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Supplementary Data Table S1: general characterisctic of the different cohorts of patients Supplementary Data Table S2: characterisctic of Population A Supplementary Data Table S3: characterisctic of Population B Supplementary Data Table S4: characterisctic of Population C Supplementary Data Table S5: characterisctic of Population D Supplementary Data Table S6: Primer sequences Supplementary Data Table S7: Differentially expressed transcripts between IgLV3-21 + and - patients Supplementary Data Table S8: differentially expressed genes between IgLV3-21 + and - patients Supplementary Data Table S9: differentially expressed genes between IgLV3-21 + and - patients. Restricted to absolute fold-change > 1.5 and protein coding genes only Supplementary Data Table S10 Gene set Enrichment analysis between IgLV3-21 positive and negative patients

Supplementary Data Figure S1. Determination of IgLV3-21 positivity by real-time PCR. Supplementary Data Figure S2: CLL-OS analysis including only CLL-related deaths. Supplementary Data Figure S3. Validation of the poor prognosis associated with IgLV3-21 usage in population C. Supplementary Data Figure S4. Validation of the poor prognosis of IgLV3-21 usage in population D. Supplementary Data Figure S5: prognostic impact of IgLV3-21 usage in the different CLL-IPI subgroups. Supplementary Data Figure S6. Immunogenetic features of IgLV3-21 patients. Supplementary Data Figure S7. CXCR4 surface expression is downregulated in IgLV3-21 patients.

Purpose: Unmutated (UM) immunoglobulin heavy chain variable region (IgHV) status or IgHV3-21 gene usage is associated with poor prognosis in chronic lymphocytic leukemia (CLL) patients. Interestingly, IgHV3-21 is often co-expressed with light chain IgLV3-21, which is potentially able to trigger cell-autonomous BCR-mediated signaling. However, this light chain has never been characterized independently of the heavy chain IgHV3-21.Experimental Design: We performed total RNA sequencing in 32 patients and investigated IgLV3-21 prognostic impact in terms of treatment-free survival (TFS) and overall survival (OS) in 3 other independent cohorts for a total of 813 patients. IgLV3-21 presence was tested by real-time PCR and confirmed by Sanger sequencing.Results: Using total RNA sequencing to characterize 32 patients with high-risk CLL, we found a high frequency (28%) of IgLV3-21 rearrangements. Gene set enrichment analysis revealed that these patients express higher levels of genes responsible for ribosome biogenesis and translation initiation (P < 0.0001) as well as MYC target genes (P = 0.0003). Patients with IgLV3-21 rearrangements displayed a significantly shorter TFS and OS (P < 0.05), particularly those with IgHV mutation. In each of the three independent validation cohorts, we showed that IgLV3-21 rearrangements—similar to UM IgHV status—conferred poor prognosis compared with mutated IgHV (P < 0.0001). Importantly, we confirmed by multivariate analysis that this was independent of IgHV mutational status or subset #2 stereotyped receptor (P < 0.0001).Conclusions: We have demonstrated for the first time that a light chain can affect CLL prognosis and that IgLV3-21 light chain usage defines a new subgroup of CLL patients with poor prognosis. Clin Cancer Res; 24(20); 5048–57. ©2018 AACR.

Some studies date the origins of US intercollegiate football—and, by extension, the modern game of American football—back to a soccer-style game played between Princeton and Rutgers universities in 1869. This article joins with others to argue that such a narrative is misleading and goes further to clarify the significance of two “international” fixtures in 1873 and 1874, which had a formative and lasting impact on football in the United States. These games, contested between alumni from England’s Eton College and students at Yale University, and between students at Canada’s McGill University and Harvard University, combined to revolutionize the American football code. Between 1875 and 1880, previous soccer-style versions of US intercollegiate football were replaced with an imported, if somewhat modified, version of rugby football. It was the “American rugby” that arose as a result of these transnational exchanges that is the true ancestor of the gridiron game of today.

The early twentieth century was a time when the US public consciousness recognized an increasing association between their political leaders and sports and athleticism. With an exceptional precedent for this connection set by Theodore Roosevelt (1901–09), his replacement as US president would inevitably find it hard to keep pace. In the modern-day popular consciousness, Roosevelt’s immediate successor, William Howard Taft (1909–13), is often noted more for his obesity than for his physical athleticism or sporting prowess. Yet, as this article shows, as Taft moved closer to the White House, the contemporary US press increasingly associated him with sports, and at least the pursuit of physical fitness. In a post-Rooseveltian America, a rise to national political prominence demanded a portrayal of a president’s links to sports and athleticism, even in the unlikeliest of candidates.

The supply of blood in many low- and middle-income nations in Sub-Saharan Africa (SSA) does not meet the patient care needs. Lack and delay of blood transfusion cause harm to patients and slow the rate of progress in other parts of the health system. Recognizing the power of implementation science, the BLOODSAFE Program was initiated which supports three SSA research study teams and one data coordinating center (DCC) with the goal to improve access to safe blood transfusion in SSA.The study team in Ghana is focusing on studying and decreasing iron deficiency in blood donors and evaluating social engagement of blood donors through different approaches. The study team in Kenya is building a "vein to vein" workflow model to elucidate and devise strategies to overcome barriers to blood donation and improve infrastructural components of blood product production and use. The Malawi team is studying the infectious disease ramifications of blood donation as well as blood donor retention strategies aimed at blood donors who commence their donation career in secondary schools.Together the project teams and the DCC work as a consortium to support each other through a shared study protocol that will study donor motivations, outcomes, and adverse events across all three countries. The BLOODSAFE Program has the potential to lead to generalizable improvement approaches for increasing access to safe blood in SSA as well as mentoring and building the research capacity and careers of many investigators.

Unlike many leading studies of sport history in the United States, such as Steven Riess's Sport in Industrial America, 1850–1920 (1995), Kenneth Cohen spends relatively little time exploring the mo...

Review of "The Good Occupation: American Soldiers and the Hazards of Peace, Susan L. Carruthers (2016) Cambridge, MA and London: Harvard University Press, 386 pp., ISBN 978-0-67454-570-0, h/bk, £21.95".

Background and AimsPancreatic ductal adenocarcinoma (PDAC) is characterised by advanced disease stage at presentation, aggressive disease biology and resistance to therapy resulting in extremely poor five-year survival MethodsGenome-wide epigenetic mapping of DNA modifications 5-hydroxymethylcytosine (5mc) and 5-hydroxymethylcytosine (5hmc) using oxidative bisulphite sequencing (oxBS). Bioinformatics using iCluster and mutational profiling to identify overlap with transcriptional signatures in FFPE from resected patients and confirmation in vivo.ResultsWe find that more aggressive squamous-like PDAC subtypes result from epigenetic inactivation of loci including GATA6 that promote differentiated classical-pancreatic subtypes. We show that squamous-like PDAC transcriptional subtypes are associated with greater loss of 5hmc due to reduced expression of the 5mc-hydroxylase TET2. Furthermore, we find that SMAD4 directly supports TET2 levels in the pancreas and classical-pancreatic tumors and loss of SMAD4 expression is associated reduced 5hmc, GATA6 and squamous-like tumors. Importantly, enhancing TET2 stability using Metformin and VitaminC/ascorbic acid (AA) restores 5hmc and GATA6 levels, reverting squamous-like tumor phenotypes and WNT-dependence in vitro and in vivo.ConclusionsWe identify epigenetic deregulation of pancreatic differentiation as an underpinning event behind the emergence of transcriptomic subtypes in PDAC. Our data shows that restoring epigenetic control increases biomarkers of classical-pancreatic tumors and raises the possibility that combination of Vitamin C and Metformin may prolong survival in patients with squamous-like pancreatic cancer.

BACKGROUND & AIMS: Pancreatic ductal adenocarcinoma (PDAC) is characterized by advanced disease stage at presentation, aggressive disease biology, and resistance to therapy, resulting in an extremely poor 5-year survival rate of

Manual wheelchair users exhibit pain related to repetitive and demanding shoulder activities of daily living. Wearable sensing systems like force measurement gloves can provide insights about upper extremity loading in the community and home environments. We calibrated and evaluated the accuracy of a novel force measurement glove with a body-worn data logger. The device was calibrated with loads of 0-800 N applied to the palmar surface of the glove. Calibration conditions were tested that varied the stiffness of the material in the glove, the temperature, and the curvature of the force applicator. Calibration equations from each condition were evaluated by comparing the glove’s force prediction with the output of an instrumented wheelchair rim during propulsion and weight relief exercises. The force measurement glove detected 72.7% of 355 propulsion peaks and had a strong linear correlation with the instrumented rim force measurements (r=0.80). The most accurate calibration equation was constructed using data from all conditions, with an RMS force measurement error of 64.7 N and 31.7 N for weight relief exercise and propulsion, respectively. The force measurement glove design described here may serve as a useful tool for detection of loading events and relative magnitude changes.

Most studies of President Theodore Roosevelt address his “southern strategy” to revive the Republican Party’s fortunes in a region where it was effectively shut out by 1900. This essay revi...

Background: Cardiovascular disease (CVD) is the number 1 killer of women in the United States, yet few younger women are aware of this fact. CVD campaigns focus little attention on physicia...

This corrects the article DOI: 10.1038/leu.2017.177., info:eu-repo/semantics/published

Chronic lymphocytic leukaemia (CLL) is characterised by considerable clinical and biological heterogeneity, with specific recurrent genomic alterations, includingTP53mutations, deletions of chromosome 17p, and IgHV mutational status, impacting on response to chemo-immunotherapy and targeted agents. Consequently, diagnostic screening for these predictive biomarkers is recommended in both national and international clinical guidelines. Current conventional methods, including fluorescentin-situhybridisation and Sanger sequencing, exhibit shortcomings in terms of cost, speed and sensitivity, and even second-generation sequencing methods encounter technical limitations imposed by short-read lengths and bio-informatics analysis. The MinION platform from Oxford Nanopore Technologies generates exceptionally long (1-100kbp) read lengths in a short period of time and at low cost, making it a good candidate for diagnostic testing. In this paper, we present a nanopore-based CLL-specific screening assay, to simultaneously screen for bothTP53mutations and del17p13.1, as well as determining the IgHV mutation status for a single patient in one sequencing run. We sequenced 11 CLL patients and were able to generate a full diagnostic dataset for all. We identified somatic SNVs and indels in the coding region ofTP53, and demonstrate that, following error correction of the data, we could accurately define the somatically hypermutated IgHV region in all patients. We also demonstrated the ability of the MinION platform to detect large-scale genomic deletions through low-coverage whole-genome sequencing. We conclude that nanopore sequencing has the potential to provide accurate, low-cost and rapid diagnostic information, which could be applied to other cancer types.

The 100 000 Genome Project aims to develop a diagnostics platform by introducing whole genome sequencing (WGS) into clinical practice. Samples from patients with chronic lymphocytic leukaemia were subjected to WGS. WGS detection of single nucleotide variants and insertion/deletions were validated by targeted next generation sequencing showing high concordance (96·3%), also for detection of sub-clonal variants and low-frequency TP53 variants. Copy number alteration detection was verified by fluorescent in situ hybridisation and genome-wide single nucleotide polymorphism array (concordances of 86·7% and 92·9%, respectively), confirming adequate sensitivity by WGS. Our results confirm that WGS can provide comprehensive genomic characterisation for clinical trials, drug discovery and, ultimately, precision medicine.

The immunoglobulin heavy-chain variable region gene (IgHV) mutational status is considered the gold standard of prognostication in chronic lymphocytic leukemia (CLL) and is currently determined by Sanger sequencing that allows the analysis of the major clone. Using next-generation sequencing (NGS), we sequenced the IgHV gene from two independent cohorts: (A) 270 consecutive patient samples obtained at diagnosis and (B) 227 patients from the UK ARCTIC-AdMIRe clinical trials. Using complementary DNA from purified CD19+CD5+ cells, we demonstrate the presence of multiple rearrangements in independent experiments and showed that 24.4% of CLL patients express multiple productive clonally unrelated IgHV rearrangements. On the basis of IgHV-NGS subclonal profiles, we defined five different categories: patients with (a) multiple hypermutated (M) clones, (b) 1 M clone, (c) a mix of M-unmutated (UM) clones, (d) 1 UM clone and (e) multiple UM clones. In population A, IgHV-NGS classification stratified patients into five different subgroups with median treatment-free survival (TFS) of280(a), 131(b), 94(c), 29(d), 15(e) months (P0.0001) and a median OS of397(a), 292(b), 196(c), 137(d) and 100(e) months (P0.0001). In population B, the poor prognosis of multiple UM patients was confirmed with a median TFS of 2 months (P=0.0038). In conclusion, IgHV-NGS highlighted one quarter of CLL patients with multiple productive IgHV subclones and improves disease stratification and raises important questions concerning the pre-leukemic cellular origin of CLL.