Introduction {#Sec1}
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Duplication of genes followed by diversification is a common process shaping the evolution of plant species by natural and artificial (breeding) selection^[@CR1]^. Genes can be duplicated by different mechanisms, including tandem duplication, transposon-mediated duplication, segmental duplication, or in the most extreme form by whole-genome duplication (WGD) or polyploidization. WGD is common in the evolutionary history of many wild and cultivated plant species. Different terms have been used frequently to describe short- and long-range genomic duplication and genome structural variation (SV), a term originally defined in reference to insertions, deletions and inversions greater than 1 kb in size^[@CR2]--[@CR4]^. In contrast to small-scale insertion-deletion (InDel) polymorphisms, which are generally defined as small insertions or deletions of a few nucleotides (up to 50 bp), SV in the size range of genes (up to a few kb) can give rise to copy number variation (CNV) or presence/absence variation (PAV). The latter is an extreme form of CNV where fragments in the size range of genes are missing from the genomes of some investigated genotypes.

Genes affected by duplications, InDels, CNVs and PAVs in diploid and polyploid plant species have been linked to local adaptation of wild populations^[@CR5]^ and to important agronomical traits in crops^[@CR1],[@CR3],[@CR6]--[@CR8]^ for example flowering time and vernalization requirement in oilseed rape^[@CR9],[@CR10]^ and wheat^[@CR11]^, abiotic stress tolerance in wheat^[@CR12],[@CR13]^ and biotic stress tolerance in tobacco^[@CR14]^. However, the strong impact of CNV and other forms of SV in polyploid crop genomes on evolution and trait selection was not recognized until the last few years, when recognition of their relevance was facilitated by large-scale genotyping including long-range sequencing technologies in large breeding populations of numerous crops^[@CR15]^. One recent genotyping technology which is particularly suitable for detection of long-range SV is Bionano optical genome mapping using nano-channel arrays^[@CR16]^. This method involves imaging of high-molecular weight, fluorescently-labeled DNA molecules and creation of large restriction maps represented as stretches of light and dark regions (resembling a barcode), which then can be aligned to an *in silico* generated optical map of a reference genome assembly. A key factor distinguishing this approach from previous technologies for SV analysis is that the DNA molecules are not sheared, thus enabling the capture of long-range genomic information stretching up to several hundred kilobases. In combination with accurate genome assemblies even for strongly complex polyploid crop genomes, optical mapping opens new avenues for dissection of genomic rearrangements associated with traits relevant for commercial plant breeding.

Oilseed rape (*Brassica napus*) is a recent polyploid crop species originating from the inter-specific hybridization between the two diploid progenitor species, *B. oleracea* and *B. rapa*. Due to high levels of homoeology between the two progenitor subgenomes, widespread structural rearrangements are a common phenomenon within the rapeseed genome^[@CR17]--[@CR19]^, while its ancestral hexaploid progenitor genomes already carried intensive structural and functional modifications through long-term genome fractionation and evolution^[@CR20]--[@CR23]^.

This study focuses exclusively on gene PAV in oilseed rape breeding populations and evaluates the biological relevance and prevalence of gene PAV associated with resistance to a common fungal disease of oilseed rape, Verticillium stem striping caused by *Verticillium longisporum*. By genetic mapping of genome-wide SNP/SNaP markers^[@CR24]^, along with short-read Illumina sequencing in combination with long-range optical mapping, we demonstrate that inclusion of presence-absence polymorphisms in quantitative trait locus (QTL) mapping strategies enables also reliable identification of gene PAV with a putative role in *V. longisporum* resistance. The results provide a valuable example for the importance of pangenomic gene variation for breeding of a key trait in a major polyploid crop.

Results {#Sec2}
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QTL detection for *V. longisporum* resistance {#Sec3}
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Analysis of raw genotype data from the Brassica 60k SNP array^[@CR25],[@CR26]^ for a doubled haploid population ExR53-DH was performed for 244 genotypes. Two genetic maps were produced, one using only SNP markers (the "SNP map") and one using SNP plus SNaP markers^[@CR24]^ (the "SNaP map"), respectively. Comparison of the SNP and SNaP maps revealed that large chromosomal regions were not covered in the SNP map (e.g. for chromosome A03 compare Fig. [1a,b](#Fig1){ref-type="fig"}). Surprisingly, QTL mapping using the SNaP map increased the number of detectable QTL from 5 to 17 (Supplementary Table [S1](#MOESM2){ref-type="media"}), with substantially increased LOD scores also indicating a dramatic increase in QTL detection power when including SNaP marker data. Furthermore, the map resolution and precision across QTL intervals were considerably increased by inclusion of SNaP markers. Interestingly, some QTL detected only in the SNaP map contained only SNP markers within the QTL confidence interval (e.g. q23k-BP-A1-1, q23k-BP-A3-2, q23k-BP-A3-2, q23k-BP-A5-1), while other QTL spanned intervals containing only SNaP markers (e.g. q23k-BP-C2-1). Many SNaP markers clustered in groups, spanning large regions up to chromosome scale, while other SNaP markers were located within blocks of SNP markers (Supplementary Table [S1](#MOESM2){ref-type="media"}).Figure 1Comparison of co-localizing (CoLOC) QTL positions (in Mbp anchored to Darmor-*bzh*) obtained by QTL mapping in the biparental ExR53-DH population using maps produced with SNP markers only (light green, hatched) or SNP plus SNaP markers (dark green, hatched), and by GWAS in a NAM panel with 5 subpopulations using maps produced with SNP markers only (light blue, solid) or SNP plus SNaP markers (dark blue, solid). Only QTL above a threshold or LOD \> 3 and −log (p-value) \> 3 were included in the figure. Red boxes indicate QTL regions overlapping in biparental QTL and GWAS.

Additionally, a subset of subpopulations from crosses of a common elite oilseed rape parent with five synthetic *B. napus* parents was selected based on segregation of parental lines for *V. longisporum* resistance from the *B. napus* nested association mapping (NAM) panel described by Snowdon *et al*.^[@CR27]^. GWAS including both SNP and SNaP markers increased QTL detection power and detected a total of 41 significant marker-trait associations (Supplementary Fig. [S2](#MOESM1){ref-type="media"}, Supplementary Table [S2](#MOESM2){ref-type="media"}). Most of the additional detected QTL harboured only SNaP markers.

Co-localizing resistance QTL in diverse genetic backgrounds {#Sec4}
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Using only SNP markers, comparison of QTL detected by biparental QTL mapping and by GWAS revealed no co-localizing QTL (Fig. [1](#Fig1){ref-type="fig"} in light green and light blue, Supplementary Tables [S1](#MOESM2){ref-type="media"} and [S2](#MOESM2){ref-type="media"}). In contrast, adding SNaP markers to the QTL analyses revealed a strong increase to 5 co-localizing QTL (Fig. [1](#Fig1){ref-type="fig"} in dark green and dark blue, Table [1](#Tab1){ref-type="table"}) harbouring 2 to 90 genes per confidence interval (Supplementary Table [S3](#MOESM2){ref-type="media"}). Sizes of QTL intervals for *V. longisporum* resistance showed differences between the biparental QTL mapping and the multi-parental GWAS (Supplementary Tables [S1](#MOESM2){ref-type="media"} and [S2](#MOESM2){ref-type="media"}). Generally, smaller QTL intervals are expected in a NAM-GWAS approach, as higher numbers of recombinations are expected from crosses involving multiple non-related parents. On the other hand, QTL intervals in a GWAS mapping approach can only be measured for markers which can be positionally anchored, whereas biparental mapping can also consider marker loci which can be genetically, but not physically anchored. This led to some co-localizing resistance QTL with smaller confidence intervals observed in the biparental mapping compared to the NAM-GWAS (e.g. QTL on chromosomes A05 and C05, Table [1](#Tab1){ref-type="table"}).Table 1Comparison of QTL locations detected by biparental QTL mapping in ExR53-DH and by GWAS in a NAM panel using SNP and SNaP markers.chromosomeQTL ID in biparental populationStart position of QTL interval in ExR53-DHStop position of QTL in ExR53-DHSize of QTL interval\
(bp)QTL ID in\
NAM\
populationStart position of QTL LD block in NAM populationStop position of QTL LD block in NAM populationSize of LD block\
(bp)chrA03q23k-BP-A3-17,963,05911,419,4763,456,417q22k-NAM-A3-210,075,38810,458,202382,814chrA05q23k-BP-A5-22,357,5352,473,365115,830q22k-NAM-A52,384,1532,808,636424,483chrC05q23k-BP-C5-1andq23k-BP-C5-23,670,200and3,688,1153,672,842and3,949,6172,642and261,502q22k-NAM-C5-23,089,1323,698,279609,147chrC08q23k-BP-C8801,92513,488,67512,686,750q22k-NAM-C812,201,74912,596,542394,793

Gene ontology and enrichment analysis for genes underlying resistance QTL {#Sec5}
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Gene ontology enrichment analysis of the biparental population revealed five enriched GO terms in 17 QTL regions, with highest significance (topgoFisher score) attributable to the terms 'chitin catabolic process' and 'response to biotic stimulus' (Supplementary Table [S4](#MOESM2){ref-type="media"}). In the NAM population eight enriched terms for genes harboured in 28 QTL regions were mainly related to cell growth. In the co-localizing QTL sections six enriched GO terms were identified. A total of 144 genes are harboured within the confidence intervals of the 5 co-localizing resistance QTL (CoLOC1 to CoLOC5, Supplementary Table [S5](#MOESM2){ref-type="media"}). The GO terms for the 144 genes harboured in the 5 co-localizing QTL intervals reflect annotations from just 8 genes (labelled in green in Supplementary Table [S5](#MOESM2){ref-type="media"}) related to cell-wall modification (expansins) and pathogen defence (defensins) on chromosome A05 (CoLOC2) and selenium binding on chromosome C08 (CoLOC5). One gene each returned annotation terms containing 'response to stress' and 'systemic acquired resistance', respectively. For four genes an annotation term 'defense response to fungus' or 'response to symbiotic fungus' was assigned, mostly based on plant defensin genes, whereas seven genes returned annotations containing the term 'cell wall', mostly based on expansin genes and pectin esterase-like protein genes.

Both long and short-range PAV associate with *V. longisporum* resistance {#Sec6}
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To validate if consecutively mapped SNaP markers can help to reliably detect deletions within gene-range size associated with otherwise invisible QTL, resistance-associated QTL detected in the ExR53-DH population were physically located in the *B. napus* Darmor-*bzh* reference genome and the corresponding sequences were compared with optical genome maps. *De novo* assembly for parental lines Express 617 and R53 was performed using 300 Gb (\~250x coverage) from Express 617 and 140 Gb (\~116x coverage) of Bionano data, respectively. DNA molecules from both genotypes exhibited a very high N50 \> 180 kbp, ensuring that long-range genomic information was covered (Supplementary Table [S6](#MOESM2){ref-type="media"}). One nick label was detected for every 10,000 bp of the molecules, also indicating a uniform coverage and a high SV detection power. The final assembly comprised 1,368 and 1,331 optical maps with N50 values of 234 kb and 235 kb, respectively. The total lengths of the optical mapping assemblies were 978 Mb for Express617 and 874 Mb for R53. The high molecule sizes, along with the total size of the assemblies close to the predicted genome size for *B. napus*, indicate a good assembly quality suitable for reliable detection of long-range SV. Large-scale deletions were consistently detected by consecutively anchored SNaP markers in the ExR53-DH genetic map as well as by optical mapping data (Supplementary Fig. [S1](#MOESM1){ref-type="media"}). Optical maps enabled accurate detection of small to medium-size deletions and insertions in the size range of genes (from 3 to 5 kb).

The 17 detected QTL regions harboured between 3 and 21 SNP and/or SNaP markers. In total, 122 markers were contained within QTL regions and 72 were anchored to the Darmor-*bzh* reference genome (Supplementary Table [S7](#MOESM2){ref-type="media"}). All regions were investigated for long-range structural variants within QTL regions in parental genomes by analyzing the optical map data and comparing the data with SNP/SNaP marker patterns in the segregating population. Short to medium -range structural variation (deletion \<5 kb) were confirmed for 16 out of 18 (89%) SNaP marker positions in the two parents (Supplementary Table [S7](#MOESM2){ref-type="media"}), suggesting that genetically anchored consecutive SNaP markers can reliably detect short to medium-range presence/absence polymorphism associated with *V. longisporum* resistance.

Figure [2](#Fig2){ref-type="fig"} shows an example for comparison of genetic mapping from the DH population, reference anchoring of SNP and SNaP markers and optical mapping data of two parents for chromosome A03, which harbours two additional QTL not detectable using the SNP map. One of the QTL detectable only with the SNaP map, q23k-BP-A3-1, harboured 4 SNaP markers, whereas another, QTL q23k-BP-A3-2, harboured 14 SNP markers within the confidence interval (Supplementary Table [S7](#MOESM2){ref-type="media"}). The QTL q23-BP-A3-1 is localized in a region with long consecutively ordered stretches of SNaP markers. For the parental line Express 617, both QTL regions were covered in the optical maps (Fig. [2c](#Fig2){ref-type="fig"}), whereas for the synthetic *B. napus* parent R53 about 6 Mb overlapping the QTL region q-23k-BP-A3-1 was not covered. The region not covered in R53 corresponds to a 13.8 cM interval on the genetic SNaP map (3.456 Mb), with the resistance allele contributed by Express 617 (Fig. [2a,b](#Fig2){ref-type="fig"}). This lack of optical map alignments in QTL q23k-BP-A3-1, combined with segregating, consecutively anchored SNaP markers in the segregating DH population, confirms the deletion of this large chromosomal region in parental line R53 and suggests that this deletion is involved in resistance expression. In contrast, the second QTL, q23k-BP-A3-2, is located in a region on chromosome A03 with long consecutively mapped stretches of SNP markers. Flanking the QTL region, only isolated SNaP markers were mapped. The isolated SNaP markers detected close to the QTL thus probably represent short-range PAV, potentially down to even single-nucleotide level. Nevertheless, saturating the genetic map on chromosome A03 by adding SNaP markers facilitated the detection of QTL q23k-BP-A3-2, whereas no QTL could be mapped in this region using only the SNP map (Fig. [2a](#Fig2){ref-type="fig"}, Supplementary Table [S1](#MOESM2){ref-type="media"}). This example demonstrates that the addition of SNaP markers for genetic mapping was causal for a higher detection power of *V. longisporum* resistance QTL associated with both presence/presence as well as presence/absence polymorphisms.Figure 2Genetic and physical localisation of biallelic SNP and presence/absence SNaP markers in two *V. longisporum* resistance QTL on chromosome A03 in the ExR53-DH population and its parents. (**a**) Genetic linkage maps showing positions of biallelic SNP (black) and SNaP (red) markers. Green lines connect consensus markers between the different map versions. (**b**) Positions of SNP probe sequences anchored by BLASTn to the Darmor-*bzh* reference sequence. (**c**) Optical Bionano genome maps (blue) of the two parental lines Express617 and R53 aligned to the Darmor-*bzh* reference sequence (green). The pink lines connect marker positions flanking QTL regions in the SNaP map, the physical map and the optical maps (**c**). No resistance QTL were detected using the genetic SNP map.

Gene PAV is a key determinant of *V. longisporum* resistance {#Sec7}
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In order to identify putatively absent genes from regions associated with *V. longisporum* disease resistance in the NAM panel, we combined Illumina 60 K SNP chip array genotyping and Illumina resequencing data for the six NAM parental lines with GWAS data from the segregating NAM population. Illumina resequencing confirmed medium to long-range presence/absence variation in the respective parents for 17 QTL regions (Supplementary Table [S6](#MOESM2){ref-type="media"}). 60% of the 28 detected resistance QTL were affected by medium to long-range PAV in the NAM population.

Figure [3](#Fig3){ref-type="fig"} shows an example of the analyses for the SNaP marker Bn-A03-p10964394, which is associated with *V. longisporum* resistance within the QTL interval q22k-NAM-A3-2 (Supplementary Table [S8](#MOESM2){ref-type="media"}). This region CoLOC1 (Supplementary Table [S3](#MOESM2){ref-type="media"}) also overlaps with QTL q23k-BP-A3-1 detected in the biparental population (Fig. [3a](#Fig3){ref-type="fig"}). Comparison of the marker-trait segregation in the NAM panel with marker and resequencing data for the six parental genotypes confirmed the expected pattern and revealed a putatively deleted interval within a part (8%) of the QTL region of the susceptible parent. This deletion corresponds to a 30 kb region containing 4 protein-coding genes (Fig. [3c](#Fig3){ref-type="fig"}, red box) within the entire QTL interval of 382 kb containing a total of 90 genes (Fig. [3b](#Fig3){ref-type="fig"}). However, from 5 markers (3 SNaPs, 2 SNPs) within the LD block of the co-localizing QTL interval CoLOC1, only the SNaP marker Bn-A03-p10964394 within the deleted region in R53 is significantly associated with *V. longisporum* resistance, suggesting that presence/absence polymorphism is involved in resistance. Based on the resequencing data of the parental lines, two to four genes are affected by deletions within the QTL interval (Fig. [3c](#Fig3){ref-type="fig"}), namely BnaA03g21190D (coding for an uncharacterized protein), BnaA03g21200D (coding for a skp1-like protein involved in ubiquitin-dependent protein catabolic processes), BnaA03g21210D (coding for aquaporin pip1-2 protein involved in transporter activities), BnaA03g21220D (coding for an uncharacterized protein) and BnaA03g21230D (coding for an ATP-dependent helicase brm-like protein involved in DNA binding).Figure 3Comparison of SNP/SNaP marker polymorphism, sequence coverage and gene content at the co-localizing QTL CoLOC1 on *B. napus* chromosome A03. (**a**) Positions of QTL for *V. longisporum* resistance in the NAM panel (purple) and in the biparental mapping population ExR53-DH (green). (**b**) Chromosome interval and annotated genes in the QTL region in the *B. napus* Darmor-*bzh* v4.1 reference genome. (**c**) Allele patterns of reference-anchored markers in parents of six NAM subpopulations along with resequencing coverage data in the six parents. Red boxes indicate confirmed segmental deletions involving genes. Failed alleles represent SNaP (absence) alleles in susceptible parents.

SNP/SNaP marker patterns within *V. longisporum* QTL regions were also compared with parental whole-genome sequence coverage in the two parents of the biparental population ExR53-DH. Complete or partial presence/absence events were reconfirmed by resequencing coverage analysis of the two parents for 12 out of 18 reference-anchored SNaP markers (67%) and for 5 out of 6 QTL regions harbouring one or several reference-anchored SNaP markers (83%, Supplementary Table [S9](#MOESM2){ref-type="media"}). The 2.5 cM QTL interval q23k-BP-A3-1 spans 3,45 Mb on the Darmor-*bzh* reference, containing 683 genes (Supplementary Table [S9](#MOESM2){ref-type="media"}). Part of this region is shown in detail in Fig. [2](#Fig2){ref-type="fig"} as co-localizing QTL region CoLOC1. Within CoLOC1 a total of 382 kb, containing 90 genes, overlaps in the biparental and multi-parental QTL analyses (see above, Supplementary Table [S3](#MOESM2){ref-type="media"} and Fig. [3](#Fig3){ref-type="fig"}).

Using genomic resequencing data from 52 accessions^[@CR28]^, including the six NAM parents investigated in our study, we tested the software package SGSGeneLoss^[@CR29]^ for gene loss calling and classification of genes as PAV genes or non-PAV genes. Parameters of 5% minimum gene size coverage (lostCutoff = 0.05) and minimum coverage of two reads for a gene (minCov = 2) was used by Hurgobin *et al*.^[@CR19]^ to call a gene as present in a genotype. However, using these parameters for the 6 NAM parents used in our study, and comparing gene loss calling with PCR amplification data for 11 gene fragments from two genomic regions on chromosome A03 and C04 (Gabur *et al*.^[@CR24]^, Fig. [3](#Fig3){ref-type="fig"} and Supplementary Fig. [S3](#MOESM1){ref-type="media"}) revealed an inaccurate gene fragment loss classification in 9 of 66 cases (86% accuracy) and no PAV was called for the chromosome C04 regions. This suggests that the chosen parameters were not stringent enough in the six genotypes to distinguish genuine gene loss from data processing noise due to misalignments. Hence, gene loss was accordingly underestimated using these SGSGeneLoss program parameters (and using Darmor-*bzh* v.4.1 as a reference). We therefore developed a customized pipeline which calibrated gene loss calling parameters based on PCR wet lab data and also included the gene size in the calculations to reduce further alignment biases. This procedure increased accuracy of gene loss calling to 96%. Both methods were then used for gene loss calling in the six NAM parents, Express617 and R53, firstly across the whole genome and then within identified QTL regions. Using our modified approach, 49% of total genes (49,701 of 101,039) were estimated to be affected by gene loss and 51% of genes within the 28 QTL regions (1,334 of 2,601) were estimated to be affected by gene loss (PAV genes) between the 6 NAM parents. Between Express 617 and R53, 23% of total genes (23,772 of 101,039) and 23% of genes within 17 QTL regions (590 of 2,646) were estimated to be affected by gene loss. A total of 144 genes were harboured in the 5 QTL intervals which co-localized between the biparental and multi-parental QTL mapping. Of these, 74 (52%) in the NAM panel and 37 (26%) in the ExR53-DH panel were estimated to be affected by PAVs. The genes estimated to be affected by PAV overlap in 20% of cases within the 5 co-localizing QTL regions between the two populations (Supplementary Table [S10](#MOESM2){ref-type="media"}).

For all genes including PAV genes (144) within the co-localizing QTL regions, 30% of genes have no annotation (43 out of 144). This could suggest that some of the gene models are bioinformatics artefacts. However, from the 43 not annotated genes, 38 (92%) had expression in pan-transcriptome data from He *et al*.^[@CR30]^ or in RNASeq data from the ExR53-DH population (mock or *V. longisporum*-infected, Supplementary Table [S11](#MOESM2){ref-type="media"}). This suggests that the lack of gene annotation does not necessarily mean that the gene models are incorrect. 12% of all gene models in Darmor-*bzh* could not be annotated using Blast2GO. This is in contrast to the 30% of genes we found in the co-localizing QTL regions which could not be annotated using Blast2GO. Thus, the prevalence of these transcript-validated genes in QTL regions which cannot be annotated from public databases seems to be not a random phenomenon.

Discussion {#Sec8}
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The reanalysis and integration of SNP array data, short-range Illumina sequencing data and long-range Bionano optical mapping data with QTL data provided new insights into the importance of gene PAV for disease resistance expression against Verticillium stem striping in oilseed rape. QTL analyses including presence/absence markers increased the power of detection for *V. longisporum* resistance in two *B. napus* breeding population. This type of variation (genic or nongenic) was previously reported to be associated with quantitative resistance against two other major fungal disease of oilseed rape 24 suggesting that it is a common and prevalent phenomenon in *B. napus*.

The recent allopolyploid crop species *B. napus* shows a strong abundance of SV and genomic rearrangements^[@CR18],[@CR31]^. In maize, Beló *et al*.^[@CR32]^ used comparative genomic hybridization arrays for detailed genome-wide analysis of SV, and a more recent study discovered some degree of CNV between 100 analyzed lines across more than 90% of the maize genome^[@CR33]^. However, to date it is unknown how widespread this phenomenon is in other, older polyploid crop species and it is unclear for *B. napus* and most crops how many genes, what kind of genes and what traits are predominantly affected. Particularly for complex polyploid crops, considerably higher numbers of diverse, high-quality reference assemblies are required before reliable pangenomic analysis of genome-wide gene PAV becomes possible. Hurgobin *et al*.^[@CR19]^ classified 38% of all *B. napus* genes in a diversity set of 53 genotypes as PAV-affected, including genes involved in important agronomical traits. In the NAM parental lines, Express617 an R53 we classified 49% and 23% of all genes as PAV-affected by using a PCR-calibrated gene loss bioinformatics pipeline. As five of the 6 NAM parents are resynthesized from exotic *Brassica* species a high gene loss could be expected which are known to exhibit an elevated ratio of rearrangements and SVs^[@CR17],[@CR18]^. Surprisingly the percentage of genes affected by PAV was similar within the whole genome, the *V. longisporum* QTL for both populations suggesting that the prevalence of gene PAV is a general phenomenon randomly distributed throughout the genome and not restricted to *V. longisporum* resistance or other traits. For gene cluster, gene families and LRR genes involved in qualitative disease resistance it has documented that gene PAV is multiple times higher than on average within the entire genome^[@CR19]^. However, this seems not to apply for quantitative resistance where diverse genes involved in complex pathways influence resistance responses.

Different mixtures of *V. longisporum* lineages/pathotypes were used for disease resistance screening in the two different *B. napus* mapping populations. Comparison of QTL regions detected by including SNaP markers between the biparental and the multi-parental populations revealed a considerable number of common QTL. This indicates broad-spectrum, lineage/pathotype-independent resistance reactions in genetically diverse germplasm which are of great interest for commercial resistance breeding. Thus we focused for a more detailed analysis on these 5 co-localizing QTL regions, which harbour 2 to 90 genes each within the co-localizing section of the QTL intervals. Based on the quantitative genetic nature of the disease resistance, we assumed that single specialized genes involved in very different biological functions from each QTL interval will not contribute to *V. longisporum* resistance, but rather a number of genes from common biosynthesis pathways or with common biological functions from one or all QTL intervals. To prioritize candidate PAV genes putatively involved in broad-range resistance expression we performed a gene ontology enrichment analysis for all QTL-associated genes, separately for both populations, and also for the genes from the 5 colocalizing QTL intervals. The enriched GO terms for the genes from the QTL regions were quite different between the two populations, being mainly related to chitin and cell-wall metabolism for the biparental ExR53-DH population and mainly related to cell growth for the multi-parental NAM population. This difference is not unexpected, as the disease resistance screenings are known to be highly susceptible to the environment and slightly different mixes of *V. longisporum* lineages/pathotypes were used for inoculation of the two populations. However, the result might also indicate that common as well as different resistance mechanisms are activated by different pathotypes.

Within the 144 genes from the co-localizing QTL putatively involved in pathotype-independent resistance, we found a number of genes coding for selenium-binding proteins, for plant defensin proteins and for expansin proteins have been found to be affected by PAV. In *A. thaliana*, expression of selenium binding proteins is tightly linked to detoxification processes related to oxidative stress^[@CR34]^. Plant defensins are major components of the innate immune system of plants, are involved in the cell wall integrity signaling pathway and often show a potent, broad-spectrum antifungal activity^[@CR35]^. The antifungal protein RsAFP2 from *Raphanus sativus*, a close relative of *B. napus*, has been described to exhibit antifungal activitiy against the fungus *V. dahliae*, which is closely related to *V. longisporum*^[@CR36]^. Furthermore, a synthetic defensin expressed in *A. thaliana* has also been shown to exhibit antifungal activity against *V. dahliae*^[@CR37]^. Expansins mediate cell wall-loosening and down-regulation of an expansin-like protein in *A. thaliana* that has been shown to increase resistance against nectotrophic fungi^[@CR38]^. This suggest that genes affected by PAV and involved in cell wall integrity and signaling at the cell wall surface are key components of *V. longisporum* broad-spectrum resistance in *B. napus*.

We found that in the majority of cases gene presence was associated with resistance against *V. longisporum*. However, interestingly one of the strongest QTL, q23k-BP-C2-3 (R^2^ = 23%) was mapped using SNaP markers only. In this case, the absence alleles were inherited by the parent R53 and associated with resistance. Bionano Optical Mapping confirmed the deletion of a region containing 70 genes in the parent R53. This phenomenon has been rarely reported for quantitative disease resistance, but has previously been shown for Sclerotinia stem rot in *B. napus*^[@CR24]^ and for three other fungal pathogens in *Medicago truncatula*^[@CR39]^.

Surprisingly, we found another QTL region containing a total of 7 nucleotide binding site-leucine-rich repeat (NLR) resistance genes (TIR-NBS-LRR) to be affected by PAV and to harbour a QTL on chromosome C09 for *V. longisporum* resistance in the NAM population in this study. This region was also described by Samans *et al*.^[@CR18]^ to be part of two gene clusters of 14 and 8 TIR-NBS-LRR genes on *B. napus* chromosome C09, which are frequently deleted in natural *B. napus* compared to synthetic *B. napus* accessions. NBS-LRR genes are frequently described to be involved in monogenic disease resistance. For NLR resistance genes it is well known that they are often organized in clusters or tandem repeats in a number of plant species and crops and numerous studies have shown that fitness costs can lead to multiplication and deletion of gene family members, such as in *A. thaliana*^[@CR40],[@CR41]^ and in *B. napus*^[@CR17]^. *V. longisporum* resistance in the NAM and ExR53-DH population is quantitatively inherited. However, similar co-segregation of TIR-NBS-LRR genes with QTL have been described for other crops, e.g. for soybean^[@CR42]^, barley^[@CR43]^, potato^[@CR44]^ which might result from evolution and local genome diversification of genes involved in qualitative and quantitative disease resistance mechanisms.

The prioritization of these candidate PAV genes mainly involved in cell wall integrity, growth and modification is consistent with our earlier findings that QTL for the concentration of soluble simple phenylpropanoids, which are putative precursors and degradation products of cell-wall modifications and cross-linking, are co-localizing with major resistance QTL in the biparental ExR53-DH population. In addition, the concentrations of some of these cell wall-associated compounds are significantly correlated with *V. longisporum* resistance^[@CR45]^. Further functional characterization of PAV genes from QTL regions may help to improve our understanding of disease resistance mechanisms and to improve fungal resistance in *B. napus* by exploitation for in future plant breeding programs.

Materials and Methods {#Sec9}
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Phenotyping for *Verticillium longisporum* resistance in *B. napus* {#Sec10}
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Resistance phenotyping was conducted in the greenhouse at Georg August University Göttingen, Germany. In order to represent a broad range of pathogenicity traits occurring in oilseed rape fields, a spore suspension mixture of *V. longisporum* isolates VL43 (lineage A1/D1, North Germany), VLS3 (lineage A1/D1, Sweden) and PD589 (lineage A1/D3, Japan)^[@CR46],[@CR47]^ with a density of 1 × 10^6^ spores/ml concentrations was used to inoculate the 200 NAM lines applying the root-dipping method^[@CR48]^ in four experiments. Each experiment included 20 inoculated and 20 control plants for each tested genotype. Rating of symptoms was done weekly over a 4-week period, using the 1--9 disease scoring scale described by Eynck *et al*.^[@CR48]^. Resistance screening for the biparental population ExR53-DH was performed similarly using a mixture of isolates VL40 and VL43 (both lineage A1/D1) with 202 DH lines as described in detail by Obermeier *et al*.^[@CR45]^.

High molecular weight DNA isolation for optical mapping {#Sec11}
-------------------------------------------------------

High molecular weight (HMW) DNA isolation was carried out for Express617 and R53 according to the IrysPrep™ Plant Tissue-Nuclei protocol provided by Bionano Genomics. Young leaves (approximately 2 grams) were harvested from dark-treated rapeseed plants. The harvested leaves were immediately fixed with 2% formaldehyde followed by homogenization in isolation buffer containing PVP-10, BME and Triton X-100. The isolated nuclei were then purified on Percoll cushions. Purified nuclei were further embedded in an agarose matrix. Agarose plugs were further subjected to proteinase K treatment followed by rigorous washings steps. Finally, HMW DNA was recovered by melting the plugs using GELase™ (Epicentre) treatment. An additional drop dialysis step was performed to ensure ultra-clean DNA. High molecular weight DNA was further subjected to sequence-specific nick-labeling using the IrysPrep™ Labeling-NLRS protocol provided by Bionano Genomics. HMW DNA was subjected to digestion by the single-stranded nicking endonuclease *Nt.BspQI* (recognition site GCTCTTC). The nicks created by *Nt.BspQI* were then repaired using fluorophore-labeled nucleotides. Nicked and labeled single DNA molecules were subsequently loaded onto an IrysChip for imaging on the Bionano Genomics Irys system.

Bio-informatics analysis for Bionano optical mapping data {#Sec12}
---------------------------------------------------------

DNA molecule images generated from the Irys system were computationally translated into single-molecule optical maps. These single molecules were then assembled into consensus maps using the dedicated IrysSolve pipeline (v5134) provided by Bionano Genomics. An *in silico* optical map was generated for the Darmor-*bzh* v. 4.1 ref. ^[@CR17]^ using Knickers v1.5.5 and was used to calculate noise parameters for the final assembly. Optical map assemblies from Express617 and R53 were finally aligned to the Darmor-*bzh* reference using the runCharacterize script provided by Bionano Genomics, with the settings published by Kawakatsu *et al*.^[@CR49]^. The alignment was imported into Bionano IrysView (v2.5.1.29842) software for visualizing and detecting structural variations between Express617, R53 and the Darmor-*bzh B. napus* reference genome.

Global RNA-Seq analysis for confirmation of gene expression {#Sec13}
-----------------------------------------------------------

Two contrasting lines from the mapping population ExR53-DH, DH41 (partially resistant genotype) and DH94 (susceptible genotype), were grown for four weeks after inoculation with *V. longisporum*- (isolate VL43) and mock-inoculation in two independent experiments under different environmental conditions (optimal, drought stress and heat stress). For each mock and *V. longisporum*-inoculated plants, pooled hypocotyl samples from 20 plants/line were harvested 28 days after inoculation. In total, 16 samples were analyzed and total RNA was extracted. Sequencing libraries were produced by service provider LGC Genomics GmbH (Berlin, Germany) and in total 1,022 Million 100 bp pair-end raw reads were obtained by Illumina HiSeq 2000 3′end sequencing. Alignment of reads to the *B. napu*s reference genome Darmor-*bzh* v4.1 and all statistical analysis were performed using CLC Genomics Workbench version 9.0 (QIAGEN Bioinformatics CLC bio, Aarhus, Denmark).

Genetic mapping and QTL analysis {#Sec14}
--------------------------------

244 DH lines from the F1 of the cross Express617 × R53 and the two parents were analyzed with the Brassica 60 k Illumina Infinium array^[@CR26]^. DNA was extracted from leaves using the CTAB method^[@CR50]^ and array genotyping assays were outsourced to TraitGenetics (Seeland, Germany). SNP calls with \>85% failed calls across all 244 genotypes were removed from further analyses. Also, SNP probe calls with \>90% or \<10% of a single allele across the population were removed, leaving a total of 22,064 SNP markers. From these, 4,654 SNP probes (21.1%) showed a segregation pattern with one allele displaying a failed call and 17,410 SNP probes showed a normal biallelic segregation. A "SNaP map" was created from all quality-filtered 22,064 markers (2,714 marker bins), including biallelic and presence/absence polymorphisms, while a "SNP map" was created using only the 17,410 markers showing presence/presence polymorphism (2,176 marker bins). Genetic maps were created using the software MSTMap^[@CR51]^ applying the kosambi distance function and a cut-off *p* value of 10^−30^. QTL analyses were performed using the software QGene 4.3.9 and 4.4.0^[@CR52]^ applying composite interval mapping with a scan interval of 1 milliMorgan and automatic forward cofactor selection. Mean normalized AUDPC values from the four *V. longisporum* greenhouse resistance screenings were used as trait input data^[@CR45]^.

Genome-wide association studies {#Sec15}
-------------------------------

The NAM panel was also genotyped using the 60 K Illumina Infinium Brassica SNP array as described above and data was filtered according to Gabur *et al*.^[@CR24]^. Using the Darmor-*bzh* reference v4.1 we anchored 28,073 SNP markers by BLASTn using CLC Genomics Workbench v. 9.0 (Qiagen Bioinformatics). The SNP map contained 18,068 markers, the SNaP map contained 21,695 markers. Association analyses were conducted using the R package GenABEL^[@CR53]^. A mixed linear model approach that increases detection power^[@CR54]^ was adjusted for population stratification by including the kinship matrix and the first two principal components as covariates^[@CR55]^. Stringent significance cutoff values were set at a false discovery rate (FDR) correction of 10%^[@CR56]^. To reduce the type II error rate, we also captured the SNP-trait associations for disease resistance using an arbitrary threshold of −log10(p) ≥ 3.

Linkage disequilibrium (LD) analysis and haplotype construction {#Sec16}
---------------------------------------------------------------

Whole genome linkage disequilibrium (LD) was calculated using the squared allele-frequency correlations (r^2^) between pairs of SNPs. Only markers with a maximum of 10% missing data and MAF ≥ 0.05 were included in the analysis. Haplotype patterns were assessed for SNP and SNaP markers that showed significant marker trait association at the adjusted Bonferroni threshold of −log10(P-value) ≥ 4.33. Haplotype blocks were defined with the confidence interval method described by Gabriel *et al*.^[@CR57]^ in Haploview version 4.2^[@CR58]^ and visualized with the R package LDheatmap^[@CR59]^.

Resequencing and coverage analysis {#Sec17}
----------------------------------

The Illumina 250 bp paired-end raw sequencing data for the two parents of the biparental population, Express617 and R53, was described previously by Stein *et al*.^[@CR60]^, while Illumina 100 bp paired-end raw sequencing data for the 6 NAM parents was described by Schmutzer *et al*.^[@CR29]^. Sequences were aligned to the Darmor-*bzh* v. 4.1 reference using CLC Genomics Workbench v. 9.0 (Qiagen Bioinformatics). Genes from QTL regions were classified as affected or not affected by presence/absence variation based on coverage analysis of the WGS data from the parental lines. Coverage differences were calculated using the bedtools software package v. 2.27.0 with multiBamCov. A minimum cutout threshold of 1.5 aligned reads was used to differentiate between gene presence and absence. The threshold was selected using PCR data for calibration available for the six NAM parental lines from Gabur *et al*.^[@CR24]^. Additionally, we used the SGSGeneLoss v0.1 described by Golicz *et al*.^[@CR30]^. The visualization in Fig. [1](#Fig1){ref-type="fig"} was performed using the software MapChart v.2.3, while Fig. [3](#Fig3){ref-type="fig"} was generated using CLC Genomics Workbench 9 track lists. Identification of homoeologous exchanges (HE) was performed using the method described by Samans *et al*.^[@CR18]^. Further analysis was partly performed by using the R package 'gsrc'^[@CR61]^.

Gene ontology enrichment analyses {#Sec18}
---------------------------------

To produce gene ontology information for all *B. napus* genes, 101,039 Darmor-*bzh* peptide sequences (Brassica_napus.annotation_v5.pep.fa.gz) were downloaded from the website: <http://www.genoscope.cns.fr/brassicanapus/data/> and were used as input for Blast2Go v. 4.1.9. The R package topGO v1.0^[@CR62],[@CR63]^ was used for gene ontology enrichment analysis.

Supplementary information
=========================

 {#Sec19}

Supplementary figures S1, S2 and S3. Supplementary Tables S1 to S11.

**Publisher's note** Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

These authors contributed equally: Iulian Gabur and Harmeet Singh Chawla.

Supplementary information
=========================

is available for this paper at 10.1038/s41598-020-61228-3.

We thank Jutta Schaper (Georg August University Goettingen) for excellent technical assistance in resistance phenotyping, Matthias Frisch (Justus Liebig University Giessen) for advice in statistical analyses, Philip Howard and Andrew Leitch (Queen Mary University of London) for processing HMW DNA samples and analysis on a Bionano Genomics Irys System for optical mapping. Iulian Gabur acknowledges funding through a scholarship from the DAAD. Funding was provided by the German Federal Ministry of Food and Agriculture (BMEL grant 28-1-45.051-10) and from the German Federal Ministry of Education and Research (BMBF grant 031A325 for the German-French Plant-KBBE consortium GEWIDIS and grant 0315964 for the national consortium PreBreedYield).

C.O. and R.S. designed the research. A.v.T. and D.T.L. produced the phenotyping data. H.S.C. performed the Optical mapping and bioinformatics analysis. D.T.L. performed the RNASeq experiment. I.G. performed genetic and other bioinformatics analyses. I.G., H.S.C. and C.O. analyzed the data. I.G. and C.O. wrote the original draft; all authors discussed and approved the final manuscript.

The authors declare no competing interests.


Polyphosphazene-based nanocarriers for the release of agrochemicals and potential anticancer drugs.
The synthesis and characterisation of novel polyphosphazene nanocarriers, based on hydrophilic polyalkylene oxide Jeffamine M1000 and hydrophobic steroids with a glycinate linker for pH-controlled release of diosgenin and two brassinosteroids (DI31 and S7) with agrochemical and potential anticancer activity, is hereby described. Polyphosphazenes carrying approximately 17 wt% of DI31 or S7 self-assembled in water to form 120-150 nm nanoaggregates, which showed an excellent plant growth effect on radish cotyledons due to sustained delivery of approximately 30% of the agrochemicals after 4 days. Cytotoxic evaluation showed that all polymers carrying steroids and Jeffamine M1000 resulted in strong to moderate toxicity to MCF-7 cancer cells and were non-toxic to primary human lung fibroblast cells at 0.1 to 0.025 mg mL-1. Thus, DI31 and S7 bearing polymers applied at 10-4 to 10-6 mg mL-1 for delivery of recommended DI31 or S7 quantities to crops should be harmless to humans. Particularly, DI31 and S7 bearing polymers with strong cytotoxicity on MCF-7 and non-toxicity on primary human lung fibroblasts, good cell uptake after 6 hours, proper hydrodynamic sizes between 100 and 200 nm, and slow sustained release of cytotoxic drugs (DI31, S7) in acidic conditions might potentiate their accumulation in cancer tissues with good antitumour effects and minor side effects. These results demonstrated that preparation of brassinosteroid bearing polymers is a promising strategy for the preparation of better agrochemicals with reduced pollutant impact on sustainable agriculture and potential anticancer formulations based on analogues of brassinosteroids.


Who do Liverpool need to sign to retain the title? Keep up with all the Reds news with our free daily newsletter Sign up Thank you for subscribing We have more newsletters Show me See our privacy notice Invalid Email

Liverpool are 'lucky' to have Jurgen Klopp as manager, says Reds legend Steven Gerrard.

The Anfield great returned to Merseyside to train with his old club during the MLS off-season, and was impressed with what he saw from the German.

Klopp took over from Brendan Rodgers after an indifferent start to the season for Liverpool, and has endured a run of up-and-down results since taking over.

He oversaw a 6-0 demolition of Aston Villa on Sunday, and Gerrard is hopeful he can be the man to bring the success the club has craved in recent years.

In pictures: Aston Villa 0-6 Liverpool

"I'm a huge fan, I was a big fan at Dortmund and I think we're very lucky to have him at Liverpool," the LA Galaxy midfielder told CNN.

"I struck up a bit of a relationship with him. I'm hoping with my fingers crossed that as a Liverpool fan he can get us trophies and bring us some success."

Gerrard has also spoken about his post-retirement plans, including the possibility of him entering coaching and possibly even management when he finally hangs up his boots.

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He is doing his badges, but admits there is a lot of uncertainty about which path he will follow.

"As far as my coaching, management dreams and aspirations are concerned they're better parked until I retire," he added.

"I don't know if I'll be good enough and I don't know what roles are going come my way in the future."


Maintaining a clean home isn't an easy task, and whether you're holding down the bachelor pad or making the job of cleaning up after the family a little easier for everyone, one of the most effective tools to keep your place clean is a solid vacuum cleaner.

To make life around your humble abode a bit more enjoyable, we've assembled a list of the absolute best vacuums that are currently available on the market – you'll find something that will help you with virtually every type of messy situation.

Even if you're not the one who runs your new machine, you can bet your wife or girlfriend will love you for the upgrade!

Also, we'll constantly be updating this list to ensure the best ones are available – so keep coming back to see what we've discovered.

The Bissell CleanView vacuum is a lightweight vacuum that can get things clean in the first pass. It uses the cyclonic system to keep long lasting powerful suction. You can empty the tank easily thanks to the quick release tank.

Dirt Devil Simpli-StikThe Dirt Devil Simpli-Stik is a versatile 3 in 1 vacuum cleaner that can be used as a stic vac, handheld vac, and a utility vac. The vacuum is very lightweight, making it easy for carrying around the house or shop. It measures 7 x 11 x 45 inches.

The Dirt Devil Dynamite Plus vacuum cleaner weighs less than 8 lbs and can clean carpets and bare floors. It has a HEPA filter and an easy to empty container. You can clean carpets and bare floors with ease. You can move easily around your house without needing to unplug it to move it to another room thanks to the 25 ft cord.

Today only. Amazon.com offers the Shop-Vac 9633400 6.5-Peak HP Ultra Pro Series 12-Gallon Wet or Dry Vacuum with Detachable Blower for just $89 with FREE shipping. That's 60% off list price and the best price we've seen on this model all year.

This Shop Vac is a 6.5-Peak HP Ultra Pro Series 12-Gallon Wet or Dry Vacuum with Detachable Blower. Great for any workshop. It has a large on/off switch making it easy to turn it on or off while you're holding something or need to reach over and start it while you're far away from it. This vacuum can switch from suck to blow easily and comes with a blower nozzle.

The PowerSmith PAVC101 10 Amp Ash Vacuum is ideal for cleaning up warm and cool ash out of fireplaces. It can also do wood stoves, pellet stoves, and bbq grills. It has a replaceable fire-resistant filter that can filter out the finest dust and ash.

The Vacmaster is a detachable blower wet/dry vacuum with a 12 gallon canister and 5 peak HP. This vacuum manufacturer has a really cool name and a vacuum that can do it all. You can detach the top part of the vacuum and turn it into a leaf blower to get those messy jobs done quicker.

If you’ve ever known a guy that’ll help you out of a jam and still be that cool James Bond sort of character in the background of your life, then you’ve obviously met our Johnny “G Money” Garcia.

Born and raised in Orange County, California, Mr Garcia (if you please) specializes in deals on Man Toys, Gaming, Cars, Outdoors, and Grooming on Faveable. With extensive experience as a web developer, Johnny is a detail freak. He’ll spend hours combing through hundreds of sites and pages to share the latest, coolest men stuff around.

In his spare time, you’ll catch Johnny hunting down travel deals and driving something that could possibly be mistaken for a Batmobile.

It’s what you didn’t know you were missing

We spend thousands of hours researching and testing products to create the most authoritative product ranking on the web. Over 5 million users and counting have trusted our rankings and product reviews.


Q:

Jackson JSON lib how to specify start of the element to deserialize

I am using jackson to deserialize a json data. I am using objectmapper to de-serialize the data and want to ignore unknown properties using mapper.configure(DeserializationConfig.Feature.FAIL_ON_UNKNOWN_PROPERTIES, false);
Is there a way in Jackson to specify the start of the element which I want to deserialize? My JSON data has some more elements in it but I am interested only in some element to deserialize.

A:

Object structure has to match JSON structure, so oftentimes it is most convenient to just use wrapper classes if necessary. Something like:
class Response {
  public Thing value;
}
class Thing {
  public String name;
}

so even if you just wanted "name", you would do something like:
Response resp = mapper.readValue(jsonInput, Response.class);
String name = response.value.name;

If so, you can omit defining properties you don't care about; or define them and not use.


NRSP

The tireless Tom Harris, executive director of the NRSP (Not Really Science People), has discovered the right-wingy Alberta website Free Dominion, “The Voice of Principled Conservatism.” Harris popped up on FD a week ago, pleading for $10 donations that might help him argue that his mysteriously funded astroturf group is actually a grassroots organization with wide public support.

Here 's a bit of pathos, the lonely lament of a couple of public relations has-beens who can no longer get anyone (other than Financial Post Editor Terence Corcoran) to answer their phone calls.

The mournful victims here are Dr. Tim Ball and Tom Harris (right), the nominal brains behind the Natural Resources Stewardship Project (NRSP, or Not Really Science People). In this article, printed in the business pages of the National Post, Ball and Harris complain that, even among Canadian Conservatives, “open discussion of the vast uncertainties (my emphasis) in climate change science has been declared off-limits.” Which is to say, the Tories no longer invite industry-funded climate-change deniers like Tim Ball to their policy sessions.

Climate denier, retired geography professor and chair of the Natural Resources Stewardship Project (NRSP or Not Really Science People), Dr. Tim Ball, has been less in evidence lately: perhaps Canada's biggest media company (CanWest Global) is reticent to publish his meanderings after being added to the lawsuit against University of Lethbridge Professor (and defender of truth) Dan Johnson.

But apparently Ball is still chatting with the converted, spreading disinformation and (this is juicy) claiming that we at the DeSmogBlog are tracking his mail.

We have received this excerpt from a ClimateSceptics listserv which you can only access with an account. (We don't have one, but it would appear from the anonymous hits in our inbox that at least a couple of actual skeptics do.)

Natural Resources Stewardship Project (think: Not Really Science People) Executive Director Tom Harris is back in the only venue that seems willing to give him a soapbox, the “news” site Canada Free Press. And he's up to the same tawdry tricks as ever.

Ball and Harris go on the attack against Canada's Johanne Gélinas, Canada's Commissioner of Environment and Sustainable Development. Gélinas, whose job is to audit “how well the federal government is meeting its environmental and sustainable development commitments,” earned the NRSP's wrath by releasing a report in September, suggesting that the last federal adminstration had done an appalling job dealing with climate change. Ball and Harris were particularly offended that Gelinas said this:

"Fossil-fuel companies have spent millions funding anti-global-warming think tanks, purposely creating a climate of doubt around the science. DeSmogBlog is the antidote to that obfuscation." ~ BRYAN WALSH, TIME MAGAZINE


Synthesis and amnesia-reversal activity of a series of 7- and 5-membered 3-acylamino lactams.
A series of 3-(acylamino)-epsilon-caprolactams and 3-(acylamino)-2-pyrrolidinones was synthesized. Some of these compounds reversed at different degrees electroconvulsive shock- and Scopolamine-induced amnesia, using a step-through passive avoidance in mice. Classical nootropic drugs, i.e., Aniracetam, Oxiracetam, and Piracetam, were used as reference compounds. Within the analyses of data performed, we introduced a new parameter, the confrontation index (CI), which is a function of Mann-Whitney's U statistic. The CI permits a common scale of activity of substances to be generated, independently of probabilistic hypotheses, with higher scores representing higher activities. The most active compounds were characterized by the formylamino and [3-(trifluoromethyl)benzoyl]amino groups in the 3-position of the ring. None of the substances assayed showed any effect on spontaneous behavior and neurovegetative system.


Gene therapy includes any one or more of: the addition, the replacement, the deletion, the supplementation, the manipulation etc. of one or more nucleotide sequences in, for example, one or more targeted sites—such as targeted cells. If the targeted sites are targeted cells, then the cells may be part of a tissue or an organ. General teachings on gene therapy may be found in Molecular Biology.
By way of further example, gene therapy also provides a means by which any one or more of: a nucleotide sequence, such as a gene, can be applied to replace or supplement a defective gene; a pathogenic gene or gene product can be eliminated; a new gene can be added in order, for example, to create a more favourable phenotype; cells can be manipulated at the molecular level to treat cancer or other conditions—such as immune, cardiovascular, neurological, inflammatory or infectious disorders; antigens can be manipulated and/or introduced to elicit an immune response—such as genetic vaccination.
In recent years, retroviruses have been proposed for use in gene therapy. Essentially, retroviruses are RNA viruses with a life cycle different to that of lytic viruses. In this regard, when a retrovirus infects a cell, its genome is converted to a DNA form. In other words, a retrovirus is an infectious entity that replicates through a DNA intermediate. More details on retroviral infection etc., are presented later on.
As mentioned above, retroviruses have been proposed as a delivery system (otherwise known as a delivery vehicle or delivery vector) for inter alia the transfer of a NOI, or a plurality of NOIs, to one or more sites of interest. The transfer can occur in vitro, ex vivo, in vivo, or combinations thereof. When used in this fashion, the retroviruses are typically called retroviral vectors or recombinant retroviral vectors. Retroviral vectors have even been exploited to study various aspects of the retrovirus life cycle, including receptor usage, reverse transcription and RNA packaging.
In a typical recombinant retroviral vector for use in gene therapy, at least part of one or more of the gag, pol and env protein coding regions may be removed from the virus. This makes the retroviral vector replication-defective. The removed portions may even be replaced by a NOI in order to generate a virus capable of integrating its genome into a host genome, but wherein the modified viral genome is unable to propagate itself due to a lack of structural proteins. When integrated in the host genome, expression of the NOI occurs—resulting in, for example, a therapeutic effect. Thus, the transfer of a NOI into a site of interest is typically achieved by: integrating the NOI into the recombinant viral vector; packaging the modified viral vector into a virion coat; and allowing transduction of a site of interest—such as a targeted cell or a targeted cell population.
It is possible to propagate and isolate retroviral vectors (e.g., to prepare suitable titres of the retroviral vector) for subsequent transduction of, for example, a site of interest by using a combination of a packaging or helper cell line and a recombinant vector.
In some instances, propagation and isolation may entail isolation of the retroviral gag, pol and env genes and their separate introduction into a host cell to produce a “packaging cell line”. The packaging cell line produces the proteins required for packaging retroviral RNA. However, when a recombinant vector carrying a NOI and a psi region is introduced into the packaging cell line, the helper proteins can package the psi-positive recombinant vector to produce the recombinant virus stock. This is commonly known as a “producer cell”. The vector can be used to infect cells to introduce the NOI into the genome of the cells. The recombinant virus whose genome lacks all genes required to make viral proteins can infect only once and cannot propagate. Hence, the NOI is introduced into the host cell genome without the generation of potentially harmful retrovirus. A summary of the available packaging lines is presented in “Retroviruses”.
The design of retroviral packaging cell lines has evolved to address the problem of inter alia the spontaneous production of helper virus that was frequently encountered with early designs. As recombination is greatly facilitated by homology, reducing or eliminating homology between the vector and gag/pol has reduced the problem of helper virus production.
More recently, packaging cells have been developed in which the gag/pol and env viral coding regions and the viral vector are carried on separate expression plasmids that are independently transfected into a packaging cell line so that three recombinant events are required for wild type viral production.
Transient transfection can be used to make vectors. In this regard, transient transfection has been used if the vector or retroviral packaging components are toxic to cells. Components typically used to generate retroviral vectors include a plasmid encoding the Gag/Pol proteins, a plasmid encoding the env protein and a plasmid containing a NOI. Vector production involves transient transfection of one or more of these components into cells containing the other required components. If the vector encodes toxic genes or genes that interfere with the replication of the host cell, such as inhibitors of the cell cycle or genes that induce apoptosis, it has proved difficult to generate stable vector-producing cell lines, thus transient transfection can be used to produce the vector before the cells die. However, the aforementioned technique can be problematic in the sense that the titre levels are not always satisfactory, it is difficult to make large batches of virus, and safety tests must be performed on each small batch.
In view of the toxicity of some HIV proteins, e.g., the HIV protease—which can make it difficult to generate stable HIV-based packaging cells —HIV vectors are usually made by transient transfection of vector and helper virus. Some workers have even replaced the HIV env protein with that of vesicular stomatis virus (VSV). A drawback, however, with this approach is that the VSV-G protein is quite toxic to cells.
Thus, and as indicated, retroviral vectors are used extensively in biomedical research and for gene therapy. Current methods for the production of retroviral vectors make use of the fact that the two roles of the wild-type retrovirus genome, that is protein encoding and as a template for new genome copies, can be de-coupled. Protein that is required for the assembly of new virus particles and for enzyme and regulatory functions can be produced by non-genome sequences in, for example, a mammalian packaging cell line. A genome sequence lacking the protein encoding functions is provided, so that the resulting retroviral vector particles are capable of infecting but not of replicating in a target cell. The genome sequence can also be designed for delivery and integration of a therapeutic gene. Standard methods for producing murine leukaemia virus (MLV)-based vectors, for example, include use of stably engineered cell lines expressing the gag-pol and env genes (the packaging components) of MLV. These will package a compatible retroviral vector genome introduced by transfection with an appropriate plasmid. An alternative method for producing HIV based vectors, for example, involves simultaneous transient transfection of gag-pol, env, and vector genome plasmids into suitable cells.
Although the principles of these systems are well understood, in practice the re-constructed virus assembly system often fails to generate the quantity of vector particles required in practice for use in gene therapy. Retroviral vector particles are generally harvested by removing supernatant from a culture of particle-producing cells. The resulting suspension may be concentrated with respect to the vector particles, using physical methods, but only to a limited degree as problems such as aggregation and damage tend to arise. Thus, it may only be possible to concentrate a suspension of vector particles by up to 100-fold.
The same issues arise when trying to produce a recombinant protein which is potentially harmful to the host cell in which it is being expressed. One approach that has been considered is to use an inducible system; however, this does not overcome the basic problem of toxicity, inducible production is only transient and problems have arisen from “leaky” promoters.
Vectors based on human immunodeficiency virus type 1 (HIV-1) offer a means for the delivery of therapeutic transgenes into a wide variety of cell types, both dividing and non-dividing. HIV-1 based vectors have commonly been produced either transiently or using packaging cell lines in which vector production is induced. Until recently no stable packaging cells to continuously produce high titre HIV-1 vectors were available, because it had been difficult to stably express large amounts of HIV-1 gag-pol. Furthermore, it has also proven to be difficult to continuously produce the rhabdovirus vesicular stomatitis virus G protein (VSV-G), which is most commonly used to replace HIV-1's own envelope proteins and ‘pseudotype’ HIV-1 vector particles.


Flag of Harrisburg, Pennsylvania

The flag of Harrisburg, Pennsylvania, consists of a blue field, with the city emblem centered in the middle of the flag. The emblem should not be confused with the city seal. The emblem consists of a white keystone with a red border, and incorporates the capitol dome which denotes the city's significance as the state capital of the Commonwealth of Pennsylvania. The current flag came into use in the early 1990s. Harrisburg received the All-America City Award given by the National Civic League in 1984-85 and 1990. An older version of the city’s flag incorporated the city emblem with a dark blue field and yellow border.

References

Category:Flags of cities in Pennsylvania
Flag
Category:1990s establishments in Pennsylvania


<?php
/**
 * @link https://craftcms.com/
 * @copyright Copyright (c) Pixel & Tonic, Inc.
 * @license https://craftcms.github.io/license/
 */

namespace craft\errors;

use yii\base\Exception;

/**
 * InvalidFieldException represents an exception caused by accessing a field handle that doesn’t exist.
 *
 * @author Pixel & Tonic, Inc. <support@pixelandtonic.com>
 * @since 3.5.12
 */
class InvalidFieldException extends Exception
{
    /**
     * @var string The invalid field handle.
     */
    public $handle;

    /**
     * Constructor.
     *
     * @param string $handle The field handle
     * @param string|null $message The error message
     * @param int $code The error code
     * @param \Throwable|null $previous The previous exception
     */
    public function __construct(string $handle, string $message = null, int $code = 0, \Throwable $previous = null)
    {
        if ($message === null) {
            $message = "Invalid field handle: $handle";
        }

        parent::__construct($message, $code, $previous);
    }

    /**
     * @return string the user-friendly name of this exception
     */
    public function getName(): string
    {
        return 'Invalid field';
    }
}


Adams v. Howerton

Adams v. Howerton, 673 F.2d 1036 (9th Cir. 1982), cert. denied, 458 U.S. 1111 (1982) is a decision from the United States Court of Appeals for the Ninth Circuit that held that the term "spouse" refers to an opposite-sex partner for the purposes of immigration law and that this definition met the standard at the time for rational basis review. It was the first U.S. lawsuit to seek recognition of a same-sex marriage by the federal government.

Lawsuit
In 1975, Richard Frank Adams, an American citizen, and Anthony Corbett Sullivan, an immigrant from Australia, were one of several same-sex couples that received marriage licenses from the clerk of Boulder County, Colorado. Adams then petitioned the Immigration and Naturalization Service (INS) for classification of Sullivan as an immediate relative of Adams, on the basis that they were married under Colorado law. The petition was initially denied, with a letter stating that "[Adams and Sullivan] have failed to establish that a bona fide marital relationship can exist between two faggots." A revised letter was later sent, explaining that "[a] marriage between two males is invalid for immigration purposes and cannot be considered a bona fide marital relationship since neither party to the marriage can perform the female functions in marriage." After the INS petition was ultimately denied, they filed suit against the INS in the United States District Court for the Central District of California, alleging that their marriage was valid under both Colorado law and immigration law, and that failure to recognize its validity violated the Fifth Amendment's substantive due process and equal protection components.

The district court rejected the plaintiffs' claims, noting that "Congress in its immigration statutes is not obligated to follow the law of the place where the marriage was contracted". The court also held that under Colorado law marriage is a union of "a man and a woman", agreeing with the state attorney general's opinion, and also noted that "[t]he Colorado statutes don't specifically allow, nor do they specifically prohibit, marriages between persons of the same sex." It also held that "[t]he legal protection and special status afforded to marriage (being defined as an union of persons of different sex) has historically ... been rationalized as being for the purpose of encouraging the propagation of the race."

Regarding the constitutional claims, it held that if federal law governs, then "the constitutional challenge is rejected as insubstantial", because "Congress has virtually plenary power in immigration matters and is not bound by otherwise applicable equal protection requirements". If state law governs, the court held that "the Colorado state law which rejects a purported marriage between persons of the same sex does not violate the due process or the equal protection clause of the federal constitution", on the basis that the Supreme Court's decision in Baker v. Nelson, 409 U.S. 810, 34 L.E.2d 65, 93 S Ct 37 (1972), "is of paramount importance because a state court judgment prohibiting two people of the same sex from marrying each other was appealed to the U. S. Supreme Court on various constitutional grounds, including due process and equal protection. The High Court dismissed the appeal for want of a substantial federal question. Such a dismissal is an important adjudication on the merits." The court rejected the counterargument "that some persons are allowed to marry and their union is given full recognition and constitutional protection even though the above stated justification—procreation—is not possible" because "if the classification of the group who may validly marry is overinclusive, it does not affect the validity of the classification".

Appeal
On appeal, a three-judge panel of the Ninth Circuit affirmed the District Court's opinion on alternative grounds on February 25, 1982. They decided that it was unnecessary for them to determine if the Adams-Sullivan union qualified as a marriage under Colorado law. They noted that it was clear that "Congress did not intend the mere validity of a marriage under state law to be controlling" and that "even though two persons contract a marriage valid under state law and are recognized as spouses by that state, they are not necessarily spouses for purposes of section 201(b)." They held that "[t]he term marriage ordinarily contemplates a relationship between a man and a woman" (internal quotations and citations omitted) and that "[t]he term spouse commonly refers to one of the parties in a marital relationship so defined. Congress has not indicated an intent to enlarge the ordinary meaning of those words. In the absence of such a congressional directive, it would be inappropriate for us to expand the meaning of the term spouse for immigration purposes" (internal quotations and citations omitted) and that "Congress intended that only partners in heterosexual marriages be considered spouses under section 201(b)."

In reviewing the constitutionality of the law, they rejected the plaintiffs' claim that strict scrutiny was required, on the grounds that "Congress has almost plenary power to admit or exclude aliens". They held that "Congress's decision to confer spouse status under section 201(b) only upon the parties to heterosexual marriages has a rational basis and therefore comports with the due process clause and its equal protection requirements. There is no occasion to consider in this case whether some lesser standard of review [than rational basis] should apply." They found it unnecessary to determine if Congress's justification "is because homosexual marriages never produce offspring, because they are not recognized in most, if in any, of the states, or because they violate traditional and often prevailing societal mores. In any event, having found that Congress rationally intended to deny preferential status to the spouses of such marriages, we need not further 'probe and test the justifications for the legislative decision.'"

The plaintiffs petitioned for a writ of certiorari to the United States Supreme Court, which denied the petition.

Sullivan appealed his deportation order arguing that it would cause him "extreme hardship". The Ninth Circuit rejected that argument on September 30, 1985, in a 2-1 decision in the case of Sullivan v. INS authored by Circuit Judge Anthony Kennedy, later an Associate Justice of the U.S. Supreme Court.

The plaintiffs
Adams was born in Manila in the Philippines. His family moved to the United States when he was 12, and he grew up in Long Prairie, Minnesota. He studied liberal arts at the University of Minnesota. Adams became a naturalized U.S. citizen in 1968 and was living in Los Angeles, California, in 1971 when he met Anthony Corbett "Tony" Sullivan, an Australian citizen who was visiting the U.S. on a tourist visa. They were one of six gay couples granted marriage licenses by County Clerk Clela Rorex in Boulder, Colorado on April 21, 1975. They exchanged vows in a religious ceremony as well.

To publicize their lawsuit, they appeared on the Phil Donahue Show where, according to their attorney, "people in the audience said some pretty nasty things. But they withstood it all because they felt it was important to speak out."

After losing Sullivan's appeal of his deportation order in 1985 and being denied Adams' request for residency by Australia, in 1985 they couple traveled in Europe for a year. Adams later said: "It was the most difficult period because I had to leave my family as well as give up my job of 18 1/2 years. It was almost like death". They returned to the U.S., lived in Los Angeles, and avoided high-profile activism that might attract the attention of immigration authorities. Adams worked for a law firm as an administrator until his retirement in 2010. After retirement, Adams and Sullivan made some appearances at events supporting same-sex marriage.

Adams died at his Los Angeles home on December 17, 2012, after a short illness. Sullivan survived him. On April 21, 2014, on their 39th wedding anniversary, Sullivan filed a motion with the Los Angeles Field Office of U.S. Citizenship and Immigration Services (USCIS) to reopen and reconsider his late husband's petition for a marriage-based green card which that office had denied.

Limited Partnership, a documentary telling the couple's story, was released by Tesseract Films in 2014.

On January 5, 2014, the USCIS approved Adams' immigrant visa petition filed in 1975 on behalf of his husband. Sullivan received his green card in April 2016.

United States v Windsor
The decision of the U.S. Supreme Court in United States v. Windsor in June 2013 established that the federal government treats the term spouse as gender-neutral. On July 1, 2013, Janet Napolitano, director of the Department of Homeland Security directed the U.S. Citizenship and Immigration Services to "review immigration visa petitions filed on behalf of a same-sex spouse in the same manner as those filed on behalf of an opposite-sex spouse."

References

External links
Adams v. Howerton, U.S. District Court, February 25, 1980, accessed March 26, 2015
Adams v. Howerton, Ninth Circuit Court of Appeals, February 25, 1982, accessed March 26, 2015
Blythe Wygonik, "Refocus on the Family: Exploring the Complications in Granting the Family Immigration Benefit to Gay and Lesbian United States Citizens", 45 Santa Clara L. Rev. 493 (2005), accessed December 24, 2012
Robert Barnes, "40 years later, story of a same-sex marriage in Colo. remains remarkable", Washington Post, April 18, 2015, accessed April 18, 2015

Category:United States substantive due process case law
Category:United States same-sex union case law
Category:1982 in LGBT history
Category:1982 in United States case law
Category:United States Court of Appeals for the Ninth Circuit cases


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The demonstration of the inferior sternal cleft using three-dimensional reconstruction: a case report.
Congenital sternal cleft is a rare disorder in which there is a gap in the midline of the anterior chest wall between the two halves of the sternum. Typically, the contour of the mediastinal structures can be seen beneath the skin. It is rare and the exact incidence is not known. It results from failure of fusion of the two lateral mesodermal sternal bars by 8 weeks of gestation. Most cases are diagnosed shortly after birth and are reported only rarely in adults. We report here one of the congenital major chest wall deformities; inferior sternal cleft is rarely seen, associated with sternal and costal variations in a 22-year young man.


The purpose of this study is to determine whether amino acid feedings, provided to advanced-stage cancer patients, can prevent or reduce the normal weight loss (muscle atrophy) associated with the disease and treatment of the disease. Our results will establish whether cancer patients' muscle responds in a similar fashion to that of normal volunteers, by increasing the amount of lean muscle and preventing the loss of muscle in response to amino acid supplementation.


A new wheelchair has been designed to make life easier for individuals living with disabilities in developing regions. The SafariSeat, from London-based Uji, is aimed at being affordable, easy to repair with inexpensive parts, and able to be manufactured and repaired locally.

Like the Freedom Chair and Mountain Trike, the SafariSeat was conceived for use on rough terrain. Where it differs from those examples, though, is in having four wheels instead of three and a focus on affordability.

The project came about in 2014 after an accident left Uji founder Janna Deeble in a wheelchair temporarily. Deeble, who grew up in Kenya, felt the experience helped him to better understand the difficulties that are faced by wheelchair users people in developing countries, who may have to deal with rough terrain without adequate equipment.

To begin, the team researched the way such individuals live and the day-to-day issues they face. It was decided that the chair should be made out of bike parts, as these would be affordable, easy to work with and widely available.

Levers on each side are pumped with the arms to propel the SafariSeat via a leveraged mechanism Uji

The chair is designed in such a way that it can be assembled without the need for tools. Levers on each side are pumped with the arms to propel the chair via a leveraged mechanism. Gripping the levers at the bottom gives the sense of a high gear for use at speed over flat ground, while gripping them at the top provides more leverage for increased torque over rough ground.

A suspension system allows the chair to keep all four wheels in contact with the ground for stability while moving over rough terrain. The seat, meanwhile, adjusts its shape as the chair moves, helping to reduce the potential for pressure sores and stimulating blood-flow.

Uji plans to make the blueprints for the SafariSeat freely available, meaning that anyone will be able to build one. It also plans to include workarounds so that the chair can still be constructed even if not all of the required components are available.

A Kickstarter crowdfunding campaign for the SafariSeat is underway. Pledges from £200 will see a SafariSeat gifted to an individual whom it could benefit.

The video below is the Kickstarter pitch for the SafariSeat.

Source: Kickstarter


Q:

how to bin numbers quickly in python with multiple conditions

I want to bin the figures based on different ranges with my own definition.
lambda is easy but what if the condition is more than 2. I used for if but it does not change anything
country = pd.DataFrame({'COUNTRY':['China','JAPAN','KOREA', 'USA', 'UK'],
               'POPULATION':[1200,2345,3400,5600,9600],
               'ECONOMY':[86212,11862,1000, 8555,12000]})

for x in country.POPULATION:
if x < 2000:
    x = 'small'
elif x >2000 and x <=4000:
    x='medium'
elif x > 5000 and x <=6000:
    x='big'
else:
    'huge'

I hope the data can return the 'small', 'medium', etc. according to the range.

A:

I would use np.select with multiple conditions:
conditions = [
    country['POPULATION'] < 2000,
    ((country['POPULATION'] > 2000) & (country['POPULATION'] <= 4000)),
    ((country['POPULATION'] > 5000) & (country['POPULATION'] <=6000))
]

choices = [
    'small',
    'medium',
    'big'
]

# create a new column or assign it to an existing
# the last param in np.select is default
country['new'] = np.select(conditions, choices, 'huge')

  COUNTRY  POPULATION  ECONOMY     new
0   China        1200    86212   small
1   JAPAN        2345    11862  medium
2   KOREA        3400     1000  medium
3     USA        5600     8555     big
4      UK        9600    12000    huge


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July 12, 2014

Looks can be deceiving
It fit well and I love the fabric. My only complaint about this top is that it looks like a full complete wrap top. It's not. You have to wear a shirt under this top otherwise when you sit or move a certain way it will show your stomach. The "wrap" is completely open except for a small stitch on the top holding it together. I had to use some fashion tap to hold down the rest of the top or wear a shirt underneath it.
May 22, 2014

Don't put this top in the washer.
This was a great, comfortable top... but only for the one and only time I got to wear it. The label says machine washable. However, after one time in the washer and line-dried, there was pilling all over. I can't even wear it again. If you purchase this top, do yourself a favor and hand wash it.
May 11, 2014

Great top!
This top fits great and looks great! I bought it for work and am very pleased.
April 30, 2014

Great top, looks great! Just need to be careful at times because it a wrap dress, parts of the body can be seen.
April 30, 2014

Springtime cute
Love this top. It runs a little big and the draped front is open…it is not sewn. It is definitely meant to be worn with a cami underneath. Great for spring and summer wear-very breathable.
April 29, 2014

Loved it, but Short
I bought this shirt in two colors. I love the fit and the way the shirt lays. The only thing that could have made it any better would have been a slightly longer length. It sits right at the top of the hip and can sneak up a little.
April 7, 2014

good basic...
While there is an opening between the layers in this shirt, it really isn't noticeable if you have it arranged correctly. I already own a navy cami so I wasn't concerned. This top fits exactly as it looks in the picture, and isn't TOO "blouson-y." Personally I was very happy to find something that actually adheres to my shape and isn't box-like. Looks good with a statement necklace.
March 22, 2014


Characterization of monoclonal antibodies directed against swine leukocytes.
Hybridomas were produced from fusions of the SP2/0 mouse myeloma with splenic cells from: 1) an outbred Sprague Dawley rat immunized with swine peripheral blood mononuclear (PBM) cells; 2) a (CBA/NDub X BALB/c Dub) F1 mouse immunized with concanavalin A (Con A) activated swine PBM cells and 3) a (BALB/c Dub X C3H/He Dub) F1 mouse immunized with swine thymocytes. The resulting supernatants were screened by a microcytotoxicity assay for activity against swine PBM cells. Four hybridomas (MSA1, MSA2, MSA3 and MSA4) were selected, cloned and characterized by their cell reactivity and effect on mitogenic assays. MSA1 and MSA2 belong to the rat IgG2b subclass. MSA3 and MSA4 are of the mouse IgG2a subclass. These monoclonal antibodies reacted in the following manner: MSA1 with monocytes, granulocytes, red blood cells and bone marrow cells; MSA2 with subset of T cells; MSA3 with B cells and subsets of T cells and monocytes (class II molecule) and MSA4, a pan-T cell reagent (E-rosette receptor). The involvement of the various cell types reactive to the different monoclonal antibodies in the mitogenic response of swine PBM cells to Con A, phytohemagglutinin (PHA) or pokeweed mitogen (PWM) was investigated by cellular depletion with monoclonal antibody plus complement. Cellular depletion of PBM cells with the following monoclonal antibodies plus complement treatment resulted in: MSA1, almost total reduction in the mitogenic response to low doses of Con A or PWM; MSA2, partial reduction in the proliferative responses to any concentration of Con A, PHA or PWM; MSA3, partial reduction in proliferative responses to low concentrations of Con A or PWM and 4) MSA4, total elimination of any proliferative response to Con A, PHA or PWM.


A unique treatment program at Fort Campbell is changing the lives of soldiers recovering from brain injuries. One of the patients receiving treatment, Sgt. 1st Class Michael Hall, was in Afghanistan onMore >>

A unique treatment program at Fort Campbell is changing the lives of soldiers recovering from brain injuries.More >>

A year after learning of her brain tumor, a Middle Tennessee woman is sharing her incredible story of survival. It's something Dianna Milam has done dozens of times, but singing at church now has evenMore >>

A year after learning of her brain tumor, a Middle Tennessee woman is sharing her incredible story of survival.More >>

A unique camp in Middle Tennessee camp is helping children with autism learn important life skills. Communication is a big struggle for kids with autism, but this summer they're getting the opportunityMore >>

A unique camp in Middle Tennessee camp is helping children with autism learn important life skills.More >>

Breast cancer can be a tough diagnosis to work through, and even when the cancer is gone the fight isn't always over. Last summer, Janetta Elmore had a mastectomy after being diagnosed with breast cancer. AfterMore >>

Breast cancer can be a tough diagnosis to work through, and even when the cancer is gone the fight isn't always over.More >>

As the holiday weekend approaches, there are some important reminders to stay safe when you're out on the water at the lake or river. Last year, there were 171 boating accidents in Tennessee, and manyMore >>

As the holiday weekend approaches, there are some important reminders to stay safe when you're out on the water at the lake or river.More >>

NASHVILLE, TN (WSMV) -

A new state law takes effect Jan. 1 that aims to protect Tennessee's student athletes from brain damage.

The concussion legislation forces schools to adopt guidelines to educate kids, their parents, coaches and school staff about the dangers of head injuries in sports like football.

"We see it all the time: Kids go back in a game they shouldn't have gone back into," said Dr. Andrew Gregory with Vanderbilt University Sports Medicine.

Gregory has been vocal about his support of the legislation and said because there's no central governing body for youth sports in Tennessee, there's actually more support for adult athletes than young athletes.

Under the new law, schools cannot allow injured students to continue playing until they've been cleared by a medical professional.

The law also includes provisions that would require students to leave a sporting event if they show concussion symptoms.

Supporters of the new law hope educating young athletes will aid them in making important decisions.

"If you ask athletes why they were concussed after being concussed, one of the reasons is they didn't know it was a concussion, so they didn't know they shouldn't go back in a game," Gregory said.


Q:

Hashcode from reflection?

I have got
interface Module {
    List<String> parse();
}

There are several implementations of the interface and in application I would like to have a HashSet<Module> which ensures there is only one instance of each module. For this I need a proper hashCode for each class. The proper implementation of hashCode would be the one that returns the same constant for various instances of one module class but different one for different modules.
To create a nice design solution I would like to compute the hashCode from name of the module like:
public int hashCode() {
    return ConcreteModule.class.getName().hashCode();
}

but this code would be the same for every implementation of interface... My idea was to create an abstract module which implements the hashcode but is it possible to reach the name of the class which extends this abstract class?
Like:
public abstract class AbstractModule implements Module {
    public int hashCode() {
        // get the class name of class extending the abstract module
        // and return hashcode of its string name
    }
}

and then
public class ConcreteModule extends AbstractModule implements Module {
    // implementation of parse() no need to create hashcode for each module
}

Would you suggest to create hashCodes for each module or is it possible to create something I am trying? Any suggestions or advices are welcome. Thanks in advance

A:

You could simply implement it like the following:
@Override
public final int hashCode() {
    return this.getClass().getName().hashCode();
}

@Override
public final boolean equals(Object other) {
    if (other == this) {
        return true;
    }
    if (other == null) {
        return false;
    }
    return other.getClass().equals(this.getClass());
}

But I'm not sure it's the best idea. Moreover, you can't guarantee that all implementations of the interface extend your abstract class.
You could simply maintain a Map<Class<? extends Module>, Module>. Each module implementation would add itself to your registry, and if the class of the module is already in the map, you would simply reject or ignore the new module instance.


Q:

Geoserver and Geofence - using custom rules

I've checked all the options of the rule creation in GeoFence Embedded (integrated with GeoServer) and GeoFence Standalone web application. The available options for building data access rules seem to be limited to: username, user's role, service name, request name, workspace and layer. The additional option is to limit the access with the geographical area.
Is there a way, using GeoServer and GeoFence, to make rules more specific, as in the following examples:

Restricting access to a group of features from a layer, which belong to the selected bigger layer (show only cities from a selected state)
Using CQL expressions in GeoFence rules with system variables (showing only features from a layer which have in one attribute the username of the logged-in, using GeoFence authenticator, user)

A:

Rules can also filter by the caller's IP address, as described here:
https://github.com/geoserver/geofence/wiki/Main-concepts#rules

you can restrict the set of returned features by using a CQL filter. Once you define a layer in a rule, the "detail" button will become active. Pressing it, a dialog will be displayed where various settings can be set for the rule. Among these, there is a textarea where you can enter the CQL filter. More info here: https://github.com/geoserver/geofence/wiki/Main-concepts#constraints.
the CQL string is reported as-is to GeoServer, and no parsing or token substitution is performed on it.


JooE recently revealed on the latest episode of Happy Together that Yoo Jae Suk is the reason for her success and popularity lately!

She said that after appearing on Infinity Challenge as a surprise guest, she bumped into Yoo Jae Suk he remembered and greeted her.

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1. Field of the Invention
This invention relates to a motor drive system used with industrial robots, processing machines, and the like.
2. Description of the Prior Art
Conventional motor drive systems use sensors to detect the rotation of a motor, and with the detection signals from the sensors as feedback signals a host controller generates a command signal to control the rotational position and speed of the motor based on the feedback signals. A drive object, such as a robot arm or the like, is shifted to a target position, at a desired speed, by the motor being subjected to the above feedback control.
In such a system, it is desirable that the characteristics of the motor driving circuit, power source, etc, be modifiable to cope with the user's various requirements.
The host controller, which provides commands to the system, includes various types of controllers or controller functions, such as providing command values in the form of digital signals, providing command values in the form of analog signals, etc. It is, therefore, desirable that the motor drive system be connectable to these various host controllers.
The sensor for detecting motor revolutions may be an optical resolver or a magnetic resolver. Generally, the optical resolver is of high precision and high resolution, but, is expensive. On the other hand, the magnetic resolver is relatively inexpensive, but is inferior in precision and resolution as compared to the optical resolver. Because of the differences in characteristics, the optical resolver and the magnetic resolver are usually used selectively according to whether precision or cost is most desired.
Also, a different motor drive system is used for the optical resolver and the magnetic resolver.
According to the embodiment, the motor drive system to be used with the sensor must be selected properly each time so as to cope with the particular sensor to be used therewith. The motor drive system is originally expensive. Thus, if the entire motor drive system has to be replaced to handle different applications, the costs will be disadvantageously expensive.
For driving a robot arm on the motor drive system, there may occur a case where the absolute rotational position of the motor must be identified so as to ensure the position of the arm, etc. However, an absolute type sensor is expensive to install originally. Accordingly it is desired to have a system which is inexpensive and readily usable for detecting the absolute rotational position.
On the other hand, in case an incremental type sensor is used, the origin of the rotational position will be required. An origin detection means which is moderate in cost and high in precision is preferable.
As mentioned, the motor drive system has many desired requirements However, no motor drive system satisfying all of the desired requirements concurrently has yet been realized.


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