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MINI REVIEW article
Sec. Plant Breeding
A Concise Review on Multi-Omics Data Integration for Terroir Analysis in Vitis vinifera
- 1Environmental Epigenetics and Genetics Group, Plant Research Centre, School of Agriculture, Food and Wine, University of Adelaide, Glen Osmond, SA, Australia
- 2The Waite Research Institute, The School of Agriculture, Food and Wine, The University of Adelaide, Glen Osmond, SA, Australia
Vitis vinifera (grapevine) is one of the most important fruit crops, both for fresh consumption and wine and spirit production. The term terroir is frequently used in viticulture and the wine industry to relate wine sensory attributes to its geographic origin. Although, it can be cultivated in a wide range of environments, differences in growing conditions have a significant impact on fruit traits that ultimately affect wine quality. Understanding how fruit quality and yield are controlled at a molecular level in grapevine in response to environmental cues has been a major driver of research. Advances in the area of genomics, epigenomics, transcriptomics, proteomics and metabolomics, have significantly increased our knowledge on the abiotic regulation of yield and quality in many crop species, including V. vinifera. The integrated analysis of multiple ‘omics’ can give us the opportunity to better understand how plants modulate their response to different environments. However, ‘omics’ technologies provide a large amount of biological data and its interpretation is not always straightforward, especially when different ‘omic’ results are combined. Here we examine the current strategies used to integrate multi-omics, and how these have been used in V. vinifera. In addition, we also discuss the importance of including epigenomics data when integrating omics data as epigenetic mechanisms could play a major role as an intermediary between the environment and the genome.
Grapevine is one of the most economically important fruit crops, and it is largely used for wine production (FAO, 2012). Most the chemical compounds that give its unique characteristics to wine are synthesized during berry development (Conde et al., 2007). However, fruit/wine composition is strongly influenced by the interactions between the plant’s genome and the local growing conditions (including the vine management system), and the oenological practices of each winery (Figure 1), which could explain why it is so difficult to replicate a wine from a region outside that area.
FIGURE 1. A conceptual view of some of the terroir factors that contribute to vine balance, fruit composition and wine quality. ∗Vine genome/s refers to the possibility of the scion and rootstock used in a vineyard could be from different varieties/species (Modified from: Smart et al., 1985 and reproduced with permission from the copyright holder).
Terroir is defined as the interactions between the plants, the environment and human factors (Gladstones, 2011) and it is frequently used to relate wine sensory attributes to its geographic origin (Van Leeuwen and Seguin, 2006). Although the relevance of terroir is still under debate (Anesi et al., 2015), a better understanding of how the environment affects grape berry composition can have a significant effect on viticulture. To achieve such an understanding, it is necessary to identify the elements that drive terroir and analyze the interaction between them and the grapevine.
Terroir has been long studied, through the characterization of the different environmental factors affecting berry composition and wine quality, and climate exerts the strongest effect on berry composition (Robinson et al., 2012). Soil physicochemical properties as well have been identified as an influential factor defining the uniqueness of berry composition by vines grown in a specific climate (Cheng et al., 2014; Zerihun et al., 2015). Grapevine microbiome community may play an important role determining wine quality (Burns et al., 2015; Bokulich et al., 2016). Efforts have been made to study the grapevine microbiome landscape in relation to the vegetative growth cycle of the plant (Pinto et al., 2014), the post-harvest treatment of berries (Salvetti et al., 2016) and provenance (Bokulich et al., 2016) (For a review on microbiome analysis see Ibrahim and Kumar, 2017). Less work has been done to elucidate the molecular mechanisms involved in the plant response to terroir. A strategy to better understand the genome and environment interaction is to use ‘omics’ technologies. Omics refers to high throughput technologies that generate a large amount of data for each sample, allowing a deeper insight of the mechanisms regulating biological systems.
Analysis of Terroir Effect on Grape Composition Using Transcriptomics
Using transcriptomics is possible to study the grape’s complete set of RNA transcripts encoded by the genome using high throughput methods (Hale et al., 2005). Dal Santo et al. (2013) performed gene expression analysis in a single Corvina clone cultivated in 11 different vineyards for three consecutive years. Samples strongly clustered by season, known as a vintage effect, rather than common environmental conditions. However, the genes that showed more variation in expression between years were those involved in secondary metabolism, (mainly the biosynthesis of phenylpropanoids). Only when samples from a single vintage (i.e., 2008) were analyzed, it was observed that 5% of the studied annotated coding genes were differentially regulated under different growing conditions and agronomical practices. Anesi et al. (2015) complemented this study by analyzing the transcriptome and metabolome of the same cultivar. They identified metabolites that could describe a terroir signature for each vineyard. Moreover, it was possible to correlate terroir-sensitive metabolites with changes in the transcript level of genes involved in the biosynthesis of these metabolites. Similar results were obtained by Dal Santo et al. (2016) as they identified a clear correlation between gene expression and accumulation of phenylpropanoids and flavonoids in the variety Garganega grown at four different vineyards. Small RNA profiles have been analyzed to understand the interaction between genotype and environment in the varieties Sangiovese and Cabernet Sauvignon. In silico analysis suggests that microRNAs may be involved in berry development and the accumulation of secondary metabolites (Paim Pinto et al., 2016). Transcriptional analysis of berries from different regions has also shown that transcripts from the abscisic acid (ABA) biosynthesis pathway are among the most terroir sensitive genes (Sun et al., 2015). ABA is a plant hormone that regulates important steps in plant growth and development as well as play a key role in plant biotic and abiotic stress response (Cutler et al., 2010). ABA concentrations affect anthocyanin and flavonol accumulation (Koyama et al., 2010), suggesting a possible mechanism through which the environment affects grape berry composition and wine flavor and aroma.
Analysis of Terroir Effect on Grape Composition Using Metabolomics
Metabolomics is defined as the identification and quantification of metabolites using high-throughput techniques (Cevallos-Cevallos et al., 2009). This technology can screen higher numbers of products than more traditional approaches (Pereira et al., 2006; Atanassov et al., 2009; Hong, 2011), while the use of non-targeted metabolomics approaches allows the identification of un characterized metabolites (Panighel et al., 2015). Terroir can be explored by analyzing berry metabolite composition through different analytical methods (For a review in grape and wine metabolomics see Cozzolino, 2016). Son et al. (2009) identified that differences in berry metabolomes associated to environmental regional differences (radiation and rainfall) could explain the observed differences in wine composition. Similar results were obtained by Tarr et al. (2013) who distinguished the metabolic signatures of different grapevine varieties. Metabolomic analysis has also been performed to identify chemical compounds that can be associated to regional wine quality traits (Gambetta et al., 2016, 2017). Roullier-Gall et al. (2014) assessed the metabolomics profiles of two different terroirs, which were just 2 km apart, over three vintages. Although vintage had the greatest effect in the berry’s metabolite composition, differences in fruit chemical composition associated to nearby terroirs could be detected when vintages were individually analyzed. This suggests that subtle geographical differences have a significant effect on grape/wine composition even when variability within vineyards can be relatively high (Mulas et al., 2011).
The aim of integrating multi-omic data is to reduce the gap between data generation and the ability to analyze and understand the biological mechanisms behind an organism’s response to environmental cues. The objective of multi-omic data integration is to combine different types of data to construct a model that can be used to predict complex traits and phenotypes (Figure 2). This approach also allows the identification of biomarkers and of previously unknown relationships between the datasets (Rajasundaram and Selbig, 2016). Through the integration of environmental information with genomic, epigenomic, transcriptomic, and metabolomic data, we hypothesize that it will be possible to better understand the effect of terroir at a molecular level. The use of a multi-omic approach will also help reduce the incidence of false positives generated from single source data sets (Aho, 2013; Ritchie et al., 2015). However, integration of multi-omics data is not a trivial task, because the diversity of characteristics of the data generated from the different high throughput technologies (machine sensitivity, error rate, data structure) makes its combination challenging.
FIGURE 2. Data integration workflow for four omics technologies in addition to environmental data, and fruit and wine quality data (Modified from Wanichthanarak et al., 2015 and reproduced with permission from the copyright holder).
Approaches to Datasets Integration
Analysis of large data sets from different origins has been done using two main approaches: network models (NMs) and pathway analysis (PA). Both share the basic idea of storing the data in a clear and meaningful way. NMs use concepts from mathematical graph theory, to represent biological components (e.g., genes) as nodes and their interactions (physical, genetic or functional) as their links (For a review on NM applied to plant biology see Fukushima et al., 2014). NMs are classified as homogeneous or heterogeneous depending on the number of different levels of information integrated (Gligorijević and Pržulj, 2015). Homogenous approaches integrate datasets with the same type of nodes and therefore cannot analyze the connectivity between multiple datasets simultaneously. However, complex biological questions such as the molecular regulation of fruit composition in grapevine are increasingly being addressed through the integration of multiple layers of cellular information (Wong and Matus, 2017), including but not restricted to genomics, transcriptomics, proteomics and metabolomics, using heterogeneous methods. Bayesian networks (BNs) and Kernel-based methods (KBMs) are heterogeneous approaches commonly used for data integration (Zhang, 2009; Gligorijević and Pržulj, 2015). BNs are efficient detecting relationships hidden in noisy datasets but they are computationally demanding (Gligorijević and Pržulj, 2015) and are therefore, better suited for the interrogation of small datasets in hypothesis driven questions (Gligorijević and Pržulj, 2015) (i.e., the analysis of terroir on defined pathways such as those leading to the biosynthesis of metabolites related to fruit quality). KBMs are not as computationally demanding and so can integrate large molecular, structural and phenotypic datasets (Mizrachi et al., 2017), making them ideal for data driven terroir exploratory studies, biomarker discovery or for the reclassification of previously identified drivers of quality (Qi et al., 2008).
On the other hand, pathway analysis requires well documented biochemical pathways where omics data is combined to seek overrepresented groups (Wanichthanarak et al., 2015). For example, multiple co-inertia analysis (MCIA) can detect explanatory omic features even when they are not present in all datasets (Meng et al., 2014), which makes it attractive for the integration of terroir data from different studies. Random Forest implemented for pathway analysis (Pang et al., 2006), can be used to predict fruit/wine quality traits associated to terroir integrating multi-omic and phenotypic data as shown recently for potato (Acharjee et al., 2016).
Most of these multi-omics analysis approaches are pipelines that perform task sequences which share statistical methods (Bersanelli et al., 2016). Correlation analyses are the most common approaches performed to find relationships between the omics data. Simple correlation analyses, like Pearson or Spearman correlation, are widely used for multi-omics data integration (Rajasundaram et al., 2014; Rajasundaram and Selbig, 2016). Partial least square/projections to latent structures (PLS) and its extension, orthogonal partial least square (OPLS) (Tobias, 1995) have also been used for data integration from multi-omics results. Even though their predictive power is similar, OPLS results are much easier to interpret and outliers are quickly detected. OPLS can be used as a discriminate analysis (OPLS-DA), to identify differences between the overall data properties while removing systematic variation (Kirwan et al., 2012). However, these methods provided little insight when they are used in complex biological systems (highly multicollinear systems) (Wanichthanarak et al., 2015).
Modifications to these methods have been implemented to facilitate the interpretation of the data, for example, sparse PLS (sPLS) (Chun and Keleş, 2010) can better predict phenotypes through multi-omics data integration than previous methods (Rajasundaram et al., 2014). Orthogonal 2PLS (O2PLS), capable of dealing with unrelated systematic variation between datasets (Bouhaddani et al., 2016), has been successfully used for data integration of transcriptomics and metabolomics results from aspen under different light treatments (Bylesjö et al., 2007). Srivastava et al. (2013) used orthogonal projections to latent structures (OnPLS), an extension of O2PLS, to integrate transcriptomics, proteomics, and metabolomics data to construct a model that could identify biological relevant events in the oxidative stress response in poplar.
Data Integration in V. vinifera
In plant science, most of data integration of omics results comes from model plants; however, there is an increase in publications on multi-omics data integration in V. vinifera. One of the first publications in multi-omics data integration in V. vinifera was the work of Zamboni et al. (2010). Integrating transcriptome, proteome and metabolome data, they identified stage specific biomarkers for berry development. Data integration was performed using two strategies, one hypothesis driven (i.e., a hypothesis was tested) and the other hypothesis free (i.e., discovery driven), in both cases principal component analysis (PCA), O2PLS and O2PLS-DA were used.
Using five different omics technologies and correlation analysis (PCA and Pearson correlation) together with biochemical pathway analysis (KEGG, PlantCyC and VitisCyC), Ghan et al. (2015) could differentiate biochemical characteristics from five different cultivars. Moreover, Anesi et al. (2015) studied the terroir effect in V. vinifera cultivar Corvina in seven different sites over a 3 years period using metabolome and transcriptome data. Using correlation analyses (PCA, PLS-DA and O2PLS-DA) they could identify a terroir signature in the berry metabolome composition for each growing site. Network analyses have been recently adopted to integrate grapevine multi omics results (Wong and Matus, 2017). For example, Palumbo et al. (2014) using network-based methods, identified “fight-club” nodes (genes with negatively correlated profiles) that may be relevant for the control of berry transition between development and ripening.
There are also online resources available that can help analyze omic data from V. vinifera. For example, VitisNet (Grimplet et al., 2009, 2012) offers manually annotated molecular networks (16,000 genes and 247 networks) where omics data can be loaded to visualize changes in the transcriptome, proteome and metabolome for a given experiment. VTCdb (Wong et al., 2013) is a gene co-expression database for V. vinifera that allows exploring transcription regulation. With more than 29,000 genes (95% of the predicted grapevine transcriptome) to query co-expression networks, VTCdb offers the possibility to analyze the transcriptional network of grapevine development, metabolism and stress response. VitisCyc (Naithani et al., 2014) is a grapevine metabolic pathway database that also allows omics data to be uploaded (transcriptome, proteome and metabolome) and to analyze changes in metabolic networks in each experiment. VESPUCCI (Moretto et al., 2016) is a manually annotated gene expression compendium exploratory tool that can be used to investigate grapevine’s gene expression patterns.
Phenotypic Plasticity Through Epigenetic Modifications
Epigenetics is the study of heritable phenotypes that occur through modifications that alter DNA activity without modifying its basic nucleotide structure (Feil and Fraga, 2012). Many epigenetic mechanisms, acting in an interactive and redundant fashion (Grant-Downton and Dickinson, 2005; Berger et al., 2009), have been described to date, with DNA methylation probably being the best-studied of all (Rapp and Wendel, 2005). DNA methylation affects chromatin condensation in a rapid and reversible manner (Grativol et al., 2012). In turn, the regional level of chromatin condensation affects the transcriptional state of nearby genomic features such as genes and transposable elements (Zhang et al., 2006). Global changes in DNA methylation associated to local environments can be analyze using a myriad of methods (Kurdyukov and Bullock, 2016). Bisulfite modification of genomic DNA combined with whole genome sequencing (BS-Seq) is the gold standard for methylation analysis because it can assess an entire methylome with single base resolution (Krueger et al., 2012). However, due to their lower cost, other approaches such as next generation sequencing following the capture of the methylated fraction of the genome or its fragmentation using methylation sensitive restriction enzymes (Bock et al., 2010; Li et al., 2010; Kitimu et al., 2015) are better suited to study large number of samples. Both generate quantitative and qualitative information of the methylation status of a reduced but significant representation of the total genome.
Environmental signals are one of the elements that can have a major effect in modifying the DNA methylation patterns leading to gene expression changes that ultimately affect the plant phenotype (Feil and Fraga, 2012). The idea that the environment could modify the epigenetic status, and these modifications passed to the offspring (Tricker et al., 2013) or maintained as epigenetic memory on long lived organisms (Latzel et al., 2016), has attracted attention from scientists studying mechanisms involved in adaptation to local environments (Consuegra and Rodríguez López, 2016) and how these could be used to enhance crop performance (Rodríguez López and Wilkinson, 2015). There are many reported examples of how the environment affects the epigenome in natural environments and how epigenetic variations in plant populations could help to overcome the lack of genetic diversity (Fonseca Lira-Medeiros et al., 2010; Verhoeven et al., 2010).
One of the most well-known examples in which the environment affects the phenotype through epigenetic modifications is vernalization (Feil and Fraga, 2012). Through this process, plants in temperate regions mitigate the deleterious effects of low winter temperatures on flower and fruit development by breaking dormancy only after having been exposed to a cold period (Kumar et al., 2016). Unusual environmental conditions during dormancy such as high winter temperatures have been shown to exert a negative effect on fruit quality and yield on perennial crops requiring a vernalization period (Sugiura et al., 2012). Recent work in apple shows how methylation and expression levels of key genes involved in flowering and fruit set are modified by the level of chill received during bud dormancy (Kumar et al., 2016), indicating that the environmentally induced changes observed in fruit quality could be regulated by DNA methylation.
Together these studies suggest that the environment can have a long lasting phenotypic effect in plants through epigenetic changes without the need for genetic variation, and that epigenetic mechanisms could be working as intermediaries between environmental variation and the plant genome, and in this way, potentially contributing to plant phenotypic plasticity. Moreover, this mechanism could give plant populations a way of adapting to the local growing conditions (Platt et al., 2015; González et al., 2016). However, to our knowledge, almost all epigenetic studies done in V. vinifera have focused on the identification of commercial clones (Imazio et al., 2002; Schellenbaum et al., 2008; Ocaña et al., 2013) and on the assessment of in vitro culture on the epigenome (Baránek et al., 2015), there is, therefore, a lack of information of how the environment affects a grapevine’s epigenome and to what extent this interaction affects fruit quality. Until now, there are no studies looking at the epigenome to understand the control of gene expression in V. vinifera and how environmental signals can change the regulation of metabolic pathways through epigenetic modifications. In our view, the inclusion of epigenomic data on the analysis of the terroir effect will not only increase the resolution of analysis but will also help us to understand the regulatory mechanisms behind the observed differences.
There is no doubt that the elements affecting grapevine growth and fruit composition are complex and multifarious. While the concept of terroir is widely discussed, the underlying mechanisms remain somewhat enigmatic. However, with the recent parallel development of omics technologies and of statistical approaches for their integration, we are reaching a point where it may be possible to overcome this challenge. The geographic delimitation of a terroir is the first challenge to overcome before its molecular characterization. This delimitation could be achieved 1. Empirically, based in the number of significantly different environmental subregions present in the study or/and 2. based on the traditionally defined wine regions. Moreover, the masking effect that environmental inter-annual variations can have over single year measurements demands the incorporation of data from multiple seasons to be able to determine terroirs with enough confidence. Ideally such seasons should be, from a weather perspective, variable within the range characteristic for the region of study to be able to capture its “normal terroir.”
Understanding how the genome, environment and viticulture practices interact to affect fruit quality will allow us the opportunity to implement agricultural practices aimed to obtain the desired fruit characteristics for every climate/cultivar combination (Jones and Davis, 2000), leading to more efficient use of resources and better management of vineyards. In addition, grape growers can maximize the terroir effect on the grapevine to highlight the uniqueness of their vineyards ultimately increasing their industrial competitiveness. We propose that the integration of multi-omic and environmental datasets will contribute to a better understanding of the drivers of the terroir effect in grapevine. Moreover, multiple dataset data integration will increase our understanding of the molecular mechanisms involved in the regulation of multifactorial genome by environment interactions. Finally, it is increasingly recognized that plants are involved in complex interactions their soil and epiphytic microbiomes, which can affect their phenotype (Mueller and Sachs, 2015). The ‘omics’ era gives us the ability to explore the nature and consequences of biotic/abiotic interactions and so, a future challenge will be to bring the concept of the holobiont (the plant host plus its microbiomes) into the analysis of terroir and its effect on grapevine growth and fruit composition.
PF, TC, CC, and CR designed and wrote the manuscript.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
PF was supported by Australian Grape and Wine Authority (AGWA) and University of Adelaide Graduate Research Scholarships. CR is supported by a University of Adelaide Research Fellowship. This study was funded through a Pilot Program in Genomic applications in Agriculture and Environment Sectors jointly supported by the University of Adelaide and the Australian Genome Research Facility Ltd.
Acharjee, A., Kloosterman, B., Visser, R. G. F., and Maliepaard, C. (2016). Integration of multi-omics data for prediction of phenotypic traits using random forest. BMC Bioinformatics 17(Suppl. 5), 180. doi: 10.1186/s12859-016-1043-4
Anesi, A., Stocchero, M., Dal Santo, S., Commisso, M., Zenoni, S., Ceoldo, S., et al. (2015). Towards a scientific interpretation of the terroir concept: plasticity of the grape berry metabolome. BMC Plant Biol. 15, 191. doi: 10.1186/s12870-015-0584-4
Atanassov, I., Hvarleva, T., Rusanov, K., Tsvetkov, I., and Atanassov, A. (2009). Wine metabolite profiling: possible application in winemaking and grapevine breading in Bulgaria. Biotechnol. Biotechnol. Equip. 23, 1449–1452. doi: 10.2478/V10133-009-0011-9
Baránek, M., Èechová, J., Raddová, J., Holleinová, V., Ondrušíková, E., and Pidra, M. (2015). Dynamics and reversibility of the DNA methylation landscape of grapevine plants (Vitis vinifera) stressed by in vitro cultivation and thermotherapy. PLoS ONE 10:e0126638. doi: 10.1371/journal.pone.0126638
Bersanelli, M., Mosca, E., Remondini, D., Giampieri, E., Sala, C., Castellani, G., et al. (2016). Methods for the integration of multi-omics data: mathematical aspects. BMC Bioinformatics 17(Suppl. 2), S15. doi: 10.1186/s12859-015-0857-9
Bock, C., Tomazou, E. M., Brinkman, A. B., Müller, F., Simmer, F., Gu, H., et al. (2010). Quantitative comparison of genome-wide DNA methylation mapping technologies. Nat. Biotechnol. 28, 1106–1114. doi: 10.1038/nbt.1681
Bokulich, N. A., Collins, T. S., Masarweh, C., Allen, G., Heymann, H., Ebeler, S. E., et al. (2016). Associations among wine grape microbiome, metabolome, and fermentation behavior suggest microbial contribution to regional wine characteristics. MBio 7:e00631-16. doi: 10.1128/mBio.00631-16
Bouhaddani, S. E., Houwing-Duistermaat, J., Salo, P., Perola, M., Jongbloed, G., and Uh, H.-W. (2016). Evaluation of O2PLS in Omics data integration. BMC Bioinformatics 17(Suppl. 2), S11. doi: 10.1186/s12859-015-0854-z
Burns, K. N., Kluepfel, D. A., Strauss, S. L., Bokulich, N. A., Cantu, D., and Steenwerth, K. L. (2015). Vineyard soil bacterial diversity and composition revealed by 16S rRNA genes: differentiation by geographic features. Soil Biol. Biochem. 91, 232–247. doi: 10.1016/j.soilbio.2015.09.002
Bylesjö, M., Eriksson, D., Kusano, M., Moritz, T., and Trygg, J. (2007). Data integration in plant biology: the O2PLS method for combined modeling of transcript and metabolite data. Plant J. 52, 1181–1191. doi: 10.1111/j.1365-313X.2007.03293.x
Cevallos-Cevallos, J. M., Reyes-De-Corcuera, J. I., Etxeberria, E., Danyluk, M. D., and Rodrick, G. E. (2009). Metabolomic analysis in food science: a review. Trends Food Sci. Technol. 20, 557–566. doi: 10.1016/j.tifs.2009.07.002
Cheng, G., He, Y.-N., Yue, T.-X., Wang, J., and Zhang, Z.-W. (2014). Effects of climatic conditions and soil properties on Cabernet Sauvignon berry growth and anthocyanin profiles. Molecules 19, 13683–13703. doi: 10.3390/molecules190913683
Chun, H., and Keleş, S. (2010). Sparse partial least squares regression for simultaneous dimension reduction and variable selection. J. R. Stat. Soc. Ser. B. Stat. Methodol. 72, 3–25. doi: 10.1111/j.1467-9868.2009.00723.x
Conde, C., Silva, P., Fontes, N., Dias, A. C. P., Tavares, R. M., Sousa, M. J., et al. (2007). Biochemical changes throughout grape berry development and fruit and wine quality. Food 1, 1–22. doi: 10.1186/s12864-016-2660-z
Cutler, S. R., Rodriguez, P. L., Finkelstein, R. R., and Abrams, S. R. (2010). Abscisic acid: emergence of a core signaling network. Annu. Rev. Plant Biol. 61, 651–679. doi: 10.1146/annurev-arplant-042809-112122
Dal Santo, S., Fasoli, M., Negri, S., D’Incà, E., Vicenzi, N., Guzzo, F., et al. (2016). Plasticity of the berry ripening program in a white grape variety. Front. Plant Sci. 7:970. doi: 10.3389/fpls.2016.00970
FAO (2012). FAOSTAT. Available at: http://faostat.fao.org/site/339/default.aspx [accessed August 3, 2016].
Fonseca Lira-Medeiros, C., Parisod, C., Fernandes, R. A., Mata, C. S., Cardoso, M. A., and Ferreira, P. C. (2010). Epigenetic variation in mangrove plants occurring in contrasting natural environment. PLoS ONE 5:e10326. doi: 10.1371/journal.pone.0010326
Gambetta, J. M., Cozzolino, D., Bastian, S. E. P., and Jeffery, D. W. (2016). Towards the creation of a wine quality prediction index: correlation of Chardonnay juice and wine compositions from different regions and quality levels. Food Anal. Method 9, 2842–2855. doi: 10.1007/s12161-016-0467-9
Gambetta, J. M., Schmidtke, L. M., Wang, J., Cozzolino, D., Bastian, S. E. P., and Jeffery, D. W. (2017). Relating expert quality ratings of Australian chardonnay wines to volatile composition and production method. Am. J. Enol. Vitic. 68, 39–48. doi: 10.5344/ajev.2016.16058
Ghan, R., Van Sluyter, S. C., Hochberg, U., Degu, A., Hopper, D. W., Tillet, R. L., et al. (2015). Five omic technologies are concordant in differentiating the biochemical characteristics of the berries of five grapevine (Vitis vinifera L.) cultivars. BMC Genomics 16, 946. doi: 10.1186/s12864-015-2115-y
González, A. P., Chrtek, J., Dobrev, P. I., Dumalasová, V., Fehrer, J., Mráz, P., et al. (2016). Stress-induced memory alters growth of clonal offspring of white clover (Trifolium repens). Am. J. Bot. 103, 1567–1574. doi: 10.3732/ajb.1500526
Grativol, C., Hemerly, A. S., and Ferreira, P. C. G. (2012). Genetic and epigenetic regulation of stress responses in natural plant populations. Biochim. Biophys. Acta 1819, 176–185. doi: 10.1016/j.bbagrm.2011.08.010
Grimplet, J., Cramer, G. R., Dickerson, J. A., Mathiason, K., Van Hemert, J., and Fennell, A. Y. (2009). VitisNet: “Omics” integration through grapevine molecular networks. PLoS ONE 4:e8365. doi: 10.1371/journal.pone.0008365
Grimplet, J., Van Hemert, J., Carbonell-Bejerano, P., Díaz-Riquelme, J., Dickerson, J., Fennell, A., et al. (2012). Comparative analysis of grapevine whole-genome gene predictions, functional annotation, categorization and integration of the predicted gene sequences. BMC Res. Notes 5:213. doi: 10.1186/1756-0500-5-213
Ibrahim, K. S., and Kumar, N. S. (2017). “Methods for microbiome analysis,” in Translational Bioinformatics and Its Application, eds D.-Q. Wei, Y. Ma, W. C. S. Cho, Q. Xu, and F. Zhou (Dordrecht: Springer), 269–298. doi: 10.1007/978-94-024-1045-7_12
Imazio, S., Labra, M., Grassi, F., Winfield, M., Bardini, M., and Scienza, A. (2002). Molecular tools for clone identification: the case of the grapevine cultivar ‘Traminer.’ Plant Breed. 121, 531–535. doi: 10.1046/j.1439-0523.2002.00762.x
Kitimu, S. R., Taylor, J., March, T. J., Tairo, F., Wilkinson, M. J., and Rodríguez López, C. M. (2015). Meristem micropropagation of cassava (Manihot esculenta) evokes genome-wide changes in DNA methylation. Front. Plant Sci. 6:590. doi: 10.3389/fpls.2015.00590
Koyama, K., Sadamatsu, K., and Goto-Yamamoto, N. (2010). Abscisic acid stimulated ripening and gene expression in berry skins of the Cabernet Sauvignon grape. Funct. Integr. Genomics 10, 367–381. doi: 10.1007/s10142-009-0145-8
Kumar, G., Rattan, U. K., and Singh, A. K. (2016). Chilling-Mediated DNA methylation changes during dormancy and its release reveal the importance of epigenetic regulation during winter dormancy in apple (Malus × domestica Borkh.). PLoS ONE 11:e0149934. doi: 10.1371/journal.pone.0149934
Li, N., Ye, M., Li, Y., Yan, Z., Butcher, L. M., Sun, J., et al. (2010). Whole genome DNA methylation analysis based on high throughput sequencing technology. Methods 52, 203–212. doi: 10.1016/j.ymeth.2010.04.009
Mizrachi, E., Verbeke, L., Christie, N., Fierro, A. C., Mansfield, S. D., Davis, M. F., et al. (2017). Network-based integration of systems genetics data reveals pathways associated with lignocellulosic biomass accumulation and processing. Proc. Natl. Acad. Sci. U.S.A. 114, 1195–1200. doi: 10.1073/pnas.1620119114
Moretto, M., Sonego, P., Pilati, S., Malacarne, G., Costantini, L., Grzeskowiak, L., et al. (2016). VESPUCCI: exploring patterns of gene expression in grapevine. Front. Plant Sci. 7:633. doi: 10.3389/fpls.2016.00633
Mulas, G., Galaffu, M. G., Pretti, L., Nieddu, G., Mercenaro, L., Tonelli, R., et al. (2011). NMR analysis of seven selections of Vermentino grape berry: metabolites composition and development. J. Agric. Food Chem. 59, 793–802. doi: 10.1021/jf103285f
Naithani, S., Raja, R., Waddell, E. N., Elser, J., Gouthu, S., Deluc, L. G., et al. (2014). VitisCyc: a metabolic pathway knowledgebase for grapevine (Vitis vinifera). Front. Plant Sci. 5:644. doi: 10.3389/fpls.2014.00644
Paim Pinto, D. L., Brancadoro, L., Dal Santo, S., De Lorenzis, G., Pezzotti, M., Meyers, B. C., et al. (2016). The influence of genotype and environment on small RNA profiles in grapevine berry. Front. Plant Sci. 7:1459. doi: 10.3389/fpls.2016.01459
Palumbo, M. C., Zenoni, S., Fasoli, M., Massonnet, M., Farina, L., Castiglione, F., et al. (2014). Integrated network analysis identifies fight-club nodes as a class of hubs encompassing key putative switch genes that induce major transcriptome reprogramming during grapevine development. Plant Cell 26, 4617–4635. doi: 10.1105/tpc.114.133710
Pang, H., Lin, A., Holford, M., Enerson, B. E., Lu, B., Lawton, M. P., et al. (2006). Pathway analysis using random forests classification and regression. Bioinformatics 22, 2028–2036. doi: 10.1093/bioinformatics/btl344
Panighel, A., De Rosso, M., Dalla Vedova, A., and Flamini, R. (2015). Putative identification of new p-coumaroyl glycoside flavonoids in grape by ultra-high performance liquid chromatography/high-resolution mass spectrometry. Rapid Commun. Mass Spectrom. 29, 357–366. doi: 10.1002/rcm.7115
Pereira, G. E., Gaudillere, J. P., van Leeuwen, C., Hilbert, G., Maucourt, M., Deborde, C., et al. (2006). 1H NMR metabolite fingerprints of grape berry: comparison of vintage and soil effects in Bordeaux grapevine growing areas. Anal. Chim. Acta 563, 346–352. doi: 10.1016/j.aca.2005.11.007
Platt, A., Gugger, P. F., Pellegrini, M., and Sork, V. L. (2015). Genome-wide signature of local adaptation linked to variable CpG methylation in oak populations. Mol. Ecol. 24, 3823–3830. doi: 10.1111/mec.13230
Qi, Y., Suhail, Y., Lin, Y.-Y., Boeke, J. D., and Bader, J. S. (2008). Finding friends and enemies in an enemies-only network: a graph diffusion kernel for predicting novel genetic interactions and co-complex membership from yeast genetic interactions. Genome Res. 18, 1991–2004. doi: 10.1101/gr.077693.108
Rajasundaram, D., Runavot, J.-L., Guo, X., Willats, W. G. T., Meulewaeter, F., and Selbig, J. (2014). Understanding the relationship between cotton fiber properties and non-cellulosic cell wall polysaccharides. PLoS ONE 9:e112168. doi: 10.1371/journal.pone.0112168
Ritchie, M. D., Holzinger, E. R., Li, R., Pendergrass, S. A., and Kim, D. (2015). Methods of integrating data to uncover genotype-phenotype interactions. Nat. Rev. Genet. 16, 85–97. doi: 10.1038/nrg3868
Robinson, A. L., Adams, D. O., Boss, P. K., Heymann, H., Solomon, P. S., and Trengove, R. D. (2012). Influence of geographic origin on the sensory characteristics and wine composition of Vitis vinifera cv. Cabernet Sauvignon wines from Australia. Am. J. Enol. Vitic. 63, 467–476. doi: 10.5344/ajev.2012.12023
Roullier-Gall, C., Lucio, M., Noret, L., Schmitt-Kopplin, P., and Gougeon, R. D. (2014). How subtle is the “terroir” effect? Chemistry-related signatures of two “climats de Bourgogne”. PLoS ONE 9:e97615. doi: 10.1371/journal.pone.0097615
Salvetti, E., Campanaro, S., Campedelli, I., Fracchetti, F., Gobbi, A., Tornielli, G. B., et al. (2016). Whole-Metagenome-Sequencing-Based community profiles of Vitis vinifera L. cv. Corvina berries withered in two post-harvest conditions. Front. Microbiol. 7:937. doi: 10.3389/fmicb.2016.00937
Son, H.-S., Hwang, G.-S., Kim, K. M., Ahn, H.-J., Park, W.-M., Van Den Berg, F., et al. (2009). Metabolomic studies on geographical grapes and their wines using 1H NMR analysis coupled with multivariate statistics. J. Agric. Food Chem. 57, 1481–1490. doi: 10.1021/jf803388w
Srivastava, V., Obudulu, O., Bygdell, J., Löfstedt, T., Rydén, P., Nilsson, R., et al. (2013). OnPLS integration of transcriptomic, proteomic and metabolomic data shows multi-level oxidative stress responses in the cambium of transgenic hipI- superoxide dismutase Populus plants. BMC Genomics 14:893. doi: 10.1186/1471-2164-14-893
Sun, R., He, F., Lan, Y., Xing, R., Liu, R., Pan, Q., et al. (2015). Transcriptome comparison of Cabernet Sauvignon grape berries from two regions with distinct climate. J. Plant Physiol. 178, 43–54. doi: 10.1016/j.jplph.2015.01.012
Tarr, P. T., Dreyer, M. L., Athanas, M., Shahgholi, M., Saarloos, K., and Second, T. P. (2013). A metabolomics based approach for understanding the influence of terroir in Vitis Vinifera L. Metabolomics 9, 170–177. doi: 10.1007/s11306-013-0497-x
Tobias, R. D. (1995). “An introduction to partial least squares regression,” in Proceedings of the Twentieth Annual SAS Users Group International Conference, (Cary, NC: SAS Institute Inc.), 1250–1257.
Tricker, P. J., López, C. M. R., Gibbings, G., Hadley, P., and Wilkinson, M. J. (2013). Transgenerational, dynamic methylation of stomata genes in response to low relative humidity. Int. J. Mol. Sci. 14, 6674–6689. doi: 10.3390/ijms14046674
Verhoeven, K. J. F., Jansen, J. J., van Dijk, P. J., and Biere, A. (2010). Stress-induced DNA methylation changes and their heritability in asexual dandelions. New Phytol. 185, 1108–1118. doi: 10.1111/j.1469-8137.2009.03121.x
Wong, D. C. J., and Matus, J. T. (2017). Constructing integrated networks for identifying new secondary metabolic pathway regulators in grapevine: recent applications and future opportunities. Front. Plant Sci. 8:505. doi: 10.3389/fpls.2017.00505
Wong, D. C. J., Sweetman, C., Drew, D. P., and Ford, C. M. (2013). VTCdb: a gene co-expression database for the crop species Vitis vinifera (grapevine). BMC Genomics 14:882. doi: 10.1186/1471-2164-14-882
Zamboni, A., Di Carli, M., Guzzo, F., Stocchero, M., Zenoni, S., Ferrarini, A., et al. (2010). Identification of putative stage-specific grapevine berry biomarkers and omics data integration into networks. Plant Physiol. 154, 1439–1459. doi: 10.1104/pp.110.160275
Zerihun, A., McClymont, L., Lanyon, D., Goodwin, I., and Gibberd, M. (2015). Deconvoluting effects of vine and soil properties on grape berry composition. J. Sci. Food Agric. 95, 193–203. doi: 10.1002/jsfa.6705
Zhang, A. (2009). “Data fusion in the analysis of protein interaction networks,” in Protein Interaction Networks: Computational Analysis (New York, NY: Cambridge University Press), 243–257. Available at: http://assets.cambridge.org/97805218/88950/frontmatter/9780521888950_frontmatter.pdf
Keywords: multi-omics, environment, Vitis vinifera, data integration, epigenetics, transcriptomics, metabolomics
Citation: Fabres PJ, Collins C, Cavagnaro TR and Rodríguez López CM (2017) A Concise Review on Multi-Omics Data Integration for Terroir Analysis in Vitis vinifera. Front. Plant Sci. 8:1065. doi: 10.3389/fpls.2017.01065
Received: 08 February 2017; Accepted: 02 June 2017;
Published: 20 June 2017.
Edited by:Giovanni Battista Tornielli, University of Verona, Italy
Reviewed by:Pablo Carbonell-Bejerano, Instituto de las Ciencias de la Vid y del Vino (ICVV), CSIC, Spain
Stefania Pilati, Fondazione Edmund Mach, Italy
Copyright © 2017 Fabres, Collins, Cavagnaro and Rodríguez López. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Carlos M. Rodríguez López, firstname.lastname@example.org

domain: agronomy


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Browsing History by Title
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(Wheaton College (Norton, Mass.), 2014)"This is a study of women who attended Wheaton College during the 1940s through the early 1960s. It tries to understand how real college women interacted with political and cultural influences that existed in America during ...

domain: library


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St. Luke s Hospital lost $7.6 million from the business of caring for patients last year as fewer money-making surgical procedures were done and other critical conditions hampered the Maumee hospital s bottom line.
To stanch the bleeding, St. Luke s is recruiting doctors to replace those who have retired or left to help increase business. Only the most essential capital purchases are being made. And while no one has been laid off, employees are taking time off without pay as needed.
Still, losses are expected to continue into next year, with St. Luke s returning to profitability in 2010, said David Oppenlander, the hospital s vice president and treasurer.
There s no panic whatsoever from the senior executive level, said Mr. Oppenlander, adding that hourly employees are more skittish because of scheduling changes.
We re not skimping on patient care or quality. That s our focus, he said.
An annual look at hospital financial performance found St. Luke s in the most troubled position last year, although St. Charles Mercy Hospital in Oregon also was in the red. St. Charles $1.8 million operating loss, however, was folded into parent Mercy Health Partners, which had an overall operating profit of $17.8 million locally.
To determine how area hospitals fared financially last year, The Blade obtained the most recent Internal Revenue Service forms filed by three nonprofit entities, St. Luke s, Mercy, and ProMedica Health System, which generally take nearly a year to become public.
The Blade also gathered other statistics from those three health-care providers as well as from the University of Toledo Medical Center. As a public institution, the former Medical College of Ohio doesn t file the same IRS annual forms, called 990s and filed by all nonprofit organizations, but keeps public financial records.
All Toledo-area hospital operators say this year and next will be tough because of the troubled economy, but they and other hospitals nationwide are adjusting their businesses as a result. All but St. Luke s, for example, have laid off employees this year.
Last year, ProMedica fared the best financially, with an operating profit of $50.2 million, giving it a 3.39 percent operating margin. (Operating profit is the amount of money hospitals make from treating people; operating margin is a profitability measure computed from revenues as well as costs related to patient care.)
Nonprofit hospitals nationally have hit operating margins of 3 percent to 5 percent for awhile, but this year s results likely will be a percentage point below what hospitals estimated, said Nick Hilger, senior vice president in the health systems group for Ingenix, an information and consulting firm.
Hospital managers need to be on top of managing cash flow, reimbursement contracts, and other business aspects, Mr. Hilger said.
Not everything in the hospital is about nurses and about needles and syringes, he said.
ProMedica s chief executive, Alan Brass, last year continued to have the highest total compensation among local heads of hospitals and their systems. He received nearly $1.9 million in salary and other perks last year, and as the economy worsened, Mr. Brass recently agreed to stay on through 2010 instead of retiring next June.
Leaping to the No. 2 position last year among top executives, meanwhile, was Frank Bartell III, St. Luke s now retired president and chief executive.
While Mr. Bartell s 2007 salary was about $365,000, his total compensation last year was nearly $1.9 million. Mr. Bartell received a $1.5 million lump-sum payment from a supplemental retirement plan that had accumulated money over several years.
St. Luke s last year spent money on doctor recruitment and invested in a multiyear electronics medical records program that includes helping physician offices acquire software, said Mr. Oppenlander, the vice president and treasurer.
It also started treating more patients without insurance as people lost health-care coverage. St. Luke s is providing charity care or giving discounts to those with enough income to afford partial payments, he said.
We understand there s a lot of need in the community, and we re trying to be there as best we can, Mr. Oppenlander said.
Moody s Investors Service recently downgraded St. Luke s bond rating for $11.2 million of outstanding revenue bonds issued by Maumee, although it remains investment grade. Last month, Moody s revised its outlook for the entire non-for-profit health-care sector to negative because of damage from the credit crisis and bad economy.
Nationwide, weakened financial markets both have hurt hospital investments and the ability to issue bonds, and patient volumes have flattened or dropped as people put elective surgeries and diagnostic tests on hold, said Moody s analyst Lisa Martin, a senior vice president.
St. Luke s has been affected by those nationwide economic conditions as well as local competitive pressures, and losses are magnified because the Maumee hospital is small and not affiliated with other providers, she said.
The larger the hospital, the easier it is to absorb, Ms. Martin said.
St. Luke s is committed to staying independent, and it has a strong balance sheet and long-term plan, Mr. Oppenlander said. It has renegotiated a reimbursement contract with Anthem Blue Cross and Blue Shield that will start in July so customers with that insurance can seek covered care there, which along with other measures will help the hospital return to profitability, he said.
Looking at local hospitals on an individual basis, ProMedica s Flower Hospital in Sylvania again had the highest operating margin last year at nearly 10 percent.
Among local Mercy hospitals, all of which are part of Catholic Healthcare Partners in Cincinnati, St. Anne Mercy Hospital in Toledo had the highest operating margin at nearly 5 percent. Mercy s overall operating margin last year was 2.07 percent, which is within the hospital operator s annual goal.
St. Charles, which had an operating loss last year, lost a reimbursement contract with Anthem in 2006. That contract has been renegotiated, and St. Charles operations have improved this year, said Samantha Platzke, Mercy s chief financial officer.
For the fiscal year ending June 30, UTMC had an operating margin of 3 percent after operating profit came in at $7.6 million, more than $2.5 million higher than budgeted.
Contact Julie M. McKinnon at:firstname.lastname@example.org 419-724-6087.
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domain: news


Letter: Moynihan — initiative and follow-through
Kevin Moynihan will be a very effective first selectmen for New Canaan. His experience on the Town Council has shown that he takes the initiative and follows through on projects. New Canaan needs to move forward on several projects that Kevin is very interested in, including commuter parking and cell phone coverage.
I have known Kevin and his family since they moved to New Canaan over 30 years ago. I've worked with Kevin on some projects and can attest that he has always been diligent and conscientious in all of his community service work.
I heartily endorse Kevin Moynihan for first selectman.


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david painter realtor
Although mortgage rates are rising, demand for a limited supply of homes appears strong, according to the SC Realtors Association.
Some buying, the trade group said in its August report, might be triggered by speculation that rates will climb further.
“The dream of homeownership is very much intact, but buyers should be prepared with competitive offers, since every measure of market health is pointing upwards,” the report by the Columbia-based association said.
Some keys to what will happen in the housing market include:
- Whether the Federal Reserve will alter its policy toward stimulus tapering.
- Tepid, but positive labor market growth.
- Consumer sentiment swayed by gas prices, stock market shifts and global economics.
Overall, new listings in the state increased 5.9% to 8,982 units compared with August 2012 levels, while pending sales rose 6.7% to 5,404. Inventory levels shrank 6.6% to 46,827 units.
The overall median sales price rose 3.3% to $155,000 compared with August 2012 numbers, while the number of days on the market dropped 13.6% to 111 days.At the end of the month, the state housing market had about a nine-month supply of available units. The strongest demand for housing was in single-family homes, where inventory shrank 5.4%.
Among the state’s major metro markets, Charleston Trident recorded a 22.4% increase in sales for August to 1,256 units. Meanwhile, the median price in Charleston climbed 9.6% to $219,000 in August compared with $199,865 for the same month in 2012, and the average number of days on the market dropped 18.7% to 72. Greater Columbia reported a year-over-year increase of 19.4% in August sales to 947 homes, while the median price slipped 1% to $143,000. The average number of days on market for Columbia dropped 19.9% to 96.
In Greater Greenville SC real estate, sales rose 18.1% to 921 homes compared with 780 for August 2012. Meanwhile, the median price rose 5.4% to $165,000 compared with $156,500 for August 2012. The average number of days on market dropped 12.9% to 84.
David Painter – Realtor Keller Williams Greenville SC
Article: Staff Report scbiznews.com
According to KW research, the top three updates sellers make in preparation for listing their house are fresh paint, new flooring, and updated light fixtures. Surprisingly, sellers spend more than two times as much money updating their house while they live in it than they do getting it ready for sale. To find out which updates matter most, contact David Painter, Keller Williams Greenville SC Realtor (864) 616-1608.

domain: finance


This year, CPAC came at a critical time for our nation.
Joined by conservatives from across the globe, CPAC was a major success that capitalized on a unique moment in history where it seems that socialism has surrounded us on all sides. In just a few short years under the Biden administration, free speech has been branded as violence, and liberty has been reduced to an enemy of public health. As America sat idly by, our education system, institutions, and agencies fell into entitled, self-loathing socialists.
Ultimately, every right, freedom, and ounce of Americanism has been rendered unimportant in the face of an insatiable socialist movement being further propelled by the Biden presidency. This is why at CPAC this year, we were focused on Protecting America Now.
While in the past, conservatives might have been content keeping to themselves, being motivated by a dead notion of “live and let live,” this is no longer a winning strategy. So, from day one, CPAC this year had to be different.
Instead of just the typical self-righteous anger directed at the Left, Americans on the CPAC stage were focused on providing solutions, instilling knowledge, and providing unique insights to advance the tactical response to Protecting America and her founding principles.
Rep. Jim Jordan, for instance, kicked off CPAC with a provocative conversation on the weaponization of the federal government and his plan of attack to hold them accountable. Chaya Raichik, the creator of ‘Libs of Tik Tok,’ also took center stage to expose the mental corrosion of people victimized by the Left’s woke ideology and explained that with an open media, the Left practically destroys itself.
Conservatives on the CPAC stage veered away from the status quo talking points on grievances and moved to provide courage and unique perspectives. This fresh and action-oriented perspective was achieved by including new voices, entrepreneurs and community rabble-rousers.
The most enjoyable part of CPAC was the reliably leftist media complaining about the lack of establishment Republicans on the stage. Establishment Republicans are good at one thing, following, and they will mostly follow when we show strength and lead. What the media failed to show the public is that CPAC this year had an unmatched lineup of high-caliber individuals, including nine members of the senate, chairmen of four of the most powerful House committees; Jim Jordan, Mark Green, James Comer, and Jason Smith, two former world leaders; Jair Bolsonaro, the former president of Brazil, and President Donald J. Trump and an impressive list of future leaders.
With no lack of talent on the CPAC stage, attendees got more out of this year’s CPAC than ever before.
But the media’s attacks on CPAC targeted not only the attendance (CPAC stopped selling tickets midway through the conference because of capacity concerns) of the event itself but also the speakers onstage. After Michael Knowles of the Daily Wire gave a compelling speech to protect our children and the preservation of truth, the media was quick to twist his words and claimed that Mr. Knowles was calling for the genocide of trans people. In reality. They are always desperate to discredit anyone who questions the ‘freedom’ that comes with prescribing to our children the same hormone blockers used to chemically castrate sex offenders.
It seems that the fight to protect women and children is now a responsibility uniquely held by the Right. Proudly accepting this obligation, CPAC hosted twelve-time All-American Swimmer, Riley Gains, who was simply there, to tell the truth: Men should not compete in women’s sports. And just as, if not more notable, de-trans activist Chloe Cole joined us to share her story and why children must be protected from the harms of the Left’s radical gender ideology.
When it came to the many issues surrounding culture, children, and ideology on the CPAC stage, the prevailing line of thought was that if the Left does not want a rational conversation, then we move to legislation at any level, anywhere.
CPAC this year was also packed with a strong international presence, including delegations from Japan, Hungary, Italy, Israel, and Australia on top of regular attendees. Notably, many Brazilians came out in droves Saturday to see Jair Bolsonaro give a captivating speech on the proliferation of socialism and corruption in his home country of Brazil. Speaking Portuguese, Jair’s best statements were immediately met with almost unruly cheers from the Brazilian crowd, and then once more when his electric statements were translated.
With our Constitution under attack, basic rules of society in question, and our international position in contention, CPAC this year provided the platform for the best leaders among us to begin making the changes conservatives need. At CPAC, we have set a better conservative agenda for the future and have set the stage for Protecting America Now. While the media and the Left will remain committed to bashing the conference and attempting to destroy this effective organization, conservatives can find respite in the fact that if we are making them upset, we definitely did something right.
• Matt Schlapp is CPAC chairman and former White House political director under President George W. Bush.


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A DECADE after Pasminco’s lead and zinc smelter closed at Boolaroo and as the land is being prepared for redevelopment, the Newcastle Herald and Macquarie University collected more than 130 soil and dust samples from homes and public spaces in surrounding suburbs. The analysis revealed alarming levels of contamination remained, despite a government-approved Lead Abatement Scheme.
FEARS about the health impacts of black slag were raised two years before Lake Macquarie City Council stopped using it in roads and parks.
A report from the Newcastle Herald archives reveals council was concerned about slag posing “possible health risks” as far back as May, 1992.
It did not stop using it until August, 1994, citing concerns that heavy metals could leach into surrounding soil.
Despite assurances at the time from Pasminco management that the slag was not dangerous, council engaged private consultants to test samples in 1992.
Lake Macquarie State MP Greg Piper, speaking as a councillor at the time, expressed concern about possible high-lead levels in slag.
“I don’t want to panic people but I think if they have slag in their backyard I think that it is best for them to be aware that there might be a problem with it,” Mr Piper said in 1992.
“And certainly not to have their children playing with it, and perhaps consider having it top-dressed, especially if we find out there are contaminated levels associated with it.”
At the time the council said it was working with the Environment Protection Authority and Department of Health investigating the matter.
The Cockle Creek smelter was producing about 80,000 tonnes of slag each year, with the majority of it mixed with dried sewage sludge and coke fines to produce an artificial soil.
It was then used in landscaping mounds around the factory.
Reacting to council’s 1994 decision to stop using the black slag in its projects, Pasminco’s then environmental services manager Sharon Howes said the council had used the slag for more than 20 years as fill at Speers Point Park and in other land reclamation projects.
Ms Howes described the move as an "overreaction".
Boolaroo Action Group spokesman and retired environmental-health council officer Jim Sullivan said council knew it was a problem for years before it stopped delivering it to homes.
‘‘It was horrendous really, what was happening with the slag,’’ Mr Sullivan said.
‘‘People were filling up box trailers for their backyards.’’
The council has previously insisted it was ‘‘unaware of any research or data on the human or environmental health impacts of Pasminco slag’’ until 2003.
The EPA issued a remediation order for the Pasminco site in 2003, saying it was ‘‘contaminated in particular with lead, cadmium and zinc in such a way as to present a significant risk of harm’’.
Lake Macquarie mayor Jodie Harrison said council funded research into the health impacts of black slag in 2001. It found lead in the slag could affect human health if it was ingested or dust inhaled.
"In most cases, slag deposits in Lake Macquarie City present a low health risk if the slag is covered," she said. "Black slag is generally managed on-site, either by reburying the material or adding topsoil to a minimum depth of 200mm and establishing grass cover."
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text:
The new 3,500m2 two-storey purpose-built student association building provides the student body with facilities including a Performance Theatre, Function Centre, Social Spaces, Club Spaces, Bars, UCSA office and external entertainment areas including an amphitheatre and outdoor kitchen.
- VAE were awarded the project on a construct only basis, however due to inherent problems with the design that was issued for construction the functionality of the building would have been compromised (including architectural & acoustic) this led to VAE taking on a vast amount of design work in order to ensure a fully functioning building solution.
- The team managed to produce the above positive result whilst having a significant lack of full design control which meant many challenges including time delays with approvals etc.
- Due to documents not being coordinated between structure, architecture and services prior to project commencement VAE took on full design level co- ordination. VAE have received praise from the client, stating that the project would not have been successful without VAE’s input.

domain: design


The happiest place is on Earth is also one of the best places to ring in the New Year. The 2020 festivities at the Walt Disney World Resort and Epcot Center include huge dance parties and fireworks displays that run on both December 30 and December 31.
At the Magic Kingdom Park in Orlando, Florida, there are dance parties happening in Frontierland and Tomorrowland. Then there are the Fantasy in the Sky Fireworks at 6 p.m. and midnight. Meanwhile, in Epcot Center, there is a countdown in each country’s respective pavilion as it hits midnight in different timezones around the world. There is also live music at the America Gardens Theatre and a midnight countdown to 2020 and New Year’s Eve fireworks display at the World Showcase Lagoon.
You can catch all the action in the live streams below. The Monday, December 30 stream starts at 6 p.m. ET/3 p.m. PT, with a second 11:40 p.m. ET/8:40 p.m. PT show later in the night. The Tuesday, December 31 stream also begins at 6 p.m. ET/3 p.m. PT, with a second show ringing in the new year just before midnight.
There are all kinds of New Year’s packages and parties at the Walt Disney World resorts, including dancing on the boardwalk and a fine dining experience at Narcoossee’s, the restaurant at Disney’s Grand Floridian Resort & Spa. All the details can be found at the Disney World website.
New Year’s Eve marks the end of a calendar year on the Gregorian calendar, the calendar first introduced in 1582 by Pope Gregory XIII, after whom it is named. Most countries celebrate the final day the year with parties, social gatherings, festivals, and/or fireworks. It goes by many names the world over, including Hogmanay in Scotland, Calennig in Wales, Baharu in Indonesia and Malaysia, Silvester in many European countries, Reveillon in France, Portugal, and Brazil, Kanun Novodgo Goda in Russia, and Omisoka in Japan.
Kiritimati, Tonga, and New Zealand are some of the first places to celebrate New Year’s Eve because they are located just west of the International Date Line, while the U.S.’ Baker Island is one of the last places to celebrate because it is just east of the International Date Line. Interestingly, because of the way the International Date Line jogs around a bit, Kiritimati is actually east of Baker Island.
In the United States, New Year’s is traditionally celebrated with parties and “drops,” the most famous of which is the ball drop held in New York City’s Times Square. But there are dozens of other “drops” held across the country, including a conch drop in Key West, Florida; a peach drop in Atlanta; an Indy car drop in Indianapolis; an acorn drop in Raleigh, North Carolina; a moon pie drop in Mobile, Alabama; a fleur-de-lis drop in New Orleans, a “Glowtato” drop in Boise, Idaho; and a tortilla chip drop in Tempe, Arizona, which is tied in to the Tostitos Fiesta Bowl game of college football.
New York also rings in the new year with a “Midnight Run” around Central Park that includes a fireworks show. Other fireworks displays around the country include shows at the Disney theme parks, the Las Vegas strip, and the Chicago “Chi-Town Rising” event.


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4x4 Sprinters are very capable and versatile vehicles, however many Sprinters have stiff factory ride over small bumps and extreme swaying on uneven ruts. Van Compass has developed a custom tuned FOX shock kit to improve the Sprinter’s handling both on and off the trail. Our custom tuned Fox shocks smooth out the harshness experienced on washboard roads and sharp, abrupt impacts. The bounce experienced over large bumps is reduced and the side to side sway over uneven surfaces is minimized. Each set of Fox shocks are specifically tuned per application. This kit is recommended for light, high roof crew and cargo vans.
Fits for Sprinter vans with and without body lift !
This kit includes:
Simple installation with basic hand tools.

domain: hobby


text:
Police release images of suspected sorority burglar
SEATTLE -- The Seattle Police Department has released three surveillance-camera images of the man suspected of breaking into a University of Washington sorority house last week.
According to the police report for the incident, one of the Sigma Kappa sorority house's approximately 96 residents was working on her computer in her room around 3:20 a.m. March 7 when she left to use the restroom. When she got back, the suspect was reportedly standing in her bedroom with her blanket over his head.
Unsure if the suspect was a friend of someone who lived in the house, the woman asked for her blanket back. According to the report, the man threw her blanket down and ran away.
Police originally stated no one was inside the sorority in the 4500 block of 22nd Avenue Northeast when the suspect entered. That is not the case.
It appears the suspect entered the house through a second-floor window, using a ladder left behind by a repair company. A door was also found propped open by a sponge, indicating the suspect may have made multiple trips in and out of the house.
According to the report, the suspect rode away on a bicycle and was unable to be found by an extensive search.


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Last week, I finished Kathy Van Riper’s autobiography called, A Race Worth Running. A friend had given me the book as a birthday present last year and I put off reading it for obviously a good year. I am not sure if that was God timing, but I think I got more out of it now–being a year later–as I have been through a bit more experiences in my physical body, such as the blood clot. I was so emotional reading the book that after I finished I was talking with my mom and just started crying (a variety of things), but I pointed to the book on the couch and said, “I don’t know why this book has made me so emotional.” Mom looked at me and said, “Probably because you can relate to what she went through.”
I thought about this. It is probably true. If you compared my life to Kathy’s life, there are some extreme differences: she was married, had two kids, battled an extreme case of breast cancer for ten years, and lived in CA her whole life–and yet we also had similarities: We both loved running (I can’t physically anymore, but love walking!); we both had treatments on the same chemo called Avastin (although she went through ten other different chemo treatments; I have only been through two.) We both had a blood clot with all the blood thinner shots and pills that follow; we both lost our hair; we both had radiation; we both were (and I still am) supported by an amazing circumference of family and friends who love endlessly and share support during the good and bad times; we both had to switch hospitals and doctors after years of care offered by another one; and we both have faith. Kathy’s faith is now complete. I am still on the journey.
Kathy’s “life verse” was Hebrews 12:1-2,
Therefore, since we are surrounded by so great a cloud of witnesses, let us also lay aside every weight, and sin which clings so closely, and let us run with endurance the race that is set before us, 2 looking to Jesus, the founder and perfecter of our faith, who for the joy that was set before him endured the cross, despising the shame, and is seated at the right hand of the throne of God.
I put in bold the phrase that she quoted most often in her journal entries that were at the end of the book. Kathy was a runner. Unlike myself who ran cross-country in high school but hated the races, Kathy thrived in races–anything from 5k, 10k to marathons. Whether you run or not, running as if in a race is almost the strength and perseverance mind-set for life: “[Forgetting] what lies behind and straining forward to what lies ahead, I press on toward the goal for the prize of the upward call of God in Christ Jesus.” Philippians 3:13b-14.
When I ran cross-country in high school, training was what got me through the races. For training, we would run on a variety of things during the week: grass, sidewalks through town, and other times, run up the mountain. I remember this one place…not quite sure where exactly in the mountains…but all of a sudden after you “warmed up” running the road, it just turned into a straight shot up the mountain. Rocks everywhere, so steep. It was literally a “breath-taking” view! 🙂 Coach always encouraged us to never stop. Never. So even though I was bringing up the rear end of the team while running slower than most people’s walking speed, I never stopped. After you got over that huge steep hill, things leveled out. Sure, it was still a mountain, but it was more like the hiking paths and less gruesome. Even after all this training, sometimes during the races I would want to give up. But then I would remember and say to myself, “Pull yourself together. You trained harder than this.” And I would not stop. Mental determination. The finish line was my goal. And that was the best part, because as you near the end you start to hear the cheers from everyone at the finish line. You pick up the pace, knowing that the finish line was just ahead. You forgot about the rest of the race…how much pain or how many people passed you. You strived to the end with all strength left in you. The verse that was so special to Kathy fit her life–her personality, her faith, and her race against cancer. The common phrase for people who have died from cancer is, “They lost the fight.” Indeed, our lives are worth fighting for but after reading Kathy’s book…I would not say that she lost the fight. Rather, she finished her race.
“Pressing on towards the goal”–I have shared Kathy’s favorite verse. I would like to share with you now my favorite verse. It is from II Corinthians 12:9-10,
But he said to me, “My grace is sufficient for you, for my power is made perfect in weakness.” Therefore I will boast all the more gladly of my weaknesses, so that the power of Christ may rest upon me. For the sake of Christ, then, I am content with weaknesses, insults, hardships, persecutions, and calamities. For when I am weak, then I am strong.
I had started running in the 7th grade…slacked off here and there, and in college, I started going to the gym every night. Not much kept me from the gym each night in college. I think it was my own way of “pressing on” during those times. I loved it. Something happened after I graduated, I stopped exercising every night and lost a little determination for a bit. I lived close to some different parks in my neighborhood and started walking there. I loved seeing the season changes and talked to God while I walked a few miles. My old job was also connected to a mall which had an indoor mile lap and I would often walk during my lunch breaks. Please understand me when I say this…as this pertains to my life and mental determination: When I either give up exercising, or can’t exercise from health reasons, I see a change. Like my attitude changes about how I see things in life…big struggles seem even harder, and even little things seem so hard. I told my mom about Kathy’s life during our conversation that night I finished the book–how she would go on mile runs a few days after chemo treatments. I said, “I don’t think I could do that,” as if it made me feel guilty for not getting out for walks lately or getting a few other things done that were stacking up in my room (which it sort of did). Mom said I didn’t have to. We were all made different.
Then I thought back to times in Denver. I worked Thursday-Saturday eight hour shifts. The weeks of chemo I worked Thursday, chemo Friday and then back to work Saturday-Sunday. Regardless of how much I tried to talk to work about changing this schedule, it never did. For months this happened. I think that is where my favorite verses really set in…”When I am weak, then I am strong,” because it was almost my reminder of my semester in college when I was on chemo pills. I don’t know how I got out of bed then, or on the weekends of work after chemo. Definitely not on my own…but on God’s strength.
The thing is, most people look at me and say the exact thing of what I said about Kathy’s life, “I don’t think I could do that.” And I would tell you, “You don’t have to. We are all made different.” God gives me strength to endure, just like he gave his strength to Kathy. He gives it according to our need. I think that is what makes life so special. We all have our own race–our own life worth running…enduring through pain, encouraging others and in return receiving it, rejoicing with those who have crossed the finish line, and striving for what lies ahead.

domain: literature


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As a political prisoner, Tajuddin Ahmad wrote from prison many letters to his wife Zohra and to his eldest daughter Reepi, who had just begun to read and write. Sadly, all of these letters were either lost or destroyed during the Bangladesh liberation war. Among the letters, which he wrote to his relatives and acquaintances, only a few have survived.
Tajuddin Ahmad’s letter to his relative Shafiuddin Khan from Maymansingh prison.1966.
Tajuddin Ahmad’s letter to Shafiuddin Khan, after the liberation of Bangladesh. 1972.
Tajuddin Ahmad’s letter to Hazrat Mia from Maymansingh prison.1966.

domain: history


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Monsieur Ibrahim is a story about a young Jewish boy in Paris who meets an old Muslim Turkish grocery store owner. The film touches the themes of friendship and love as the old man is a father figure to the boy as he teaches him of the Koran. The film is based on a book of the popular book by the French author Eric-Emmanuel Schmitt.
Flowers in colour.
A tomato is planted, harvested and sold at a supermarket, but it rots and ends up in the trash. Ends? No. Isle of Flowers follows it up until its real end, among animals, trash, women and children. And then the difference between tomatoes, pigs and human beings becomes clear.
A little flower film by Gaston Velle.
Up until now, marriage has been smooth sailing for Jennifer Jones. But after suffering a miscarriage, she begins to question her happiness and whether the spark in her marriage has dimmed. She shrugs it off as just being the 7-year itch and gives her husband, Freddy, who is content in their relationship, the benefit of the doubt. Things take an unexpected turn when Jennifer catches Freddy in a few unassuming lies. And when a sudden accident throws Freddy into a coma, Jennifer is left to pick up the pieces and reconcile everything for herself. With Freddy in hospital, a larger story begins to unfold - something that might just be Jennifer's worst nightmare - prompting her to question if she ever really knew the man she was married to.
After a quarrel Pedro walks out of the cafe leaving his fiancée (Stacia Napierkowska) alone. Heartbroken, she visits the flower garden and there, flowers metamorphose into women and begin to dance.
Later in his career, Ozu started becoming increasingly sympathetic with the younger generation, a shift that was cemented in Equinox Flower, his gorgeously detailed first color film, about an old-fashioned father and his newfangled daughter.
Flower shop owner Poppy enters the annual holiday floral show for a chance to win the grand prize but standing in her way of victory is Garrett, the underappreciated protégé of florist extraordinaire, Blair. While Poppy and Garrett’s rivalry heats up on the flower show floor, an undeniable romance begins amidst a whimsical holiday backdrop.
A couple of dudes pack a van full of beer and go on a road trip in search of sexual adventures. They get that and more.
Envoi de Fleurs is based on incidents in the life of French composer Paul Delmet. Played by popular French singing star Tino Rossi, Delmet is depicted as a man all too willing to give up personal happiness in favor of blind ambition. After carrying on a romance by correspondence with beautiful young Suzanne (Micheline Francey), Delmet is on the verge of marrying the girl. Instead, he allows himself to be talked out of leaving France to further his own career, with disastrous results for all concerned.
A man with a low IQ takes part in a scientific experiment with a mouse.
It's All Saints' Day. Bérénice criss-crosses the city. Flowers must be found for the grave. Her son Sacha has other projects. Too bad. Flowers must be found for the grave.
A flow of funny scenes inspired by paintings of René Magritte.
When Henry's wife dies, he deals with his grief by handing out flowers and wisdom to strangers he meets in the park.
A coming-of-age story set during China's Cultural Revolution. 11 year old Wang Han finds himself entangled with a fugative and struggles to understand the adult world.
Paris-Tehran. A rootless love story between Gecko, young and free, and Anahita, an Iranian woman in exile - tangled up in History and steeped in Internet, unique and spontaneous
Dr. Evan Cooper is the ideal match on paper and according to her grandmother, Rose Durham and friends, including an equally pushy wedding planner, for florist Laurel Haverford, who fears to be the last of her generation to get married. Yet she keeps dragging her heels after meeting cocky Stephen Banks, who is all wrong on paper but makes her heart leap by pre chemistry.
Three generations of a family struggle to be open with each other during a week of summer vacation at their country cottage.
Fleurs d'amour, fleurs d'amitié was a Canadian variety television series which aired on Radio-Canada in 1968.
The outward perfection of a family-run flower business hides a dark side rife with dysfunctional secrets in this darkly humorous comedy series.
A variety show with a twist: a celebrity guest get to attend their own memorial service – an entertaining send-off filled with eulogies and some gentle ribbing.
Pastel Yumi, the Magic Idol is a magical girl anime series by Studio Pierrot. It was simultaneously released as a manga by Kiyoko Arai. The fourth magical girl series created by Studio Pierrot, Pastel Yumi also appears in a feature-length OVA as well as the Majokko Club Yoningumi A-kūkan kara no Alien X OVA. It is currently streaming on Anime Sols for a potential crowd-funded DVD.
The corrupt pure-love story revolves around Kasuga Takao, a bookish boy who loves the poems of Charles Baudelaire (the original author of the poetry collection "Les Fleurs du mal" or "Flowers of Evil"). One day after school, he discovers and steals the gym clothes of Saeki Nanako, the girl he has a crush on. However, he learns that Nakamura Sawa, a girl he loathes, happens to catch him in the act. Nakamura blackmails Kasuga into a contract, or else she will reveal his secret.
Fumi Manjoume enters Kamakura's accelerated high school - Matsuoka All-Girls High School. While waiting at the Kamakura station on the day of her entrance ceremony, she runs into an old childhood friend whom she had not seen in 10 years: Akira Okudaira. As their friendship is rekindled and they start falling back into the rhythm of friends again, it starts a delicate love story...

domain: artistic


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KANSAS CITY METRO — The ice is falling, and with it comes already bad road conditions. A Winter Storm Warning is now in effect until 8pm tonight for most of the Kansas City metro.
FOX4 is working for you to show you what it’s like on the road. See what’s going on in the live blog below.
You can also check road conditions in Missouri on the MODOT Traveler Information Map.
You can check road conditions in Kansas on KanDrive.

domain: geography


2SAXY is a unique collaboration between two internationally acclaimed saxophonists, Grace Kelly and Leo P, who blend their remarkable skills and distinct styles to bring an unparalleled level of “saxiness” to the music scene. Known for their individual accomplishments, including sold-out international tours, lauded albums, and television appearances, they join forces to create a one-of-a-kind musical experience. Grace, with her impressive discography and numerous accolades, brings her inventive electro jazz-pop stylings, while Leo P, recognized for his virtuosic baritone sax playing and agile dance moves, adds his signature dynamic energy.
2SAXY’s fusion of musical talent, colorful hairstyles, and irresistible stage presence results in a captivating performance style that is as visually stimulating as it is audibly. Their videos, featuring hit parody pieces such as “Battle of the Saxes” and “Sax Wars,”, improvised musical walks, as well as original music like the fan-favorite “Fish & Chips,” have collectively attracted tens of millions of views worldwide. Together, Grace Kelly and Leo P’s unique synergy as 2SAXY continues to push the boundaries of jazz, funk, and performance art, delighting audiences with their innovative approach to the saxophone.
Please register an account to submit events and news. On Stage will review your information and let you know the next steps.


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Monaco energy supplier SMEG has reported a nearly 10% drop in energy usage across all sectors, with driving forces ranging from government awareness campaigns to consumer concern surrounding the rise in energy costs.
The rhetoric surrounding the energy crisis has spooked many people. Visions of families or the elderly huddling around a small heater in several layers of clothes, being forced to choose between eating and heating, has given society a chance to readjust the way it traditionally uses energy, with the results being somewhat surprising.
The Monegasque Electricity and Gas Company (SMEG), the energy supplier to Monaco for 133 years, has for years been collecting data on consumer usage, but until recently, the algorithms painted a picture of energy usage focused on overall consumption alone. Improvements to the system have allowed SMEG to break it down further into three sub-categories, namely customers with high, medium and low usage.
Across all three groupings, there was an almost 10% drop in energy use from October through to December 2022, compared with the statistics from the same period in 2021. The biggest consumers, such as industrial and large surface businesses, reduced their use by 9.9%; small and medium-sized companies by 8.7%; and the lowest consumers, which includes households, by 8.5%.
“It is significant,” says Sylvain Didierjean, the director of Electricity and Gas Activities at SMEG. “We clearly see that the average consumption over this period is still below the historical average.”
The drops are being accounted for by several factors: an awareness campaign that explained that the Principality could face intermittent power cuts if energy consumption remained high; the current cost of energy; and telecommuting, a change brought on by the Covid epidemic, but which has meant that companies aren’t spending as much on lighting and heating as before.
Though the signs are encouraging, and hopes are high that this shift in behaviour will be long-lasting, there are worries that when summer comes, people will forget. Monaco’s energy usage skyrockets in the summer as people turn on energy-sucking air conditioners as a way to beat the heat during the long hot days.
“We must continue to make efforts, each at their own level, because we remain in a situation of global energy shortage,” continues Didierjean. “There is also the planetary situation, which reminds us that if we limit our consumption, we preserve resources and the planet at the same time.”
But despite the positive developments, Didierjean has been keen to remind people that winter tensions still continue for the time being.
“Due to maintenance operations and power plants still under construction in Europe, we do not yet have all the means of production to ensure that the coming winters go smoothly, and therefore we will still experience tensions during the next two winters,” says the director.
Photo source: Pasquale Iovino for Unsplash


When your Infiniti FX50 was born, it was destined for a person with discerning taste who wants unique styling in their automobile. It's not a regular occurrence that you realize that you have to buy a replacement Sway Bar Link to fix your vehicle. The vehicle in your driveway symbolizes the best in class compared to all the other cars, trucks, and SUVs on the road today. Infiniti car, truck, or SUV drivers have grown accustomed to a certain level of a reputation for safety when driving their FX50 around town.
Chassis flex is the enemy of a firm and stable suspension. To ensure your suspension prevents excessive roll during cornering, Infiniti FX50 sway bar links are attached to the stabilizer bar and the control arms. Without stabilizer links or a sway bar, the chassis would lose traction and understeer into each turn. This was first tested on race cars at the beginning of the 20th century, and now sway bar links have made their way to almost every production vehicle manufactured today; including your Infiniti FX50.
Don't let a Sway Bar Link repair spell the end of your car, truck, or SUV, and don't risk lowering its resale value by installing anything but the highest quality replacement parts. At Car Parts Discount, we have live customer service members on the phone with enough expertise to help you buy the appropriate Infiniti FX50 part for your needs. Bring your FX50 back to life with the best replacement parts from the experts at Car Parts Discount. Need 2013, 2012, 2011, 2010, 2009 Infiniti FX50 Sway Bar Link parts? We've got them right here.


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DETROIT – There are ways to enjoy food over the holidays without derailing your diet.
- Holiday-proof your plan -- Stick to your usual meal time, and don’t skip meals before the big feast. Also, offer to bring a healthy dish.
- Outsmart the buffet -- Use small plates. Load up your plate with vegetables first so you aren’t tempted to choose other less healthy foods.
- Fit in the favorites -- Enjoy the foods you only get during the holidays. When eating those foods, choose them over foods you can have all year.
- Keep moving -- Don’t ditch your fitness routine over the holidays.
- Get sleep -- Getting sleep will give your blood sugar and metabolism time to reset. Plus, you’ll be less likely to overeat just because you are tired.

domain: food


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domain: entertainment


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How to play the lottery: online or in a lottery shop?
In the world of gambling, everyone has their own little fetish ritual that they can't live without. Between those who like to play online and those who prefer physical outlets, it is important to find your place and feel comfortable there, to increase your chances of winning! So, do you think that the old way of doing things can still hold its own against the many advantages offered by the new technology? Let's see what we can say about this.
Going to a lottery shop
The only advantage you can get from the classic lotto is the human side. Indeed, by going to a sales outlet to buy your ticket, you are more in contact with people. The relational aspect is therefore fully taken care of. Plus, if you win, you get to keep your winning ticket as a souvenir of that memorable day.
On the other hand, this solution also has many disadvantages, including the loss of time by queuing in front of the counters for hours on end, not to mention the obligation to travel and the inconveniences linked to the incompatibility of the opening hours with your schedule. In addition, you will have to check the latest lottery results by visiting the lottery shop in due course and this may be a bit inconvenient!
By buying your lottery ticket online
To answer the question: "why is it worth playing the lotto online?" read the passage below.
What you will probably appreciate most about the free-play game are the possibilities and options for playing, in addition to the many chances it offers you to increase your winnings. You can, for example, play the 5/90 NAP 2 lottery online and try to win the jackpot.
In addition to the fact that an electronic ticket cannot be lost, this approach allows you to know the results within one minute of the draw and gives you that feeling of security that you will get absolutely all the results without exception.
We have often heard of sad experiences of people who have won the lottery but have not been able to collect their money because they have lost their ticket or missed the draw for one reason or another! One thing is for sure, though, and that is that this is not likely to happen to you if you play the lottery online.
Moreover, and this is what is likely to convince you the most, online gaming frees you from all the constraints of lottery shops since you can play whenever and wherever you want, in the comfort of your living room, in your office, or on the beach. The key is to have a good internet connection and to know the best sites that offer online lotto.
To maximize your chances of winning, it is imperative to know how to play the Mega Jackpot.
Which one to choose?
It seems clear from our comparison that online gaming is far superior to the traditional method. Leaving aside the sentimental and social aspects, buying a lottery ticket on the internet seems to be the best solution.
For the reasons mentioned above, online lotto offers a completely new opportunity for the customer to play freely and double his chances of winning!
We also recommend to read our article about the 5 types of people who buy lottery tickets.Back to all news →


The Fifth Circuit Court of Appeals cited an expert report and statewide survey conducted by Matt Barreto and Gabriel Sanchez in finding that SB 14, the Texas voter ID law violates section 2 of the Voting Rights Act. In October 2014 the district court initially ruled against the Texas voter ID law in which Barreto and Sanchez offered an expert report and provided testimony at trial. The Latino Decisions researchers implemented a statewide survey of 2,300 eligible voters in Texas to ascertain whether or not Whites, Blacks and Latinos in Texas possessed a valid photo ID.
The survey was presented at trial by the plaintiffs alongside a database match which both concluded that Latinos and Blacks were statistically less likely than Whites to have a valid photo ID. According to expert testimony at the 2014 district court trial, Barreto and Sanchez found that 4.7% of Whites lacked an ID, 8.4% of Blacks, and 11.4% of Latinos lacked a valid photo ID.
In a comprehensive review of the lower court decision the Federal Appeals Court found that the lower court was indeed correct in concluding that the Texas voter ID law had a discriminatory impact and therefore violated the VRA. The three-judge panel issued a conclusive 49-page ruling:
“These statistical analyses of the No-Match List were corroborated by a survey of over 2,300 eligible Texas voters, which concluded that Blacks were 1.78 times more likely than Whites, and Latinos 2.42 times more likely, to lack SB 14 ID… The district court thus credited the testimony and analyses of Plaintiffs’ three experts, each of which found that SB 14 disparately impacts African-American and Hispanic registered voters in Texas….The district court did not err in concluding that SB 14 disproportionately impacts Hispanic and African-American voters…We AFFIRM the district court’s finding that SB 14 violates Section 2 of the Voting Rights Act through its discriminatory effects”
In addition to the overall finding that Blacks and Latinos faced an discriminatory impact, the Barreto and Sanchez report contained an extensive analysis of the socioeconomic and class effects involved in possession of photo ID. According to data in the Barreto-Sanchez survey Blacks and Latinos are also negatively impacted by their socioeconomic status relative to Whites. Data from the U.S. Census confirmed that Black and Latino eligible voters are more likely to be poor, and that low-income residents are ten times less likely to possess a valid photo ID. The Fifth Circuit Court of Appeals also referenced findings from the Barreto-Sanchez expert report that socioeconomic status was an important contributing factor to the discriminatory effect of the Texas voter ID law.


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As in the previous 9 years will once again be hosting Pine River Winter Pistol Leagues. Anyone interested in participating is asked to sign up prior to firing for record. League shooting will be held each Wednesday evening from 6:30 – 9:00 or later if necessary.
The League will be known as the Pine River Winter League (PRWL).
Sign up will be on November 27th, December 4th 2019 and December 11th, 2019. League fees are payable prior to shooting for record and no later than December 11th, 2019.
You do not need to be a member of the Pine River Sports Association. You may shoot either rimfire, centerfire or both. There will be eight (8) shoots and the best five (5) will be used for record and awards. To be eligible for awards a shooter must have fired at least 5 shoots for record.