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Fluorescence in-situ hybridization

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https://www.readbyqxmd.com/read/28527402/a-rare-e13a3-b2a3-bcr-abl1-fusion-transcript-with-normal-karyotype-in-chronic-myeloid-leukemia-the-challenges-in-diagnosis-and-monitoring-minimal-residual-disease-mrd
#1
M-H Duan, H Li, H Cai
Patients with chronic myeloid leukemia (CML) have a t (9;22)(q34;q11.2) or variant translocation that results in a BCR-ABL1 fusion gene. For many years, conventional karyotyping has been used as the standard diagnostic tool for t (9;22) (q34;q11.2). However, it has several limitations that may lead to failure for detecting BCR-ABL1 gene rearrangements in around 5% of all CML patients. Although reverse transcription polymerase chain reaction (RT-PCR) has evolved as a sensitive method for detecting BCR-ABL1 translocation, this method fail to detect certain BCR-ABL1 fusion transcript type, such as e13a3 (also known as b2a3), as a result of many commercially available and laboratory-developed primer sets...
May 12, 2017: Leukemia Research
https://www.readbyqxmd.com/read/28525305/clinical-impact-of-the-cell-of-origin-classification-and-the-myc-bcl2-dual-expresser-status-in-diffuse-large-b-cell-lymphoma-treated-within-prospective-clinical-trials-of-the-german-high-grade-non-hodgkin-s-lymphoma-study-group
#2
Annette M Staiger, Marita Ziepert, Heike Horn, David W Scott, Thomas F E Barth, Heinz-Wolfram Bernd, Alfred C Feller, Wolfram Klapper, Monika Szczepanowski, Michael Hummel, Harald Stein, Dido Lenze, Martin-Leo Hansmann, Sylvia Hartmann, Peter Möller, Sergio Cogliatti, Georg Lenz, Lorenz Trümper, Markus Löffler, Norbert Schmitz, Michael Pfreundschuh, Andreas Rosenwald, German Ott
Purpose To explore the prognostic impact and interdependence of the cell-of-origin (COO) classification, dual expression (DE) of MYC and BCL2 proteins, and MYC, BCL2, and BCL6 translocations in two prospectively randomized clinical trials of patients with diffuse large B-cell lymphoma (DLBCL). Patients and Methods Overall, 452 formalin-fixed paraffin-embedded samples from two prospective, randomized DLBCL trials (RICOVER-60, prospective, randomized study for patients > 60 years, all IPI groups; and R-MegaCHOEP, prospective, randomized study for patients ≤ 60 years with age-adjusted IPI 2,3) of the German High-Grade Non-Hodgkin Lymphoma Study Group were analyzed with the Lymph2Cx assay for COO classification, with immunohistochemistry for MYC and BCL2, and with fluorescent in situ hybridization for MYC, BCL2, and BCL6 rearrangements...
May 19, 2017: Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology
https://www.readbyqxmd.com/read/28523559/techniques-for-single-molecule-mrna-imaging-in-living-cells
#3
Kevin Czaplinski
Typical measurement of macromolecules in a biological sample typically averages the result over all the cells or molecules within the sample, and while these types of measurements provide very useful information, they completely miss heterogeneity among the components within the sample that could be a very important aspect of the sample's function. These techniques are also limited in their ability to examine intracellular spatial orientation of molecular activity, which is often a critical component to the regulation of biological processes, particularly in cells with unique spatial relationships, such as neurons...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28523449/expression-of-coagulation-factor-xiii-subunit-a-correlates-with-outcome-in-childhood-acute-lymphoblastic-leukemia
#4
Bettina Kárai, Zsuzsanna Hevessy, Eszter Szánthó, László Csáthy, Anikó Ujfalusi, Katalin Gyurina, István Szegedi, János Kappelmayer, Csongor Kiss
Previously we identified B-cell lineage leukemic lymphoblasts as a new expression site for subunit A of blood coagulation factor XIII (FXIII-A). On the basis of FXIII-A expression, various subgroups of B-cell precursor acute lymphoblastic leukemia (BCP-ALL) can be identified. Fifty-five children with BCP-ALL were included in the study. Bone marrow samples were obtained by aspiration and the presence of FXIII-A was detected by flow cytometry. G-banding and fluorescent in situ hybridization was performed according to standard procedures...
May 18, 2017: Pathology Oncology Research: POR
https://www.readbyqxmd.com/read/28522811/pd-l1-expression-in-xp11-2-translocation-renal-cell-carcinoma-indicator-of-tumor-aggressiveness
#5
Kun Chang, Yuanyuan Qu, Bo Dai, Jian-Yuan Zhao, Hualei Gan, Guohai Shi, Yiping Zhu, Yijun Shen, Yao Zhu, Hailiang Zhang, Dingwei Ye
Programmed death ligand-1 (PD-L1), a promising antitumor target, has proven clinical value against many malignancies. However, the PD-L1 content of Xp11.2 translocation renal cell carcinoma (Xp11.2 RCC) and its correlation with clinical outcomes remain unclear. This study aimed to investigate PD-L1 expression in Xp11.2 RCC and to assess its prognostic value. Formalin-fixed paraffin-embedded specimens from 36 adult patients that were histologically confirmed (by fluorescence in situ hybridization) were subjected to immunohistochemical analysis...
May 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28521634/chorionic-histiocytic-hyperplasia-is-associated-with-chronic-inflammatory-lesions-in-the-placenta
#6
Philip J Katzman, Leon A Metlay, Loralee A McMahon, Li LiQiong, Bin Zhang
Background Chorionic histiocytic hyperplasia (CHH) is a cellular proliferation at the base of the chorion that was identified by the authors in examining placentas for chronic chorioamnionitis (CC). This study is a retrospective review of cases diagnosed with CHH, describing its incidence alone and with associated lesions, and the immunophenotype of lesional cells. Design In this retrospective study, a laboratory information system search over a 34-month period using the key phrase "chorionic stromal" was performed...
June 2017: Pediatric and Developmental Pathology
https://www.readbyqxmd.com/read/28521575/high-frequency-of-de-novo-daz-microdeletion-in-sperm-nuclei-of-subfertile-men-possible-involvement-of-genome-instability-in-idiopathic-male-infertility
#7
Hossein Mozdarani, Pegah Ghoraeian, Sohail Mozdarani, Parvin Fallahi, Anahita Mohseni-Meybodi
The occurrence and diagnosis of Y-chromosome microdeletions, specifically deletions of the DAZ (Deleted in Azoospermia) genes are an important issue in male infertility. Screening Y chromosome microdeletion is mainly done using polymerase chain reaction (PCR) on blood leukocytes. However, there is some evidence indicating that presence of DAZ in somatic cells might not be indicative of its presence in the germ cell lineage. Therefore, a total of 130 men with poor semen quality were examined for presence of DAZ microdeletion in their leukocytes...
May 19, 2017: Human Fertility: Journal of the British Fertility Society
https://www.readbyqxmd.com/read/28521466/extranodal-marginal-zone-lymphoma-of-the-uterine-cervix-with-concomitant-copy-number-gains-of-the-malt1-and-bcl2-genes-a-case-report
#8
Tomoko Takimoto, Saori Maegawa, Hiroshi Tatsumi, Hisao Nagoshi, Yoshiaki Chinen, Yuji Shimura, Tsutomu Kobayashi, Shigeo Horiike, Shigeo Nakamura, Jo Kitawaki, Junya Kuroda, Masafumi Taniwaki
Extranodal marginal zone lymphoma (EMZL) of mucosa-associated lymphoid tissue (MALT) of the uterus is rare, and the etiology, pathophysiology and cytogenetic features remain unknown at present. The present study reports a case of a 71-year-old female with EMZL of the uterine cervix that was 80 mm in diameter and invaded directly into the rectal serosa. Complete remission was successfully induced by 6 courses of immunochemotherapy with rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisolone...
May 2017: Oncology Letters
https://www.readbyqxmd.com/read/28521363/a-cisplatin-derivative-tetra-pt-bpy-as-an-oncotherapeutic-agent-for-targeting-alt-cancer
#9
Xiao-Hui Zheng, Xin Nie, Yiming Fang, Zepeng Zhang, Yingnan Xiao, Zongwan Mao, Haiying Liu, Jian Ren, Feng Wang, Lixin Xia, Junjiu Huang, Yong Zhao
Background: In approximately 15% of human cancers, telomere length is maintained independently of telomerase by the homologous recombination (HR)-mediated alternative lengthening of telomeres (ALT) pathway. Whether the ALT pathway can be exploited for therapeutic treatment remains unknown. The purpose of this study is to develop oncotherapeutic agent to target ALT cancers. Methods: Surface plasmon resonance assay, antibody to G-quadruplex, and fluorescence in situ hybridization (FISH) were used to discover Tetra-Pt(bpy), a cisplatin derivative that specifically targets telomeric G-quadruplex...
October 1, 2017: Journal of the National Cancer Institute
https://www.readbyqxmd.com/read/28520527/open-label-multicenter-phase-ii-study-of-ceritinib-in-patients-with-non-small-cell-lung-cancer-harboring-ros1-rearrangement
#10
Sun Min Lim, Hye Ryun Kim, Jong-Seok Lee, Ki Hyeong Lee, Yun-Gyoo Lee, Young Joo Min, Eun Kyung Cho, Sung Sook Lee, Bong-Seog Kim, Moon Young Choi, Hyo Sup Shim, Jin-Haeng Chung, Yoon La Choi, Min Jeong Lee, Maria Kim, Joo-Hang Kim, Siraj M Ali, Myung-Ju Ahn, Byoung Chul Cho
Purpose ROS1 rearrangement is a distinct molecular subset of non-small-cell lung cancer (NSCLC). We investigated the efficacy and safety of ceritinib in patients with ROS1-rearranged NSCLC. Patients and Methods We enrolled 32 patients with advanced NSCLC who tested positive for ROS1 rearrangement by fluorescent in situ hybridization. Ceritinib 750 mg was administered once daily. The primary end point was objective response rate. The secondary end points were disease control rate; duration of response; progression-free survival; overall survival; toxicity; and concordance among fluorescent in situ hybridization, immunohistochemistry, and next-generation sequencing...
May 18, 2017: Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology
https://www.readbyqxmd.com/read/28518171/fish-flow-a-protocol-for-the-concurrent-detection-of-mrna-and-protein-in-single-cells-using-fluorescence-in-situ-hybridization-and-flow-cytometry
#11
Riccardo Arrigucci, Yuri Bushkin, Felix Radford, Karim Lakehal, Pooja Vir, Richard Pine, December Martin, Jeffrey Sugarman, Yanlin Zhao, George S Yap, Alfred A Lardizabal, Sanjay Tyagi, Maria Laura Gennaro
We describe a flow-cytometry-based protocol for intracellular mRNA measurements in nonadherent mammalian cells using fluorescence in situ hybridization (FISH) probes. The method, which we call FISH-Flow, allows for high-throughput multiparametric measurements of gene expression, a task that was not feasible with earlier, microscopy-based approaches. The FISH-Flow protocol involves cell fixation, permeabilization and hybridization with a set of fluorescently labeled oligonucleotide probes. In this protocol, surface and intracellular protein markers can also be stained with fluorescently labeled antibodies for simultaneous protein and mRNA measurement...
June 2017: Nature Protocols
https://www.readbyqxmd.com/read/28516197/optimized-immunohistochemistry-using-the-d5f3-antibody-provides-a-reliable-test-for-identification-of-alk-positive-lung-adenocarcinomas
#12
Gian Luca Rampioni Vinciguerra, Stefania Scarpino, Benedetto Pini, Claudia Cippitelli, Flavio Fochetti, Luigi Ruco
We used optimized immunohistochemistry (IHC) with the D5F3 antibody for detection of tumours in a prospective study of 307 pulmonary adenocarcinomas. Cases positive by IHC (1+, 2+, 3+) were further investigated by fluorescent in situ hybridization (FISH). Of 307 cases, 22 (7.2%) were moderately intensely positive (2+/3+); 18 of these (82%) were also positive by FISH. Of the four IHC-positive/FISH-negative cases, one was unsuitable for FISH and three had abnormalities of the ALK gene. All cases with weak reactivity with D5F3 (1+) were FISH-negative...
May 17, 2017: Virchows Archiv: An International Journal of Pathology
https://www.readbyqxmd.com/read/28515389/ca-2-in-hybridization-solutions-for-fluorescence-in-situ-hybridization-facilitates-the-detection-of-enterobacteriaceae
#13
Shin Haruta, Takao Iino, Moriya Ohkuma, Ken-Ichiro Suzuki, Yasuo Igarashi
Fluorescence in situ hybridization (FISH) has been employed to identify microorganisms at the single cell level under a microscope. Extensive efforts have been made to improve and extend the FISH technique; however, the development of a widely applicable protocol is a continuing challenge. The present study evaluated the effects of divalent cations in the hybridization solution on the FISH-based detection of various species of bacteria and archaea with rRNA-targeted probes. A flow cytometric analysis after FISH with a standard hybridization buffer detected positive signals from less than 30% of Escherichia coli IAM 1264 cells...
May 18, 2017: Microbes and Environments
https://www.readbyqxmd.com/read/28515301/the-influenza-a-virus-ns1-protein-promotes-efficient-nuclear-export-of-unspliced-viral-m1-mrna
#14
Carina F Pereira, Eliot K C Read, Helen M Wise, Maria J Amorim, Paul Digard
Influenza A virus mRNAs are transcribed by the viral RNA-dependent RNA polymerase in the cell nucleus before being exported to the cytoplasm for translation. Segment 7 produces two major transcripts: an unspliced mRNA that encodes the M1 matrix protein and a spliced transcript that encodes the M2 ion channel. Export of both mRNAs is dependent on the cellular NXF1/TAP pathway but it is unclear how they are recruited to the export machinery or how the intron-containing but unspliced M1 mRNA bypasses the normal quality control checkpoints...
May 17, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28515100/detection-of-chromosomal-translocation-in-hematologic-malignancies-by-a-novel-dna-based-looped-ligation-assay-lola
#15
Shuko Harada, Emily Sizzle, Ming-Tseh Lin, Christopher D Gocke
BACKGROUND: Disease-defining chromosomal translocations are seen in various neoplasms, especially in lymphomas and leukemias. Translocation detection at the DNA level is often complicated by chromosomal breakpoints that are distributed over very large regions. We have developed a ligation-based assay [the looped ligation assay (LOLA)] to detect translocations from diseases with multiple widely spaced breakpoint hot spots. METHODS: Oligonucleotide sets that probe breakpoints of IGH-BCL2 (immunoglobulin heavy-apoptosis regulator) in follicular lymphoma (FL), MYC-IGH (MYC proto-oncogene, bHLH transcription factor- immunoglobulin heavy) in Burkitt lymphoma (BL) and BCR-ABL1 (RhoGEF and GTPase activating protein-ABL proto-oncogene 1, non-receptor tyrosine kinase) in chronic myelogenous leukemia (CML) were designed...
May 17, 2017: Clinical Chemistry
https://www.readbyqxmd.com/read/28513875/incidence-clinicopathologic-features-and-fusion-transcript-landscape-of-translocation-renal-cell-carcinomas
#16
Xiao-Tong Wang, Qiu-Yuan Xia, Xiao-Jun Zhou, Qiu Rao
The recent article entitled "Incidence, clinicopathologic features, and fusion transcript landscape of translocation renal cell carcinomas" by Marion Classe and colleagues has been accepted in January 2017 by Histopathology(1) . The authors aimed to describe the landscape of fusion transcript in a large single institution series of interphase fluorescence in situ hybridization (FISH) confirmed translocation renal cell carcinomas (RCCs). They also identified two new fusion partners, GRIPAP1 and MED15, in Xp11 translocation RCCs...
May 17, 2017: Histopathology
https://www.readbyqxmd.com/read/28513868/her2-assessment-in-locally-advanced-gastric-cancer-comparing-the-results-of-using-2-primary-tumour-blocks-versus-all-primary-tumour-blocks
#17
Chen Xu, Yalan Liu, Dongxian Jiang, Xiaowen Ge, Ying Zhang, Jieakesu Su, Haiying Zeng, Jie Huang, Yuan Ji, Jun Hou, Yihong Sun, Kuntang Shen, Tianshu Liu, Yingyong Hou, Jing Qin
AIMS: HER2 is currently the only biomarker used to select eligible patients with advanced gastric cancer (GC) for targeted therapy. To verify the value of dual-block HER2 assessment and to explore whether increasing the block number is more beneficial, we carried out this randomized prospective cohort study by comparing dual-block and all-block HER2 assessment in resected specimens of GC. METHODS AND RESULTS: 549 resected GC specimens were randomly enrolled into two cohorts: a dual-block group (n=274) with two primary tumor blocks tested, and an all-block group (n=275) with all primary tumor blocks tested...
May 17, 2017: Histopathology
https://www.readbyqxmd.com/read/28511854/acot1-is-a-sensitive-indicator-for-ppar%C3%AE-activation-after-perfluorooctanoic-acid-exposure-in-primary-hepatocytes-of-sprague-dawley-rats
#18
Hui Liu, Jianshe Wang, Nan Sheng, Ruina Cui, Yitao Pan, Jiayin Dai
Perfluorooctanoic acid (PFOA) is one of the most commonly detected and persistent perfluoroalkyl substances (PFASs) found in the environment. We found that cell viability and intracellular oxidant stress increased in primary rat hepatocytes exposed to PFOA (100μM PFOA, 24h), and mitochondrial superoxide increased from 6.25μM PFOA treatment group. To screen for sensitive indicators in mRNA level, we investigated global transcriptome profile alteration after PFOA exposure using RNA-sequencing (RNA-seq) in primary rat hepatocytes, and identified 177 gene transcripts (158 upregulated, 19 downregulated) as significantly changed after exposure to 100μM of PFOA for 24h (fold change ≥2, FDR<0...
May 13, 2017: Toxicology in Vitro: An International Journal Published in Association with BIBRA
https://www.readbyqxmd.com/read/28511041/biofilm-formation-and-granule-properties-in-anaerobic-digestion-at-high-salinity
#19
M C Gagliano, S B Ismail, A J M Stams, C M Plugge, H Temmink, J B Van Lier
For the anaerobic biological treatment of saline wastewater, Anaerobic Digestion (AD) is currently a possibility, even though elevated salt concentrations can be a major obstacle. Anaerobic consortia and especially methanogenic archaea are very sensitive to fluctuations in salinity. When working with Upflow Sludge Blanket Reactor (UASB) technology, in which the microorganisms are aggregated and retained in the system as a granular biofilm, high sodium concentration negatively affects aggregation and consequently process performances...
May 10, 2017: Water Research
https://www.readbyqxmd.com/read/28510806/role-of-core-needle-biopsy-in-primary-breast-lymphoma
#20
Giulia Bicchierai, Luigi Rigacci, Vittorio Miele, Icro Meattini, Diego De Benedetto, Valeria Selvi, Simonetta Bianchi, Lorenzo Livi, Jacopo Nori
Primary breast lymphoma (PBL) may mimic carcinoma clinically and also mammographic and ecographic distinction is subtle. Despite its rarity, incidence of PBLs has increased over the last four decades and continues to increase for younger women and for some subtypes, and for this reason it is increasingly important to achieve a preoperative pathological diagnosis using core needle biopsy (CNB) or fine-needle aspiration cytology (FNA). The aim of this retrospective study was to report our single-center experience in CNBs performed for histological diagnosis of PBL compared to FNA...
May 16, 2017: La Radiologia Medica
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