(P)H oxidase (NOX) activity were measured. Activation of the mitogen-activated protein kinases (MAPKs) MEK/ERK1/2, p38 and JNK; phosphoinositol-3-kinase (PI3K)/Akt, and nuclear factor-kappa B (NF-κB) were detected by Western blot analysis or immunofluorescence staining. AM4 incubation induced intracellular ROS generation accompanied by increase in NOX activity, which was further increased by Na + /H + exchange-1 (NHE1)-dependent inhibition but was prevented by inhibition of NOX or mitochondria complex I. AM4 also induced phosphorylation of MEK/ERK1/2, p38 MAPK, PI3K/Akt, and nuclear translocation of NF-κB, and all these effects, except for p38 MAPK phosphorylation, were abolished by inhibition of ROS. ROS-dependent PI3K/Akt activation, which mediates NF-κB nuclear translocation, was inhibited by protein tyrosine kinase (PTK) inhibitors and NHE1-specific inhibitor. All inhibitors of NHE, ROS, PTK, PI3K or NF-κB further decreased AM4-induced cell viability. Acid loading in the presence of NHE1-dependent protection induced ROS generation by activating NOX and mitochondria complex I, which stimulated PTK/PI3K/Akt/NF-κB-dependent survival in EEC. Our data indicate that normal EEC initially respond to acid loading through intrinsic survival activation. This article is protected by copyright. All rights reserved

Description: Osteolytic bone diseases are characterized by excessive osteoclast formation and activation. Protein kinase C (PKC)-dependent pathways regulate cell growth, differentiation and apoptosis in many cellular systems, and have been implicated in cancer development and osteoclast formation. A number of PKC inhibitors with anti-cancer properties have been developed, but whether they might also influence osteolysis (a common complication of bone invading cancers) is unclear. We studied the effects of the PKC inhibitor compound, GF109203X on osteoclast formation and activity, processes driven by receptor activator of NFκB ligand (RANKL). We found that GF109203X strongly and dose dependently suppresses osteoclastogenesis and osteoclast activity in RANKL-treated primary mouse bone marrow cells. Consistent with this GF109203X reduced expression of key osteoclastic genes, including cathepsin K, calcitonin receptor, tartrate resistant acid phosphatase (TRAP) and the proton pump subunit V-ATPase-d2 in RANKL-treated primary mouse bone marrow cells. Expression of these proteins is dependent upon RANKL-induced NF-κB and NFAT transcription factor actions; both were reduced in osteoclast progenitor populations by GF109203X treatment, notably NFATc1 levels. Furthermore, we showed that GF109203X inhibits RANKL-induced calcium oscillation. Together, this study shows GF109203X may block osteoclast functions, suggesting that pharmacological blockade of PKC-dependent pathways has therapeutic potential in osteolytic diseases. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The sesquiterpene lactone Parthenolide (PN) exerted a cytotoxic effect on MDA-MB231 cells, a triple-negative breast cancer (TNBC) cell line, but its effectiveness was scarce when employed at low doses. This represents an obstacle for a therapeutic utilization of PN. In order to overcome this difficulty we associated to PN the suberoylanilide hydroxamic acid (SAHA), an histone deacetylase inhibitor. Our results show that SAHA synergistically sensitized MDA-MB231 cells to the cytotoxic effect of PN. It is noteworthy that treatment with PN alone stimulated the survival pathway Akt/mTOR and the consequent nuclear translocation of Nrf2, while treatment with SAHA alone induced autophagic activity. However when the cells were treated with SAHA/PN combination, SAHA suppressed PN effect on Akt/mTOR/Nrf2 pathway, while PN reduced the prosurvival autophagic activity of SAHA. In addition SAHA/PN combination induced GSH depletion, fall in Δψm, release of cytochrome c, activation of caspase 3 and apoptosis. Finally we demonstrated that combined treatment maintained both hyperacetylation of histones H3 and H4 induced by SAHA and down-regulation of DNMT1 expression induced by PN. Inhibition of the DNA-binding activity of NF-kB, which is determined by PN, was also observed after combined treatment. In conclusion, combination of PN to SAHA inhibits the cytoprotective responses induced by the single compounds, but does not alter the mechanisms leading to the cytotoxic effects. Taken together our results suggest that this combination could be a candidate for TNBC therapy. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Genomic and trascriptomic profiling has recently contributed details to the characterization of luminal B breast cancer. We explored the contribution of anthropometric, metabolic and molecular determinants to the multifaceted heterogeneity of this breast cancer subtype, with a specific focus on the association between body mass index (BMI), pre-treatment fasting glucose, hormone receptors and expression of human epidermal growth factor receptor 2 (HER2). Extensively annotated specimens were obtained from 154 women with luminal B breast cancer diagnosed at two Italian comprehensive cancer centres. Participants' characteristics were descriptively analyzed overall and by HER2 status (positive vs negative). BMI (〈25 vs ≥25), pre-treatment fasting glucose (〈median value of 94 mg/dl vs ≥94) and percentage of hormone receptors were tested for association with HER2 expression in regression models. In univariate models, BMI, fasting glucose and, at a lesser extent, percentage of estrogen receptors (ER) were significantly and inversely associated with HER2 expression (OR: 0.32, 95%CI: 0.16-0.66; 0.43, 0.23-0.0.82; 0.96, 0.94-0.97, respectively). The multivariate models confirmed the protective role of BMI and ER on HER2 expression, with luminal B HER2 positive patients being significantly less frequent among women within the highest category of BMI and percentage expression of ER compared with their counterparts (OR: 0.22, 95%CI: 0.09-0.53; 0.95, 0.93-0.97). In conclusions, BMI and percentage of ER representation are inversely associated with HER2 expression in luminal B breast cancers. Upon confirmatory findings, this might help identify patient subgroups who may best benefit from the use of interventions targeting insulin resistance in well depicted breast cancer scenarios. This article is protected by copyright. All rights reserved

Description: Histone deacetylases (HDACs) play a crucial role in the regulation of gene expression through remodeling of chromatin structures. However, the molecular mechanisms involved in this event remain unknown. In this study, we sought to examine whether HDAC inhibition-mediated protective effects involved HDAC4 sumoylation, degradation, and the proteasome pathway. Isolated neonatal mouse ventricular myocytes (NMVM) and H9c2 cardiomyoblasts were subjected to 48 hours of hypoxia (H) (1% O 2 ) and 2 hours of reoxygenation (R). Treatment of cardiomyocytes with trichostatin A (TSA) attenuated H/R-elicited injury, as indicated by a reduction of lactate dehydrogenase (LDH) leakage, an increase in cell viability, and decrease in apoptotic positive cardiomyocytes. MG132, a potent proteasome pathway inhibitor, abrogated TSA-induced protective effects, which was associated with the accumulation of ubiquitinated HDAC4. NMVM transduced with adenoviral HDAC4 led to an exaggeration of H/R-induced injury. TSA treatment resulted in a decrease in HDAC4 in cardiomyocytes infected with adenoviral HDAC4, and HDAC4-induced injury was attenuated by TSA. HDAC inhibition resulted in a significant reduction in reactive oxygen species (ROS) in cardiomyoblasts exposed to H/R, which was attenuated by blockade of the proteasome pathway. Cardiomyoblasts carrying wild type and sumoylation mutation (K559R) were established to examine effects of HDAC4 sumoylation and ubiquitination on H/R injury. Disruption of HDAC4 sumoylation brought about HDAC4 accumulation and impairment of HDAC4 ubiquitination in association with enhanced susceptibility of cardiomyoblasts to H/R. Taken together, these results demonstrated that HDAC inhibition stimulates proteasome dependent degradation of HDAC4, which is associated with HDAC4 sumoylation to induce these protective effects. This article is protected by copyright. All rights reserved

Description: Ecological Applications, Volume 21, Issue 4, Page 1085-1095, June 2011.

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Description: Ecological Applications, Volume 21, Issue 4, Page 1105-1119, June 2011.

Description: Ecological Applications, Volume 21, Issue 4, Page 1039-1054, June 2011.

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Description: Ecological Applications, Volume 21, Issue 4, Page 1037-1038, June 2011.

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Description: Endothelial cells play a major role in the initiation and perpetuation of the inflammatory process in health and disease, including their pivotal role in leukocyte recruitment. The role of pro-inflammatory transcription factors in this process has been well-described, including NF-κB. However, much less is known regarding transcription factors that play an anti-inflammatory role in endothelial cells. Myocyte enhancer factor 2 C (MEF2C) is a transcription factor known to regulate angiogenesis in endothelial cells. Here, we report that MEF2C plays a critical function as an inhibitor of endothelial cell inflammation. Tumor necrosis factor (TNF)-α inhibited MEF2C expression in endothelial cells. Knockdown of MEF2C in endothelial cells resulted in the upregulation of pro-inflammatory molecules and stimulated leukocyte adhesion to endothelial cells. MEF2C knockdown also resulted in NF-κB activation in endothelial cells. Conversely, MEF2C overexpression by adenovirus significantly repressed TNF-α induction of pro-inflammatory molecules, activation of NF-κB, and leukocyte adhesion to endothelial cells. This inhibition of leukocyte adhesion by MEF2C was partially mediated by induction of KLF2. In mice, lipopolysaccharide (LPS)-induced leukocyte adhesion to the retinal vasculature was significantly increased by endothelial cell-specific ablation of MEF2C. Taken together, these results demonstrate that MEF2C is a novel negative regulator of inflammation in endothelial cells and may represent a therapeutic target for vascular inflammation. This article is protected by copyright. All rights reserved

Description: Ecological Applications, Volume 0, Issue 0, Ahead of Print.

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Description: Ecological Applications, Volume 21, Issue 7, Page 2478-2486, October 2011.