Molecular markers associated with relevant agronomic traits could significantly reduce the time and cost involved in developing new sugarcane varieties. Previous sugarcane genome-wide association analyses (GWAS) have found few molecular markers associated with relevant traits at plant-cane stage. The aim of this study was to establish an appropriate GWAS to find molecular markers associated with yield related traits consistent across harvesting seasons in a breeding population. Sugarcane clones were genotyped with DArT (Diversity Array Technology) and TRAP (Target Region Amplified Polymorphism) markers, and evaluated for cane yield (CY) and sugar content (SC) at two locations during three successive crop cycles. GWAS mapping was applied within a novel mixed-model framework accounting for population structure with Principal Component Analysis scores as random component.
A total of 43 markers significantly associated with CY in plant-cane, 42 in first ratoon, and 41 in second ratoon were detected. Out of these markers, 20 were associated with CY in 2 years. Additionally, 38 significant associations for SC were detected in plant-cane, 34 in first ratoon, and 47 in second ratoon. For SC, one marker-trait association was found significant for the 3 years of the study, while twelve markers presented association for 2 years. In the multi-QTL model several markers with large allelic substitution effect were found. Sequences of four DArT markers showed high similitude and e-value with coding sequences of Sorghum bicolor, confirming the high gene microlinearity between sorghum and sugarcane.
In contrast with other sugarcane GWAS studies reported earlier, the novel methodology to analyze multi-QTLs through successive crop cycles used in the present study allowed us to find several markers associated with relevant traits. Combining existing phenotypic trial data and genotypic DArT and TRAP marker characterizations within a GWAS approach including population structure as random covariates may prove to be highly successful. Moreover, sequences of DArT marker associated with the traits of interest were aligned in chromosomal regions where sorghum QTLs has previously been reported. This approach could be a valuable tool to assist the improvement of sugarcane and better supply sugarcane demand that has been projected for the upcoming decades.
Sugarcane, the highest tonnage crop among cultivated plants, plays a substantial role in the global economy. Nowadays, this crop has gained great importance not only for its traditional use as food (80 % of world's sugar is produced from sugarcane) but also for ethanol and biomass production. The production of alternative energy sources as well as the establishment of the biorefinery concept has also increased sugarcane world demand rapidly . In order to supply this continuous increasing requirement, the development of new varieties with high biomass and sugar yield is essential.
The modern sugarcane cultivars are interspecific hybrids derived essentially from early crosses between Saccharum officinarum (2n = 80, x = 10), a species with high sugar content stalks, and Saccharum spontaneum (2n = 40–128, x = 8), a wild and vigorous species resistant to several sugarcane diseases. The initial interspecific hybrids were repeatedly backcrossed to S. officinarum clones or to other hybrids in order to recover high sugar content, a process known as "nobilization". These modern cultivars are highly polyploid and often aneuploid, with chromosome numbers ranging from 100 to 130 . Due to this genetic complexity, the application of both conventional and molecular breeding is a challenge in sugarcane.
Most of sugarcane production regions have their own breeding programs to develop and improve local varieties adapted to their specific environments and agricultural practices. Developing a new sugarcane variety takes on average 12 years . Molecular markers associated with relevant agronomic traits could significantly reduce the time and cost involved in developing new varieties because they could aid in selecting the best parents as well as accelerating the rate of genetic gain in the breeding program. In that sense, association mapping has become widely used to identify molecular markers associated with relevant traits in several crops [4–9]. This method is based on the linkage disequilibrium (LD) between molecular markers and quantitative trait loci (QTL) . The resolution and applicability of association mapping depends on the extent of LD within the population under consideration. The breeding history of sugarcane, consisting of a strong foundation bottleneck followed by a small number of cycles of intercrossing and vegetative propagation, suggest that LD should be extensive, thus a high density of markers may not be needed to detect marker–trait associations . In 1999 , and more recently in 2008 , the persistence of high LD in modern sugarcane cultivars was confirmed.
The forces generating and/or conserving LD are those that produce allele frequency changes, i.e. population stratification, genetic relatedness, selection, mutation, genetic drift and linkage . With the exception of linkage, all the genetic forces may cause false positive correlation between markers and traits in population-based association mapping approaches. The effects of a structured population in association mapping studies have been well documented and identified as one of the main causes of spurious associations [14–16]. For that reason and considering the often complex relationships among genotypes in breeding populations, it is extremely important to control for population structure in order to effectively decrease type I error rates (i.e. false positives) . For this purpose, a range of statistical methodologies have been developed that include some sort of population or relatedness control using mixed models [16–19].
In addition to controlling for population structure, the availability of both accurate phenotypic data and molecular markers distributed across the genome are critical requirements for the success of association mapping. One of the advantages of this mapping method for plants compared to classical QTL analysis based on balanced mapping populations is that association mapping allows the use of historical phenotypic data sets collected by the breeding programs . Typically, this data come from multiple trials across different environments and years, therefore, statistical analysis such as mixed models are necessary to obtain phenotypic values that best represent the performance of each genotype. Malosetti et al.  extended the standard phenotypic analysis of multiple trials by mixed models to arrive at models suitable for association mapping by introducing marker genotype information as random covariates to model the correlation between genotypes.
The recently developed technology of DArT in sugarcane  makes it possible to have genome-wide scans of this genetically complex crop, capturing genomic profiles with many thousands of polymorphic markers of several kinds (INDELs, SNPs, methylation changes) . Another molecular marker system recently developed that could also be convenient to detect markers associated with desirable traits is Target Region Amplification Polymorphism (TRAP). These dominant markers enable the identification of polymorphisms in coding regions involved in specific pathways as sucrose metabolism or drought tolerance among others [21, 22].
Information of the marker sequences for DArT is available and could be anchored to the sugarcane genome if sequenced. Several efforts are still ongoing in order to sequence the sugarcane genome which has a high genetic complexity due to its ploidy level. However, considering that i) sugarcane monoploid genome estimated on 930 Mb is similar to the sorghum genome (2n = 2x = 10) estimated on 730 Mb ; ii) sugarcane and sorghum both belong to the Poaceae family and the same sub-tribu Saccharinae, and iii) their high degree of colinearity [24, 25]; the available sequence of sorghum genome becomes an important tool for the analysis of regions of interest in sugarcane.
The goal of this research was to establish an appropriate genome-wide association analysis (GWAS) tool in a sugarcane breeding population, and to find molecular markers associated with high yield of both biomass and sugar stable through successive crop cycles. Therefore, a GWAS mapping within a mixed-model framework following Malosetti et al.  was used. Spurious associations were minimized while the power to detect true associations was maximized by considering the possible population structure. A Principal Component Analysis (PCA) from a genotype data set was performed  and values obtained from the significant axes for each genotype were used as covariates in the model. In contrast with others sugarcane GWAS studies reported earlier involving yield related traits [27, 28] where analyzes were conducted at plant-cane stage, the novel methodology to analyze multi-QTLs through successive crop cycles used in the present study allowed us to find several markers associated with relevant traits. Results highlighted that this approach could be a valuable tool to assist the improvement of sugarcane and better supply the sugar and biomass demand that has been projected for the upcoming decades.
where σ G 2 is the genetic variance, σ ε 2 the residual variance and r the number of replicates .
DNA was extracted from frozen leaf tissue following the Diversity Arrays Technology (DArT) Pty Ltd (Yarralumla, Australia) protocol . The quality and quantity of DNA were verified on a 0.8 % agarose gel. All clones were genotyped using DArT  and TRAP markers [21, 22]. DArT genotyping of the population mapping was carried out by DArT Pty Ltd with the Sugarcane High Density 1.0 array. This service involves two methods of complexity reduction (both based on PstI-based methyl filtration) against the array containing 7680 probes. TRAP genotyping was carried out according to  with minor modifications. All PCR reactions were carried out in our lab and performed in a Bio-Rad My clycler Termalcycler (Hercules, CA, USA) in 5 μl reaction containing 50 ng DNA sample, 10X reaction buffer (Fermentas, Spain, EU), 2.5 mM MgCl2 (Fermentas), 0.088 mM of each dATP, dTTP and dGTP, 0.072 mM of dCTP, 0.16 μM of each primer (Table 2), and 0.5 U of Taq DNA polymerase (Fermentas). Different concentrations of Cy5.5-dCTP (GE Healthcare, Buckinghamshire, UK) were included in the reaction depending on the primer combination (Table 2). Amplifications were performed by initially denaturing the template DNA at 94 °C for 2 min, followed by five cycles at 94 °C for 45 s, 35 °C for 45 s, and 72 °C for 1 min, 35 cycles at 94 °C for 45 s, 50 °C for 45 s, and 72 °C for1 min, and a final extension step at 72 °C for 7 min. Loading dye was added and 0.3 μl PCR products were separated on a 25 cm polyacrylamide gel (Amersham Biosciences) (0.25 mm thick) in a LI-COR 4300 DNA Analyzer (LICOR Biosciences, Lincoln, NE, USA) according to manufacturer's instructions. Images were captured with slow scan laser at 700 nm and analyzed with the SAGATM software (LICOR Biosciences). The product sizes were determined by comparison with molecular weight marker LI-COR IRDye 50–700 bp Size Standard (LICOR Biosciences). TRAP markers, classified as 1 (presence) or 0 (absence), and the binary data from DArT were used for association analysis. All markers with a minor allele frequency (MAF) lower than 0.1 were excluded from the GWAS analysis.
All polymorphic DArT and TRAP markers scored on the 88 sugarcane accessions were used to estimate genetic relationship among clones. Genetic dissimilarities between all pairwise combinations of clones were calculated using the Dice index . Then, a Neighbor Joining tree was built from the matrix of pairwise dissimilarities using the Darwin software V.5.0.158 .
In order to detect and correct for population structure, a PCA was carried out using a subset of 107 DArT markers. All the available markers were not included in this analysis mainly because using the same markers to estimate population structure and then including them in the model to test for an association could create a dependency among terms in the model absorbing some of the QTL effects . The markers used for PCA were sampled according to their position on different Linkage Groups of the Homology Groups of a sugarcane map recently published .
where Y is the phenotypic means vector (i.e. BLUEs from field analysis), X is the incidence matrix of molecular markers, β is the vector of parameters related to the simple regression of the markers on the phenotypes, Q are the eigenvectors of the significant axes of the PCA matrix, υ is a vector of predicted values of population structure, and e is the vector of random errors. The PCA scores were used in the model as random components following  and . Modeling population structure as random effects not only does the relatedness matrix capture population structure, but also encodes a wider range of structures, including cryptic relatedness and family structure [36, 47, 48]. The significant PC axes included in the model were determined with the Tracy-Widom statistic . The analyses were performed using R-code developed by the author's with modifications from the emma  and GAPIT  packages and recently published  using the R software 3.0.0. The code will be uploaded to the R-Cran repository as mmQTL package . Briefly, a two-step approach was followed to arrive to a multi-QTL model. First, a marker-by-marker scan of the genome was conducted to identify significant marker-trait associations with a false-discovery rate (FDR) (α = 0.05) to control for multiple testing. Since a large number of significant marker-trait associations were found, and to report the more relevant QTL, a second pruning of markers with a more stringent FDR P-value (0.01) was conducted. Second, all significant markers were fitted in a single final multi-QTL model adding markers at a time in a stepwise-forward selection manner to control for residual QTL and to identify QTL following [52–54]. The Wald statistic with a liberal P-value < 0.01 following [19, 36] was used for this model.
QQ-plots assuming a uniform distribution of P-values under the null-hypothesis of no-QTL (i.e., Schwederand Spjøtvoll plots; ) were used to evaluate the models. Briefly, the observed P-values values are plotted against the expected theoretical values (i.e. cumulative density function) for a uniform distribution. This is standard methodology to evaluate the models ability to control for spurious association [17, 36, 56]. These analyses were also performed in R statistical software.
Sequences from sugarcane DArT markers significantly associated with CY or SC at least in 2 years of study and DArT markers significantly associated with a trait in the multi-QTL model that resulted in highest Allelic Substitution Effect (ASE) were used to determine their similarity and position on the sorghum genome. This was conducted by using BLASTN 2.2.22  on non-redundant databases of sorghum sequences with different algorithms. First, "Megablast" was employed to identify query sequences. In the cases where no significant similarity was found, a second algorithm "Discontiguous megablast" was chosen since it uses an initial seed that ignores some bases and is intended for cross-species comparisons. Finally, when no significant similarity was found using the second algorithm, BLAST was performed using "blastN".
The 88 sugarcane clones used in this study were phenotyped by SCBP-EEAOC for CY and SC during 2009, 2010 and 2011 and genetically characterized by DArT and TRAP markers. The BLUE values obtained with the adjusted model, described above, were 48 to 85 t ha-1 for CY and 9.2 to 10.9 % for SC (Table 3 and Additional file 2). The genetic correlations observed between years for CY were 0.60 for 2009 and 2010, 0.78 for 2010 and 2011, and 0.50 between 2009 and 2011. Meanwhile, genetic correlations observed between years for SC were 0.40 for 2009 and 2010, 0.72 for 2010 and 2011, and 0.46 between 2009 and 2011. There were low correlations between CY and SC across years (-0.06, -0.24 and -0.14 for 2009, 2010 and 2011, respectively), being only significant (P-value <0.05) correlation among CY 2010 and SC 2010 (Additional file 3). Results of broad-sense heritability for both trait and location are presented in Table 4. CY was under strong genetic control, since estimates of broad-sense heritability were high, ranging from 0.51 to 0.84. Estimates of H2 for SC were also high (from 0.55 to 0.80), with the only exception for SC 2010 with a moderate value of H2 of 0.30. This high estimates of heritability indicated that the field trials produced good-quality data for the association study.
Out of the 7680 probes evaluated in the DArT array, 1642 markers were informative (i.e. polymorphic, with a MAF higher than 0.10). Out of the 177 TRAP markers evaluated, only 103 markers were included in the GWAS and 74 were excluded because the MAF was lower than 0.1. Among the 1642 informative DArT markers, 258 were mapped on the recently published sugarcane genetic map .
Diversity analysis using all the informative TRAP and DArT markers revealed no particular structure in the mapping population (Fig. 1 and Additional file 4; http://dx.doi.org/10.5061/dryad.mv88m). The most closely related clones (parent–descendant or full-sib) were grouped in the same area of the neighbor-joining tree. However, they do not form outstanding branches. Surprisingly, there were two exceptions where full-sib clones were located in different branches, i.e. TUC 02-38 and TUC 02-37 whose genealogical records indicate that they are descendant from the same parents; and TUC 03-32 that would be full-sib with TUC 03-31, TUC 03-33, TUC 03-37 and TUC 04-4, and grouped separately from the rest. At the most distant branch, located at the lower right portion of the tree, grouped LCP 85-384 and most of the clones derived from this variety. At the lower center position of the tree, clones derived from HOCP 85-845 were grouped. Then, at the lower left portion of the tree, TUCCP 77-42 and clones derived from this variety were located. On the other hand, the first three axes of the PCA using 107 DArT markers distributed across the sugarcane genome were significant following the Tracy-Widom statistic. The PCA scores for each genotype at each axes were included as random covariates in the GWAS model to model the variance-covariance matrix among genotypes. The first two axes explained 7.47 and 4.99 % of the total variation, respectively (Fig. 2). The first axis could be associated to filial relations; where two groups seems associated to LCP 85-384 offspring (right side of the PC1 axis) and non-LCP 85-384 offspring (left side of the PC1 axis). At PC2 level, TUCCP 77-42 variety was distant from the rest of the genotypes. Results showed at Fig. 2 are congruent with those previously mentioned in Fig. 1, since clone descendant from LCP 85-384 were detached from the rest of genotypes.
GWAS analysis was conducted by using 1638 discrete markers (1535 DArT and 103 TRAP). QQ-plots of P-values showed that population structure was properly accounted for by using a stratified selection of markers to correct for population structure as random effect (Additional file 5). In the present study, 43, 42 and 41 markers significantly associated (FDR α = 0.01) with CY in 2009 (cane plant), 2010 (first ratoon) and 2011 (second ratoon), respectively, were found. In addition, 38, 34 and 47 significant marker-trait associations for SC were detected, in 2009 (cane plant), 2010 (first ratoon) and 2011 (second ratoon), respectively (Additional file 6). Certain stability across crop-cycles was observed since twenty markers were found to be associated with CY in 2 years of study, being the coincidence between 2010 and 2011 (first and second ratoon) more frequent. For SC, one marker-trait association was found significant for the 3 years of study, while twelve markers presented association for 2 years. These association were also more frequent when 2010 and 2011 years were involved (Table 5). Mostly markers associated with one trait were not associated with the other; however, four markers were associated with both traits (M54 for CY-2010, CY-2011 and SC-2011; M58 for CY-2010, CY-2011 and SC-2011; M173 for CY-2010, SC-2010 and SC-2011; and, M188 for CY-2010, SC-2010 and SC-2011).
A multi-QTL model by year was constructed with markers significantly associated with each trait. Considering the 3 years, 23 markers were significant in the multi-QTL for CY while 21 remained significant in the multi-QTL for SC (Table 6). For CY, markers M100, M120, M140, M200 and M202 had allelic substitution effect (ASE) larger than 8.33 t ha-1. For SC, M28, M51 and M171 had ASE larger than 0.70 %. Marker M64 was detected in more than 1 year in the multi-QTL model (SC 2010 and 2011). The effect of this marker was the same in the 2 years of association and 57 % of the genotypes analyzed had the favorable allele for this marker.
The 27 available sequences of DArT markers significantly associated with a trait in at least 2 years of study were blasted to the sorghum genome sequence database (Table 5). When the sequences of sugarcane DArT markers were analyzed, three of them were found to present the same nucleotide sequence. This was useful as internal control because genotypes presented the same configuration (absence or presence) for markers with the same sequence. Most of alignments involved sequences of hypothetical proteins of sorghum showing a high identity and low e-values. Noticeably M120 showed a high identity (94 %) with a sorghum sequence located on chromosome 6 with an e-value of 0, indicating that there is no probability of alignments with scores equivalent to or better in a database search by chance.
Similarly, sequences of DArT markers significantly associated with a trait in the multi-QTL model that resulted in highest ASE value (M100, M120 and M140 for CY; M28, M51, M64 and M171 for SC) were blasted to the sorghum genome sequence database. Results are shown in Table 7, markers M120 and M64 were not included since they were already shown in Table 5. Other markers with highest ASE value that were not blasted since no sequence information is available, were TRAP markers M200 and M202, which derive from T15 and T17 amplifications respectively (see Table 2). Interestingly, some DArT markers, mainly associated with SC, showed high identity with an alpha kafirin protein that it is involved in the storage of nutritious substrates.
In the last decade, several approaches tested in plant genetics have allowed the precise identification of "desirables" alleles at molecular level. In the most recent years, the development of association mapping for this purpose has gained large importance. In this work, association mapping was used in sugarcane to identify molecular markers associated with both sugar and biomass yields. The quantitative nature of both traits and the polyploid genome of this crop make the use of association mapping a great challenge compared to other studies conducted for crops with less complex genomes. Even considering that, in the present study we were able to detect QTL for both traits, which are consistent across harvesting seasons.
Several studies suggest that the use of elite germplasm could be useful for association mapping [58–60], although there are only a few approaches conducted with this type of population in plant crops (see  for a review). In order to take advantage of the available large phenotypic data accumulated from replicated field experiments over locations and years for the SCBP-EEAOC, association mapping was conducted over accessions of its current elite breeding pool (genotypes of the advanced yield trails). All genotypes characterized in the present study were planted and evaluated at the same time obtaining both balanced data across environments and an extensive phenotyping. Therefore, although population sizes are relatively small, the high quality extensive phenotyping provides a reasonable foundation for the GWAS study. Small population sizes would result in decreased power to detect QTL [61, 62] and increased false-positive rate [63, 64]. However, assembling large populations in sugarcane could be a challenge mainly because of the phenotyping requirements and the relative size of the breeding programs. Furthermore, exploring diverse germplasm not adapted to local conditions and with strong population structure could hinder the QTL detection due to the additional challenge of modeling such population structure . Additionally, the mapping approach (i.e., candidate-gene or genome-wide), the relatedness of the individuals, the extent of LD, and the number of markers will determine the optimal population size in GWAS studies . Finally, population sizes close to 100 have been used elsewhere as a first approach to QTL mapping in other species [66–70]. Recent sugarcane GWAS studies included 189 and 183 individuals [27, 71]. However, since the experimental population in the present study was a representative sample of the population to which inference is desired, it is expected that the information obtained from the association study will be useful and readily applicable to local crop improvement .
The presence of subpopulations in the mapping population creates a challenge for association studies. Several methods have been proposed for dealing with false positives related to population structure [16–18]. In that sense, many studies conducted especially with small datasets and diploid organisms implemented the method proposed in the freely available software Structure . However, in the case of sugarcane considering its complex polyploid genome, several assumptions are not fulfilled for the use of Structure; therefore, the applicability of this algorithm may be limited in sugarcane . For example, in a previous study in sugarcane , when population structure was taken into account by using Structure, arbitrary subpopulations of the genotypes were observed; however, as there were no clear discontinuities in the population, this algorithm failed to conclusively group the population . In the present study a GWAS mapping was applied within a mixed-model framework according to  and . Spurious associations were controlled while the power to detect true associations was maximized by using a PCA as a random component to control for population structure [19, 36, 46–48]. When PCA as a random component is included in the analysis, the large population structure is captured with the first few axes that account for most of the variation while the more subtle relationships among individuals are captured by the remaining significant axes.
Population structure was inferred with an independent set of markers to avoid dependency among terms in the model and to prevent the structure from absorbing the QTL effects from the model [44, 46]. A sub-set of available markers to infer population structure has been used in other studies , including sugarcane . Gouy et al.  used a sub-set of the available markers to ensure genome coverage and avoid over-representation of genomic regions. The sugarcane DArT-based map recently published  was used to sample independent markers of each linkage group. Furthermore, QQ-plots of P-values showed that population structure was properly accounted for by using a stratified selection of markers to correct for population structure as random effects (Additional file 5). On the other hand, using a random selection of markers without accounting for marker position failed to properly account for population structure (data not shown). Additionally, the grouping observed at PC1 has a biological interpretation, reflecting genetic variation among progeny (Fig. 2). For instance, the right-hand side of the plot includes cv. LCP 85-384 and its progeny; while the left-hand side represents the remaining genotypes. This was also found in other studies, where LCP 85-384 was genetically more distant to modern varieties [74–76]. Cultivar LCP 85-384 is a BC4 derived line of S. spontaneum US 56-15-8 and therefore have a strong wild genetic component . TUCCP 77-42, another variety with a strong wild genetic component (BC1 of S. spontaneum SES 147B), was distant from the rest of the genotypes at PC2 level (Fig. 2). These results showed enough evidence of the ability of these few markers (107) used in the PCA to reveal the genetic background of genotypes. Furthermore, most of the structure found in these genotypes seems to come from subtle kinship relationships more than large-scale population structure. Our method properly accounted for these relationships.
Association studies are becoming a popular strategy for unraveling the genetic underlying complex traits. The first association mapping studies conducted in sugarcane have focused on genome-wide approaches attempted at looking for associations between disease resistance and molecular markers [27, 71, 73, 77, 78]. Few reports were found [27, 28] involving associations between molecular markers and traits related with cane and sucrose yield and/or yield components. Wei et al.  conducted a study where field-data for cane yield (t ha-1) and commercially extractable sucrose content were obtained in plant-cane. However, one of the major concerns in order to find markers contributing to yield during several ages is the repeatability of the marker–trait associations across harvesting seasons (mainly for ratoons). Another study including sucrose yield and yields components, among other traits, was carried out by Gouy et al. , obtaining plant-cane phenotypic data from trials planted during different season or years and locations. However, only a few marker-trait associations were detected for the traits analyzed. In that sense, in the present study several markers (20) were found to be associated with CY in at least 2 years, being more frequent the coincidence among first and second ratoon. Sequences of four markers (M58, M54, M97 and M120) showed very high similitude and low e-value with coding sequences of Sorghum bicolor. Sequences of M58 and M54 were found to be the same, and they presented high identity and low e-value with a sequence located on chromosome 9 from Sorghum bicolor, where QTLs for plant height and tiller number were previously found . Sequence of M97 was located on chromosome 2 of Sorghum bicolor, were QTLs for stem diameter and plant height were previously reported  and validated later . Regarding M120 marker, whose effect was 8.70 t ha-1 in the multi-QTL model, the sequence of this marker presented high similitude with a Sorghum bicolor sequence located on chromosome 6, where QTLs for stem biomass yield, plant height and tiller number were reported in different studies (see ).
For SC, one marker-trait association was found significant for the 3 years of study. This marker, M64, showed high identity (99 %) and low e-value (6,00 E-143) with a sequence located in chromosome 3 from Sorghum bicolor, where several QTL related to sugar content were previously reported (Glucose content ; Sugar content ; Brixº, Juice Sugars g L-1 and Juice Sucrose g L-1 ) and validated . Moreover, in our multi-QTL model, M64 showed the same marker effect (-0.48 %) in two consecutive years (2010 and 2011), indicating that a negative selection for this marker could increase SC.
The highly conserved sequences found in the sorghum genome confirm the usefulness of this database to study regions of interest in sugarcane genome. Sequence of marker M120 presented 407 identical nucleotides with an e-value of zero, suggesting that this is a region shared by sugarcane and sorghum. Other sugarcane sequence markers were also significantly similar to sorghum, which is in agreement with previous studies [24, 25, 83] that reported a high gene microlinearity between sorghum and sugarcane.
Even though this GWAS study is mostly focused on exploring the entire genome with DArT makers, also TRAP markers that targeted to coding regions were employed. This information resulted useful in finding regions controlling traits of interest since three of the 103 TRAP markers used for association analysis were significantly associated with CY in 2 years of study.
It is important to highlight the challenge in finding strong marker-trait associations in complex polyploid species using dominant markers. It is well known that this type of markers are less informative than co-dominant ones, especially in polyploids, because copies of homologous chromosomes "dilute" the polymorphisms. When the markers are evaluated with a binary system, they are scored as 0 for the absence of the allele, or 1 for the presence of at least one copy of the allele. This constitutes one intrinsic limitation of the method that is associated with overlooking ploidy level . In that sense, further research need to be conducted to investigate the establishment of associations between continuous data obtained from DArT markers and allele dosage, instead of binary data. This would probably increase the number of markers associated with characteristics of interest. Moreover, considering that the study was carried out on a panel of sugarcane varieties and elite lines, most favorable alleles would probably be fixed; however, no single variety has all favorable alleles giving an opportunity to accumulate those alleles and thus achieve crop improvement.
This study demonstrated that association mapping in elite germplasm seems to have a clear potential for improving sugarcane, especially for complex traits such as CY and SC, for which measurements are costly and time consuming. Combining existing phenotypic trial data and genotypic DArT and TRAP marker characterizations within an LD approach using PCA as a random component to control for population structure may prove to be highly successful to find molecular markers significantly associated with the measured traits. Two aspects were key to obtain the results shown here: the high quality of phenotypic data from the EEAOC-SCBP collected in successive crop cycles and under the same environmental conditions for all genotypes; and the adequate selection of markers to be used in the analysis of population structure, since the choice of markers that do not adequately reflect the presence of such structure could hinder the detection of QTLs of interest. Additionally, sequences of DArT marker associated with trait of interest were aligned in chromosomal regions where sorghum QTLs has been previously reported. The whole role of these regions will need to be further investigated.
Even though the small size of the population could affect the power of the GWAS and increase false positive rate , findings reports here must be considered early evidence about the genome regions and markers associated with the genetic control of yield-related characteristics in sugarcane and should be further validated.
ASE, allelic substitution effect; CONICET, Consejo Nacional de Investigaciones Científicas y Técnicas; CY, cane yield; DArT, Diversity Arrays Technology; EEAOC, Estación Experimental Agroindustrial Obispo Colombres; GWAS, genome-wide association; IVT, Infield Variety Trials; LD, linkage disequilibrium; MAF, minor allele frequency; MINCYT, Ministerio de Ciencia, Tecnología e Innovación Productiva; PCA, principal component analysis; QTL, quantitative trait loci; SC, sugar content; SCBP-EEAOC, sugarcane breeding program of Estación Experimental Agroindustrial Obispo Colombres; TRAP, target region amplification polymorphism.
We thank Natalia Ovejero for technician support in TRAP markers development in LICOR system.
Ministerio de Ciencia, Tecnología e Innovación Productiva (MINCYT, Argentina) provided funding for Sugarcane DArT markers. Estación Experimental Agroindustrial Obispo Colombres (EEAOC) and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) provided the financial support of the studies conducted in this project. MPF and APC are CONICET members; JR is CONICET fellow.
The datasets supporting the conclusions about phylogenetic data of this article are available in the Dryad Digital Repository [http://dx.doi.org/10.5061/dryad.mv88m]. Other relevant data are within the paper and its Supporting Information files.
JR carried out the molecular genetic studies, performed the statistical analysis of multi-QTL association mapping, the comparative mapping analysis, participated in the sequence alignment and drafted the manuscript. LG participated in the design of the study, performed the statistical analysis of multi-QTL association mapping. MFP generated molecular data and critically revised the manuscript. SO and MIC generated phenotypic data and performed the statistical analysis of these. EMP participated in the sequence alignment. MIC, BW and APC conceived of the study, and participated in its design and coordination. APC revised the manuscript. All authors read and approved the final manuscript.
The R Foundation for statistical Computing. http://www.r-project.org/. Accessed 16 June 2016.
Plant DNA Extraction Protocol for DArT. http://www.diversityarrays.com/sites/default/files/pub/DArT_DNA_isolation.pdf. Accessed 16 June 2016.


There seems to be a clash between generations in the workplace. Elmore suggests the most important quality each generation must exhibit in order for all generations to flourish in their work.
In whatever role you find yourself serving in the church, the power of speaking in front of people is significant. Dr. Geiger reminds us to not neglect this powerful tool.
If you are looking for some really helpful teaching tips as a young adult ministry group leader, Ken Braddy has excellent thoughts in his blog. This article encourages leaders to put groups into a circle instead of rows, which is one of the easiest ways to begin to build community in a group.


This is my first plane (just the beginning).
Seb , have been watching but no more updates? What a great idea ,how have you glued it together ? Looks a very quick way to build . Electric or IC ? How will you cover or not at all ? Geoff.
Your reply asking for more info was on Oct. 2014. Not sure he is still there. That looked like an awfully BIG Foamy. With all that large flat areas between any structure I don't think that could fly. It would set up way too much vibration.
Too bad it he was brand new to the hobby and it was his first project. I always have suggested NEW people look through our Club members, or Flyers where they are, to try and find a match, and ask if they would help them. Other Flyers always do help. It is important new modelers see their first project go together and to see their frist project fly, sucess! That's what builds confidence to continue.
That was nice work for a first start.


Have been used my Fedora 9 desktop for half a year, I really got bored with the default Fedora Themes, especially the default icons theme, so I tried to look for a better one on the whole net, and I was so excited when I found the unofficial tango icon theme. It's high quality and looks very cool. The Tango team is just the contributor of GNOME icons themes, so their icons have a good visual metaphor for Nautilus and other GNOME modules.
And I want to recommend some sites that help you to customise your Fedora (or other Linux distro) desktop, one is http://www.gnome-look.org (for KDE users, go to http://www.kde-look.org), you all know this site 🙁 Another one I'd like to introduce to you is the GNOME Art http://art.gnome.org/, and tell you the truth, the official Tango icon theme is just downloaded from there. I found a cool GNOME wallpaper too, which I will set it as my wallpaper this night.
I enjoy Tangoish icons much more too.


It's difficult to say with any certainty what the drone industry will look like in a few years time. Industry reports and predictions tend to vary, but most are extremely bullish on the potential growth of drones in business. Some are more sleptical on how widely they will be adopted given the current legislative landscape, among other things.
What we can say with confidence is that the industry doesn't look like it's going anywhere soon. This is especially the case in the photography sector, with aerial photography and video being the most popular use of drones today.
So what is it about being a drone pilot that makes it a job of the future?
One of the most appealing aspects of becoming a drone pilot is the flexibility. In an economy in which more and more people are taking on flexible work and becoming self-employed, it makes sense to try and earn some extra cash doing what you love.
Being a full-time pilot is difficult while the industry is in its early stages. But there's plenty of money and experience to be earned all the same.
The reason that so many projections are bullish about the industry's future prospects is the assumption that more and more businesses will adopt drone technology in one way or another.
So that might include real estate agencies increasingly using drones to shoot marketing videos for properties, or media companies searching for a cheaper way to grow their catalogue of aerial video.
And that's before you explore industries such as construction, oil and gas and agriculture, in which aerial sensors and automated data gathering are starting to gain traction.
In a perfect world, it's easy to imagine autonomous drones darting about, delivering packages, carrying out inspections and performing tasks without any supervision. Maybe one day they will even ferry us from place to place.
But in the meantime, the growing industry will need pilots behind the controls. Although automation is in many ways the holy grail, safety concerns, public perception and strict regulations mean that it's a long way off, despite recent advances in the technology.
For that reason drone pilots will be in demand for the forseable future.
There aren't many industries that are so closely entwined with emerging technologies. Becoming a drone pilot gives you the opportunity to work alongside artificial intelligence, advanced robotics and computer vision. Not to mention the chance to see firsthand the progress of industry pioneers such as DJI and Intel.
We humans have always been obsessed with capturing moments. From the cave paintings of prehistoric man to the first video ever recorded. Becoming a drone pilot is the next evolution in that journey.
Interested in becoming a professional drone pilot but not sure where to start? Let us help! We provide the training and resources you need to land a professional drone pilot job. Get started today!


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text:
Q: PCB Board design for H-Bridge TLE5206 I would like to make PCB board for TLE 5206-2G (check the datasheet).
It is TO263 package.
Basically, what i dont know is how to transfer heat out of the IC. I saw few PCB's that has little copper square with grid of holes in it. Is that what is used for taking out the heat?
Datasheet (page 21) says: Metal surface min. X=7.25, Y=6.9. Is that refers to heat transfer surface ? Will plated holes enhanced heat transfer ?

A: These sorts of devices (surface mount with exposed metal) are meant to be directly coupled to heavy copper on the PCB to get the heat out of the device.
Usually this involves a multilayer PCB, with a bunch of vias that connect copper on several layers to the device metal. The vias not only channel heat through the PCB by conduction, they also ensure that the device is well soldered with good copper-to-copper contact.

redpajama_set_name: RedPajamaStackExchange


text:
De Van Hauwermeirenmolen is een voormalige watermolen in het tot de Oost-Vlaamse gemeente Wichelen (deelgemeente Schellebelle) behorende gehucht Bruinbeke, gelegen aan de Watermolenstraat 28.

Deze watermolen op de Molenbeek van het type turbinemolen (voorheen: onderslagmolen) fungeerde als oliemolen en korenmolen.

Geschiedenis
Hier lag een klooster van Norbertinessen, het Klooster van Tussenbeke. Dit heeft bestaan van 1258 tot 1783 en bezat twee windmolens en een watermolen. De watermolen bestond al in 1570 en fungeerde toen als oliemolen. Tussen 1840 en 1850 werden de huidige gebouwen opgetrokken, maar het complex heeft een oudere kern. Later werd de molen omgebouwd tot korenmolen en werd het onderslagrad door een turbine vervangen. De molen fungeerde nog tot 1972. In 1994 werd de molen met omgeving beschermd als dorpsgezicht.

Gebouw
Het betreft een complex bestaande uit een molenaarshuis en aangebouwd een molenhuis. Verder is er nog de sluis en de turbine. De maalstoel is nog aanwezig. Aan de overkant van de beek ligt de voormalige oliemolen waarin zich nog de aandrijfmotor bevindt.

Hauwermeirenmolen
Onroerend erfgoed in Wichelen

redpajama_set_name: RedPajamaWikipedia


text:
< Friday, October 25, 2019 >
1:00 PM Talk with Potter Shumpei Yamaki Clay Artists of San Diego will present a special after12:00 PM with Potter Shumpei Yamaki at The Studio Door.
Yamaki is visiting San Diego for a show art Entoten Gallery on Saturday. The artist will share his experiences as an artist, give a digital presentation and do a live demo in the gallery. Free to the Public.
The Studio Door
3867 Fourth Avenue
2:00 PM Book Signing: J. Elke Ertle November 9 will mark the 30th anniversary of the Fall of the Berlin Wall. As the world prepares to commemorate this historic event,J. Elke Ertle, will discuss the factors that led to the construction of the Berlin Wall in 1961 and the dynamics that brought it down in 1989.
Elke, born and raised in West Berlin, was fifteen years of age on that summer day in August when construction of the Wall first began. She vividly recalls how West Berliners, whose city was no more than a tiny capitalist island within a large communist sea, were terrified of winding up behind the Iron Curtain.
Elke will end the discussion by reading a brief excerpt from her memoir, "Walled-In: A West Berlin Girl's Journey to Freedom" that describes the panic, the doom, the confusion, the lack of reliable information and the rumors of collusion, under-the-table deals and nuclear war that swept thought the city.
San Carlos Public Library
7265 Jackson Dr.
6:00 PM Film & Discussion About Race, Mental Health & Community As part of the Community, Arts and Resistance and Challenging Conversations series, the UC San Diego Institute of Arts and Humanities invites you to a special documentary film screening and discussion.
Minding the Gap, a film by Bing Liu, follows three young men who bond across racial lines to escape volatile families in their hometown. Ten years later, unsettling revelations force them to reckon with their families and each other.
7:00 PM Music: Clave Dura Clave Dura is the combined talents of 9 professional musicians. Their fusion of Salsa, Cumbia, Merengue & Bachata delivers the authentic sounds of Puerto Rico, Cuba & Colombia. This energetic ensemble brings a sound that
7:00 PM Opera: Aida Loyalty to country, love, and betrayal collide when the pharaoh
7:30 PM SDIFF Film: Selfie + Shorts: Shine, Beauty Several years after an unarmed teenager was shot by police in the Traiano neighborhood of Naples, filmmaker Agostino Ferrente traveled to the gang-ravaged neighborhood to make a documentary about the lives of its young people
8:00 PM Stage: Handbagged Step through the palace doors and into the ring with the Queen and the Iron Lady. Liz and Maggie. Born 6 months apart, Queen Elizabeth II and Prime Minister Margaret Thatcher were from two different worlds. One destined to rule, the other elected to lead. But when the gloves came off, which one had the upper hand? FOUR of San Diego

redpajama_set_name: RedPajamaCommonCrawl


text:
The role of Information Sharing Agreements in tackling corruption and doping in sport
Published 04 June 2018 By: Danielle Sharkey, Hannah Kent
Information Sharing Agreements have recently been making the news, with the Austrian Play Fair Code and Sportradar and UEFA and ESSA joining forces,1 and the news on Wednesday 30 May 2018 that the Council of Europe and UEFA have signed a Memorandum of Understanding.2
Accordingly, this article examines what Information Sharing Agreements are, specifically looking at:
how they work;
the impact of General Data Protection Regulation (GDPR);
who uses them in UK sport,
recent developments; and
a conclusion as to their effectiveness.
Get access to this article and all of the expert analysis and commentary at LawInSport
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This work was written for and first published on LawInSport.com (unless otherwise stated) and the copyright is owned by LawInSport Ltd. Permission is granted to make digital or hard copies of this work (or part, or abstracts, of it) for personal use provided copies are not made or distributed for profit or commercial advantage, and provided that all copies bear this notice and full citation on the first page (which should include the URL, company name (LawInSport), article title, author name, date of the publication and date of use) of any copies made. Copyright for components of this work owned by parties other than LawInSport must be honoured.
Tags: Anti-Corruption | Anti-Doping | Gambling Act 2005 | General Data Protection Regulation | Information Sharing Agreements | Integrity | International Standard | Macolin Convention | Protection of Privacy and Personal Information | Sports Betting Intelligence Unit (SBIU) | UK Anti-Doping (UKAD) | United Kingdom (UK) | WADA Code | World Anti-Doping Agency (WADA)
INTERPOL Integrity in Sport Bi-Weekly Bulletin - 20 March - 3 April 2018
Why sport needs a unified approach to sanctions for corruption offences
From corruption & scandal to reform: How the Brazilian Olympic Committee overhauled its governance model
Danielle Sharkey
Danielle is an Associate in the Sports Team at Charles Russell Speechlys LLP. Previously she spent eight years as legal advisor to the British Horseracing Authority (BHA) where her focus was on regulatory, governance, data protection and disciplinary matters. Danielle prosecuted a number of British horseracing's high profile corruption and doping cases during her time with the BHA.
Hannah is an Associate in the Commercial Dispute Resolution team at Charles Russell Speechlys LLP where she specialises in sport, media and general commercial disputes, and has completed a secondment to The FA. She acts for a range of clients, including individuals, large corporates and national governing bodies, and has advised clients involved in a number of different sports.


text:
Drawa (česky Dráva, ) je 186 kilometrů dlouhá řeka v západním Polsku, pravý přítok Noteće. Pramení v Západopomořanském vojvodství ve výšce 212 m n. m. u vesnice Zdroje jižně od města Połczyn-Zdrój. Teče převážně na jih a vlévá se do Noteće u města Krzyż Wielkopolski ve Velkopolském vojvodství. Leží na ní mimo jiné druhé nejhlubší polské jezero, Drawsko, a Drawieński Park Narodowy. Rozloha povodí Drawy je 3 296 čtverečních kilometrů.

Odkazy

Reference

Externí odkazy

Povodí Warty
Řeky ve Velkopolském vojvodství
Řeky v Západopomořanském vojvodství


text:
Do you like to babble about football, me too!
Parting gift for Scudamore, parting slap for fans
What has happened to Mourinho?
Blooding the Youngsters
Liverpool firing again
Ronaldo Returns to Rape Utd
The Mourinho Show is Back
The Premier League is back!
Author: The Babbler
I am a huge fan of the English Premier League and want to share my opinions about what goes on in the most competitive and entertaining league in the world!
'The Special One' is back!
22nd November 2019 ~ The Babbler	~ Leave a comment
The world of football rarely ceases to amaze and this has certainly been another amazing week. With the oddly timed sacking of Pochettino, 'The Special One' has stepped into the void. It almost feels like Spurs had been in contact with Mourinho for some time before the sacking but wanted to make sure he'd take the job before getting rid of Pochettino. After all it could be deemed a step down for Mourinho, who has won more major trophies in his career than Spurs have in their history, and with no billionaire owner to throw the cash around Spurs don't seem like a good fit.
Mourinho usually guarantees success wherever he's been. Even during his ill-fated time at Man Utd he still won 2 trophies but are his methods still relevant and is his management style still effective? He's lucky to have inherited a good squad, albeit one lacking in confidence. Mourinho will undoubtedly revert to type though and some of the 'piano players' in the squad could find their place under threat from 'piano lifters' as he implements his pragmatic style. Will the Spurs fans be happy with this after watching some fantastic free-flowing football in recent years? Going from that to grinding out results playing ugly football can be quite a culture shock for some but if the results improve I'm sure most fans won't care how they play.
The burning question everybody will be asking is how will Mourinho fit in at Spurs? They always work within a tighter budget than the rest of the top 6, with a net spend of just £100 million in 5 years. They also prefer to buy young players with potential. Mourinho on the other hand likes to buy ready-made established players, who tend to cost a lot of money, especially in the current transfer climate. Are Spurs going to smash the piggy bank for him or will he be forced to work within the constraints of the club structure and adapt his style. All these questions remain unanswered but it is a fascinating thought. One thing we know for sure, it will be pure drama all the way.
UnVARgettable Weekend in the FA Cup
29th January 2019 29th January 2019 ~ The Babbler	~ Leave a comment
Another exciting weekend of FA Cup football is behind us and we can safely say that the decision to only have VAR available at the Premier League grounds was somewhat questionable.
At the Millwall v Everton match they had their own unofficial version of VAR, it's a pity that the referee team didn't make better use of it though. Apparently you can't overturn a decision based on a giant replay of the incident seen by thousands inside the stadium, but it is OK to overturn the decision if it's an official VAR system.
Is there no common sense in football these days? Surely if a member of the referee team sees something that leads to a wrong decision he should alert the referee. How can he go on knowing a wrong decision was taken regardless of the manner in which he received that information. In this instance it was a massive game changing incident allowing Millwall to draw level late in the game with a clearly illegitimate goal. Instead the 4th official just signaled vigorously for someone to turn the replay off.
Another Premier League team to crash out of the Cup was Spurs, their 2nd cup exit in a week after their defeat by Chelsea in the League Cup semi-final. Despite the different circumstances from the Everton game both Premier League teams deserved to leave the competition based on their performances alone.
Spurs obviously have problems with squad depth with Kane and Ali injured and Son on international duty. If they had played if they were available is another question. For the spurs players and fans I imagine this Cup exit will be less painful than the League Cup exit. Especially after being 1-0 up against Chelsea from the first leg. Having one foot in the final and then having it taken away again could have played a part in their poor FA Cup exit also.
One team prancing their way to the 5th round is Man Utd after their 8th straight win since Ole Gunnar Solskjaer took after a month ago. In his first few games Man Utd may not have dispatched all the teams with ease but they've kept winning and that will breed confidence throughout the squad. With away wins at Spurs and Arsenal, they have their tails up. They are playing with a much better flow than a few months ago and seem a more unpredictable outfit, with all the forward players contributing. I can see them going on to win the FA Cup now, with Man City being the only team that could stop them.
Man Utd must go to Chelsea in the next round but should still play with the same nothing to loose attitude as slight underdogs. Chelsea on the other hand may feel under pressure and have some problems scoring goals at the moment. Higuain is considered to be the solution to their problem. Personally I'm always skeptical about big name strikers coming to the Premier league so late in their carriers. Shevchenko, Falcao and Morientes to name but a few who have arrived late and flopped. Sari has worked with Higuain before so maybe that familiar face will be the difference between Higuain and those aforementioned strikers.
This years FA Cup has been something special so far with the lower league teams providing a lot of value with several cupsets. There are only 7 Premier League teams left and many of them playing away to lower league opposition in the next round. There could be a few more cupsets to come.
Liverpool's ghost of Christmas past
4th January 2019 4th January 2019 ~ The Babbler	~ 1 Comment
It has been well documented over the past few weeks that on 8 of the past 10 seasons, the team at the top of the table at Christmas have went on to win the league. The 2 exceptions are Liverpool of course. In the 08/09 under Rafa Benitez and then again 13/14 under Brendan Rodgers.
In the 08/09 season Liverpool faded somewhat in the second half after Benitez's infamous 'Facts' rant. In the 13/14 season Liverpool were still top with 3 games to go, only to choke and hand the league to Man City. The devastating loss to Chelsea along with that monumental collapse 3-0 up at Crystal Palace ended Liverpool's chances that year.
Liverpool could look back on both those occasions and learn something. Quite simply they need to keep calm on and off the pitch. In 08/09 Benitez clearly lost his cool trying to play mind games with Ferguson and as a result the whole team suffered. In 13/14 the players lost their cool on the pitch during that 3rd last game against Mourinho's bus-parking, slow-moving Chelsea. Liverpool were trying a little too hard to win that game when a draw along with 2 wins from the last 2 games would have secured the league.
Despite their defeat to Man City last night Liverpool have no need to panic and with Jurgen Klopp at the helm and this group of players they are much better equipped to deal with the pressure. In both 08/09 and 13/14 Liverpool had an excellent first 11 but beyond that it was very limited. It was almost inevitable that those players would not be able to keep up that level of performance the whole season with no backup. Liverpool have a huge squad this time. They have players who can make a big impact from the bench, and with several players still to come back from injury they are in much better shape than those previously failed attempts to win the league.
The problem for Liverpool this year is they are not up against just one great manager and squad but two. Tottenham are just 6 points behind Liverpool and must also be considered contenders for the league. They have won the same amount of matches in the league as Man City which is quite impressive. Their only negative point is they've lost the other 5 matches failing to rescue a draw yet. Grinding out a few wins or draws from difficult situations is something Liverpool and City have both managed this season and it is also seen as a sign that a team has got that extra grit and determination needed to become champions of England. If Spurs can correct that in the 2nd half of the season they will play a very important part in the title race.
With many non Liverpool fans hoping this is the start of a blip in form, and Man City's big players coming back from injury, and Spurs form improving, the 2nd half of the season is shaping up to be a cracker with a few more twists and turns to come. Can Liverpool finally rid themselves of that ghost of Christmas past? We will find out in May.
Man Utd falling into the footballing abyss
17th December 2018 ~ The Babbler	~ Leave a comment
The rate at which Man Utd appear to be falling into the footballing abyss is quite alarming. It just goes to show how quickly things can change in football.
Man Utd finished last season relatively unchallenged in 2nd place. Although the standard of football was already not of the most desirable variety, Mourinho was still getting the results. This year however things are not going the same way. Bad performances and bad results are making for a very unhappy camp.
It's difficult to see Mourinho keeping his job much longer but Man Utd are not really the panic sacking sort of club. In the post Ferguson era they have usually given their managers almost full seasons before getting rid of them. The other problem they have is who is even available right now that could come in and straighten the team out. So the question is do they continue with Mourinho till the end of the season with morale seemingly at an all time low, or do they get rid now and bring in a temporary manager or promote someone from within.
In contrast to Man Utd, Arsenal seem to be on the up despite their loss to Southampton at the weekend. Their first loss in 22 games, since they started the season with 2 defeats. Last season they found themselves in a similar situation to Man Utd's this season. With bad performances and results, they took the plunge and finally got rid of Wenger. However when you look at their match day squad, there are not that many changes to the squads of Wenger. A perfect example of how a fresh set of eyes (Unai Emery) can lift a talented set of players and give them a new direction and purpose.
A fresh set of eyes is surely something Man Utd could do with right now. When you look at their squad, it's unbelievable to think that with the talent on offer, they find themselves in this situation. Mourinho's 3rd season syndrome seems to be striking again and if someone doesn't address the situation soon, the rot that is setting in, could be widespread by the end of the season.
Liverpool on the other hand are like an unstoppable train recently. Qualification for the last 16 of the Champions League and top of the Premier League. Every player seems to be buzzing right now and very eager to make an empression, whether it's from the bench or the starting line up. The same cannot be said for Man Utd. Klopp trusts his players and clearly let's them know. Take Nathaniel Clyne for example. It was his first start of the season and he had a very solid game. If you have the backing of your manager, that can give your confidence a great boost. Clyne didn't seem fazed despite the big occasion or the lack of recent playing time. Mourinho could certainly take a leaf out of Klopp's book on this topic.
Liverpool… Top?
8th December 2018 ~ The Babbler	~ Leave a comment
If you had have told me when Liverpool were 1-0 down to Burnley on Wednesday night, that they would be top come Saturday evening, I can't say I would have believed you.
Somehow they've managed to find themselves at the summit of the Premier League despite the relentless pace of Man City. On the inside I'm dancing but my head is telling me, haven't we been here before?
They've kept up with Man City with several grinded results so far and avoiding defeat against the 3 next best teams in the league. Their brand of football this season has not always been befitting of a league winning team but their defence certainly has. The addition of Alisson has been a huge success already. He has come up with some vital saves at crucial times in matches and his distribution has been excellent, leading to several breakaway goals for Liverpool this season. The central defensive partnership of van Dijk and Gomez has been an absolute rock at the back. Something Liverpool have been missing for quite some time.
The midfield however is not as strong as it has been in recent years. Ironically I think that the amount of options they have in midfield has weaked it slightly. With Henderson as captain Klopp probably feels obliged to play him when injury free despite his lack of form so far this season. With Milner's age Klopp possibly feels he needs rested more than others, but not something I would agree with due to his great form when he plays. Keita and Lallana have been somewhat injured and unable to get a consistent run in the team. Fabinho and Shaqiri have recently been given their chance and have given Klopp a headache with some good performances. The one almost ever-present and consistently well performing player is Wijnaldum. Due to his role, which mainly involves adapting to whichever 2 players he lines up along side, he has flown under the radar but has this season become an absolutely vital part of Liverpool's midfield 3.
Of course I'm playing Liverpool's chances of winning the league down. It is afterall only December and there's a lot of football to go but it's certainly exciting times for Liverpool fans. The next month however will severely test Liverpool's resolve in both the Champions League and Premier League. With vital home games to Napoli, Man Utd and Arsenal coming up in December and starting the new year with an away trip to Man City we will know more about Liverpool's title and Champions League credentials. Let's hope for the sake of the Premier League, they can keep up the pace.
For Klopp Sake
2nd December 2018 ~ The Babbler	~ 1 Comment
As a Liverpool fan it's difficult to criticise Klopp, but let's be honest if Mourinho had have run on to the pitch in the way Klopp did during the Everton match, he would be getting butchered by the fans and press right now. It seems Klopp can just do no wrong.
I cannot defend Klopp's actions but I can offer some opinions as to why he gets away with this sort of thing when Mourinho wouldn't. Personality and respect. These 2 traits come across very differently in the 2 managers. Klopp always appears very personable and open in his press conferences which makes him very likeable. Mourinho on the other hand seems more closed off and distant.
Any form of criticism Mourinho receives, he takes it very personally and sees it as disrespectful. With all his achievements in the game, he feels he should be immune from criticism but football is a fickle sport and people have short memories. Klopp can handle the criticism a little better usually giving more insight into the situation or trying to turn the criticism into a positive. Klopp never gets frustrated during interviews even if he receives some stupid questions unlike Mourinho.
All of these factors combined make Klopp a very likeable character in the world of football and therefore somewhat immune to criticism. He is also more likely to get away with some controversial acts such as running onto the pitch during a match. Something Mourinho would get lambasted for. In this particular incident Klopp is lucky the opposition manager Marco Silva and his players didn't view the incident as disrespectful or react angrily which may help Klopp's case if the FA come calling.
Let's not let this incident take away from what was another great weekend in the Premier League. 3 big derbys today and 3 big wins. Spurs continue to blow hot and cold on the big matches this season. They've had 2 big wins against Man Utd and Chelsea but also convincing defeats to Liverpool and Arsenal now. After the first 2 matches of the season ended in defeat for Arsenal, they've been on an uphill roller-coaster ever since, quietly creeping back up the table and now look very capable of finishing in the top 4 or better.
After Jordan Pickford's horrendous error gifted Liverpool a somewhat undeserved 96th minute victory in the Merseyside derby, Liverpool stay in touching distance behind Man City with just a 2 point difference. Despite the small gap at the top of the table Man City still appear to be on a different footballing planet to the rest of the Premier League. They managed to cruise to a relatively comfortable 3-1 win against a determined Bournemouth without their 3 most influential players, Aguero, de Bruyne and Silva although the latter did appear as a substitute.
Man Utd also stumbled to another draw against a very poor Southampton side, or is it the other way around.
A touchdown for Mourinho
28th November 2018 28th November 2018 ~ The Babbler	~ Leave a comment
José Mourinho is an absolute master in the art of distraction and provides us football fans with an endless means of commentary.
Was anybody really supprised when he scored a touchdown last night with that crate of water bottles? His team had just been saved by a classic Fellaini goal in Fergie time but Mourinho still managed to steal the show. What people are mostly talking about now is that crate throwing incident. Realistically this is just papering over the cracks though as this was Man Utd's first home goal in the Champions League this season in 3 matches.
Fan frustration is starting to show at Old Trafford with a few empty seats when the match started and many empty by the time the 90th minute began. A sign of the times that the fans are getting fed up with the lack of exciting football being played at the 'Theatre of dreams'. The trouble for the fans that want rid of Mourinho is how can you fire a manager who wins one week and draws the next with a few sporadic defeats. That inconsistency is probably what is keeping Mourinho in the job oddly enough.
The other major point in the match last night was that David de Gea save to keep the score at 0-0. That guy continues to bail out his team time and time again giving them that opportunity to draw or even win these matches. One can only wonder what shape Man Utd would be in without him. He really is a world class keeper and Man Utd are lucky that he's sticking around dispite the downturn in fortunes.
Dispite the lack of goals in the Champions League this year Man Utd have still managed to qualify for the last 16 with a match to spare. Something Mourinho has never failed to do in 14 seasons in the Champions League. Something he pointed out after last nights match. He is of course a 2 times winner of the competition and a further 6 times semi-finalist. The historical stats look great for Mourinho but currently reality doesn't and time will tell if constant masking of the problems will keep Mourinho in this job beyond this season.
Diving for Gold
26th November 2018 ~ The Babbler	~ Leave a comment
During the Premier League match last Saturday between Brighton and Leicester there were 2 cases of diving, one was punished and the other was rewarded. If there's anything in football I cannot have, it is diving. It is a scourge of the modern game.
The first case of diving was when James Maddison, already on a yellow card, tried to to skip through some players in the box hung out a leg and fell to the ground. It was a clear and deliberate dive and thankfully the referee had the strength and conviction to show him a 2nd yellow and send him off. He later apologised on Twitter claiming he was waiting on contact which never came but that's not good enough. How can you honestly play football when your mind is somewhere else waiting on some contact so you can throw yourself to the floor. How can this be in the forefront of your thought's? What ever happened to being brave, playing on and just taking the hits if and when they come.
The reaction from his own team mates is also concerning, most of them opting for the 'unlucky you got caught' pat on the back. His team mates should be livid with him and give him a good clap round the ear. He left them for the remaining 62 minutes of the match to play with 10 men.
The other case of diving was when Leicester won a penalty. At first I also thought it was a foul but on further inspection having seen the incident from a side on angle it's more of a dive than a foul. It's less clear cut than the Maddison incident but you can see that Kelechi Iheanacho is already falling long before the contact arrives. In this case there is some contact and a slightly wild tackle in the box but the dive came before the contact so in my opinion this is not a penalty.
The punishment for diving is also far to lenient. How can a yellow card for cheating be sufficient. There's no deterrent whatsoever for diving and unless the authorities grow a set and get tough on diving it will never go away and probably only get worse. All these so called experts on TV are barely raising an eyebrow at the alarming frequency with which we see new cases of diving. It's so common now that some are seeing it as part of the game but I'm sorry I cannot and will not accept that. Diving is cheating and should be outlawed from the game with harsher punishments. Diving should be a straight red card with a heafty fine and ban included. Only then might we see some changing attitudes towards diving from the players.
The Premier League have today confirmed that departing executive chairman Richard Scudamore will receive a £5m bonus with a £250,000 wip round from all 20 Premier League clubs.
Maybe it's just me but this seems ubsurd. Fans are left bemused as ticket prices continually rise, club merchandise is very expensive and Premier league TV subscriptions increase annually, but clubs can just come up with £250,000 at the drop of a hat.
Of course £250,000 is really just pocket money for most Premier league clubs but it's the principle of the matter. OK, the guy did a good job over the many years he worked there, but that's why he was getting paid £2.5m per year. He's also staying on with the Premier League on an advisory capacity, which probably brings it's own 'modest' salary.
The Premier League has grown exponentially over the past 20 years and Scudamore has surely played his part in all that but let's be honest on the reasons we watch. These billionaire owners have came along and brought some of the best players in the world with them. We watch it because of the Aguero's, Salah's and Hazard's, but also because the league still has that typical tough English core that we all love. None of this really has anything to do with Scudamore and the growth of the Premier league was inevitable.
If Scudamore has any shred of decency he should take that money and give it to charity or pump it into a worthy project in need of investment. With the rise of the Premier league, grass roots football has taken a hit over the years. More people are going to the pubs to watch football on Saturday afternoons on some dodgy foreign channel instead of playing for their local clubs. This seems like a worthy project for Scudamore to invest his £5m in. Give back a little from the area you slowly chipped away from all those years.
12th November 2018 ~ The Babbler	~ 1 Comment
It could be one of a number of things or maybe all of them combined but the once self proclaimed 'Special One' has surely lost his edge.
Mourinho is a subject that I never get tired of talking about. One of the greatest managers and pantomime villains of all time. When he's managing in the Premier League I just love to ridicule him but when he's not there I actually miss him. Of course if he had have managed a Liverpool team to a Premier League title my opinion of him would be very different.
Maybe years of fighting back against the press has finally worn him out. Mourinho always launches a ferocious defence of himself and in the past his players but his tenure at Man Utd has shown a different side to him, often hanging his own players out to dry. This is something the Mourinho of old was not known for and loyalty and unity were key aspects of his previous squads. It was almost like an impenetrable machine charging toward all those league titles in various leagues around Europe.
Are his tactics dated? It's not easy to understand why his tactics are not appearing to work any more. Maybe with the emergence of many young foreign managers who are coming into the game with fresh ideas and the willingness to take a risk, football is becoming more dynamic across the league and less predictable for managers to know how their opponent is going to play. This could be something Mourinho has failed to adapt to. Sticking with his pragmatic style of sitting back and playing on the counter for a club like Man Utd just doesn't suit and the fans are getting frustrated as it's a world away from the swashbuckling football of the Ferguson era.
Are the players not good enough? With the money Mourinho has spent it's difficult to say this is the problem. There is more than enough talent in that squad to challenge for the league under normal circumstances. With Pep Guardiola's Man City however these are not normal circumstances and anything less is being highly scrutinised. Of course the players may not be applying themselves correctly to the Mourinho's style. Sitting back and soaking up pressure is not the same as letting the opposition play and that's exactly what Utd did against City. There was no pressure from the Utd players the entire game. Even the last 10 minutes when 2-1 down Utd were playing like they were 2-1 up. They desperately needed to get the ball back but instead sat back and allowed City to orchestrate probably the best team goal of the season already to end any chance in this game.
Whether it's his personality, his tactics or his players it's still down to him. He needs stop worrying what people think of him and find the strength to re-evaluate his methods and adapt to the modern game. If it's his players then he needs to get tough on them, they clearly aren't putting in the effort on the pitch and that's obvious to see.


Edited 1 time(s). Last edit at 04/09/2019 12:55PM by Monte.
Obviously a very rewarding weekend.
So many places to hunt and not enough time in the week !
Good, Better, Best . . . . .
Re: Good, Better, Best . . . . .
Change of Venue ? Change of plans . . .
Re: Change of Venue ? Change of plans . . .
Picking it up then dumping it out.


Horrendous statistics! White Ribbon Day and the ongoing associated activities have a deep meaning for The Royal Australian Air Force, who are serious about becoming White Ribbon accredited. WGCDR Stuart Scott tells us just how serious they are, and how they're going about achieving their aims… and just as important… why!


We can use lists and CSS to create a reflected group of photos in a photo gallery.
The following example places a copy of the original photo under each photo..
The following example inverts the reflected photos.
The following example places a gradient opacity overlay on the reflected images.


When is the Australasian Permaculture Convergence (APC13) on?
The APC runs from October 2-5, 2016 with tours, workshops and courses to follow. Attendance and participation is open to all people who have undertaken courses in permaculture and who adhere to the ethics, practices and principles that permaculture is built on.
Why should permies get along?
APC13 is an opportunity for everyone to share their permaculture journey, their research into resilient and productive systems, and have fruitful discussions about the future directions of permaculture in the world. There is also a packed schedule with some of the most renowned permaculture practitioners presenting on a wide range of topics.
What's in a name: Is it Convergence or Conference?
Historically permaculture enthusiasts have organised a Convergence, with his term better defining the gathering of like-minded minds. Most other organisations offer Conferences, and in the early years, permaculture did have a Conference which was often an open day for the general public. The Convergence was seen as for only Permaculture Design Course (PDC) graduates.
To save the confusion, APC13 in Perth will be offering a Permaculture expo, called PermaFest. This is the one day "conference" where guest speakers will talk about different aspects of permaculture to a largely uninitiated public. We also intend to have hands-on workshops and displays. We anticipate some information booths of appropriate businesses and static displays of interesting projects and case studies.
The Convergence is meant for those of us who have done a Permaculture Design Course and in light of the growing number of Accredited Permaculture Training graduates we need to include those who may have done at least CIII.
We would encourage anyone who is planning to come to APC13 and has not undertaken a PDC, to do so as soon as they can. Some Perth-based permaculture teachers will be offering a two week PDC prior to the Convergence, and we encourage all other teachers in other states and territories to offer these as well. Send in your course details and we will promote these on the APC13 website.
It's going to take a lot of CO2 to get me there… what should I do?
Air travel is recognised as one of the highest producers of carbon dioxide, and so contributes significantly to an individual's greenhouse gas emissions. Most airlines offer customers a chance (and choice) to pay a little more to offset their travel. Unfortunately most people seem to ignore this offer and I also suspect the (small) cost increase in an airfare is really well below the true cost.
While it would be great if all permaculture practitioners automatically considered accepting the carbon offset addition to a fare, it may not be possible to do so for a large number of reasons. While all such schemes are registered by the Australian Government and the funds raised are given to legitimate organisations for tree planting and other endeavours, Perth is donating part of a participant's Convergence fee to local organisations to enable worthwhile food-based and other revegetation projects to be funded.
You can go online and find any number of calculators to work out the carbon produced by travel from one city to another and to also give you some estimate of cost. We intend find ways in which we can reduce everyone's carbon footprint to attend the Convergence at Swanleigh, which is about five minutes from the airport and half-an-hour from the city.
How often have you heard that we should practice what we preach? There has always been contention by Convergence attendees that each group should obtain local, freshly grown produce and to grow as much food as possible to feed participants. In our current age of OHS and safety this is not always possible or easy. Caterers have to, by law, be able to document where food comes from and to ensure high public safety. For example, if someone came down with food poisoning then the Department of Health or some other similar government body would insist on a paper trail of where the food came from and what steps were put in place to ensure a low risk of contamination.
You may now be able to appreciate that while growing food for a Permaculture Convergence is the ideal, there will be obstacles to overcome to make even part of this possible. Nevertheless, Perth is committed to work with the caterers to make their job easier to source good, local organic food, and to supply what condiments and other treats we can. We will be growing food and we intend to make this available to accredited suppliers and to liaise with growers to ensure the best possible outcomes for everyone. Furthermore we will be self-catering for the Advanced courses that follow in Rockingham, so we have got food plants in the ground now.
Several tours are offered after the main program of APC13. Only best practice properties have been selected, and each tour will showcase different aspects of sustainable living practices. You will be able to visit urban sites in and around Perth, see small acreage properties in the hills, or take a two day tour to the south-west or up to Geraldton.
What other fun stuff can I do over in WA?
There will be enough courses and workshops to wet anyone's appetite, and these will be presented by internationally recognised teachers. Robina McCurdy starts a few day before the PermaFest with a three day course on Localising Food Facilitation Training. Just after the Convergence David Holmgren offers a one-day workshop on Future Scenarios. Over the weekend that follows he is presenting a two-day Advanced Principles course. At the same time, Robina McCurdy is facilitating Fast-tracking Urban Permaculture and Cecilia Macaulay is presenting an Advanced Life Design Workshop: Permaculture for the inside – for the first time in Western Australia.
Two five-day Advanced courses follow. An Advanced Teaching course with Rowe Morrow and Robin Clayfield is offered concurrently with an Advanced Design course with Graham Bell and Ross Mars.
Any final words to sum up?
Find out more by visiting our website www.apc13.org and join us on facebook to keep up-to-date with the program and events. You can also volunteer for the event here.


Until the 19th century this side altar was under the patronage of Alba's Cantone family, but by 1872 it had been transferred to the Biglino family. A 16th-century Late-Mannerist-style altarpiece in painted and gilded wood can still be seen on the altar table. It serves as a frame for a large panel, adapted to the space and painted in tempera, signed and dated under the Virgin Mary's mantle with an inscription of doubtful authenticity: "Macrinvs faci.t 1508". This is clearly a work by the celebrated Renaissance artist Macrino d'Alba (active 1495-1513, dead by 1528).
The painting shows the Madonna adoring the Child, Saint Joseph, Saint Nicholas of Tolentino, Saint Augustine the bishop, Saint Jerome and music-playing angels against a background of classical ruins.
The original destination of the picture (which might have initially had an arched shape) is not entirely clear. However, it clearly features the venerables favoured by the Augustinians, which, in addition to the dating, suggests that it came from the church of San Bartolomeo at the religious house of Santa Maria della Consolazione in Alba. The abbey was located outside the city, but after war left it in ruins, in 1556 the Hermits of Saint Augustine abandoned it and moved to this ecclesiastical complex.


text:
RetailView Retail Category
American Eagle Plans To Close A Number Of Stores By The End Of The Year
By StyleDemocracy
By StyleDemocracyView Author PostsSep 10th 2020
Dolphin Mall
This year has proved to be a tough one for retailers across the globe. With mandatory closures of physical locations in the first half of the year, we've seen companies both big and large choose to shutter a number of storefronts.
Among the growing list — which includes retailers like Gap, Aldo Shoes, and J.Crew, to name a few — is American Eagle.
During a recent Q2 result call, American Eagle announced that it would be closing 40 to 50 stores by the end of the year.
According to the call transcripts, the company has carefully selected the stores that will shutter based on "lease tenure, mall profile, proximity to other stores, and customer engagement levels." Based on the learnings, American Eagle plans on selecting even more locations to close. There are 500 locations with leases up for negotiation in the next two years, so those locations will be the main focus for closures.
Instagram/@americaneagle
At this time, there is no clear communication about which locations will shutter, but according to the brand's website, American Eagle has over 1,000 locations in the United States, Canada, Mexico, China and Hong Kong.
Luckily, it doesn't seem like the closures mean that the company is struggling.
The company currently has 90% of its stores open worldwide and has reset the organization in what they think puts them in a better position for sustained success. During the second quarter, Mike Mathias, American Eagle's Executive Vice President and Chief Financial Officer, said that the company has made progress in inventory and expense management and capital efficiency, with more changes are underway. Additionally, the company's online channel continues to perform well.
Aerie has been a top performer for the company.
Of the brand, Mathias said, "we had a five-year plan to get to $1 billion and we're pretty much on that plan at the end of this year." Additionally, the company plans to expand its athleticwear brand OFFLINE with its own standalone stores, which American Eagle thinks it can grow into a $16 billion brand.
Instagram/@aerie
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text:
package dom.reportes;

import java.util.ArrayList;
import java.util.List;

import net.sf.jasperreports.engine.JRDataSource;
import net.sf.jasperreports.engine.JRException;
import net.sf.jasperreports.engine.JRField;

public class IndicacionesDataSource implements JRDataSource
{
		private List<ReporteIndicaciones> listaIndicaciones = new ArrayList<ReporteIndicaciones>();
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			} else if ("medicamento".equals(jrf.getName())) {
				valor = listaIndicaciones.get(indiceIndicacionesActual).getMedicamento();
			} else if ("comotomarlo".equals(jrf.getName())) {
				valor = listaIndicaciones.get(indiceIndicacionesActual).getComotomarlo();
			} else if ("doctor".equals(jrf.getName())) {
				valor = listaIndicaciones.get(indiceIndicacionesActual).getDoctor();
			}else if ("matricula".equals(jrf.getName())) {
				valor = listaIndicaciones.get(indiceIndicacionesActual).getDoctor();
			}
			return valor;
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		public boolean next() throws JRException {
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}

redpajama_set_name: RedPajamaGithub


Carrie Ann Fondi, who has been quilting since 1992, is accomplished in the unique art form of Hawaiian Quilting, which evolved in the islands in the early 1800's. She has studied from many master Hawaiian quilters. As Hawaiian quilting became a full-time passion, she created Aloha Quilt Designs in 2010 to sell her Hawaiian quilt patterns and to promote the art of Hawaiian quilting. Her designs are inspired by the beauty of nature, and from her love of the Hawaiian Islands.


Marion Edward Hines 77, of Clarksville, died at 11:02 a.m. March 18, 2019 at Paris Regional Medical Center. A gathering of family and friends will be held from 6-8 p.m. Thursday, March 21, 2019 in the chapel of Fry & Gibbs Funeral Home. Burial will be at 2 p.m. Friday, March 22, 2019 at Lane's Chapel Cemetery in Red River County with Rev. Charles Brown officiating.
Marion was born August 16, 1941 in Clarksville to Hester and Alice Barta Hines. He had worked for the City of Paris Water Department, the railroad tie yard, and for local tree services. He was a member of the First Baptist Church of Annona.
He is survived by his wife, Judy Griffin of Clarksville; eight children, Angela Janette Hines and James "Jim" Edward Hunt III of Paris, Mary Charolette Jackson and Jerry Dean Jackson of Paris, Alice Amanda Swan and Jeffery Woodrow Moore of Sumner, Samantha Diane Totty and Benny Hulse of Paris, Tina Louise Fields of Clarksville, Donna Lenael George and Lewis Justin George of Spencerville, Oklahoma, Bessie "Becky" Ann Fields of Sumner, and Gary Wayne Fields of Paris; fourteen grandchildren, Brandi, Cody, Jeremy, Cheyenne, Mike, Justin, Madison, Nikki, Clayton, Jasmine, Kansas, Travis, Briar, and Charlie Jo; ten great-grandchildren; two brothers, George and Floyd Hines; and numerous nieces and nephews. He was preceded in death by his parents and two brothers, Calvin and Cloyce Hines.
Online condolences may be made to the Hines family by visiting www.fry-gibbs.com.