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https://www.readbyqxmd.com/read/29036167/development-and-validation-of-a-protocol-for-optimizing-the-use-of-paraffin-blocks-in-molecular-epidemiological-studies-the-example-from-the-hpv-ahead-study
#1
Marisa Mena, Belen Lloveras, Sara Tous, Johannes Bogers, Fausto Maffini, Nitin Gangane, Rekha Vijay Kumar, Thara Somanathan, Eric Lucas, Devasena Anantharaman, Tarik Gheit, Xavier Castellsagué, Michael Pawlita, Silvia de Sanjosé, Laia Alemany, Massimo Tommasino
Worldwide use of formalin-fixed paraffin-embedded blocks (FFPE) is extensive in diagnosis and research. Yet, there is a lack of optimized/standardized protocols to process the blocks and verify the quality and presence of the targeted tissue. In the context of an international study on head and neck cancer (HNC)-HPV-AHEAD, a standardized protocol for optimizing the use of FFPEs in molecular epidemiology was developed and validated. First, a protocol for sectioning the FFPE was developed to prevent cross-contamination and distributed between participating centers...
2017: PloS One
https://www.readbyqxmd.com/read/29032825/line-1-retrotransposon-mediated-dna-transductions-in-endometriosis-associated-ovarian-cancers
#2
Zhouchunyang Xia, Dawn R Cochrane, Michael S Anglesio, Yi Kan Wang, Tayyebeh Nazeran, Basile Tessier-Cloutier, Melissa K McConechy, Janine Senz, Amy Lum, Ali Bashashati, Sohrab P Shah, David G Huntsman
OBJECTIVE: Endometrioid (ENOC) and clear cell ovarian carcinoma (CCOC) share a common precursor lesion, endometriosis, hence the designation endometriosis associated ovarian cancers (EAOC). Long interspersed nuclear element 1 (LINE-1 or L1), is a family of mobile genetic elements activated in many cancers capable of moving neighboring DNA through 3' transductions. Here we investigated the involvement of specific L1-mediated transductions in EAOCs. METHODS: Through whole genome sequencing, we identified active L1-mediated transductions originating within the TTC28 gene in 34% (10/29) of ENOC and 31% (11/35) of CCOC cases...
October 9, 2017: Gynecologic Oncology
https://www.readbyqxmd.com/read/29030614/retinal-degeneration-protein-3-rd3-in-normal-human-tissues-novel-insights
#3
Sheeja Aravindan, Dinesh Babu Somasundaram, Kwok Ling Kam, Karthikeyan Subramanian, Zhongxin Yu, Terence S Herman, Kar-Ming Fung, Natarajan Aravindan
The 195-amino-acid-long human Retinal Degeneration Protein 3 (RD3) is critical in the regulation of guanylate cyclase (GC) signaling and photoreceptor cell survival. Recently, we identified significant loss of RD3 in high-risk neuroblastoma and the influential role of RD3 in tumor progression. However, the functional characterization of RD3 in tumor systems has been hampered by the dearth of information on its localization in normal tissue and by the lack of antibodies suitable for staining FFPE tissue, primarily due to the inaccessibility of the epitopes...
October 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29029636/gene-expression-modules-in-primary-breast-cancers-as-risk-factors-for-organotropic-patterns-of-first-metastatic-spread-a-case-control-study
#4
Katherine Lawler, Efterpi Papouli, Cristina Naceur-Lombardelli, Anca Mera, Kayleigh Ougham, Andrew Tutt, Siker Kimbung, Ingrid Hedenfalk, Jun Zhan, Hongquan Zhang, Richard Buus, Mitch Dowsett, Tony Ng, Sarah E Pinder, Peter Parker, Lars Holmberg, Cheryl E Gillett, Anita Grigoriadis, Arnie Purushotham
BACKGROUND: Metastases from primary breast cancers can involve single or multiple organs at metastatic disease diagnosis. Molecular risk factors for particular patterns of metastastic spread in a clinical population are limited. METHODS: A case-control design including 1357 primary breast cancers was used to study three distinct clinical patterns of metastasis, which occur within the first six months of metastatic disease: bone and visceral metasynchronous spread, bone-only, and visceral-only metastasis...
October 13, 2017: Breast Cancer Research: BCR
https://www.readbyqxmd.com/read/29024724/paxgene-fixation-enables-comprehensive-metabolomic-and-proteomic-analyses-of-tissue-specimens-by-maldi-msi
#5
Christian Urban, Achim Buck, Jens T Siveke, Florian Lordick, Birgit Luber, Axel Walch, Michaela Aichler
An alcohol-based non-crosslinking tissue fixative, PAXgene Tissue System, has been proposed as alternative fixation method to formalin, providing superior and morphological preservation. To date, metabolites have not been assessed in PAXgene-fixed tissues. The study focuses on a comparison between PAXgene and standard formalin fixation for metabolomic analysis by MALDI mass spectrometry imaging. Therefore, fifty-six samples from seven mice organs were fixed with PAXgene (PFPE) or formalin (FFPE), embedded in paraffin, and processed to a tissue microarray...
October 9, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29024530/fna-smears-of-pancreatic-ductal-adenocarcinoma-are-superior-to-formalin-fixed-paraffin-embedded-tissue-as-a-source-of-dna-comparison-of-targeted-kras-amplification-and-genotyping-in-matched-preresection-and-postresection-samples
#6
Christopher P Hartley, Aparna M Mahajan, Suzanne M Selvaggi, William M Rehrauer
BACKGROUND: The current study was conducted to compare DNA yield, including normalization to nuclear area, DNA amplification functionality, and detection of KRAS mutations between matched fine-needle aspiration (FNA) specimens and pancreatic resections diagnostic of pancreatic ductal adenocarcinoma. METHODS: A retrospective sample of 30 matched single FNA smears and macrodissected formalin-fixed, paraffin-embedded (FFPE) curls (2 5-μm curls) were compared by measuring the following: nuclear area (via digital image analysis), DNA yield (via NanoDrop spectrophotometry and Quantus fluorometry), and polymerase chain reaction threshold cycles for KRAS amplifications...
October 12, 2017: Cancer
https://www.readbyqxmd.com/read/29024035/the-role-of-human-papillomavirus-in-p16-positive-oral-cancers
#7
Simone Belobrov, Alyssa M Cornall, Richard J Young, Kendrick Koo, Christopher Angel, David Wiesenfeld, Danny Rischin, Suzanne M Garland, Michael McCullough
BACKGROUND: The aim of the present study was to identify the presence and frequency of human papillomavirus (HPV) nucleic acid in p16 positive oral squamous cell carcinomas (OSCC), to assess if the virus were transcriptionally active, and to assess the utility of p16 overexpression as a surrogate marker for HPV in OSCC. METHODS: Forty-six OSCC patients treated between 2007 and 2011 with available formalin-fixed paraffin embedded (FFPE) specimens were included. Twenty-three patients were positive for p16 by immunohistochemistry (IHC) and these were matched with 23 patients with p16 negative tumours...
October 11, 2017: Journal of Oral Pathology & Medicine
https://www.readbyqxmd.com/read/29022941/genome-wide-mapping-of-dnase-i-hypersensitive-sites-in-rare-cell-populations-using-single-cell-dnase-sequencing
#8
James Cooper, Yi Ding, Jiuzhou Song, Keji Zhao
Increased chromatin accessibility is a feature of cell-type-specific cis-regulatory elements; therefore, mapping of DNase I hypersensitive sites (DHSs) enables the detection of active regulatory elements of transcription, including promoters, enhancers, insulators and locus-control regions. Single-cell DNase sequencing (scDNase-seq) is a method of detecting genome-wide DHSs when starting with either single cells or <1,000 cells from primary cell sources. This technique enables genome-wide mapping of hypersensitive sites in a wide range of cell populations that cannot be analyzed using conventional DNase I sequencing because of the requirement for millions of starting cells...
November 2017: Nature Protocols
https://www.readbyqxmd.com/read/29021820/introduction-of-p16-ink4a-as-a-surrogate-biomarker-for-hpv-in-women-with-invasive-cervical-cancer-in-sudan
#9
Hina Sarwath, Devendra Bansal, Nazik Elmalaika Husain, Mahmoud Mohamed, Ali A Sultan, Shahinaz Bedri
BACKGROUND: Cervical cancer is the fourth most common cancer in women worldwide with highest incidence reported in Eastern Africa in 2012. The primary goal of this study was to study the expression of p16(INK4a) in squamous cell carcinoma (SCC) of the cervix by immunohistochemistry (IHC) and determine relation with clinico-pathological parameters. This study further explored the correlation of p16(INK4a) immunostaining with another proliferation marker, Ki-67 and to study if human papillomavirus (HPV) IHC can be used as a marker for detection of virus in high-grade dysplasia...
2017: Infectious Agents and Cancer
https://www.readbyqxmd.com/read/29020284/performance-of-targeted-fungal-sequencing-for-culture-independent-diagnosis-of-invasive-fungal-disease
#10
Carlos A Gomez, Indre Budvytiene, Allison J Zemek, Niaz Banaei
Background: Identification of fungi causing invasive fungal disease (IFD) is critical for guiding antifungal therapy. We describe the performance and clinical impact of a targeted panfungal PCR amplicon sequencing assay for culture-independent diagnosis of IFD. Methods: Between January 2009 and September 2016, 233 specimens, consisting of fresh and formalin-fixed paraffin embedded (FFPE) tissues, and sterile body fluids with known diagnosis of IFD based on reference method results (n=117), and specimens with negative fungal culture, but with microscopic and ancillary findings indicative of IFD (n=116), were included...
August 17, 2017: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
https://www.readbyqxmd.com/read/29019127/dige-saturation-labeling-for-scarce-amounts-of-protein-from-formalin-fixed-paraffin-embedded-ffpe-tissue
#11
Paul Dowling
In this chapter, we describe the utility of difference gel electrophoresis (DIGE) as a proteomics platform for the global detection of expressed proteins in formalin-fixed paraffin-embedded (FFPE) tissues and its use for biomarker discovery/identification of proteins that may contribute to cancer development and progression. Formalin fixation and paraffin embedding of tissue is the standard processing methodology practiced in pathology laboratories worldwide, resulting in a highly stable form of tissue that is easily stored due to its inherent stability at room temperature...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29018799/truseq-based-gene-expression-analysis-of-formalin-fixed-paraffin-embedded-ffpe-cutaneous-t-cell-lymphoma-samples-subgroup-analysis-results-and-elucidation-of-biases-from-ffpe-sample-processing-on-the-truseq-platform
#12
Philippe Lefrançois, Michael T Tetzlaff, Linda Moreau, Andrew K Watters, Elena Netchiporouk, Nathalie Provost, Martin Gilbert, Xiao Ni, Denis Sasseville, Madeleine Duvic, Ivan V Litvinov
Cutaneous T-cell lymphomas (CTCLs) are a heterogeneous group of malignancies with courses ranging from indolent to potentially lethal. We recently studied in a 157 patient cohort gene expression profiles generated by the TruSeq targeted RNA gene expression sequencing. We observed that the sequencing library quality and depth from formalin-fixed paraffin-embedded (FFPE) skin samples were significantly lower when biopsies were obtained prior to 2009. We also observed that the fresh CTCL samples clustered together, even though they included stage I-IV disease...
2017: Frontiers in Medicine
https://www.readbyqxmd.com/read/28991373/novel-enriched-pathways-in-superficial-malignant-peripheral-nerve-sheath-tumors-mpnst-and-spindle-desmoplastic-melanomas-sdm
#13
George Jour, Nicole K Andeen, Rami Al -Rohil, Phyu P Aung, Meenakshi Mehrotra, Dzifa Duose, Benjamin Hoch, Zolt Argenyi, Rajyalakshmi Luthra, Ignacio I Wistuba, Victor G Prieto
Superficial malignant peripheral nerve sheath tumor (MPNST) is a rare, soft tissue neoplasm that shares morphological features and some molecular events with spindle and desmoplastic melanoma (SDM). Herein we sought to identify molecular targets for therapy using targeted RNA/DNA sequencing and gene expression of key immunological players. DNA and RNA from formalin-fixed, paraffin-embedded (FFPE) tissue were extracted and processed. Massive high-throughput deep parallel sequencing was performed using the Oncomine comprehensive panel enabling detection of relevant SNVs, CNVs, gene fusions, and indels from 143 unique genes on the Ion torrent sequencer for clinical trial research programs...
October 9, 2017: Journal of Pathology
https://www.readbyqxmd.com/read/28985579/metformin-and-insulin-impact-on-clinical-outcome-in-patients-with-advanced-hepatocellular-carcinoma-receiving-sorafenib-validation-study-and-biological-rationale
#14
Andrea Casadei Gardini, Luca Faloppi, Serena De Matteis, Francesco Giuseppe Foschi, Nicola Silvestris, Francesco Tovoli, Vincenzo Palmieri, Giorgia Marisi, Oronzo Brunetti, Umberto Vespasiani-Gentilucci, Giuseppe Perrone, Martina Valgiusti, Anna Maria Granato, Giorgio Ercolani, Giulia Negrini, Emiliano Tamburini, Giuseppe Aprile, Alessandro Passardi, Daniele Santini, Stefano Cascinu, Giovanni Luca Frassineti, Mario Scartozzi
PURPOSE: In 2015, we published a study on a small series of patients with hepatocellular carcinoma (HCC) treated chronically with metformin for type II diabetes mellitus (DM2) who showed a poorer response to sorafenib. The aim of the present study was to validate the prognostic significance of metformin in HCC patients treated with sorafenib, providing a biological rationale for the mechanism of resistance to sorafenib in patients on chronic metformin therapy, and to clarify the role of sirtuin-3 (SIRT-3), a protein involved in metabolic diseases and acknowledged as a tumour suppressor in HCC, in this resistance...
October 3, 2017: European Journal of Cancer
https://www.readbyqxmd.com/read/28984675/feasibility-of-prostate-paxgene-fixation-for-molecular-research-and-diagnostic-surgical-pathology-comparison-of-matched-fresh-frozen-ffpe-and-pfpe-tissues
#15
Gunilla Högnäs, Kati Kivinummi, Heini M L Kallio, Reija Hieta, Pekka Ruusuvuori, Antti Koskenalho, Juha Kesseli, Teuvo L J Tammela, Jarno Riikonen, Joanna Ilvesaro, Saara Kares, Pasi P Hirvikoski, Marita Laurila, Tuomas Mirtti, Matti Nykter, Paula M Kujala, Tapio Visakorpi, Teemu Tolonen, G Steven Bova
Advances in prostate cancer biology and diagnostics are dependent upon high-fidelity integration of clinical, histomorphologic, and molecular phenotypic findings. In this study, we compared fresh frozen, formalin-fixed paraffin-embedded (FFPE), and PAXgene-fixed paraffin-embedded (PFPE) tissue preparation methods in radical prostatectomy prostate tissue from 36 patients and performed a preliminary test of feasibility of using PFPE tissue in routine prostate surgical pathology diagnostic assessment. In addition to comparing histology, immunohistochemistry, and general measures of DNA and RNA integrity in each fixation method, we performed functional tests of DNA and RNA quality, including targeted Miseq RNA and DNA sequencing, and implemented methods to relate DNA and RNA yield and quality to quantified DNA and RNA picogram nuclear content in each tissue volume studied...
October 3, 2017: American Journal of Surgical Pathology
https://www.readbyqxmd.com/read/28978092/native-glycan-fragments-detected-by-maldi-ft-icr-mass-spectrometry-imaging-impact-gastric-cancer-biology-and-patient-outcome
#16
Thomas Kunzke, Benjamin Balluff, Annette Feuchtinger, Achim Buck, Rupert Langer, Birgit Luber, Florian Lordick, Horst Zitzelsberger, Michaela Aichler, Axel Walch
Glycosylation in cancer is a highly dynamic process that has a significant impact on tumor biology. Further, the attachment of aberrant glycan forms is already considered a hallmark of the disease state. Mass spectrometry has become a prominent approach to analyzing glycoconjugates. Specifically, matrix-assisted laser desorption/ionisation -mass spectrometric imaging (MALDI-MSI) is a powerful technique that combines mass spectrometry with histology and enables the spatially resolved and label-free detection of glycans...
September 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28978063/predictive-value-of-molecular-subtyping-in-nmibc-by-rt-qpcr-of-erbb2-esr1-pgr-and-mki67-from-formalin-fixed-tur-biopsies
#17
Johannes Breyer, Ralph Markus Wirtz, Wolfgang Otto, Mark Laible, Kornelia Schlombs, Philipp Erben, Maximilian Christian Kriegmair, Robert Stoehr, Sebastian Eidt, Stefan Denzinger, Maximilian Burger, Arndt Hartmann
Expression of ESR1, PGR, HER2 and Ki67 is important for risk stratification and therapy in breast cancer. Hormone receptor expression can also be found in MIBC, reflecting luminal and basal subtypes of breast cancer. Thus the purpose was to investigate on the mRNA expression of the aforementioned markers and their prognostic value in pT1 bladder cancer. Retrospective analysis of clinical data and Formalin-Fixed Paraffin-Embedded tissues (FFPE) of patients with stage pT1 NMIBC who underwent transurethral resection of the bladder was performed...
September 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28975726/muscle-fiber-diameter-assessment-in-cleft-lip-using-image-processing
#18
Mohammad Faisal Jamal Khan, Julian Little, Luigi Abelli, Peter Anthony Mossey, Luca Autelitano, Tapas Chandra Nag, Michele Rubini
OBJECTIVE: To pilot investigation of muscle fiber diameter (MFD) on medial and lateral sides of the cleft in 18 infants with cleft lip with or without cleft palate (CL/P) using image processing. MATERIAL AND METHODS: Formalin-fixed paraffin embedded (FFPE) tissue samples from the medial and lateral sides of the cleft were analyzed for MFD using an image processing program (ImageJ). For within-case comparison a paired student's test was performed. For comparisons between classes an unpaired t-test was used...
October 4, 2017: Oral Diseases
https://www.readbyqxmd.com/read/28972045/pharmacogenetic-analysis-of-the-uk-mrc-magic-trial-association-of-polymorphisms-with-toxicity-and-survival-in-patients-treated-with-perioperative-ecf-chemotherapy
#19
Elizabeth Smyth, Shenli Zhang, David Cunningham, Andrew Wotherspoon, Richie Soong, Clare Peckitt, Nicola Valeri, Matteo Fassan, Massimo Rugge, Alicia Fc Okines, William Allum, Sally Stenning, Matthew Nankivell, Ruth E Langley, Patrick Tan
PURPOSE: Germline polymorphisms may affect chemotherapy efficacy and toxicity. We examined the effect of polymorphisms in drug metabolism and DNA repair genes on pathological response rates, survival, and toxicity for patients randomised to surgery alone or perioperative ECF chemotherapy in the MRC MAGIC trial. EXPERIMENTAL DESIGN: DNA was extracted from non-tumor resection FFPE blocks. ERCC1, ERCC2, XRCC1, DYPD, and OPRT SNPs were evaluated using Sequenom, GSTP1, GSTT1 deletion and TYMS (TS) 5' 2R/3R using multiplex PCR...
October 2, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28967989/staining-of-frozen-and-formalin-fixed-paraffin-embedded-tissues-with-metal-labeled-antibodies-for-imaging-mass-cytometry-analysis
#20
Qing Chang, Olga Ornatsky, David Hedley
This unit describes protocols for labeling tissue sections using combinations of metal-tagged antibodies and an iridium-containing DNA intercalator for analysis by imaging mass cytometry. Imaging mass cytometry (IMC) allows the labeling of up to 40 individual markers simultaneously using antibody cocktails. We discuss labeling of both cryostat sections and sections from formalin-fixed, paraffin-embedded (FFPE) tissue blocks. The protocols are similar to those used for optical microscopy techniques, while allowing much higher complexity of analysis...
October 2, 2017: Current Protocols in Cytometry
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