Resin ducts (RDs) are features present in most conifer species as defence structures against pests and pathogens; however, little is known about RD expression in trees following fire injury. This study investigates changes in RD size and density in fire scars of Douglas fir (Pseudotsuga menziesii) and western larch (Larix occidentalis) as a means to evaluate the ecophysiological significance of traumatic resinosis for tree defence and survival.
Over the past century an ongoing establishment of Scots pine (Pinus sylvestris L.), sometimes at accelerating rates, is noted at three studied Lithuanian peat bogs, namely Ker?plis, R?kyva and Aukštumala, all representing different degrees of tree coverage and geographic settings. Present establishment rates seem to depend on tree density on the bog surface and are most significant at sparsely covered sites where about three-fourth of the trees have established since the mid-1990s, whereas the initial establishment in general was during the early to mid-19th century. Three methods were used to detect, compare and describe tree establishment: (1) tree counts in small plots, (2) dendrochronological dating of bog pine trees, and (3) interpretation of aerial photographs and historical maps of the study areas. In combination, the different approaches provide complimentary information but also weigh up each other's drawbacks. Tree counts in plots provided a reasonable overview of age class distributions and enabled capturing of the most recently established trees with ages less than 50years. The dendrochronological analysis yielded accurate tree ages and a good temporal resolution of long-term changes. Tree establishment and spread interpreted from aerial photographs and historical maps provided a good overview of tree spread and total affected area. It also helped to verify the results obtained with the other methods and an upscaling of findings to the entire peat bogs. The ongoing spread of trees in predominantly undisturbed peat bogs is related to warmer and/or drier climatic conditions, and to a minor degree to land-use changes. Our results therefore provide valuable insights into vegetation changes in peat bogs, also with respect to bog response to ongoing and future climatic changes.
Sphingosine-1-phosphate (S1P) mediates several cytoprotective functions of HDL. apoM acts as a S1P binding protein in HDL. Erythrocytes are the major source of S1P in plasma. After glomerular filtration, apoM is endocytosed in the proximal renal tubules. Human or murine HDL elicited time- and dose-dependent S1P efflux from erythrocytes. Compared with HDL of wild-type (wt) mice, S1P efflux was enhanced in the presence of HDL from apoM transgenic mice, but not diminished in the presence of HDL from apoM knockout (Apom(-/-)) mice. Artificially reconstituted and apoM-free HDL also effectively induced S1P efflux from erythrocytes. S1P and apoM were not measurable in the urine of wt mice. Apom(-/-) mice excreted significant amounts of S1P. apoM was detected in the urine of mice with defective tubular endocytosis because of knockout of the LDL receptor-related protein, chloride-proton exchanger ClC-5 (Clcn5(-/-)), or the cysteine transporter cystinosin. Urinary levels of S1P were significantly elevated in Clcn5(-/-) mice. In contrast to Apom(-/-) mice, these mice showed normal plasma concentrations for apoM and S1P. In conclusion, HDL facilitates S1P efflux from erythrocytes by both apoM-dependent and apoM-independent mechanisms. Moreover, apoM facilitates tubular reabsorption of S1P from the urine, however, with no impact on S1P plasma concentrations.
The outer segments of cones serve as light detectors for daylight color vision, and their dysfunction leads to human blindness conditions. We show that the cone-specific disruption of DGCR8 in adult mice led to the loss of miRNAs and the loss of outer segments, resulting in photoreceptors with significantly reduced light responses. However, the number of cones remained unchanged. The loss of the outer segments occurred gradually over 1 month, and during this time the genetic signature of cones decreased. Reexpression of the sensory-cell-specific miR-182 and miR-183 prevented outer segment loss. These miRNAs were also necessary and sufficient for the formation of inner segments, connecting cilia and short outer segments, as well as light responses in stem-cell-derived retinal cultures. Our results show that miR-182- and miR-183-regulated pathways are necessary for cone outer segment maintenance in vivo and functional outer segment formation in vitro.
Fire scars have been widely used as proxies for the reconstruction of fire history; however, little is known about the impact of fire injury on wood anatomy. This study investigates changes in tracheid and ray traits in fire scars of Douglas fir (Pseudotsuga menziesii), western larch (Larix occidentalis) and ponderosa pine (Pinus ponderosa), and discusses their ecophysiological implications for tree recovery from fire.
Dysfunctional microRNA (miRNA) networks contribute to inappropriate responses following pathological stress and are the underlying cause of several disease conditions. In pancreatic ? cells, miRNAs have been largely unstudied and little is known about how specific miRNAs regulate glucose-stimulated insulin secretion (GSIS) or impact the adaptation of ? cell function to metabolic stress. In this study, we determined that miR-7 is a negative regulator of GSIS in ? cells. Using Mir7a2 deficient mice, we revealed that miR-7a2 regulates ? cell function by directly regulating genes that control late stages of insulin granule fusion with the plasma membrane and ternary SNARE complex activity. Transgenic mice overexpressing miR-7a in ? cells developed diabetes due to impaired insulin secretion and ? cell dedifferentiation. Interestingly, perturbation of miR-7a expression in ? cells did not affect proliferation and apoptosis, indicating that miR-7 is dispensable for the maintenance of endocrine ? cell mass. Furthermore, we found that miR-7a levels are decreased in obese/diabetic mouse models and human islets from obese and moderately diabetic individuals with compensated ? cell function. Our results reveal an interconnecting miR-7 genomic circuit that regulates insulin granule exocytosis in pancreatic ? cells and support a role for miR-7 in the adaptation of pancreatic ? cell function in obesity and type 2 diabetes.
Correct regulation of insulin secretion by the pancreas is crucial for organismal function and survival. The AMPK-related kinase SIK2 (salt-inducible kinase 2) is now shown to be stabilized in pancreatic ?-cells following glucose stimulation, leading to improved systemic glucose homeostasis by regulating cellular calcium flux and insulin secretion.
Recent studies have reported that competitive endogenous RNAs (ceRNAs) can act as sponges for a microRNA (miRNA) through their binding sites and that changes in ceRNA abundances from individual genes can modulate the activity of miRNAs. Consideration of this hypothesis would benefit from knowing the quantitative relationship between a miRNA and its endogenous target sites. Here, we altered intracellular target site abundance through expression of an miR-122 target in hepatocytes and livers and analyzed the effects on miR-122 target genes. Target repression was released in a threshold-like manner at high target site abundance (?1.5 × 10(5) added target sites per cell), and this threshold was insensitive to the effective levels of the miRNA. Furthermore, in response to extreme metabolic liver disease models, global target site abundance of hepatocytes did not change sufficiently to affect miRNA-mediated repression. Thus, modulation of miRNA target abundance is unlikely to cause significant effects on gene expression and metabolism through a ceRNA effect.
In this study we aimed to establish an unbiased automatic quantification pipeline to assess islet specific features such as ?-cell area and density per islet based on immunofluorescence stainings. To determine these parameters, the in vivo protein expression levels of TMEM27 and BACE2 in pancreatic islets of 32 patients with type 2 diabetes (T2D) and in 28 non-diabetic individuals (ND) were used as input for the automated pipeline. The output of the automated pipeline was first compared to a previously developed manual area scoring system which takes into account the intensity of the staining as well as the percentage of cells which are stained within an islet. The median TMEM27 and BACE2 area scores of all islets investigated per patient correlated significantly with the manual scoring and with the median area score of insulin. Furthermore, the median area scores of TMEM27, BACE2 and insulin calculated from all T2D were significantly lower compared to the one of all ND. TMEM27, BACE2, and insulin area scores correlated as well in each individual tissue specimen. Moreover, islet size determined by costaining of glucagon and either TMEM27 or BACE2 and ?-cell density based either on TMEM27 or BACE2 positive cells correlated significantly. Finally, the TMEM27 area score showed a positive correlation with BMI in ND and an inverse pattern in T2D. In summary, automated quantification outperforms manual scoring by reducing time and individual bias. The simultaneous changes of TMEM27, BACE2, and insulin in the majority of the ?-cells suggest that these proteins reflect the total number of functional insulin producing ?-cells. Additionally, ?-cell subpopulations may be identified which are positive for TMEM27, BACE2 or insulin only. Thus, the cumulative assessment of all three markers may provide further information about the real ?-cell number per islet.
As the evidence for human induced climate change becomes clearer, so too does the realization that its effects will have impacts on numerous environmental and socio-economic systems. Mountains are recognized as very sensitive physical environments with populations whose histories and current social positions often strain their capacity to accommodate intense and rapid changes to their resource base. It is thus essential to assess the impacts of a changing climate, focusing on the quantity of water originating in mountain regions, particularly where snow and ice melt represent a large streamflow component as well as a local resource in terms of freshwater supply, hydropower generation, or irrigation. Increasing evidence of glacier retreat, permafrost degradation and reduced mountain snowpack has been observed in many regions, thereby suggesting that climate change may seriously affect streamflow regimes. These changes could in turn threaten the availability of water resources for many environmental and economic systems, and exacerbate a range of natural hazards that would compound these impacts. As a consequence, socio-economic structures of downstream living populations would be also impacted, calling for better preparedness and strategies to avoid conflicts of interest between water-dependent economic actors. This paper is thus an introduction to the Special Issue of this journal dedicated to the European Union Seventh Framework Program (EU-FP7) project ACQWA (Assessing Climate Impacts on the Quantity and Quality of WAter), a major European network of scientists that was coordinated by the University of Geneva from 2008 to 2014. The goal of ACQWA has been to address a number of these issues and propose a range of solutions for adaptation to change and to help improve water governance in regions where quantity, seasonality, and perhaps quality of water may substantially change in coming decades.
This paper gives a synthesis of this special issue on the sensitivity to climate change of the main bio-physical processes in the Hindukush-Karakoram-Himalayas. It also describes the impacts on the water resources with a special focus on the Ganges. Consequences of changes in water resources and possible adaptation measures for different sectors are discussed.
To set the stage of this special issue this paper gives a short introduction to the sensitivity to climate change of the main bio-physical processes in the Hindukush-Karakoram-Himalayas. It also describes the socio-economic setting of the Ganges basin in northern India as the main focal point of the impact and adaptation studies in this special issue.
Polar ice core records attest to a colossal volcanic eruption that took place ca. A.D. 1257 or 1258, most probably in the tropics. Estimates based on sulfate deposition in these records suggest that it yielded the largest volcanic sulfur release to the stratosphere of the past 7,000 y. Tree rings, medieval chronicles, and computational models corroborate the expected worldwide atmospheric and climatic effects of this eruption. However, until now there has been no convincing candidate for the mid-13th century "mystery eruption." Drawing upon compelling evidence from stratigraphic and geomorphic data, physical volcanology, radiocarbon dating, tephra geochemistry, and chronicles, we argue the source of this long-sought eruption is the Samalas volcano, adjacent to Mount Rinjani on Lombok Island, Indonesia. At least 40 km(3) (dense-rock equivalent) of tephra were deposited and the eruption column reached an altitude of up to 43 km. Three principal pumice fallout deposits mantle the region and thick pyroclastic flow deposits are found at the coast, 25 km from source. With an estimated magnitude of 7, this event ranks among the largest Holocene explosive eruptions. Radiocarbon dates on charcoal are consistent with a mid-13th century eruption. In addition, glass geochemistry of the associated pumice deposits matches that of shards found in both Arctic and Antarctic ice cores, providing compelling evidence to link the prominent A.D. 1258/1259 ice core sulfate spike to Samalas. We further constrain the timing of the mystery eruption based on tephra dispersal and historical records, suggesting it occurred between May and October A.D. 1257.
MicroRNAs (miRNAs) are short noncoding RNAs, which bind to messenger RNAs and regulate protein expression. The biosynthesis of miRNAs includes two precursors, a primary miRNA transcript (pri-miRNA) and a shorter pre-miRNA, both of which carry a common stem-loop bearing the mature miRNA. MiR-122 is a liver-specific miRNA with an important role in the life cycle of hepatitis C virus (HCV). It is the target of miravirsen (SPC3649), an antimiR drug candidate currently in clinical testing for treatment of HCV infections. Miravirsen is composed of locked nucleic acid (LNAs) ribonucleotides interspaced throughout a DNA phosphorothioate sequence complementary to mature miR-122. The LNA modifications endow the drug with high affinity for its target and provide resistance to nuclease degradation. While miravirsen is thought to work mainly by hybridizing to mature miR-122 and blocking its interaction with HCV RNA, its target sequence is also present in pri- and pre-miR-122. Using new in vitro and cellular assays specifically developed to discover ligands that suppress biogenesis of miR-122, we show that miravirsen binds to the stem-loop structure of pri- and pre-miR-122 with nanomolar affinity, and inhibits both Dicer- and Drosha-mediated processing of miR-122 precursors. This inhibition may contribute to the pharmacological activity of the drug in man.
This paper provides a synthesis and comparison of methodologies and results obtained in several studies devoted to the impact of climate change on hydropower. By putting into perspective various case studies, we provide a broader context and improved understanding of climate changes on energy production. We also underline the strengths and weaknesses of the approaches used as far as technical, physical and economical aspects are concerned. Although the catchments under investigation are located close to each other in geographic terms (Swiss and Italian Alps), they represent a wide variety of situations which may be affected by differing evolutions for instance in terms of annual runoff. In this study, we also differentiate between run-of-river, storage and pumping-storage power plants. By integrating and comparing various analyses carried out in the framework of the EU-FP7 ACQWA project, this paper discusses the complexity as well as current and future issues of hydropower management in the entire Alpine region.
Climate records for locations across the southern slope of the Himalaya between 77°E and 91°E were selected together with discharge measurements from gauging stations on rivers draining partially-glacierised basins tributary to the Ganges, with a view to assessing impacts of climatic fluctuations on year-to-year variations of runoff during a sustained period of glacier decline. The aims were to describe temporal patterns of variation of glaciologically- and hydrologically-relevant climatic variables and of river flows from basins with differing percentages of ice-cover. Monthly precipitation and air temperature records, starting in the mid-nineteenth century at high elevation sites and minimising data gaps, were selected from stations in the Global Historical Climatology Network and CRUTEM3. Discharge data availability was limited to post 1960 for stations in Nepal and at Khab in the adjacent Sutlej basin. Strengths of climate-runoff relationships were assessed by correlation between overlapping portions of annual data records. Summer monsoon precipitation dominates runoff across the central Himalaya. Flow in tributaries of the Ganges in Nepal fluctuated from year to year but the general background level of flow was usually maintained from the 1960s to 2000s. Flow in the Sutlej, however, declined by 32% between the 1970s and 1990s, reflecting substantially reduced summer precipitation. Over the north-west Ganges-upper Sutlej area, monsoon precipitation declined by 30-40% from the 1960s to 2000s. Mean May-September air temperatures along the southern slope of the central Himalayas dipped from the 1960s, after a long period of slow warming or sustained temperatures, before rising rapidly from the mid-1970s so that in the 2000s summer air temperatures reached those achieved in earlier warmer periods. There are few measurements of runoff from highly-glacierised Himalayan headwater basins; runoff from one of which, Langtang Khola, was less than that of the monsoon-dominated Narayani river, in which basin Langtang is nested.
The metabolic syndrome is a collection of risk factors including obesity, insulin resistance and hepatic steatosis, which occur together and increase the risk of diseases such as diabetes, cardiovascular disease and cancer. In spite of intense research, the complex etiology of insulin resistance and its association with the accumulation of triacylglycerides in the liver and with hepatic steatosis remains not completely understood. Here, we performed quantitative measurements of 144 proteins involved in the insulin-signaling pathway and central metabolism in liver homogenates of two genetically well-defined mouse strains C57BL/6J and 129Sv that were subjected to a sustained high-fat diet. We used targeted mass spectrometry by selected reaction monitoring (SRM) to generate accurate and reproducible quantitation of the targeted proteins across 36 different samples (12 conditions and 3 biological replicates), generating one of the largest quantitative targeted proteomics data sets in mammalian tissues. Our results revealed rapid response to high-fat diet that diverged early in the feeding regimen, and evidenced a response to high-fat diet dominated by the activation of peroxisomal ?-oxidation in C57BL/6J and by lipogenesis in 129Sv mice.
The Himalayas are presently holding the largest ice masses outside the polar regions and thus (temporarily) store important freshwater resources. In contrast to the contemplation of glaciers, the role of runoff from snow cover has received comparably little attention in the past, although (i) its contribution is thought to be at least equally or even more important than that of ice melt in many Himalayan catchments and (ii) climate change is expected to have widespread and significant consequences on snowmelt runoff. Here, we show that change assessment of snowmelt runoff and its timing is not as straightforward as often postulated, mainly as larger partial pressure of H2O, CO2, CH4, and other greenhouse gases might increase net long-wave input for snowmelt quite significantly in a future atmosphere. In addition, changes in the short-wave energy balance - such as the pollution of the snow cover through black carbon - or the sensible or latent heat contribution to snowmelt are likely to alter future snowmelt and runoff characteristics as well. For the assessment of snow cover extent and depletion, but also for its monitoring over the extremely large areas of the Himalayas, remote sensing has been used in the past and is likely to become even more important in the future. However, for the calibration and validation of remotely-sensed data, and even more so in light of possible changes in snow-cover energy balance, we strongly call for more in-situ measurements across the Himalayas, in particular for daily data on new snow and snow cover water equivalent, or the respective energy balance components. Moreover, data should be made accessible to the scientific community, so that the latter can more accurately estimate climate change impacts on Himalayan snow cover and possible consequences thereof on runoff.
Reliable estimates of future climate change in the Alps are relevant for large parts of the European society. At the same time, the complex Alpine region poses considerable challenges to climate models, which translate to uncertainties in the climate projections. Against this background, the present study reviews the state-of-knowledge about 21st century climate change in the Alps based on existing literature and additional analyses. In particular, it explicitly considers the reliability and uncertainty of climate projections. Results show that besides Alpine temperatures, also precipitation, global radiation, relative humidity, and closely related impacts like floods, droughts, snow cover, and natural hazards will be affected by global warming. Under the A1B emission scenario, about 0.25°C warming per decade until the mid of the 21st century and accelerated 0.36°C warming per decade in the second half of the century is expected. Warming will probably be associated with changes in the seasonality of precipitation, global radiation, and relative humidity, and more intense precipitation extremes and flooding potential in the colder part of the year. The conditions of currently record breaking warm or hot winter or summer seasons, respectively, may become normal at the end of the 21st century, and there is indication for droughts to become more severe in the future. Snow cover is expected to drastically decrease below 1500-2000m and natural hazards related to glacier and permafrost retreat are expected to become more frequent. Such changes in climatic parameters and related quantities will have considerable impact on ecosystems and society and will challenge their adaptive capabilities.
Obesity and diabetes are particularly high in indigenous populations exposed to a Western diet and lifestyle. The prevalence of obesity, diabetes, hyperglycemia, dyslipidemia, and hypertension in one such population, the Micronesian island of Kosrae was described.
Expansion of functional islet ?-cell mass is a physiological process to compensate for increased insulin demand. Deficiency or pharmacological inhibition of the plasma membrane protease BACE2 enhances pancreatic ?-cell function and proliferation, and therefore BACE2 is a putative target for the therapeutic intervention under conditions of ?-cell loss and dysfunction. To gain a molecular understanding of BACE2 function, we performed a systematic and quantitative proteomic analysis to map the natural substrate repertoire of BACE2 and its homologue BACE1 in ?-cells. Loss- and gain-of-function studies of in vitro and in vivo models identified specific and functionally heterogeneous targets. Our analysis revealed non-redundant roles of BACE1/2 in ectodomain shedding with BACE1 regulating a broader and BACE2 a more distinct set of ?-cell-enriched substrates including two proteins of the seizure 6 protein family (SEZ6L and SEZ6L2). Lastly, our study provides insights into the global ?-cell sheddome and secretome, an important prerequisite to uncover novel mechanisms contributing to ?-cell homeostasis and a resource for therapeutic target and biomarker discoveries.
Insulin resistance represents a hallmark during the development of type 2 diabetes mellitus and in the pathogenesis of obesity-associated disturbances of glucose and lipid metabolism. MicroRNA (miRNA)-dependent post-transcriptional gene silencing has been recognized recently to control gene expression in disease development and progression, including that of insulin-resistant type 2 diabetes. The deregulation of miRNAs miR-143 (ref. 4), miR-181 (ref. 5), and miR-103 and miR-107 (ref. 6) alters hepatic insulin sensitivity. Here we report that the expression of miR-802 is increased in the liver of two obese mouse models and obese human subjects. Inducible transgenic overexpression of miR-802 in mice causes impaired glucose tolerance and attenuates insulin sensitivity, whereas reduction of miR-802 expression improves glucose tolerance and insulin action. We identify Hnf1b (also known as Tcf2) as a target of miR-802-dependent silencing, and show that short hairpin RNA (shRNA)-mediated reduction of Hnf1b in liver causes glucose intolerance, impairs insulin signalling and promotes hepatic gluconeogenesis. In turn, hepatic overexpression of Hnf1b improves insulin sensitivity in Lepr(db/db) mice. Thus, this study defines a critical role for deregulated expression of miR-802 in the development of obesity-associated impairment of glucose metabolism through targeting of Hnf1b, and assigns Hnf1b an unexpected role in the control of hepatic insulin sensitivity.
Circulating levels of insulin and glucagon reflect the nutritional state of animals and elicit regulatory responses in the liver that maintain glucose and lipid homeostasis. The transcription factor Foxa2 activates lipid metabolism and ketogenesis during fasting and is inhibited via insulin-PI3K-Akt signaling-mediated phosphorylation at Thr156 and nuclear exclusion. Here we show that, in addition, Foxa2 is acetylated at the conserved residue Lys259 following inhibition of histone deacetylases (HDACs) class I-III and the cofactors p300 and SirT1 are involved in Foxa2 acetylation and deacetylation, respectively. Physiologically, fasting states and glucagon stimulation are sufficient to induce Foxa2 acetylation. Introduction of the acetylation-mimicking (K259Q) or -deficient (K259R) mutations promotes or inhibits Foxa2 activity, respectively, and adenoviral expression of Foxa2-K259Q augments expression of genes involved in fatty acid oxidation and ketogenesis. Our study reveals a molecular mechanism by which glucagon signaling activates a fasting response through acetylation of Foxa2.
Defects in insulin signalling are among the most common and earliest defects that predispose an individual to the development of type 2 diabetes. MicroRNAs have been identified as a new class of regulatory molecules that influence many biological functions, including metabolism. However, the direct regulation of insulin sensitivity by microRNAs in vivo has not been demonstrated. Here we show that the expression of microRNAs 103 and 107 (miR-103/107) is upregulated in obese mice. Silencing of miR-103/107 leads to improved glucose homeostasis and insulin sensitivity. In contrast, gain of miR-103/107 function in either liver or fat is sufficient to induce impaired glucose homeostasis. We identify caveolin-1, a critical regulator of the insulin receptor, as a direct target gene of miR-103/107. We demonstrate that caveolin-1 is upregulated upon miR-103/107 inactivation in adipocytes and that this is concomitant with stabilization of the insulin receptor, enhanced insulin signalling, decreased adipocyte size and enhanced insulin-stimulated glucose uptake. These findings demonstrate the central importance of miR-103/107 to insulin sensitivity and identify a new target for the treatment of type 2 diabetes and obesity.
Decreased ? cell mass and function are hallmarks of type 2 diabetes. Here we identified, through a siRNA screen, beta site amyloid precursor protein cleaving enzyme 2 (Bace2) as the sheddase of the proproliferative plasma membrane protein Tmem27 in murine and human ? cells. Mice with functionally inactive Bace2 and insulin-resistant mice treated with a newly identified Bace2 inhibitor both display augmented ? cell mass and improved control of glucose homeostasis due to increased insulin levels. These results implicate Bace2 in the control of ? cell maintenance and provide a rational strategy to inhibit this protease for the expansion of functional pancreatic ? cell mass.
Fetal hemoglobin, HbF (?(2)?(2)), is the main hemoglobin synthesized up to birth, but it subsequently declines and adult hemoglobin, HbA (?(2)?(2)), becomes predominant. Several studies have indicated that expression of the HbF subunit ?-globin might be regulated post-transcriptionally. This could be confered by ?22-nucleotide long microRNAs that associate with argonaute proteins to specifically target ?-globin mRNAs and inhibit protein expression. Indeed, applying immunopurifications, we found that ?-globin mRNA was associated with argonaute 2 isolated from reticulocytes that contain low levels of HbF (<1%), whereas association was significantly lower in reticulocytes with high levels of HbF (90%). Comparing microRNA expression in reticulocytes from cord blood and adult blood, we identified several miRNAs that were preferentially expressed in adults, among them miRNA-96. The overexpression of microRNA-96 in human ex vivo erythropoiesis decreased ?-globin expression by 50%, whereas the knock-down of endogenous microRNA-96 increased ?-globin expression by 20%. Moreover, luciferase reporter assays showed that microRNA-96 negatively regulates expression of ?-globin in HEK293 cells, which depends on a seedless but highly complementary target site located within the coding sequence of ?-globin. Based on these results we conclude that microRNA-96 directly suppresses ?-globin expression and thus contributes to HbF regulation.
The potential benefits of using population isolates in genetic mapping, such as reduced genetic, phenotypic and environmental heterogeneity, are offset by the challenges posed by the large amounts of direct and cryptic relatedness in these populations confounding basic assumptions of independence. We have evaluated four representative specialized methods for association testing in the presence of relatedness; (i) within-family (ii) within- and between-family and (iii) mixed-models methods, using simulated traits for 2906 subjects with known genome-wide genotype data from an extremely isolated population, the Island of Kosrae, Federated States of Micronesia. We report that mixed models optimally extract association information from such samples, demonstrating 88% power to rank the true variant as among the top 10 genome-wide with 56% achieving genome-wide significance, a >80% improvement over the other methods, and demonstrate that population isolates have similar power to non-isolate populations for observing variants of known effects. We then used the mixed-model method to reanalyze data for 17 published phenotypes relating to metabolic traits and electrocardiographic measures, along with another 8 previously unreported. We replicate nine genome-wide significant associations with known loci of plasma cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides, thyroid stimulating hormone, homocysteine, C-reactive protein and uric acid, with only one detected in the previous analysis of the same traits. Further, we leveraged shared identity-by-descent genetic segments in the region of the uric acid locus to fine-map the signal, refining the known locus by a factor of 4. Finally, we report a novel associations for height (rs17629022, P< 2.1 × 10??).
Vessel chronologies in ring-porous species have been successfully employed in the past to extract the climate signal from tree rings. Environmental signals recorded in vessels of ring-porous species have also been used in previous studies to reconstruct discrete events of drought, flooding and insect defoliation. However, very little is known about the ability of diffuse-porous species to record environmental signals in their xylem cells. Moreover, time series of wood anatomical features have only rarely been used to reconstruct former geomorphic events. This study was therefore undertaken to characterize the wood anatomical response of diffuse-porous Alnus incana (L.) Moench and Betula pendula Roth to debris-flow-induced wounding. Tree microscopic response to wounding was assessed through the analysis of wood anatomical differences between injured rings formed in the debris-flow event year and uninjured rings formed in the previous year. The two ring types were examined close and opposite to the injury in order to determine whether wound effects on xylem cells decrease with increasing tangential distance from the injury. Image analysis was used to measure vessel parameters as well as fiber and parenchyma cell (FPC) parameters. The results of this study indicate that injured rings are characterized by smaller vessels as compared with uninjured rings. By contrast, FPC parameters were not found to significantly differ between injured and uninjured rings. Vessel and FPC parameters mainly remained constant with increasing tangential distance from the injury, except for a higher proportion of vessel lumen area opposite to the injury within A. incana. This study highlights the existence of anatomical tree-ring signatures-in the form of smaller vessels-related to past debris-flow activity and addresses a new methodological approach to date injuries inflicted on trees by geomorphic processes.
The architecture of natural variation present in a contemporary population is a result of multiple population genetic forces, including population bottleneck and expansion, selection, drift, and admixture. We seek to untangle the contribution of admixture to genetic diversity on the Micronesian island of Kosrae. Toward this goal, we used a complete genetic approach by combining a dense genome-wide map of 100,000 single-nucleotide polymorphisms (SNPs) with data from uniparental markers from the mitochondrial genome and the nonrecombining portion of the Y chromosome. These markers were typed in approximately 3200 individuals from Kosrae, representing 80% of the adult population of the island. We developed novel software that uses SNP data to delineate ancestry for individual segments of the genome. Through this analysis, we determined that 39% of Kosraens have some European ancestry. However, the vast majority of admixed individuals (77%) have European alleles spanning less than 10% of their genomes. Data from uniparental markers show most of this admixture to be male, introduced in the late nineteenth century. Furthermore, pedigree analysis shows that the majority of European admixture on Kosrae is because of the contribution of one individual. This approach shows the benefit of combining information from autosomal and uniparental polymorphisms and provides new methodology for determining ancestry in a population.
Studies on tree reaction after wounding were so far based on artificial wounding or chemical treatment. For the first time, type, spread and intensity of anatomical responses were analyzed and quantified in naturally disturbed Larix decidua Mill., Picea abies (L.) Karst. and Abies alba Mill. trees. The consequences of rockfall impacts on increment growth were assessed at the height of the wounds, as well as above and below the injuries. A total of 16 trees were selected on rockfall slopes, and growth responses following 54 wounding events were analyzed on 820 cross-sections. Anatomical analysis focused on the occurrence of tangential rows of traumatic resin ducts (TRD) and on the formation of reaction wood. Following mechanical disturbance, TRD production was observed in 100% of L. decidua and P. abies wounds. The radial extension of TRD was largest at wound height, and they occurred more commonly above, rather than below, the wounds. For all species, an intra-annual radial shift of TRD was observed with increasing axial distance from wounds. Reaction wood was formed in 87.5% of A. alba following wounding, but such cases occurred only in 7.7% of L. decidua. The results demonstrate that anatomical growth responses following natural mechanical disturbance differ significantly from the reactions induced by artificial stimuli or by decapitation. While the types of reactions remain comparable between the species, their intensity, spread and persistence disagree considerably. We also illustrate that the external appearance of wounds does not reflect an internal response intensity. This study reveals that disturbance induced under natural conditions triggers more intense and more widespread anatomical responses than that induced under artificial stimuli, and that experimental laboratory tests considerably underestimate tree response.
Pinpointing culprit causal variants along signal peaks of genome-wide association studies (GWAS) is challenging. To overcome confounding effects of multiple independent variants at such a locus and narrow the interval for causal allele capture, we developed an approach that maps local shared haplotypes harboring a putative causal variant. We demonstrate our method in an extreme isolate founder population, the pacific Island of Kosrae. We analyzed plasma plant sterol (PPS) levels, a surrogate measure of cholesterol absorption from the intestine, where previous studies have implicated 2p21 mutations in the ATP binding cassette subfamily G members 5 or 8 (ABCG5 or ABCG8) genes. We have previously reported that 11.1% of the islanders are carriers of a frameshift ABCG8 mutation increasing PPS levels in carriers by 50%. GWAS adjusted for this mutation revealed genomewide significant signals along 11 Mb around it. To fine-map this signal, we detected pairwise identity-by-descent haplotypes using our tool GERMLINE and implemented a clustering algorithm to identify haplotypes shared across multiple samples with their unique shared boundaries. A single 526-kb haplotype mapped strongly to PPS levels, dramatically refining the mapped interval. This haplotype spans the ABCG5/ABCG8 genes, is carried by 1.8% of the islanders, and results in a striking 100% increase of PPS in carriers. Resequencing of ABCG5 in these carriers found a D450H missense mutation along the associated haplotype. These findings exemplify the power of haplotype analysis for mapping mutations in isolated populations and specifically for dissecting effects of multiple variants of the same locus.
The Forkhead box A2 transcription factor (Foxa2/HNF-3beta) has been shown to be a key regulator of genes involved in the maintenance of glucose and lipid homeostasis in the liver. It is constitutively inactivated in several hyperinsulinemic/obese mouse models, thereby enhancing their metabolic phenotypes. Foxa2 is activated under fasting conditions but is inhibited by insulin signaling via phosphatidylinositol 3-kinase/AKT in a phosphorylation-dependent manner, which results in its nuclear exclusion. However, the mechanism and relative importance of its nuclear export has not yet been elucidated. Here we show that Foxa2 contains a functional nuclear export signal and is excluded from the nucleus via a CRM1-dependent pathway in response to insulin signaling. Furthermore, direct evidence is provided that nuclear export-defective Foxa2 is phosphorylated and inactivated by insulin in vitro and in vivo. These data demonstrate for the first time that phosphorylation itself is the main event regulating the activity of Foxa2, suggesting that export-independent mechanisms have evolved to ensure inhibition of Foxa2 under conditions in which insulin signaling is present.
The lateral hypothalamic area is considered the classic feeding centre, regulating food intake, arousal and motivated behaviour through the actions of orexin and melanin-concentrating hormone (MCH). These neuropeptides are inhibited in response to feeding-related signals and are released during fasting. However, the molecular mechanisms that regulate and integrate these signals remain poorly understood. Here we show that the forkhead box transcription factor Foxa2, a downstream target of insulin signalling, regulates the expression of orexin and MCH. During fasting, Foxa2 binds to MCH and orexin promoters and stimulates their expression. In fed and in hyperinsulinemic obese mice, insulin signalling leads to nuclear exclusion of Foxa2 and reduced expression of MCH and orexin. Constitutive activation of Foxa2 in the brain (Nes-Cre/+;Foxa2T156A(flox/flox) genotype) results in increased neuronal MCH and orexin expression and increased food consumption, metabolism and insulin sensitivity. Spontaneous physical activity of these animals in the fed state is significantly increased and is similar to that in fasted mice. Conditional activation of Foxa2 through the T156A mutation expression in the brain of obese mice also resulted in improved glucose homeostasis, decreased fat and increased lean body mass. Our results demonstrate that Foxa2 can act as a metabolic sensor in neurons of the lateral hypothalamic area to integrate metabolic signals, adaptive behaviour and physiological responses.
Altered growth and development of the endocrine pancreas is a frequent cause of the hyperglycemia associated with diabetes. Here we show that microRNA-375 (miR-375), which is highly expressed in pancreatic islets, is required for normal glucose homeostasis. Mice lacking miR-375 (375KO) are hyperglycemic, exhibit increased total pancreatic alpha-cell numbers, fasting and fed plasma glucagon levels, and increased gluconeogenesis and hepatic glucose output. Furthermore, pancreatic beta-cell mass is decreased in 375KO mice as a result of impaired proliferation. In contrast, pancreatic islets of obese mice (ob/ob), a model of increased beta-cell mass, exhibit increased expression of miR-375. Genetic deletion of miR-375 from these animals (375/ob) profoundly diminished the proliferative capacity of the endocrine pancreas and resulted in a severely diabetic state. Bioinformatic analysis of transcript data from 375KO islets revealed that miR-375 regulates a cluster of genes controlling cellular growth and proliferation. These data provide evidence that miR-375 is essential for normal glucose homeostasis, alpha- and beta-cell turnover, and adaptive beta-cell expansion in response to increasing insulin demand in insulin resistance.
MicroRNAs play important roles in animal development. Numerous conditional knockout (cKO) studies of Dicer have been performed to interrogate the functions of microRNA during mammalian development. However, because Dicer was recently implicated in the biogenesis of endogenous siRNAs in mammals, it raises the question whether the Dicer cKO defects can be attributable to the loss of microRNAs. Previously, we and others conditionally targeted Dicer and identified its critical roles in embryonic skin morphogenesis. Here, we focus explicitly on microRNAs by taking a parallel strategy with Dgcr8, encoding an essential component of the microprocessor complex that is exclusively required for microRNA biogenesis. With this comparative analysis, we show definitively that the Dicer- and Dgcr8-null skin defects are both striking and indistinguishable. By deep sequencing analysis of microRNA depletion in both Dicer- and Dgcr8-null skin, we demonstrate that most abundantly expressed skin microRNAs are dependent on both Dicer and DGCR8. Our results underscore a specific importance of microRNAs in controlling mammalian skin development.
A new method for investigating the detailed reaction and the energy absorption of trees during a rock impact was developed and applied to 15 subalpine Norway spruce (Picea abies L. Karst) trees. A wedge-shaped trolley, guided by prestressed steel wires, was mounted on a forested slope to simulate a falling rock. The trolley accelerates down the wires and hits a tree at a preselected stem height with variable energies. The tree displacements and accelerations during the impact were recorded to determine reactions and energy absorption for the stem, root-soil system, crown and the entire tree. Trees absorbed the kinetic energy of the trolley rapidly by mobilizing strain and inertia forces close to the impact location in the stem and the root-soil system. This energy was then gradually dissipated all over the tree through permanent deformations and damping. The stem assimilated more energy than the root-soil system. The trees energy absorption capacity was limited by stem-bending stresses at impact height, by shear stresses at the stem base and by lack of resistance of the root-soil anchorage. It was positively and exponentially related to stem diameter at breast height and negatively related to impact height. The field experiment enabled a physical description of how a tree reacts to a rock impact and highlighted the most important and critical components of the tree for its energy absorption. Such descriptions should help make computer simulations of rock-forest interrelations more precise and thus improve management strategies to ensure that forests can provide protection against rockfall.
After mechanical wounding, callus tissue and tangential rows of traumatic resin ducts (TRDs) are formed in many conifer species. This reaction can be used to date past events of geomorphic processes such as rockfall, debris flow and snow avalanches. However, only few points are known about the tangential spread or the timing of callus tissue and TRD formation after wounding. We analyzed 19 Larix decidua Mill. (European larch) and eight Picea abies (L.) Karst. (Norway spruce) trees that were severely damaged by rockfall activity, resulting in a total of 111 injuries. Callus tissue appeared sparsely on the cross sections and was detected on only 4.2% of the L. decidua samples and 3.6% of the P. abies samples. In contrast, TRDs were present on all cross sections following wounding and were visible on more than one-third (34% in L. decidua and 36.4% in P. abies) of the circumference where the cambium was not destroyed by the rockfall impact. We observe different reactions in the trees depending on the seasonal timing of wounding. The tangential spread of callus tissue and TRDs was more important if the injury occurred during the growth period than during the dormant season, with the difference between seasons being more pronounced for callus tissue formation than for TRD formation. We observed an intra-annual radial migration of TRDs with increasing tangential distance from the wound in 73.2% of the L. decidua samples and 96.6% of the P. abies samples. The persistence of TRD formation in the years following wounding showed that only L. decidua trees produced TRDs 2 years after wounding (10.5%), whereas P. abies trees produced TRDs 5 years after wounding (> 50%).
It has been argued that the limited genetic diversity and reduced allelic heterogeneity observed in isolated founder populations facilitates discovery of loci contributing to both Mendelian and complex disease. A strong founder effect, severe isolation, and substantial inbreeding have dramatically reduced genetic diversity in natives from the island of Kosrae, Federated States of Micronesia, who exhibit a high prevalence of obesity and other metabolic disorders. We hypothesized that genetic drift and possibly natural selection on Kosrae might have increased the frequency of previously rare genetic variants with relatively large effects, making these alleles readily detectable in genome-wide association analysis. However, mapping in large, inbred cohorts introduces analytic challenges, as extensive relatedness between subjects violates the assumptions of independence upon which traditional association test statistics are based. We performed genome-wide association analysis for 15 quantitative traits in 2,906 members of the Kosrae population, using novel approaches to manage the extreme relatedness in the sample. As positive controls, we observe association to known loci for plasma cholesterol, triglycerides, and C-reactive protein and to a compelling candidate loci for thyroid stimulating hormone and fasting plasma glucose. We show that our study is well powered to detect common alleles explaining >/=5% phenotypic variance. However, no such large effects were observed with genome-wide significance, arguing that even in such a severely inbred population, common alleles typically have modest effects. Finally, we show that a majority of common variants discovered in Caucasians have indistinguishable effect sizes on Kosrae, despite the major differences in population genetics and environment.
We present GERMLINE, a robust algorithm for identifying segmental sharing indicative of recent common ancestry between pairs of individuals. Unlike methods with comparable objectives, GERMLINE scales linearly with the number of samples, enabling analysis of whole-genome data in large cohorts. Our approach is based on a dictionary of haplotypes that is used to efficiently discover short exact matches between individuals. We then expand these matches using dynamic programming to identify long, nearly identical segmental sharing that is indicative of relatedness. We use GERMLINE to comprehensively survey hidden relatedness both in the HapMap as well as in a densely typed island population of 3000 individuals. We verify that GERMLINE is in concordance with other methods when they can process the data, and also facilitates analysis of larger scale studies. We bolster these results by demonstrating novel applications of precise analysis of hidden relatedness for (1) identification and resolution of phasing errors and (2) exposing polymorphic deletions that are otherwise challenging to detect. This finding is supported by concordance of detected deletions with other evidence from independent databases and statistical analyses of fluorescence intensity not used by GERMLINE.
Glacial lake hazards and glacial lake distributions are investigated in many glaciated regions of the world, but comparably little attention has been given to these topics in the Indian Himalayas. In this study we present a first area-wide glacial lake inventory, including a qualitative classification at 251 glacial lakes >0.01km(2). Lakes were detected in the five states spanning the Indian Himalayas, and lake distribution pattern and lake characteristics were found to differ significantly between regions. Three glacial lakes, from different geographic and climatic regions within the Indian Himalayas were then selected for a detailed risk assessment. Lake outburst probability, potential outburst magnitudes and associated damage were evaluated on the basis of high-resolution satellite imagery, field assessments and through the use of a dynamic model. The glacial lakes analyzed in the states of Jammu and Kashmir and Himachal Pradesh were found to present moderate risks to downstream villages, whereas the lake in Sikkim severely threatens downstream locations. At the study site in Sikkim, a dam breach could trigger drainage of ca. 16×10(6)m(3) water and generate maximum lake discharge of nearly 7000m(3)s(-). The identification of critical glacial lakes in the Indian Himalayas and the detailed risk assessments at three specific sites allow prioritizing further investigations and help in the definition of risk reduction actions.
Brown adipose tissue (BAT) uses the chemical energy of lipids and glucose to produce heat, a function that can be induced by cold exposure or diet. A key regulator of BAT is the gene encoding PR domain containing 16 (Prdm16), whose expression can drive differentiation of myogenic and white fat precursors to brown adipocytes. Here we show that after cold exposure, the muscle-enriched miRNA-133 is markedly downregulated in BAT and subcutaneous white adipose tissue (SAT) as a result of decreased expression of its transcriptional regulator Mef2. miR-133 directly targets and negatively regulates PRDM16, and inhibition of miR-133 or Mef2 promotes differentiation of precursors from BAT and SAT to mature brown adipocytes, thereby leading to increased mitochondrial activity. Forced expression of miR-133 in brown adipogenic conditions prevents the differentiation to brown adipocytes in both BAT and SAT precursors. Our results point to Mef2 and miR-133 as central upstream regulators of Prdm16 and hence of brown adipogenesis in response to cold exposure in BAT and SAT.
The integration of scientific knowledge about possible climate change impacts on water resources has a direct implication on the way water policies are being implemented and evolving. This is particularly true regarding various technical steps embedded into the EU Water Framework Directive river basin management planning, such as risk characterisation, monitoring, design and implementation of action programmes and evaluation of the "good status" objective achievements (in 2015). The need to incorporate climate change considerations into the implementation of EU water policy is currently discussed with a wide range of experts and stakeholders at EU level. Research trends are also on-going, striving to support policy developments and examining how scientific findings and recommendations could be best taken on board by policy-makers and water managers within the forthcoming years. This paper provides a snapshot of policy discussions about climate change in the context of the WFD river basin management planning and specific advancements of related EU-funded research projects. Perspectives for strengthening links among the scientific and policy-making communities in this area are also highlighted.
Therapeutics based on RNA interference (RNAi) have emerged as a potential new class of drugs for treating human disease by silencing the target messenger RNA (mRNA), thereby reducing levels of the corresponding pathogenic protein. The major challenge for RNAi therapeutics is the development of safe delivery vehicles for small interfering RNAs (siRNAs). We previously showed that cholesterol-conjugated siRNAs (chol-siRNA) associate with plasma lipoprotein particles and distribute primarily to the liver after systemic administration to mice. We further demonstrated enhancement of silencing by administration of chol-siRNA pre-associated with isolated high-density lipoprotein (HDL) or low-density lipoprotein (LDL). In this study, we investigated mimetic lipoprotein particle prepared from recombinant apolipoprotein A1 (apoA) and apolipoprotein E3 (apoE) as a delivery vehicle for chol-siRNAs. We show that apoE-containing particle (E-lip) is highly effective in functional delivery of chol-siRNA to mouse liver. E-lip delivery was found to be considerably more potent than apoA-containing particle (A-lip). Furthermore, E-lip-mediated delivery was not significantly affected by high endogenous levels of plasma LDL. These results demonstrate that E-lip has substantial potential as delivery vehicles for lipophilic conjugates of siRNAs.
Vessels of broad-leaved trees have been analyzed to study how trees deal with various environmental factors. Cambial injury, in particular, has been reported to induce the formation of narrower conduits. Yet, little or no effort has been devoted to the elaboration of vessel sampling strategies for retrospective injury detection based on vessel lumen size reduction. To fill this methodological gap, four wounded individuals each of grey alder (Alnus incana (L.) Moench) and downy birch (Betula pubescens Ehrh.) were harvested in an avalanche path. Earlywood vessel lumina were measured and compared for each tree between the injury ring built during the growing season following wounding and the control ring laid down the previous year. Measurements were performed along a 10 mm wide radial strip, located directly next to the injury. Specifically, this study aimed at (i) investigating the intra-annual duration and local extension of vessel narrowing close to the wound margin and (ii) identifying an adequate sample of earlywood vessels (number and intra-ring location of cells) attesting to cambial injury. Based on the results of this study, we recommend analyzing at least 30 vessels in each ring. Within the 10 mm wide segment of the injury ring, wound-induced reduction in vessel lumen size did not fade with increasing radial and tangential distances, but we nevertheless advise favoring early earlywood vessels located closest to the injury. These findings, derived from two species widespread across subarctic, mountainous, and temperate regions, will assist retrospective injury detection in Alnus, Betula, and other diffuse-porous species as well as future related research on hydraulic implications after wounding.
Adaptation is increasingly important for regions around the world where large changes in climate could have an impact on populations and industry. The Brahmaputra-Ganges catchments have a large population, a main industry of agriculture and a growing hydro-power industry, making the region susceptible to changes in the Indian Summer Monsoon, annually the main water source. The HighNoon project has completed four regional climate model simulations for India and the Himalaya at high resolution (25km) from 1960 to 2100 to provide an ensemble of simulations for the region. In this paper we have assessed the ensemble for these catchments, comparing the simulations with observations, to give credence that the simulations provide a realistic representation of atmospheric processes and therefore future climate. We have illustrated how these simulations could be used to provide information on potential future climate impacts and therefore aid decision-making using climatology and threshold analysis. The ensemble analysis shows an increase in temperature between the baseline (1970-2000) and the 2050s (2040-2070) of between 2 and 4°C and an increase in the number of days with maximum temperatures above 28°C and 35°C. There is less certainty for precipitation and runoff which show considerable variability, even in this relatively small ensemble, spanning zero. The HighNoon ensemble is the most complete data for the region providing useful information on a wide range of variables for the regional climate of the Brahmaputra-Ganges region, however there are processes not yet included in the models that could have an impact on the simulations of future climate. We have discussed these processes and show that the range from the HighNoon ensemble is similar in magnitude to potential changes in projections where these processes are included. Therefore strategies for adaptation must be robust and flexible allowing for advances in the science and natural environmental changes.
The pancreatic ?-cell surface protein Tmem27 is promotes the preservation of functional ?-cell mass. It is a selective substrate of the protease Bace2, yet the intramolecular features of Tmem27 that regulate its processing by this sheddase have not been characterized. In particular, the importance of homodimerization, glycosylation, trafficking to the plasma membrane (PM), the existence of multiple cleavage sites, and the amino acid residues that govern these features are currently unknown. Using Tmem27 mutational analysis and multiple biochemical approaches, we here show that Tmem27 dimerization is a dynamic process mediated by its intracellular cysteine residue and that prevents Tmem27 cleavage, that extracellular asparagine glycosylation is essential for Tmem27 trafficking to the PM and its processing by Bace2, that the amount of Tmem27 at the PM is proportional to its total cell levels upon glucose stimulation and Bace2 inhibition, and that the double phenylalanine motif in the Tmem27 cleavage site is an intramolecular Bace2 inhibitor. These findings define structural properties of Tmem27 that affect the susceptibility to its protease Bace2 and have implications for the efficiency with which Tmem27 and other Bace2 substrates are cleaved in normal and disease states.
Cambial injury has been reported to alter wood structure in broad-leaved trees. However, the duration and extension of associated anatomical changes have rarely been analysed thoroughly. A total of 18 young European ash (Fraxinus excelsior L.) trees injured on the stem by a spring flood were sampled with the aim of comparing earlywood vessels and rays formed prior to and after the scarring event. Anatomical and hydraulic parameters were measured in five successive rings over one-quarter of the stem circumference. The results demonstrate that mechanical damage induces a decrease in vessel lumen size (up to 77%) and an increase in vessel number (up to 475%) and ray number (up to 115%). The presence of more earlywood vessels and rays was observed over at least three years after stem scarring. By contrast, abnormally narrow earlywood vessels mainly developed in the first ring formed after the event, increasing the thickness-to-span ratio of vessels by 94% and reducing both xylem relative conductivity and the index for xylem vulnerability to cavitation by 54% and 32%, respectively. These vessels accumulated in radial groups in a 30° sector immediately adjacent to the wound, raising the vessel grouping index by 28%. The wound-induced anatomical changes in wood structure express the functional need of trees to improve xylem hydraulic safety and mechanical strength at the expense of water transport. Xylem hydraulic efficiency was restored in one year, while xylem mechanical reinforcement and resistance to cavitation and decay lasted over several years.
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