The Interlaboratory Robustness Of Next-Generation Sequencing (IRON) Study Phase II: Deep-Sequencing Analyses Of Hematological Malignancies Performed In 8,867 Cases By An International Network Involving 27 Laboratories

Introduction: Amplicon deep-sequencing using second-generation sequencing technology is an innovative molecular diagnostic technique and enables a highly-sensitive detection of mutations. As an international consortium we had investigated previously the robustness, precision, and reproducibility of 454 amplicon next-generation sequencing (NGS) across 10 laboratories from 8 countries (Leukemia, 2011;25:1840-8).

Aims: In Phase II of the study, we established distinct working groups for various hematological malignancies, i.e. acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), and multiple myeloma. Currently, 27 laboratories from 13 countries are part of this research consortium. In total, 74 gene targets were selected by the working groups and amplicons were developed for a NGS deep-sequencing assay (454 Life Sciences, Branford, CT). A data analysis pipeline was developed to standardize mutation interpretation both for accessing raw data (Roche Amplicon Variant Analyzer, 454 Life Sciences) and variant interpretation (Sequence Pilot, JSI Medical Systems, Kippenheim, Germany).

Results: We will report on the design, standardization, quality control aspects, landscape of mutations, as well as the prognostic and predictive utility of this assay in a cohort of 8,867 cases. Overall, 1,146 primer sequences were designed and tested. In detail, for example in AML, 924 cases had been screened for CEBPA mutations. RUNX1 mutations were analyzed in 1,888 cases applying the deep-sequencing read counts to study the stability of such mutations at relapse and their utility as a biomarker to detect residual disease. Analyses of DNMT3A (n=1,041) were focused to perform landscape investigations and to address the prognostic relevance. Additionally, this working group is focusing on TET2, ASXL1, and TP53 analyses. A novel prognostic model is being developed allowing stratification of AML into prognostic subgroups based on molecular markers only. In ALL, 1,124 pediatric and adult cases have been screened, including 763 assays for TP53 mutations both at diagnosis and relapse of ALL. Pediatric and adult leukemia expert labs developed additional content to study the mutation incidence of other B and T lineage markers such as IKZF1, JAK2, IL7R, PAX5, EP300, LEF1, CRLF2, PHF6, WT1, JAK1, PTEN, AKT1, IL7R, NOTCH1, CREBBP, or FBXW7. Further, the molecular landscape of CLL is changing rapidly. As such, a separate working group focused on analyses including NOTCH1, SF3B1, MYD88, XPO1, FBXW7 and BIRC3. Currently, 922 cases were screened to investigate the range of mutational burden of NOTCH1 mutations for their prognostic relevance. In MDS, RUNX1 mutation analyses were performed in 977 cases. The prognostic relevance of TP53 mutations in MDS was assessed in additional 327 cases, including isolated deletions of chromosome 5q. Next, content was developed targeting genes of the cellular splicing component, e.g. SF3B1, SRSF2, U2AF1, and ZRSR2. In BCR-ABL1-negative MPN, nine genes of interest (JAK2, MPL, TET2, CBL, KRAS, EZH2, IDH1, IDH2, ASXL1) have been analyzed in a cohort of 155 primary myelofibrosis cases searching for novel somatic mutations and addressing their relevance for disease progression and leukemia transformation. Moreover, an assay was developed and applied to CMML cases allowing the simultaneous analysis of 25 leukemia-associated target genes in a single sequencing run using just 20 ng of starting DNA. Finally, nine laboratories are studying CML, applying ultra-deep sequencing of the BCR-ABL1 tyrosine kinase domain. Analyses were performed on 615 cases investigating the dynamics of expansion of mutated clones under various tyrosine kinase inhibitor therapies.

Conclusion: Molecular characterization of hematological malignancies today requires high diagnostic sensitivity and specificity. As part of the IRON-II study, a network of laboratories analyzed a variety of disease entities applying amplicon-based NGS assays. Importantly, the consortium not only standardized assay design for disease-specific panels, but also achieved consensus on a common data analysis pipeline for mutation interpretation. Distinct working groups have been forged to address scientific tasks and in total 8,867 cases had been analyzed thus far.

Iowa Department of Transportation Major Items - Preliminary Quantity Report, June 21, 2016; With Addendums

Weekly letting report

Second Welland Canal - Book 3, Survey Map 21 - Village of Port Colborne

Survey map of the Second Welland Canal created by the Welland Canal Company showing the areas in and around Port Colborne. Identified structures associated with the Canal include Lighthouse, Pier Light, Old Lock House, Collector's Office, Harbour Master's House, Canal Boundary, Back Ditch, Reserved Back Ditch, Basin, Light-Keeper's House and Ferry Recess. The surveyors' measurements and notes can be seen in red and black ink and pencil. Local area landmarks and businesses are also identified and include Gordon's Woodyard, Welland Rail Road, Welland Railway Elevator and Proposed Elevator, W.R.R. Flour Shed, Roman Catholic Church, School House, Sandhills, Lake Erie, and the High Water Mark. Streets running parallel to Canal include King St., West St., East St., Queen St., Hamilton St., and the Road Allowance are labelled. Streets running perpendicular to Canal include Kent St., Victoria St., Adelaide St., SugarLoaf St., George St., Alexandrina St., William St., Fort Erie St., Lake Rd., and New Road to Dutch Settlement are also labelled. Property owners and leasers as well as buildings on lots are also idenitified and noted as follows: Adams estate, J. Towhig, J.C. Kerr, Mrs. Hill, S. Cooke, Mrs. Yocum, W.T. Cooke, P. Wintermute, J. Shickluna, William Cooke, J. McChesney, John Beatty, W. Robertson, John Gordon, T. Armstrong, John Harper, George Keefer, Estate of James Black, Thomas Park, N. Higgins, S. Hopkins, and L.G. Cartier. Map of the Village of Port Colborne. Being Lot No. 27 and part of Lot No. 28 in the 1st Con. Township of HUMBERSTONE. Scale 2 Chs. per Inch. land shaded in RED Owned by DEPT. Do. Do. BLUE Sold to the COUNTY of WELLAND

Portsmouth Oracle, 12 February 1814 (vol. 25, no. 21) - Newspaper

Volumes of interest were published between 1812 and 1815 with articles about the War of 1812.

The News, 21 November 1813 (no. 437) - Newspaper

Volumes of interest were published between 1812 and 1815 with articles about the War of 1812.

Letter - E.C. Schmon to Arthur A. Schmon, 21 August 1918

Eleanore Celeste is writing to Arthur from Whippang, New Jersey. She describes the beautiful woods, fields and mountains. The letter is labelled number 148.

Colón : monólogo en verso. Estrenado en el Teatro de Durango la noche del 27 de Octubre de 1887, á beneficio del Sr. Francisco Galán Rivas

UANL

L'Action universitaire – vol. 27, no 1, nov. 1960

Pour toute demande de reproduction de contenu se trouvant dans cette publication, communiquer avec l’Association des diplômés de l’UdeM.

Aproximación al pensamiento pedagógico del profesor de apoyo a la integración de los alumnos trisómicos-21 de Málaga.

Mostrar qué tipo de planificación realiza el profesor de apoyo para la integración de los trisómicos-21. Hacer evidente el proceso de decisiones interactivas del profesor de apoyo. Explicar las teorías implícitas, creencias del profesor y hacer visible el paisaje de integración de los trisómicos-21 en Málaga desde el pensamiento pedagógico del profesor de apoyo. Nueve profesores de apoyo para la integración de los trisómicos-21. La primera fase utiliza el cuestionario para la recogida de información, obteniendo una visión general de los profesores respecto a la integración. En la segunda fase se hacen fotografías que luego son comentadas por un mínimo de tres personas, entre ellas el profesor de apoyo, para un mayor acercamiento al tema de estudio. En la fase siguiente se aproxima la planificación, toma de decisiones y teorías de los profesores de apoyo, por último la investigación se centra en un solo sujeto. Cuestionario, cintas grabadas, videograma, diario de campo y fotografías. Estimulación del recuerdo, videoanálisis, observación persistente y triangulación. Los profesores de apoyo elaboran los programas de desarrollo individual junto al logopeda, el psicólogo y profesor tutor. Las decisiones interactivas que toma el profesor de apoyo están relacionadas con la planificación de los programas de desarrollo individual y se destina a actividades relacionadas con la comunicación oral, escritas y otras. Las creencias de estos profesores se basan en las posibilidades cognitivas de los trisómicos-21. Las funciones que creen desempeñar son las de procurar cambios para la integración, asesorar a los padres de los niños, que se verían más favorecidos si la Administración no estuviese alejada de los problemas de los colegios.

Agenda 21 de Málaga : hacia la ciudad sostenible.

El proyecto de elaboración de la Agenda 21 Provincial de Málaga (A21P), se formula siguiendo el llamamiento a todas las comunidades locales a través del capítulo 28 de la Agenda 21, que es el Plan de Acción de la ONU para un desarrollo sostenible en el siglo XXI, documento clave de los aprobados en la Cumbre de la Tierra sobre el Medio Ambiente y el Desarrollo Sostenible (CNUMA) que tuvo lugar en Río de Janeiro del 3 al 14 de junio de 1992. En dicho documento se reconoce el papel de las autoridades locales para iniciar y coordinar los procesos necesarios encaminados al desarrollo sostenible mediante un esquema participativo. La Diputación de Málaga expresó su adhesión a la Carta de Aalborg el 9 de mayo de 2000, lo que implicó adquirir el compromiso sobre la realización de sus objetivos sobre desarrollo sostenible aprobando un año más tarde la realización de la Agenda 21 Provincial. Se inicia así un proceso consensuado entre la administración provincial así como otras administraciones y los agentes sociales y económicos que propicie recoger propuestas y medidas con la finalidad de alcanzar un Desarrollo Sostenible, haciendo compatible la calidad ambiental con el desarrollo socioeconómico.