THE Dapu Road Tunnel, the city's oldest passage under the Huangpu River, will be closed to traffic next month for renovations after 38 years in service.
Cross-river traffic from Shanghai's Xuhui District in Puxi to Shangnan area in Pudong will be affected.
The tunnel is expected to reopen next March, today's Shanghai Morning Post reported.
More than 10 public bus routes using the tunnel will have to change their routes. Some will use the Lupu Bridge as an alternative. The traffic authority will publish details of the new routes, the report said.
Some commuters may use the second phase of Metro Line 8 which will start early next month, with the new phase connecting Shangnan and downtown Puxi.
Meanwhile, a second section for the Dapu Road Tunnel is expected to be completed by the end of the year, the Shanghai Morning Post reported.
The 1.5-billion-yuan (US$219 million) project will double the tunnel's capacity.
It is a key part of the city's 2010 World Expo traffic infrastructure.
The last renovations on the Dapu Road Tunnel were in 2004.


text:
Q: "DisableCreatedUser" is not working properly I am using a ASP.NET CreateUserWizard to registering users.
I wanted user to not automatically log after registration until they login manually.
    <asp:CreateUserWizard ID="RegisterUser" runat="server" EnableViewState="false" RequireEmail="false"
DisableCreatedUser="true" LoginCreatedUser="false" OnCreatedUser="RegisterUser_CreatedUser">

In here I have set the DisableCreatedUser="true" and LoginCreatedUser="false" in order to not logged the user after registration until they logged in as CreateUserWizard.DisableCreatedUser Property describes.
Any ideas about the reason this settings not working will be appreciate.
Thanks

A: I did that by adding FormsAuthentication.SignOut() in the OnCreatedUser. that works !!!

redpajama_set_name: RedPajamaStackExchange


text:
Christine Havrilla & Gypsy Fuzz
Award-winning Philadelphia area native Christine Havrilla just released a new album with her band, Gypsy Fuzz, called "Sunless Escapade" (March 2019). After embarking on an ambitious journey with her Gypsy Fuzz project in late 2011, she previously released a series of "Live Sessions" in 2014-2017, which included long-time fan favorites as well as new songs with her current band, Gypsy Fuzz. Recorded in numerous settings such as venues, living rooms, a recording studio with a live audience, an old church, etc…these volumes captured the spontaneous energy, soul-felt vibes and musical adventures that Christine brings to the stage, ANY stage! Christine utilizes the soft gritty texture of her vocals to express warmth and honesty listeners quickly embrace as well as her aggressive, raw, trash-style guitar playing and looping.
Havrilla, who has twelve albums, 3 EPs, and over 15 years of touring under her belt, continues to venture out under "Christine Havrilla & Gypsy Fuzz" as well as a solo artist, showcasing ALL of her music with new twists, and described as "a blend of 70's, 80's, and 90's rock delivered by the modern singer/songwriter."
Christine is known for bringing her "well-built, catchy, smart tunes" (Upstage Magazine) to audiences everywhere, touring nationally as a solo act as well as with Gypsy Fuzz. After an inspirational time of being based in Portland, OR, Christine is once again back in the Northeast and frequenting such venues as World Café Live, Steel City, Godfrey Daniels, Jammin' Java, Burlap and Bean, John and Peter's, Rockwood Music Hall, The Tin Pan, to name a few.
Read more on my website…

redpajama_set_name: RedPajamaCommonCrawl


I just missed, by all accounts, a landmark exhibition at the Yale Center for British Art in New Haven, the first major exhibition devoted to Johan Zoffany (1733-1810). It just moved to the Royal Academy in London where it will stay until June 10th 2012. Try to see it if you can. Zoffany is not well-known in the U.S. and not much better known in England. I myself have not studied him much but he was unquestionably a great master. Born in Frankfurt, studying art nearby, he traveled to Rome in 1750 where he stayed for ten years. In London he gained the patronage of the King and Queen and was popular with the Austrian Royal Family as well. He traveled widely as far as India, painting some remarkable scenes there, but he maintained his home in London where he died in 1810. The most interesting biographical detail is that he was initiated as a Freemason in 1763, thereby entering a world of esotericism. Thomas Gainsborough so admired him that he requested to be buried nearby.
Robin Simon reviewing the exhibition when it was at Yale recently wrote: "Zoffany was perhaps the most compulsive self-portraitist of all, for ever popping up within a composition, even when he was not – as he so often was – creating yet another separate painting or drawing of himself."1 As for the exhibition in London, catch it if you can. If you can't, I'll try to help by posting more on Zoffany in the near future. He's a fascinating artist even on the superficial level. Underneath I think we'll find a gold mine.


Free christmas printables: holiday wall decor ideas | on, Free christmas printables and diy holiday wall decor ideas. get tips for printing and framing, along with inspiration to create your own custom look.. 43 clever, over-the-top, ridiculous christmas decor ideas, Following on the heels of halloween, when everyone on pinterest collectively lost their minds making witches hats and jack-o-lanterns from every possible household item, i thought we'd see what pinterest is up to for christmas this year.. spoiler alert: a lot. on pinterest, there shall be no area of the house, fridge or yard that goes un-christmastified.. Christmas decorations and holiday decor | frontgate, Shop christmas decorations and holiday decor â from elegant christmas wreaths and garlands to pre-lit trees, festive outdoor displays and more â at frontgate.
Quick tricks: christmas decor paint projects, I'm so excited about today's post because i love christmas decor paint projects! whether transforming old tired christmas decor into something updated and fresh with paint, or getting creative with home accessories collecting dust in my junk closet for holiday decorating, a great paint revamp makes me so happy!. Outdoor christmas lantern planter idea for your porch, Easy, diy outdoor christmas lantern planter idea for your porch. festive and fun way to welcome your friends and family! uses led candle and fresh greenery.. Christmas decorations & holiday decor | oriental trading, Christmas decorations. celebrate the season with festive indoor and outdoor christmas decorating ideas. find unique christmas gift ideas, home decor and christmas decorations at unbeatable prices..
40+ best red christmas decor ideas and designs for 2019, Red christmas decor ideas are vibrant and joyful in a way that winter white could never be. get inspired be the best designs for 2019!. Christmas at menards®, Spread holiday cheer with christmas trees, sparkling lights, stockings hung with care with help from menards!. Christmas decorations - lowe's, Christmas decorations at lowe's. the lowe's christmas store has everything you need to decorate for christmas, from stocking holders and christmas tree ornaments to large outdoor christmas decorations.whether you prefer rustic christmas décor or a more contemporary look, we'll help you find all the trimmings..


text:
package org.shenit.tutorial.android.search;

import android.os.Bundle;
import android.support.v7.app.AppCompatActivity;

import org.shenit.tutorial.android.R;
import org.shenit.tutorial.android.TutorialUtils;

public class SearchExamplesActivity extends AppCompatActivity {

    @Override
    protected void onCreate(Bundle savedInstanceState) {
        super.onCreate(savedInstanceState);
        setContentView(R.layout.activity_search_examples);
        TutorialUtils.bind(this, R.id.search_dialog_example_button, SearchDialogExampleActivity.class);
        TutorialUtils.bind(this, R.id.search_view_ab_example_button, SearchViewActionbarExampleActivity.class);
    }
}

redpajama_set_name: RedPajamaGithub


text:
4mins Chennai's jewellers welcome Union government's move to ensure hallmarking of gold jewellery
Israel claims rocket fire from Gaza, launches airstrikes
JERUSALEM, August 19, 2014 19:33 IST
Updated: April 21, 2016 04:51 IST
In this August 18, 2014 photo, Palestinians walk on the rubble of the house of Hussam Kawasma, one of three Palestinians identified by Israel as suspects in the killing of three Israeli teenagers, after it was demolished by the Israeli Army in the West Bank city of Hebron. The Israeli military has said it had carried out a series of airstrikes across the Gaza Strip in response to renewed rocket fire. | Photo Credit: Nasser Shiyoukhi
The Israeli military accused Palestinian militants of violating a ceasefire and said it maintains "both defence and striking capabilities in order to address the renewed aggression".
The Israeli military says it has carried out a series of airstrikes across the Gaza Strip in response to renewed rocket fire. There was no immediate word on casualties from the strikes.
In a statement on Tuesday, the military accused Palestinian militants of violating a ceasefire and said it maintains "both defence and striking capabilities in order to address the renewed aggression".
The airstrikes came minutes after Tel Aviv claimed that Palestinian militants fired three rockets into Israel, breaking a temporary ceasefire and endangering negotiations in Egypt over ending a month-long war between Israel and Hamas. Authorities said the rockets landed in open areas and caused no injuries.
Related Topics International World
Israeli airstrike levels 7-storey building in Gaza
UN Security Council urges resumption of Gaza talks
Israeli airstrike kills three Hamas commanders
Hamas kills 18 suspected informers for Israel
Israeli offensive cost Gaza billions: Palestinian official
Israel cancels trains near Gaza Strip
Israel partially lifts Gaza fishing ban
Israel, Hamas agree on new five-day ceasefire
Printable version | Jan 18, 2020 2:10:48 PM | https://www.thehindu.com/news/international/world/israel-claims-rocket-fire-from-gaza-launches-airstrikes/article6332119.ece


I created this book as an exploration of significant memories and pivotal moments in my life, revolving around the symbolism of trees. Nature has always played an important role in my life, which I was able to begin to understand, and to honor, through this book. Each section of the book contains digitally altered images of trees juxtaposed with an actual leaf from that tree hand sewn between pieces of vellum. The covers of the book are made from birch bark mounted on bark paper.


text:
Q: How to go back multiple times in a directory path? I am trying to create a full path for several folders. basically there is a batch script that is called, and these are the commands that will be used inside that script.
I did this:
ECHO OFF
SET currentDirectory=%~dp0
echo %currentDirectory:~0,-15%

SET ROOT = %currentDirectory:~0,-15%
SET MNIST_DIR = %currentDirectory:~0,-1%
SET DATA = %ROOT%/data/mnist
SET BUILD= %ROOT%/Build/x64/Release
SET BACKEND=lmdb

The output of %~dp0 is N:\caffe-master\examples\mnist
so by %currentDirectory:~0,-15% I meant to access N:\caffe-master\.
However, It seems what I am doing is wrong, and the following command can not be executed!
"%BUILD%/convert_mnist_data.exe" %DATA%/t10k-images-idx3-ubyte %DATA%/t10k-labels-idx1-ubyte %EXAMPLES%\mnist_test_%BACKEND% --backend=%BACKEND%

How can I solve this problem.
note that I also tried CD /D to no avail as well. so this is what I came up with.

A: How about this:
@echo off
SET currentDirectory=%~dp0
PUSHD %CD%
CD ..
CD ..
SET MNIST_DIR=%CD%
POPD
ECHO %MNIST_DIR%
PAUSE

This generates the output N:\caffe-master\.
EDIT: By using PUSHD %CD% and POPD at the end of the script we can ensure that we will always end up in the original directory.

A: .. is a folder's parent (and . is the folder itself). So c:\windows\system32\drivers\..\.. equals c:\windows (as does c:\windows\system32\drivers\..\..\.\.\.\.\.).
Aso
No spaces on either side of the equals sign. You are setting %root %


text:
Scientific Categories
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Intraclonal heterogeneity and distinct molecular mechanisms characterize the development of t(4;14) and t(11;14) myeloma
Brian A. Walker, Christopher P. Wardell, Lorenzo Melchor, Sanna Hulkki, Nicola E. Potter, David C. Johnson, Kerry Fenwick, Iwanka Kozarewa, David Gonzalez, Christopher J. Lord, Alan Ashworth, Faith E. Davies and Gareth J. Morgan
Blood 2012 120:1077-1086; doi: https://doi.org/10.1182/blood-2012-03-412981
Brian A. Walker
Christopher P. Wardell
Lorenzo Melchor
Sanna Hulkki
Nicola E. Potter
David C. Johnson
Kerry Fenwick
Iwanka Kozarewa
Christopher J. Lord
Faith E. Davies
Gareth J. Morgan
We have used whole exome sequencing to compare a group of presentation t(4;14) with t(11;14) cases of myeloma to define the mutational landscape. Each case was characterized by a median of 24.5 exonic nonsynonymous single-nucleotide variations, and there was a consistently higher number of mutations in the t(4;14) group, but this number did not reach statistical significance. We show that the transition and transversion rates in the 2 subgroups are similar, suggesting that there was no specific mechanism leading to mutation differentiating the 2 groups. Only 3% of mutations were seen in both groups, and recurrently mutated genes include NRAS, KRAS, BRAF, and DIS3 as well as DNAH5, a member of the axonemal dynein family. The pattern of mutation in each group was distinct, with the t(4;14) group being characterized by deregulation of chromatin organization, actin filament, and microfilament movement. Recurrent RAS pathway mutations identified subclonal heterogeneity at a mutational level in both groups, with mutations being present as either dominant or minor subclones. The presence of subclonal diversity was confirmed at a single-cell level using other tumor-acquired mutations. These results are consistent with a distinct molecular pathogenesis underlying each subgroup and have important impacts on targeted treatment strategies. The Medical Research Council Myeloma IX trial is registered under ISRCTN68454111.
The t(4;14) is seen in 11% of presenting myeloma and defines a unique subtype associated with a poor clinical outcome.1 In contrast to other molecular subtypes, it is seen at lower frequencies in MGUS, suggesting that the genes deregulated by the t(4;14) rapidly push plasma cells to a more aggressive phenotype.2,3 Although FGFR3 was initially suggested to be the critical driver gene in t(4;14) myeloma, it is lost by deletion of the der(14) chromosome in 25% to 30% of cases, focusing attention on the other deregulated gene MMSET.4,5 MMSET is a member of a family of genes that have histone methyltransferase activity. Other members of the family are known oncogenes relevant to the pathogenesis of acute leukemia, providing strong support for the central importance of this gene in myeloma.6 Pathologically, MMSET has been shown to disrupt the distribution of dimethylation marks on histone H3 throughout the genome and is relevant to disease progression.7⇓–9 A specific DNA methylation pattern associated with t(4;14) samples also is thought to be pathologically relevant, although the relationship of these patterns to changes in histone methylation marks is uncertain.10
The t(4;14) alone is insufficient to lead directly to myeloma, being seen in monoclonal gammopathy of undetermined significance (MGUS) and smoldering myeloma.2,3 Therefore, it is likely that there are collaborating mutations at other sites throughout the genome and that the spectrum of these mutations will be unique to this subtype of disease. In this respect, the changes in histone and DNA methylation seen in this subset may affect both the spectrum and number of mutations. It also has been suggested that MMSET functions in DNA repair.8 Consequently, based on these arguments we hypothesized that there may be a unique mutational spectrum associated with t(4;14) myeloma.
Until recently, the optimum way of identifying collaborating mutations has been to study regions of copy number abnormality (CNA) within which there are relevant mutational events.5,11,12 Comprehensive mapping analyses show that CNA are common, the most frequent being deletions of 1p, 6q, 8p, 12p, 13q, 14q, 16q, and 17p.5,13 Mutational analysis of genes within regions of homozygous deletion has led to the discovery of key genes involved in the pathogenesis of myeloma, including FAM46C on 1p, BIRC2/BIRC3 on 11q, RB1 on 13q, TRAF3 on 14q, CYLD on 16q, and TP53 on 17p.5,14⇓⇓⇓⇓–19 Despite the success of this approach, we have not, as yet, identified a specific pattern associated with the t(4;14) cases. More recently, whole genome and whole exome–based sequencing (WES) approaches have been developed that provide a means of global mutational screening that have given us the ability to define mutations outside of regions of CNA. In addition to identifying the range of mutations present within a tumor, this technology can provide semiquantitative analysis of the size of the clonal population carrying the abnormality.20 This technical advantage allows for the characterization of the tumor substructure, whereas previous tools such as gene mapping arrays have only analyzed the predominant clone.
The first results generated from the application of next generation sequencing approaches were carried out on a mix of presentation and relapse samples, and they confirmed previous hypotheses suggesting that there are critical pathways that are deregulated leading to the malignant transformation of plasma cells.14 This study did not focus on the t(4;14) subgroup for which we understand in more detail the basis of its molecular etiology, and as such the changes associated with this group remain unknown. We have integrated genome-wide single-nucleotide polymorphism (SNP)–based mapping array data with WES derived from a series of presenting cases of t(4;14) myeloma and compared the mutations present with a group of cases defined by the t(11;14). The t(11;14) is a good comparator group because it also has a clear pathogenesis that is defined by the deregulation of cyclin D1. The aim of this work was to inform whether the collaborating mutational events in these different etiological settings are the same or different as well as how these events are acquired leading to tumor evolution and treatment resistance.
WES was performed on 22 presentation myeloma samples entered in the Medical Research Council (MRC) Myeloma IX trial consisting of 10 t(4;14) samples and 12 t(11;14) samples. All patients consented to storage and use of tissue in research studies, and the study was approved by the National Research Ethics Service. The MRC Myeloma IX trial is registered under ISRCTN68454111. CD138-positive bone marrow plasma cells were selected to a purity of more than 95%, as determined by cytospin, using magnetic-assisted cell sorting (Miltenyi Biotec). Tumor DNA and RNA were extracted using the AllPrep kit (QIAGEN), and the DNA concentration was assessed using Pico-green (Invitrogen). Nontumor DNA was extracted from a white cell pellet from peripheral blood samples using the FlexiGene kit (QIAGEN).
Libraries were prepared from tumor and nontumor DNA from the same patient and were sequenced to identify acquired single-nucleotide variations (SNVs) and indels in the tumor sample. We used 50 ng of genomic DNA to capture the exome using the SureSelect Human All Exon 50Mb target enrichment set (Agilent Technologies). We have validated this approach previously and shown it to have parity with approaches using larger starting amounts of DNA.21 In brief, 50 ng of DNA was fragmented on a Covaris E-series sonicator to a mean length of 200 to 400 bp, as assessed on a High Sensitivity DNA Bio-analyzer chip (Agilent Technologies). Fragmented DNA underwent end repair and A-tailing before ligation of Illumina adaptor sequences (Illumina). The prep underwent subsequent PCR amplification for 8 cycles, and then 250 ng of amplified product was hybridized overnight to the SureSelect Human All Exon baits (Agilent Technologies). After hybridization and washing, the captured DNA underwent a further 8 cycles of PCR. Purified libraries were sequenced on a GAIIx sequencer (Illumina), giving 76-bp paired end reads. Samples were sequenced to a median depth after deduplication of 61 ×, with 99% more than 1 × and 85% more than 20 × exomic coverage. After base calling and quality control metrics, the raw fastq reads were aligned to the reference human genome.
Exome analysis
The Genome Analysis Toolkit was used to call SNVs.22 These variant calls were recalibrated, and soft filters were applied to remove potential false-positives using dbSNP 132, HapMap 3.3, and the 1000 Genomes Project as truth sets. Variants that occurred in both the normal and tumor samples were filtered out, and the tumor-specific variants were annotated using ANNOVAR.23 Tumor-acquired indels and deleted exons also were called. Full details of data analysis are in supplemental Methods (available on the Blood Web site; see the Supplemental Materials link at the top of the online article). In addition to the identification of commonly affected genes, functional annotation enrichment analysis was used to identify commonly affected pathways.
Mutation validation
Confirmation of WES variants was performed on whole genome–amplified DNA. We amplified 50 ng of genomic DNA in a linear manner using the Repli-g kit (QIAGEN) to generate ∼ 40 μg of amplified DNA. Samples included the 22 samples that had undergone WES as well as an additional 127 samples from the MRC Myeloma IX study. Individual PCR primer sequences and conditions can be found in supplemental Methods. Mutations in NRAS, KRAS, and BRAF were confirmed on an individual sample basis.
Copy number and gene expression analysis
GeneChip Mapping 500K Array sets (Affymetrix) were performed as described previously.5,15,24 The SNP genotypes and inferred copy numbers were obtained using GTYPE and dChip. Data have been deposited into Gene Expression Omnibus under accession GSE21349.5 U133 Plus 2.0 gene expression arrays (Affymetrix) had been performed previously using total RNA from these samples. In total, 260 samples were available to determine the proportion of samples expressing that gene. Genes were placed in bins according to the proportion of samples expressing them, and then the number of variants was counted for each bin. Genes defined as having spiked expression in myeloma were defined according to the criteria outlined in a prior publication.25
Single-cell sorting and genotyping
Single cells from a patient were sorted on an FACSAria cell sorter (BD Biosciences) using a panel of antibodies known to discriminate abnormal from normal plasma cells26: anti–CD45-PB (Dako UK), anti–CD38-FITC (Caltag MedSystems), anti–CD138-PO (Caltag MedSystems), anti–CD56-PE (Caltag MedSystems), anti–CD19-PE–Cy7 (Beckman Coulter), anti–IGK-APC (Dako UK), and anti–λ-APC–Cy7 (BD Biosciences). Abnormal plasma cells were identified as CD38+, CD138+, CD45−, CD56−, CD19−, and IGK+. We sorted 270 single cells on 3 96-well plates (90 cells/plate). In parallel, lymphocytes from a healthy donor patient also were sorted to serve as wild-type control for further genotyping analysis 5 cells/plate). Finally, 1 well/plate was left empty to serve as no-template control for the analysis.
This novel approach for single-cell multiplex real-time quantitative (Q)–PCR analysis was followed (N.E.P., L. Ermini, E. Papaemmanuil, G. Vijayaraghavan, I. Titley, A. Ford, L. Kearney, M. Greaves, Multiplexing genetic analysis of single cancer cells; manuscript in preparation). In brief, single cells were sorted directly into lysis buffer. Specific (DNA) target amplification (STA) was performed before Q-PCR. This multiplex STA reaction involves the simultaneous amplification of target regions of interest using custom-designed TaqMan assays. Genotyping assays specific for the mutations of interest were custom-designed according to manufacturer's guidelines. The STA product was diluted before Q-PCR interrogation for mutations using the 96.96 dynamic microfluidic array and the BioMark HD (Fluidigm) as recommended by the manufacturer. Genotyping analysis was performed by Genotyping SNP Version 3.0.2 software (Fluidigm). Hierarchical clustering was performed using Pearson correlation and average linkage on the Rock platform.27
Background rates of mutation in the 2 molecular subgroups
After accounting for the variants in the nontumor DNA, there were, on average, 74 tumor-specific SNVs per sample (range, 54-99), including all captured sequences comprising exonic and surrounding intronic regions. There was a median of 36 exonic SNVs per sample (range, 27-64), and of these 24.5 were nonsynonymous (NS; range, 15-41). Although there was not a marked difference in the frequency of SNVs between the t(11;14) and t(4;14) groups, the number of SNVs was consistently higher in the t(4;14) samples (Table 1; supplemental Figure 3). The composition of somatic variations across both groups shows a predominance of C·G→T·A transitions (37%) and T·A→C·G transitions (20%; Figure 1A). A high proportion of C·G→T·A transitions has been noted previously14 and is because of spontaneous deamination of methyl-C residues to uracil that is subsequently replaced by thymine. Although we have shown previously that t(4;14) samples have more CpG methylation than other cytogenetic groups, in this analysis we found no significant difference in the rates of transition mutations between t(4;14) and t(11;14) samples, suggesting that this is not responsible for the pattern of mutations seen. There were in total 39 acquired indels over the 22 samples (mean = 1.77; range, 0-4). Of these indels, all were exonic and 28 (71.8%) introduced a frameshift and 11 (28.2%) were in-frame with the coding sequence. All 39 indels affected unique genes (supplemental Table 4).
Comparison of mutation rates between the t(11;14) and t(4;14) subgroups
Composition of somatic variations. (A) Transition and transversion rates in sample groups. (B) Intersection of t(4;14) and t(11;14) genes with NS SNVs and indels.
Because intragenic deletions have been reported in myeloma for key genes such as KDM6A (UTX), we investigated the prevalence of these deletions. This analysis identified 276 regions of deletion in the tumor samples, excluding physiologic immunoglobulin gene deletions. There was a median of 4 exonic deletions per sample, but 1 sample had an abnormally high number of deletions (n = 181) because of a large homozygous deletion on chromosome 17 at the keratin cluster; this deletion also was seen by mapping array.
There were significantly more mutations within genes that were not commonly expressed (P = .022, χ2 test), and there was an increasing mutation rate as the proportion of samples that did not express the gene increased (supplemental Figure 4). However, for the subset of genes expressed in more than 75% of myeloma samples, an elevated mutation rate was seen. Genes in this subset include known oncogenes (KRAS, NRAS, and MYC) and tumor suppressor genes (CYLD and FAM46C). We also examined genes with spiked expression in myeloma, and we found that PTPRG, FGFR3, ICAM1, LGMN, BMP2K, BCL6, MYC, and ROBO1 were mutated.
To gain mechanistic insight into the relationship of copy number abnormalities and mutations, we integrated the mutation data with genome mapping data. There were 102 distinct regions of copy number loss (totaling 3678 Mb) and 91 regions of copy number gain (4296 Mb) containing 33 and 48 NS-acquired SNVs, respectively. In regions of normal copy number, there were 517 mutations over 51 070 Mb of DNA. Per Mb of DNA, the mutation rate is 1/111.4 Mb in regions of loss, 1/110.3 Mb in regions of normal copy number, and 1/89.5 Mb in regions of gain. Taking into account the number of alleles available for analysis, the results are consistent with a mutation rate in regions of loss of heterozygosity of 1 SNV/111 Mb compared with 1/220 Mb in regions of normal copy number and 1/268 Mb in regions of gain, consistent with an increased mutation rate in regions of loss (Table 2). There were 33 NS-acquired mutations in regions of loss of heterozygosity (LOH) within the 22 samples. These were found in the common regions of LOH, namely, 8p, 13q, 14q, 16q, 17p, 22, and X in females. There was only 1 recurrently mutated gene within a region of LOH, and this gene was DIS3 on 13q. Other nonrecurrently mutated genes of interest in regions of LOH include MYC, CYLD, and FLT3 (Table 3).
Mutations in regions of copy number change
List of mutated genes in regions of LOH
Almost all t(4;14) cases have del(13), and using acquired SNVs on chromosome 13 as a tool, we have investigated the dynamics of the development of del(13) and the mutation of genes carried on it (Figure 2; see supplemental Figures). Mutational frequency was determined by mutant sequence reads, and copy number was determined by a combination of SNP array copy number and LOH. We show that in the majority of instances del(13) occurs first and is followed by the acquisition of a mutation, but when mutations occur first these are more likely to be NS. Samples in which uniparental disomy (UPD) and acquired mutations occur together can be used to validate this finding. Three samples had UPD and NS mutations. The first is on 1q that has LOH and is also trisomic and has 2 mutations at frequencies of 14% (FCRL5) and 27% (IL6R). The second on chromosome 7 has 2 mutations present in 49% (BUD31) and 38% (CTTNBP2) of reads. The third is on chromosome 8 and has 43% mutated reads (CSMD3). These data suggest that deletion and subsequent duplication, to create UPD, occurred before mutation, which is the order determined by our analysis of chromosome 13.
Dynamic relationship of mutation to LOH on chromosome 13. A cell with 2 copies of a given chromosome (A) can gain a mutation (star) in 1 copy (B) and subsequently delete the unaltered copy, giving rise to a mixed population of cells (C) that then, through selection, results in a clonal population of cells with a mutation and deletion (D); or alternatively, the original cell (A) could delete 1 copy of the chromosome (E) and subsequently a subpopulation mutates the remaining allele (F) that is the selected for giving rise to a clonal population with both deletion and mutation (D). Samples are shown next to the cell genotype, as defined by sequencing and copy number data, along with the mutated gene on chromosome 13. Synonymous and NS (underlined) SNVs are shown. Asterisk indicates interstitial deletion of RB1 region detected with mutation in a region of normal copy number.
DIS3 is located on chromosome 13 and is recurrently mutated in myeloma (10% reported previously and 18% in this set). We have identified 4 mutations in this gene, 1 mutation of which has been described previously, R750K, suggesting a gain in function of this gene. Interestingly, to date, mutation of DIS3 has only been seen in cases with either a t(4;14) or t(11;14). In samples with deletion of the whole chromosome mutation of DIS3 was seen as both a subclonal minority (29%) and as a majority population (69% and 81%; Table 4). In contrast, in the sample with an isolated interstitial deletion of RB1, the DIS3 mutation was present at 92%. It seems that DIS3 mutation always occurs in parallel with deletion of the RB1 region (13q14), raising the possibility that these are collaborating events. We also noted that of the 4 samples with DIS3 mutations in a previous dataset14 2 samples had del(13q) and the remaining 2 samples a t(4;14), which is highly associated with del(13q). Thus, it is possible that mutation and selection of DIS3, as a driver mutation, is dependent on deletion of 13q14.
Frequency of clonal subpopulations carrying a mutation
Recurrently mutated genes and families
We identified 28 genes with recurrent mutations, NS SNVs, and indels (Figure 3; supplemental Table 1). The most frequently mutated genes were KRAS (n = 7), DIS3 (n = 4), NRAS (n = 4), and DNAH5 (n = 3). In total, 5 genes were recurrently mutated in the t(4;14) samples (KRAS, ABCA13, DIS3, DNAH5, and PRKD2) and 11 genes in the t(11;14) samples (KRAS, NRAS, ACTG1, DIS3, EFNB2, F8, LRR1Q1, PCLO, PLD1, SEC31A, and SSPO). The genes mutated in the 2 cytogenetic groups were distinct, and of the 284 genes mutated in t(4;14) and 305 genes in t(11;14), samples only 18 (3.0%) genes were found mutated in both cytogenetic groups (Figure 1B). The majority of mutations occurred exclusively in either subgroup, and we looked at the pathways deregulated in each of the subgroups to determine whether they defined different pathobiologic routes to myeloma. Gene ontology analysis of genes disrupted in the t(4;14) group suggests an enrichment for genes involved in cytoskeleton organization, microtubule movement, and actin filament–based processes as well as chromatin organization, Table 5. The t(11;14) group showed an enrichment for protein phosphorylation, phosphate metabolism, and Ras signaling.
Association of abnormalities and mutations. Cytogenetic data repeated at the top of each column.
Pathway analysis for genes mutated in t(4;14) and t(11;14)
Mutated genes driving the different pattern in the t(4;14) group included chromatin modulating and cellular motility genes. Interstitial loss of exons of the histone demethylase KDM6A (UTX) have been described in myeloma, and we identified a mutated case in this analysis. MMSET is a histone methyltransferase gene, but in an extended analysis we did not identify an inverse relationship between MMSET and interstitial deletions of KDM6A. Other genes involved in DNA or histone modifications also were mutated and include ACTL7B, ASXL1, CHAF1B, ESCO1, KDM2A, KDM4B, KDM6A, MLL, MLL2, MLL5, MYST1, PCGF5, SETD1B, SETD6, and TOX. DNAH5, which encodes an axonemal dynein heavy chain, was mutated in 3 of the 22 cases and also was mutated in 2 cases in a previous report, accounting for 8% of myeloma samples sequenced to date. Several other genes encoding dynein-associated proteins also were mutated, including DNAH2, DNAH3, DNAH10, DNAH11, DYNC2H1, TXNDC2, CCDC154, WWC1, TRPS1, RPRG, and ODF1.
In the t(11;14) group, Ras signaling seemed prevalent, including NRAS, KRAS, and BRAF mutations that were validated by orthogonal technologies in an extended dataset, confirming the prevalence at 22%, 18%, and 4%, respectively (supplemental Figure 5). We show that the RAS pathway mutations (KRAS, NRAS, and BRAF) are mutually exclusive (Figure 3) and that deregulation of the pathway at any point is sufficient to lead to a clonal growth advantage. Interestingly, the mutations were not associated with any specific prognosis (supplemental Figure 5). In addition to these RAS–MAPK pathway variants, other RAS-related genes were found to have acquired mutations or deletions, including ARHGEF10, ARHGEF6, ARL2BP, MAP3K4, MAPKBP1, RALGAPA2, RANGAP1, RASA4, RASGRF2, and SPRED2, both in our dataset and the Broad dataset. These findings reflect the importance of deregulation of this pathway in presenting myeloma linking to the phenotypic function deregulated by these events in myeloma.
Both molecular subgroups showed deregulation of plasma cell biology and cell cycle–related genes. Genes involved in B-cell biology that were identified as being mutated include PRDM1, PAX2, PAX7, IL6R, IRF4, IL10RA, BCL6, MAX, MYC, CYLD, MALT1, BMP2K, CARD6, TNIP1, IRF5, MKI67, and LTB. Of these genes, only PRDM1, MKI67, and LTB carried recurrent mutations. Several cell cycle genes have mutations, including ATM, ATR, CCAR1, CDC14B, PTPLAD1, PTPN21, PTPRU, TP53, TACC2, FAT1, and FAT4.
Intraclonal heterogeneity at the level of mutation
Deregulation of the RAS–MAPK pathway is clearly a central feature of both groups. Importantly, despite their driver status, clones with a RAS pathway mutation were not always present in the dominant clone. The subclonal populations can be visualized by calculating the percentage of mutant reads for all acquired mutations within sequence of depth more than 30 × in a sample and adjusting for copy number and cell purity to generate a frequency of mutated cells for each mutation. These data can be used to generate kernel density plots, with peaks showing the dominant clone and any subclones that may be present. In doing this, we see that in all samples with a RAS pathway mutation, there is at least 1 subclonal population, and frequently more than 1 subclonal population (Figure 4).
Gaussian kernel density plots indicating the frequency of cells carrying all acquired mutations. Frequency is calculated by adjusting mutant allele burden by copy number of the loci mutated. Samples with mutated KRAS (A), NRAS (B), or BRAF (C) indicates that driver mutations may be in major (> 0.5) or minor (< 0.5) clone populations. Frequency of the mutated RAS pathway gene is indicated by a dotted line.
In NRAS, in the 4 samples with mutations, mutated reads were seen at frequencies varying from 16% to 48%, equating to 32% to 96% of the tumor population (Table 4). Examples of the 2 extremes are provided by samples 983 and 1890. Sample 983 has a mutation in NRAS (c.C181A) that is present in 47% of the sequencing reads, equating to 94% of the clonal cells. In contrast, sample 1890 has a mutation (c.A182G) in 17% of the sequencing reads, consistent with 34% of the clone. Similarly in the case of KRAS, we found mutated reads in 7 samples at a frequency between 10% and 36% of all reads or 20% and 72% of the tumor cell population (Table 4; Figure 4). Such intraclonal heterogeneity also was seen in KRAS where 2 samples had 2 acquired mutations each. In the first (sample 3), both SNVs were present in minor clones (10% and 24% of reads), yet both were activating KRAS mutations (c.G34C p.Q61R and c.A182G p.G12R). Thus, it is likely that these 2 mutations were present in different cell populations and had evolved independently. Conversely, in the second example (sample 879 that harbors both c.A199C p.M67L and c.A183C p.Q61H), the 2 mutations were present at similar frequencies, 70%, and are present in the same cells (both mutations are present in the same sequencing reads). However, M67L is not a known activating mutation; therefore, it is likely that the M67L mutation preceded or occurred concurrently with Q61H mutation and its clonal expansion is because of the activating mutation. BRAF was mutated in 2 samples and was also present as a major and minor clone (92% and 36% of cells, 46% and 12% of reads). The sample with only 12% of mutated reads was trisomic for chromosome 7, the location of BRAF, consistent with the mutation occurring after the chromosome became trisomic. Previous data on BRAF in myeloma reported 2.4% of samples with a V600E mutation and an additional 1.8% with a K601N.14 In our dataset, we identified 4% with the V600E mutation and no samples with the K601N mutation.
To provide further information on how the subclonal architecture at a mutational level develops, we studied a case of t(11;14) for which we had exome sequencing data and single cells stored. Exome sequencing identified 38 acquired NS mutations. On integration with copy number data from a SNP 6.0 array (Affymetrix), the frequency of each SNV in the cell population was calculated, and a mutation kernel plot was produced (Figure 5A). This identified 3 potential populations that could be summarized through analysis of 4 mutations, ATM (estimated at 95% of population), FSIP2 (57%), CLTC (24%), and GLMN (33%). These 4 mutations were picked for confirmation of clonal frequency by single-cell analysis. Genotyping for all 4 variants was performed on 270 single cells and showed the presence of the ATM mutation in 97% of single cells, FSIP2 in 59%, CLTC in 30%, and GLMN in 27% (Figure 5B). This confirmed that sequencing allelic reads can be used to accurately estimate the frequency of the mutation in the cell population. In addition to this, the single-cell analysis also indicated that all the cells contained the ATM mutation, and there were 3 subpopulations, 1 subpopulation of which only contained the ATM mutations; another subpopulation with an ATM and FSIP2 mutation; and a third subpopuation with ATM, CLTC, and GLMN mutations (Figure 5C-D).
Clonal heterogeneity determined by single cell analysis. (A) Mutant allele frequencies of 47 exonic synonymous and NS variants identified through exome sequencing. The frequencies of 4 variants are shown. (B) Proportion of cells identified with a mutated genotype for 4 genes as determined by exome sequencing allelic frequencies and single cell genotyping. (C) Hierarchical clustering of the genotypes generated from the analysis of 270 single cells. Grey indicates mutated; and black, reference. Individual populations are identified (1-4). (D) Determined cell lineages showing the proportion of tumor population and genotypes for the 4 genes in the identified populations (mutated genotypes underlined).
All myeloma passes through a MGUS phase; yet, there are well-described differences in the pathogenesis and clinical outcome of the t(4;14) and t(11;14) subgroups of myeloma.28,29 It has been estimated that the transition from MGUS to myeloma takes a median of 25 years30 during which time mutations may accumulate.10 The frequency at which the t(4;14) translocation occurs in MGUS patients is markedly lower than that seen in myeloma patients (4% vs 11%), whereas the t(11;14) occurs in relatively equal proportions (10%-14%).2,3 This difference in frequency reflects the unstable phenotype induced by the t(4;14) that decreases the time to develop myeloma and could affect mutational load at presentation. We show that t(4;14) samples at presentation have an increased mutational load compared with t(11;14) samples (median, 27 vs 23.5 NS mutations, respectively), and with increased sample sizes this trend may become significant. We can conclude that the number and spectrum of mutations necessary to be a presenting case of myeloma is similar in the 2 molecular subgroups. Because it is likely that cases with t(4;14) develop myeloma over a shorter period than those with t(11;14), we conclude that the genome of the t(4;14) cases is more unstable.
In addition, transition and transversion rates between the 2 groups were equivalent. Previously, we have shown that the t(4;14) subgroup has increased DNA hypermethylation compared with other subgroups.10 It may be expected therefore that there would be an increase in transversions in the t(4;14) group, because of the deamination of 5-methylcytosine to uracil that is replaced by thymine. However, this was not the case, but a previous study had identified an increased mutation rate at CpG islands,14 and it may be that the exome capture, which has a bias against first exons and CpG-rich sequences, has not detected this.
We incorporated our data with data from the other myeloma genome sequencing dataset that together identified 228 recurrently mutated genes in 60 samples. The most frequently mutated genes were KRAS, NRAS, DIS3, FAM46C, LRP1B, DNAH5, and WHSC1. There were some differences in the genes identified between the datasets, but this be accounted for by the techniques used (whole genome vs whole exome; WHSC1 and LRP1B) or the cytogenetic groups studied (random sampling versus translocations; DIS3) or treated and nontreated patients (FAM46C). To address the passenger or driver status of the genes identified, we used the COSMIC database to identify recurrently mutated genes in other cancer cell types. Of the 571 genes found mutated in our own dataset, 63 genes have not been identified as mutated in any cancer type. Twenty-one genes were identified as being mutated in hematologic cancers, and of these 6 genes were recurrently mutated in our dataset (NRAS, KRAS, BRAF, PRDM1, SEC31A, and PDE4DIP; supplemental Table 3).
One of the novel findings of the first myeloma genome publication was the identification of DIS3 mutations in 10% of myeloma samples. We have confirmed the presence of DIS3 mutations in myeloma, and we found that the proportion of affected samples was higher (18%). This difference is because of the samples analyzed, because DIS3 mutations have been found exclusively in t(4;14) or t(11;14) samples, of which our dataset is composed. In addition, we found that all samples with DIS3 mutations also have deletion of 13q14, the region surrounding RB1, and so deletion of 13q14 and mutation of DIS3 may be collaborating lesions. The gene of interest at 13q14 has been debated for many years and may be RB1 or 1 of the microRNAs implicated in chronic lymphocytic leukemia (miR-15a/16).31,32 It remains unclear which region(s) of chromosome 13 is important, but the discovery of yet another locus will confound the situation further and only through analyses of large datasets, comprising copy number and mutational data, will the true nature of these abnormalities be discovered. DIS3 mutations also have been found in relapse acute myeloid leukemia, known to have deletion or uniparental disomy of chromosome 13.33,34
When we compare the intersection of mutations in the 2 groups, we see that only 3% are shared. The majority of mutations in each group are distinct, suggesting that the pathologic contribution of the genes mutated in each group is different. We show that genes expressed at more than 75% have a greater rate of mutation and could explain some of the differences between the subgroups. Gene ontology analysis of genes disrupted in the t(4;14) group suggests an enrichment for genes involved in cytoskeleton organization, microtubule movement, and actin filament–based processes as well as chromatin organization (Table 5). The t(11;14) group showed an enrichment for protein phosphorylation and Ras signaling. This observation is consistent with the idea that collaborating mutations are likely to interact with the initiating events in each molecular subtypes of myeloma to hijack the normal physiologic role of plasma cells, contributing to their malignant transformation. RAS pathway mutations have been shown to occur in myeloma,35 and here we show convincingly that RAS-MAPK pathway mutations tend to be mutually exclusive, consistent with the lack of a clonal survival advantage for cells acquiring additional mutations within a pathway already deregulated.
We demonstrate that driver mutations are present as minor subclones in half of the samples. One sample was identified that contained 2 activating KRAS mutations present in different subclonal populations, comprising 20% and 48% of cells. This suggests that subclones are continually at risk of developing driver mutations that can confer a growth and survival advantage, leading to clonal dominance over time. Single-cell analysis confirmed the presence of subclonal substructure at a mutational level. In 1 example, we showed that all cells contained an ATM mutation, but 2 subpopulations existed with mutually exclusive mutations in FSIP2 and GLMN/CLTC. The mutant cell frequency plot for this sample shows 3 distinct peaks (at 0.3, 0.5, and 1.0), corresponding to the 3 main clones in this sample (mutated at ATM, ATM + FSIP2, or ATM + GLMN/CLTC). This type of subclonal structure has been described for other hematologic malignancies such as acute myeloid leukemia, acute lymphoblastic leukemia, breast cancer and prostate cancer using FISH, single cell sequencing or ultra-deep sequencing.34,36⇓⇓–39 The realization that tumor biology consists of multiple independent subclones has important ramifications for tumor progression and personalized treatment strategies.
We provide evidence that there is a distinct mutational landscape in the t(4;14) and t(11;14) groups of myeloma. After initial immortalization by the acquisition of a translocation, each group follows a distinct pathway to becoming a presenting case of myeloma. Copy number abnormalities may occur relatively early on in the pathogenesis of myeloma, and regions of LOH undergo mutation more frequently. Mutations in regions of LOH tend to be NS, pathologically relevant variants that contribute to clonal progression. The rate of mutation seems to be higher in the t(4;14) group, which may lead to their more rapid progression to myeloma. The net mutational load, however, that is required to develop presenting myeloma is consistent in the 2 groups. We demonstrate intraclonal heterogeneity and the continued acquisition of mutations within subclones that leads to clonal outgrowth and disease progression. The clonal architecture described here, with multiple subclones vying for dominance, is reminiscent of the Darwinian natural selection process with a branching, nonlinear accumulation of mutations in malignant plasma cells.
Contribution: B.A.W. designed and performed research, analyzed data, and wrote the paper; C.P.W. performed computational research, analyzed data, and wrote the paper; L.M. performed research and analyzed data; S.H. and K.F. performed research; N.E.P. developed single-cell protocols; D.C.J. and D.G. designed research; I.K. designed and performed research; C.J.L. and A.A. provided resources; F.E.D. designed research; and G.J.M. designed research, analyzed data, and wrote the paper.
Conflict-of-interest disclosure: The authors declare no competing financial interests.
Correspondence: Gareth J. Morgan, Haemato-Oncology Research Unit, Division of Molecular Pathology, The Institute of Cancer Research, 15 Cotswold Road, Sutton, United Kingdom, SM2 5NG; e-mail: gareth.morgan{at}icr.ac.uk.
The authors thank Ian Titley for expert assistance in flow cytometry sorting of single cells and Ricardo Morilla for assistance in setting up the myeloma flow panel.
This study was supported by grants from Myeloma UK and the National Institutes of Health Biomedical Research Center at the Royal Marsden Hospital as well as Children with Cancer UK.
↵* B.A.W. and C.P.W. contributed equally to this work.
There is an Inside Blood commentary on this article in this issue.
The online version of the article contains a data supplement.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 USC section 1734.
Submitted March 2, 2012.
Accepted May 3, 2012.
© 2012 by The American Society of Hematology
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DOI: https://doi.org/10.1182/blood-2012-03-412981
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Brian A. Walker, Christopher P. Wardell, Lorenzo Melchor, Sanna Hulkki, Nicola E. Potter, David C. Johnson, Kerry Fenwick, Iwanka Kozarewa, David Gonzalez, Christopher J. Lord, Alan Ashworth, Faith E. Davies, Gareth J. Morgan
Blood Aug 2012, 120 (5) 1077-1086; DOI: 10.1182/blood-2012-03-412981
Walker, B. A., Wardell, C. P., Melchor, L., Hulkki, S., Potter, N. E., Johnson, D. C., Fenwick, K., Kozarewa, I., Gonzalez, D., Lord, C. J., Ashworth, A., Davies, F. E., & Morgan, G. J. (2012). Intraclonal heterogeneity and distinct molecular mechanisms characterize the development of t(4;14) and t(11;14) myeloma. Blood, 120(5), 1077-1086. Accessed July 17, 2019. https://doi.org/10.1182/blood-2012-03-412981.
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The Ministry of Economic Affairs' evaluation on whether to apply for membership of China's proposed Asian Infrastructure Investment Bank (AIIB) was dated — and released internally — on March 18, and it noted the lack of widespread public support for such a bid.
That was just two weeks before President Ma Ying-jeou (馬英九) and the National Security Council made the decision to submit a letter of intent on Monday, one day before the deadline set for applications to join the bank as a founding member.
Remarks made by some government officials, and the Chinese Nationalist Party's (KMT) dismissal of attempts to compare it with the cross-strait service trade agreement — implicitly intimating, ironically, the latter's secretive character and the government's operation in dealing with China-related affairs, supported unquestioningly by the KMT — has exposed why it has been unable to secure public trust.
The economics ministry might say it has been wrongly blamed, as the Ministry of Finance is the authority responsible for weighing up a bid, and said it initiated an evaluation in November last year and finished integrating reports from other ministries early last month. However, although Vice Minister of Finance Wu Tang-chieh (吳當傑) denied the process was "black-boxed" as the reports "could" all be made public, discussion of it — government-led or not — was rarely seen until last week.
On the lack of discussion, it is easy to fault Taiwanese for being overly introspective when it comes to international affairs, but the government should know better after last year's Sunflower movement, especially about the contentiousness of China-related issues, and be responsible enough to include the public in the "evaluation" it claimed it has undergone or be more actively transparent.
The economics ministry made another misstep by refusing, and later denying its initial reluctance, to pass copies of its evaluation to reporters, which several financial journalists claimed to have received only after voicing complaints to an opposition lawmaker during a legislative committee meeting yesterday.
Long before the Sunflower movement, which was the culmination of long-running dissatisfaction with the KMT government, transparency is what the public has been constantly demanding with regards to negotiations with Beijing. It seems only fair to expect the government to be more sensitive to public misgiving about China-led agreements and institutions.
It turns out that the government and the KMT are as self-absorbed and obtuse as ever.
Their remarks, in response to public anger about the behind-the-door negotiations for the service trade pact, are worthy of applause, but ignored the real complaint — which has always been a call for the government's acknowledgment that the economy is never the only thing that Taiwan needs to take into account when dealing with China.


text:
The indescribable, here becomes reality: MAHLER'S EIGHTH at De Doelen, Rotterdam
MAHLER'S EIGHTH by the Rotterdam Philharmonisch conducted by Yannick Nezet-Seguin at De Doelen, Rotterdam on 23rd March 2018
Mahler's Eighth has certainly earned its place in the record books of classical music. There are over 500 people on stage, including nine (!) singers, three concert choirs, one children's choir and a large orchestra. Those sheer numbers create an unparalleled musical force.
This piece is also a challenge for the conductor: The various dynamic variations, from one end of the scale to the other, have to be coordinated and controlled, while still keeping the feeling of the music. And because there are so many people on stage, a conductor has to literally conduct, physically communicating his vision of the music so everyone on stage gets his message.
Directly before this Mahler Eighth in Rotterdam, Yannick Nezet-Seguin conducted runs of Strauss's Elektra and Wagner's Parsifal at the Met. And the influence of these two pieces was evident in his interpretation last night. The first part featured the strong, futuristic atonal edges as well as the complicated musical structures that we know from Strauss' one-act success. This music is deviously complicated, demanding a complicated musical structure like the double-fugue be transposed onto this monstrous sound apparatus. That is not an easy task for a conductor.
And like so many before him, Nezet-Seguin does not manage to completely solve these complications. In the first part, there is a lack of structure and transparency, Mahler is anything but easy-listening here. Especially in the passages where the soloists have to sing together as a group, their voices do not form a singular, well-sounding unit. That may also be because Sarah Connolly sadly had to cancel and was replaced by Michelle DeYoung. Instead, the pentecost choral Veni Creator Spiritus shines brightest in the choral numbers, where the stellar voices of Orfeon Donostiarra, Groot Omroep Koor, the Rotterdam Symphony Chorus and the Nationaal Kinderkoor can give their all. Still, there is that lack of clarity that prevents the audience from truly gaining access to this music.
At the beginning of the second part we hear the influence of Parsifal. The tempi are slow, the Poco Adagio is a clear and undisturbed sublime lake of sound. The dynamics are carefully layered throughout the orchestra, affording the brass some wonderful opportunities to shine. In the first two orchestral movements of part two, Nezet-Seguin forms a unified body of sound, that is then continually expanded in the arias of the individual singers. When Markus Werba beautifully sings Ewiger Wonnebrand, one is faintly reminded of Gurnemanz's Karfreitagszauber.
Michael Schade's tenor voice is overly pressed tonight, his timbre is slightly restricted. But that may also be due to the incredible demands that Mahler makes of the singers. The whole ensemble including Christof Fischesser and Mihoko Fujimura delivers stellar performance in the individual arias of part two.
The choirs never let up either, at times even dominating the orchestra. Especially in Jene Rosen aus den Händen, the orchestra is buried by the sheer tonal force of 4 choirs at the top of their craft. The orchestral force is also reduced because Nezet-Seguin has asked for an unusually light string section for this piece (only 10 celli?)
Indeed, the strings are remarkably subdued throughout the evening, even in their pizzicato passages towards the finale. Nezet-Seguin is constructing a different orchestral sound here, where the woodwinds and brass sections dominate and role out a grand red carpet of sound for singers and choirs. While that makes for rousing climaxes, I find that transparency and clarity are still somewhat lacking, compared to what other greats such as Pierre Boulez achieved in this piece.
But the Finale is something from another planet. Listeners and musicians alike are rewarded with splendid E-Flat major chords all around. Nezet-Seguin's intelligent handling of the organ (supporting, subdued) and the choirs' stellar efforts combine to make this finale one of the greatest musical experiences you will ever have. Beginning in a triple pianissimo, Nezet-Seguin animates all involved and manages to solve the riddle of the Eight in these final minutes.
Standing ovations after the final notes, totally deserved. Even though this interpretation could have featured more transparency and maturity at some stages, a complete performance comes together at the end, when everyone on stage joins forces. The brass is flawless and all on stage brilliantly communicate Mahler's musical vision in the final bars: The indiscribable, here it becomes reality. Yannik Eisenaecher 24th March 2018
Yannik Eisenaecher is the publisher of the blog FreshEarsClassics
← Drawn from Circus Rehearsals by Britt van Eijk at The Korzo in The Hague
BABEL: Old Masters Back from Japan at Museum Boijmans Van Beuningen in Rotterdam →


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Top 10 Ways to Stay Safe From Scams This Holiday Season
Lori Cline December 15, 2016 15-12-2016 10 minutes
The holidays are upon us, online sales are expected to go through the roof, and more U.S. consumers than ever (68 percent, or roughly 170 million people) plan to buy technological gifts this holiday season. Unfortunately, cybercriminals bank (no pun intended) on heightened shopping excitement and lowered safety practices, thus coming up with more creative ways to make their holiday a prosperous one.
It's hard to know what holiday scams are out there, let alone how to spot one; we're here to help. If you're aware of the dangers and how to spot them, it will give you better security on all your devices, and better peace of mind, as well.
1. Fraudulent Emails
Emails containing false offers for great deals, e-cards containing malware, imitation delivery statuses, and phony shipping confirmations are sent out in bulk this time of year. You'll get an email with FedEx telling you that there was a problem with your package. Or you'll open your inbox to find one or more offers that are practically impossible to ignore from a retailer you know. Or you'll see an email advising you of the shipping status of a package you order from UPS. They can all look real, and a lot of them could be bogus.
What to Do: Make sure it's a scam; there are a few ways you can spot one 5 Examples To Help You Spot A Fraud Or Fake Email 5 Examples To Help You Spot A Fraud Or Fake Email The shift from spam to phishing attacks is noticeable, and is on the rise. If there's a single mantra to keep in mind, it's this -- the number one defense against phishing is awareness. Read More . The punctuation and spelling will be poor, or you will be asked by the sender for personal information. Another way to tell is by checking the sender's email address against the sender's name.
Don't respond to a suspicious email or click on anything in it: No pictures, no links, no buttons, and no banners. If you click on any of the elements, it may activate malware and/or or provide personal information to the scammer. To identify the authenticity of the email content, open a new window and type in the retailer's address to go directly to the site.
If it's an e-card as an attachment, don't open the attachment if you don't know the sender. Be careful not to click on any links that say, "View the e-card". Doing either of these could download malware to your computer.
And never give your banking information, social security number, etc. to anyone. A reputable business would never ask you for personal information.
2. Change the Password on Your Cutlery
Image Credit: master_art via Shutterstock
Even a fork can connect to the internet these days, which means it needs a password. That fork, and any smart appliance and device (the Internet of Things), comes with a default one. If there is anything that is going to get your digital life into trouble, it's a default, hackable password.
What to Do: Change the manufacturer's default password to a password that's hard to guess and write it down. Make it a combination of letters, symbols and numbers, and change it regularly.
3. Don't Use the Same Password on All Sites
Passwords, passwords. When you sit down at your computer or you pull out your smartphone to shop, you're going to need a password for all those sites you're going to visit. Sure, having one password is convenient, but once someone hacks your password on one account, they'll reap the monetary benefits from all of them.
What to Do: Use a password manager that can be used on your smartphone as well as your computer, i.e. LastPass. This will keep track of your logins and passwords for any site you visit. You don't even have to type! A few clicks will fill everything in for you so you don't have to memorize any passwords if you don't want to.
Bonus Tip: When the site asks you if you want to store your financial details (like your credit card number) on the site to use the next time you're there, don't do it. It's yet another way for a criminal to get his hands on your personal information.
4. Enable Two-Factor Authentication
Speaking of shopping on websites, a lot of them offer two-factor authentication. This provides two layers of authenticity, not just one. The first layer of security is your login and password. Depending on the web site, the second layer of security will be something like a PIN, a second password, or even a voice print.
What to Do: Use two-factor authorization everywhere it's offered. Some types of businesses, like banks, have automatic two-factor authorization. It's a lot harder for a hacker to hack through two layers of security than just a password, so enable it whenever you can.
5. Check the Safety of the Website You're Visiting
A secure site has a closed padlock in the address bar and its URL starts with HTTPS, which means it has an SSL Certificate What Is an SSL Certificate, and Do You Need One? What Is an SSL Certificate, and Do You Need One? Browsing the Internet can be scary when personal information is involved. Read More . An SSL Certificate is a certificate designed to keep the connection more secure between you and the site's server.
What to Do: One excellent way to add some security to your browsing is to install a browser add-on called HTTPS Everywhere. Visit the website when you install it. It's designed to protect you from different types of surveillance Avoiding Internet Surveillance: The Complete Guide Avoiding Internet Surveillance: The Complete Guide Internet surveillance continues to be a hot topic so we've produced this comprehensive resource on why it's such a big deal, who's behind it, whether you can completely avoid it, and more. Read More and account hacking by automatically switching the site you are visiting from HTTP to HTTPS if the site supports it.
Always look for the little padlock in your address bar and https:// at the beginning of the URL — they show you are on a secure site. Note that the padlock placement in each browser varies a little (see below).
6. Set Up Account Alerts
You may be on top of your device's security, but cybercriminals are good at what they do. There's too much money at stake not to be. That being said, you may still get hacked.
What to Do: Set up alerts for your bank accounts. Almost all banks and most credit card companies give you the option to have an alert sent to your smartphone and/or email when there has been any activity on your account, your account is overdrawn or it drops below an certain amount (you set the amount), and more.
Bonus Tip: Go through your bank and credit card statements in detail. A lot of people kind of skim through their statements when they get them each month. Make sure you check each transaction to ensure that it's legitimate.
7. Watch the Smartphone Downloads
It's sad, but many apps aren't safe. In fact, malware is on the rise in smartphones. Downloading that new game you want to try or responding to a text you receive from an unknown sender can get you some harmful malware if you don't protect yourself.
What to Do: Don't download apps from questionable sources. If you're an Android user, use Google Play, although be forewarned that there is a small chance you can get hit by malware in Google Play, too. If there is an app that you are about to download that's questionable, use Verify Apps to scan it first (it's installed by default in Android). Apple seems to take security more seriously Smartphone Security: Can iPhones Get Malware? Smartphone Security: Can iPhones Get Malware? Malware affecting "thousands" of iPhones can steal App Store credentials, but the majority of iOS users are perfectly safe – so what's the deal with iOS and rogue software? Read More than Android as the only place you can download apps is at the App Store.
For safe texting, there's Signal. Signal provides end-to-end encryption, which turns a sender's regular text message into a secret one and can only be decoded by the recipient. Thus, every time you send a text, nobody can "see" its contents except the person you texted.
And take care of your smartphone like you do your computer; download and install an antivirus and a firewall program 10 Best Security & Privacy Apps for Smartphones & Tablets 10 Best Security & Privacy Apps for Smartphones & Tablets Security threats to smartphones and tablets are endless. Beyond the internal security options of your device, you should be using a few apps to protect your smartphone - here are some we recommend. Read More . From a reputable source, of course.
Bonus Tip: When you are installing an app, pay attention to the app needs access to. A lot of apps ask for far too much information than it needs. Don't give anyone, anywhere too much personal information.
8. Skype and Facebook and Twitter…
Image Credit: quka via Shutterstock
Is there any part of the internet that scammers have missed? The newest scams are being perpetrated through social engineering, which is just a fancy phrase for using social media to get people to cough up their personal details. And boy, is it amped up this time of year!
Facebook has false giveaway 3 Facebook Scams You Need to Watch Out for This Christmas 3 Facebook Scams You Need to Watch Out for This Christmas If you're not already, you should be concerned about how cybercriminals are using Facebook in order to scam you, especially during the Christmas period. These three scams show just how clever they can be. Read More contests and gifts, and Twitter has more spam than ever. One Skype email scam (it looks real but it's not) informs you that you have a call waiting. Click the link in the email and you will be taken to a site to download software full of adware or even worse, malicious code that can listen in on your conversations.
What to Do: See #1 in this article. Don't click on anything clickable in the email: pictures, links, buttons, and banners. If you get an email in Skype telling you have a new call, go to Skype itself — don't just click on the link. If you get a message in Facebook telling you can get a credit card pre-approved just in time for the holidays, go directly to the merchant who is offering the card. It may look like it's the real thing, but it quite possibly won't be. Check the real source. You can't be too careful.
9. False Charities
It's really sad, but criminals take advantage of people this time of year, knowing that they are more apt to give money to the needy. They falsify charity names or make new (fake) ones, and will do it through email, websites, phone calls, or even text.
What to Do: If you have never heard of the charity, go to the Better Business Bureau (BBB) and look up the name, or not give. It's that simple. You can always go to your favorite charity's website, or Google homeless shelters in your area. The point is to never give money to someone who you don't know.
10. Credit Card Skimming
We all have been advised to watch for people lurking in the shadows when we go to the ATM to get cash. Now we are being advised to watch the ATM 3 Danger Signs to Look for Each Time You Use an ATM 3 Danger Signs to Look for Each Time You Use an ATM Taking a few moments before popping your card into the ATM slot to withdraw money could be the difference between spotting it has been tampered with, and becoming penniless. But what should you look for? Read More and the gas pump when we scan our card. Around the holidays, this particular scam is perpetrated more, mainly because criminals know that more people will be using an ATM.
How does it work? Scammers will put a small device on an ATM or gas pump card reader (it fits on top of the real card reader so you can't tell it's fake unless you look for it) that will "skim" the magnetic tape to capture your PIN. This gives the scammer enough information to go shopping online or make an imitation card.
What to Do: When you're at the ATM or getting gas and go to put in your card, examine the card reader. If it looks at all different, reach out and shake or pull on it. It may pop right off. Take a quick glance at your card when you take it out of the reader. You might see glue around the edges of the card. Report it to the bank or proprietor if it's at all suspicious.
EMV (Europay, Mastercard, and Visa) is a standard of security that uses a computer chip to encrypt your card's data. If your card does not have an EMV chip and still has the magnetic strip, call your bank and ask for a new one. You should have one by now.
Stay Secure
Be diligent. Keep everything on your computer and smart phone patched and updated. Use antivirus and firewall software on your smartphone Do You Need Antivirus Apps on Android? What About iPhone? Do You Need Antivirus Apps on Android? What About iPhone? Does Android need antivirus apps? What about your iPhone? Here's why smartphone security apps are important. Read More as well as your computer. Make your passwords more difficult to guess, and make sure there is a different one for each log in that you have.
You don't want to be the victim of a cybercrime, and there is no such thing as too much security for things that are important to you.
Above all, have a wonderful, safe holiday!
Do you know of any more scams that surface more around the holidays? Have you had anyone hack you? Is there another scam that you know of that surfaces around the holidays? Let us know below.
Explore more about: ATM, Online Fraud, Online Security.
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Lori Cline 3 articles
A web content writer (fancy name for freelance writer) and a self-professed tech nerd, Lori loves anything tech and is happily obsessed with her current smart phone. And football. And chocolate.
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TasWater crews worked in conjunction with Veolia and their vac-truck to repair the connection line while limiting the disruption to customers.
An old copper pipe connecting a property to the mains had sprung a leak causing ongoing problems. As the pipe runs horizontally across the road, traffic control was on site to assist crews while a Veolia vac-truck was used to expose the underground connection.
The safety of the crew is our number one priority. As the old connection line was metal and a power line was in the nearby vicinity, a tool known as a clamp meter was used to test for any electrical currents. A clamp meter alarms when an electrical current is detected to alert our crews. Crew members would then stop work immediately and seek assistance from TasNetworks. Luckily, on this occasion, there were no currents on the pipe.
Once the old line was removed it was replaced with a poly pipe and water was the restored to the affected customer. Further work was then passed on to local contractors so the area could be reinstated to its original condition.


Inter-Continental Concerts held this special event to present a summary of their activities. At this event, the history of the organization, its purpose, products and achievements were reviewed by Shahed Mohseni Zonoozi, the founder and producer of the events. Highlights included the unveiling of the official ICC website, which contains the programs, events, music tracks and participant names. Sam, the ICC project manager, introduced the short movie titled "The Fellowship of EMU".
After the screening, members of the cast came to the stage. The evening culminated with a surprise appearance by former students through a video conference call.