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Fluorescence in-situ hybridization

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https://www.readbyqxmd.com/read/28732214/crispr-cas9-technology-based-xenograft-tumors-as-candidate-reference-materials-for-multiple-eml4-alk-rearrangements-testing
#1
Rongxue Peng, Rui Zhang, Guigao Lin, Xin Yang, Ziyang Li, Kuo Zhang, Jiawei Zhang, Jinming Li
The echinoderm microtubule-associated protein-like 4 and anaplastic lymphoma kinase (ALK) receptor tyrosine kinase (EML4-ALK) rearrangement is an important biomarker that plays a pivotal role in therapeutic decision making for non-small-cell lung cancer (NSCLC) patients. Ensuring accuracy and reproducibility of EML4-ALK testing by fluorescence in situ hybridization, immunohistochemistry, RT-PCR, and next-generation sequencing requires reliable reference materials for monitoring assay sensitivity and specificity...
July 18, 2017: Journal of Molecular Diagnostics: JMD
https://www.readbyqxmd.com/read/28732174/spatial-organization-of-fibroblast-and-spermatocyte-nuclei-with-different-b-chromosome-content-in-korean-field-mouse-i-apodemus-peninsulae-i-rodentia-muridae
#2
Tatyana V Karamysheva, Anna A Torgasheva, Yaroslav R Yefremov, Anton G Bogomolov, Thomas Liehr, Pavel M Borodin, Nikolay B Rubtsov
Korean field mouse (<i>Apodemus peninsulae</i>) shows a wide variation in the number of B chromosomes composed of constitutive heterochromatin. For this reason, it provides a good model to study the influence of the number of centromeres and amount of heterochromatin on spatial organization of interphase nuclei. We analyzed the three-dimensional organization of fibroblast and spermatocyte nuclei of the field mice carrying different number of B chromosomes using laser scanning microscopy and 3D fluorescence <i>in situ</i> hybridization...
July 21, 2017: Genome Génome / Conseil National de Recherches Canada
https://www.readbyqxmd.com/read/28731868/inflammatory-myofibroblastic-tumors-of-the-female-genital-tract-are-under-recognized-a-low-threshold-for-alk-immunohistochemistry-is-required
#3
Justine L Pickett, Angela Chou, Juliana A Andrici, Adele Clarkson, Loretta Sioson, Amy Sheen, Jessica Reagh, Fedaa Najdawi, Yoomee Kim, Denise Riley, Jayne Maidens, David Nevell, Kirsten McIlroy, Susan Valmadre, Greg Gard, Russell Hogg, John Turchini, Gregory Robertson, Michael Friedlander, Anthony J Gill
Inflammatory myofibroblastic tumor (IMT) of the female genital tract is under-recognized. We investigated the prevalence of ALK-positive IMT in lesions previously diagnosed as gynecologic smooth muscle tumors. Immunohistochemistry (IHC) for ALK was performed on tissue microarrays of unselected tumors resected from 2009 to 2013. Three of 1176 (0.26%) "leiomyomas" and 1 of 44 (2.3%) "leiomyosarcomas" were ALK IHC positive, confirmed translocated by fluorescence in situ hybridization (FISH) and therefore more appropriately classified as IMT...
July 20, 2017: American Journal of Surgical Pathology
https://www.readbyqxmd.com/read/28730668/recurrent-triploid-digynic-conceptions-and-mature-ovarian-teratomas-are-they-different-manifestations-of-the-same-genetic-defect
#4
Yassemine Khawajkie, William Buckett, Ngoc Minh Phuong Nguyen, Nawel Mechtouf, Asangla Ao, Jocelyne Arseneau, Rima Slim
Miscarriages affect 15% of clinically recognized pregnancies. Recurrent miscarriage (RM) is defined by the occurrence of at least two consecutive pregnancy losses and affects 1% to 5% of couples trying to conceive. In an attempt to categorize patients with RM and identify the mechanisms leading to their miscarriages, we first used flow cytometry to assess the ploidy of 93 products of conception (POCs) from 53 patients with RM (≥ 3 miscarriages). We identified a single patient with four triploid POCs. We then used fluorescent in situ hybridization to confirm the triploidies and fluorescent microsatellite genotyping with distal and pericentromeric markers to determine their parental origin and the mechanisms leading to their formation...
July 21, 2017: Genes, Chromosomes & Cancer
https://www.readbyqxmd.com/read/28730537/lung-cancer-as-a-paradigm-for-precision-oncology-in-solid-tumours
#5
Simon Schallenberg, Sabine Merkelbach-Bruse, Reinhard Buettner
Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death in the western world. However, the combination of molecular genotyping and subsequent systematic treatment of decoded target structures is a prime example of precision oncology in solid tumours. In this review, current targets of approved therapeutics and potential targets in clinical and preclinical trials are outlined. Furthermore, immune checkpoint inhibitors, as promising new therapeutic options, which have already been applied successfully in cases of lung cancer, are introduced...
July 20, 2017: Virchows Archiv: An International Journal of Pathology
https://www.readbyqxmd.com/read/28728292/-detection-and-clinical-significance-of-circulating-tumor-cells-in-osteosarcoma-using-immunofluorescence-combined-with-in-situ-hybridization
#6
H Q Zhang, M H Li, Z Wang, P H Lan, Y J Lu, G J Chen, L Wang
Objective: To investigate the clinical significance of detection of circulating tumor cells (CTCs) in peripheral blood from patients with osteosarcoma (OS) using the iFISH (immunofluorescence and fluorescence in situ hybridization) method. Methods: The live cells recovery rate of immune-magnetic beads was evaluated by live-cell fluorescent tracer technology. The expression of CD45 and CK18 on the cell surface of HOS and HepG2 cells was measured by flow cytometry. And the chromosome aneuploidy was detected by centromeric FISH probe CEP8...
July 23, 2017: Zhonghua Zhong Liu za Zhi [Chinese Journal of Oncology]
https://www.readbyqxmd.com/read/28726804/diagnostic-cytogenetic-testing-following-positive-noninvasive-prenatal-screening-results-a-clinical-laboratory-practice-resource-of-the-american-college-of-medical-genetics-and-genomics-acmg
#7
Athena M Cherry, Yassmine M Akkari, Kimberly M Barr, Hutton M Kearney, Nancy C Rose, Sarah T South, James H Tepperberg, Jeanne M Meck
Disclaimer: ACMG Clinical Laboratory Practice Resources are developed primarily as an educational tool for clinical laboratory geneticists to help them provide quality clinical laboratory genetic services. Adherence to these practice resources is voluntary and does not necessarily assure a successful medical outcome. This Clinical Laboratory Practice Resource should not be considered inclusive of all proper procedures and tests or exclusive of other procedures and tests that are reasonably directed to obtaining the same results...
July 20, 2017: Genetics in Medicine: Official Journal of the American College of Medical Genetics
https://www.readbyqxmd.com/read/28725026/microbiome-analysis-and-confocal-microscopy-of-used-kitchen-sponges-reveal-massive-colonization-by-acinetobacter-moraxella-and-chryseobacterium-species
#8
Massimiliano Cardinale, Dominik Kaiser, Tillmann Lueders, Sylvia Schnell, Markus Egert
The built environment (BE) and in particular kitchen environments harbor a remarkable microbial diversity, including pathogens. We analyzed the bacterial microbiome of used kitchen sponges by 454-pyrosequencing of 16S rRNA genes and fluorescence in situ hybridization coupled with confocal laser scanning microscopy (FISH-CLSM). Pyrosequencing showed a relative dominance of Gammaproteobacteria within the sponge microbiota. Five of the ten most abundant OTUs were closely related to risk group 2 (RG2) species, previously detected in the BE and kitchen microbiome...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28724850/spatiotemporal-dynamics-of-total-viable-vibrio-spp-in-a-nw-mediterranean-coastal-area
#9
Léa Girard, Sébastien Peuchet, Pierre Servais, Annabelle Henry, Nadine Charni-Ben-Tabassi, Julia Baudart
A cellular approach combining Direct Viable Counting and Fluorescent In Situ Hybridization using a one-step multiple-probe technique and Solid Phase Cytometry (DVC-FISH-SPC) was developed to monitor total viable vibrios and cover the detection of a large diversity of vibrios. FISH combined three probes in the same assay and targeted sequences located at different positions on the 16S rRNA of Vibrio and Aliivibrio members. We performed a 10-month in situ study to investigate the weekly dynamics of viable vibrios relative to culturable counts at two northwestern Mediterranean coastal sites, and identified the key physicochemical factors for their occurrence in water using a multivariate analysis...
July 19, 2017: Microbes and Environments
https://www.readbyqxmd.com/read/28724771/intracellular-colocalization-of-influenza-viral-rna-and-rab11a-is-dependent-upon-microtubule-filaments
#10
Eric Nturibi, Amar R Bhagwat, Stefanie Coburn, Mike M Myerburg, Seema S Lakdawala
Influenza A virus (IAV) consists of eight viral RNA (vRNA) segments that are replicated in the host cell nucleus and transported to the plasma membrane for packaging into progeny virions. We have previously proposed a model where sub-complexes of vRNA export from the nucleus and assemble en route to the plasma membrane. However, the role of host cytoskeletal proteins in the cytoplasmic assembly of IAV vRNA segments remains unknown. Previous studies have suggested that IAV vRNA segments transport via Rab11A-containing recycling endosomes (RE) and use both microtubules (MT) and actin...
July 19, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28724402/a-dual-colour-fluorescence-in-situ-hybridization-fish-assay-for-identifying-the-zoonotic-malaria-parasite-plasmodium-knowlesi-with-a-potential-application-for-the-specific-diagnosis-of-knowlesi-malaria-in-peripheral-level-laboratories-of-southeast-asia
#11
Jyotsna Shah, Akhila Poruri, Olivia Mark, Urmila Khadilka, Franziska Mohring, Robert W Moon, Ranjan Ramasamy
BACKGROUND: Plasmodium knowlesi is primarily responsible for zoonotic malaria in several Southeast Asian countries. Precise identification of the parasite in the blood of patients presently relies on an expensive and elaborate PCR procedure because microscopic examination of blood and other available field identification techniques lack adequate specificity. Therefore, the use of a simple and inexpensive dual-colour fluorescence in situ hybridization (FISH) assay, analogous to FISH assays recently described for Plasmodium falciparum and Plasmodium vivax, was investigated as a potential tool for identifying P...
July 19, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28723933/comparative-repeatome-analysis-on-triatoma-infestans-andean-and-non-andean-lineages-main-vector-of-chagas-disease
#12
Sebastián Pita, Francisco Panzera, Pablo Mora, Jesús Vela, Ángeles Cuadrado, Antonio Sánchez, Teresa Palomeque, Pedro Lorite
Triatoma infestans is the most important Chagas disease vector in South America. Two main evolutionary lineages, named Andean and non-Andean, have been recognized by geographical distribution, phenetic and genetic characteristics. One of the main differences is the genomic size, varying over 30% in their haploid DNA content. Here we realize a genome wide analysis to compare the repetitive genome fraction (repeatome) between both lineages in order to identify the main repetitive DNA changes occurred during T...
2017: PloS One
https://www.readbyqxmd.com/read/28719910/correlating-the-genetic-and-physical-map-of-barley-chromosome-3h-revealed-limitations-of-the-fish-based-mapping-of-nearby-single-copy-probes-caused-by-the-dynamic-structure-of-metaphase-chromosomes
#13
Fernanda O Bustamante, Lala Aliyeva-Schnorr, Jörg Fuchs, Sebastian Beier, Andreas Houben
Genetic maps are based on the recombination frequency of molecular markers which often show different positions in comparison to the corresponding physical maps. To decipher the position and order of DNA sequences genetically mapped to terminal and interstitial regions of barley (Hordeum vulgare) chromosome 3H, fluorescence in situ hybridization (FISH) on mitotic metaphase chromosomes was performed with 16 genomic single-copy probes derived from fingerprinted BAC contigs. Long genetic distances at subterminal regions translated into short physical distances, confirming that recombination events occur more often at distal regions of chromosome 3H...
July 19, 2017: Cytogenetic and Genome Research
https://www.readbyqxmd.com/read/28719462/spindle-cell-lipomas-in-women-a-report-of-53-cases
#14
Jennifer S Ko, Brianne Daniels, Patrick O Emanuel, Paul Elson, Vadim Khachaturov, Jesse K McKenney, John R Goldblum, Steven D Billings
Spindle cell lipomas (SCL) are typically tumors of the upper back/neck (shawl region) of men (80% to 90%). In general, there is a frequent tendency to restrict the diagnosis to this specific clinical scenario and a hesitancy to diagnose SCL in women. We hypothesized that SCL in women have a more varied presentation. A total of 395 SCL were diagnosed at our institution over the last 11 years. The diagnosis of SCL in women was confirmed by re-review. Immunohistochemical stains for CD34, desmin, estrogen receptor, and p16 were performed...
July 17, 2017: American Journal of Surgical Pathology
https://www.readbyqxmd.com/read/28714990/an-approach-to-suppress-the-evolution-of-resistance-in-braf-v600e-mutant-cancer
#15
Yaohua Xue, Luciano Martelotto, Timour Baslan, Alberto Vides, Martha Solomon, Trang Thi Mai, Neelam Chaudhary, Greg J Riely, Bob T Li, Kerry Scott, Fabiola Cechhi, Ulrika Stierner, Kalyani Chadalavada, Elisa de Stanchina, Sarit Schwartz, Todd Hembrough, Gouri Nanjangud, Michael F Berger, Jonas Nilsson, Scott W Lowe, Jorge S Reis-Filho, Neal Rosen, Piro Lito
The principles that govern the evolution of tumors exposed to targeted therapy are poorly understood. Here we modeled the selection and propagation of an amplification in the BRAF oncogene (BRAF(amp)) in patient-derived tumor xenografts (PDXs) that were treated with a direct inhibitor of the kinase ERK, either alone or in combination with other ERK signaling inhibitors. Single-cell sequencing and multiplex fluorescence in situ hybridization analyses mapped the emergence of extra-chromosomal amplification in parallel evolutionary trajectories that arose in the same tumor shortly after treatment...
July 17, 2017: Nature Medicine
https://www.readbyqxmd.com/read/28714966/nanoscale-imaging-of-clinical-specimens-using-pathology-optimized-expansion-microscopy
#16
Yongxin Zhao, Octavian Bucur, Humayun Irshad, Fei Chen, Astrid Weins, Andreea L Stancu, Eun-Young Oh, Marcello DiStasio, Vanda Torous, Benjamin Glass, Isaac E Stillman, Stuart J Schnitt, Andrew H Beck, Edward S Boyden
Expansion microscopy (ExM), a method for improving the resolution of light microscopy by physically expanding a specimen, has not been applied to clinical tissue samples. Here we report a clinically optimized form of ExM that supports nanoscale imaging of human tissue specimens that have been fixed with formalin, embedded in paraffin, stained with hematoxylin and eosin, and/or fresh frozen. The method, which we call expansion pathology (ExPath), converts clinical samples into an ExM-compatible state, then applies an ExM protocol with protein anchoring and mechanical homogenization steps optimized for clinical samples...
July 17, 2017: Nature Biotechnology
https://www.readbyqxmd.com/read/28713994/molecular%C3%A2-cytogenetic-study-of-de-novo-mosaic-karyotype-45-x-46-x-i-yq-46-x-idic-yq-in-an-azoospermic-male-case-report-and-literature-review
#17
Yuting Jiang, Ruixue Wang, Linlin Li, Lintao Xue, Shu Deng, Ruizhi Liu
The present study describes a 36‑year‑old male with the 45,X/46,X,i(Yq)/46,X,idic(Yq) karyotype, who suffered from azoospermia attributed to maturation arrest of the primary spermatocyte. To the best of our knowledge, this rare karyotype has not yet been reported in the literature. The results of detailed molecular‑cytogenetic studies of isodicentric (idic)Y chromosomes and isochromosome (iso)Y, which are identified in patient with complex mosaic karyotypes, are presented. The presence of mosaicism of the three cell lines 45,X, 46,X,i(Yq) and 46,X,idic(Yq) may be a contributing factor for spermatogenic failure, in addition to the instability of iso/idic Y chromosomes to pass the spermatogenesis process...
July 14, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28713251/age-related-changes-in-lateral-entorhinal-and-ca3-neuron-allocation-predict-poor-performance-on-object-discrimination
#18
Andrew P Maurer, Sarah A Johnson, Abbi R Hernandez, Jordan Reasor, Daniela M Cossio, Kaeli E Fertal, Jack M Mizell, Katelyn N Lubke, Benjamin J Clark, Sara N Burke
Age-related memory deficits correlate with dysfunction in the CA3 subregion of the hippocampus, which includes both hyperactivity and overly rigid activity patterns. While changes in intrinsic membrane currents and interneuron alterations are involved in this process, it is not known whether alterations in afferent input to CA3 also contribute. Neurons in layer II of the lateral entorhinal cortex (LEC) project directly to CA3 through the perforant path, but no data are available regarding the effects of advanced age on LEC activity and whether these activity patterns update in response to environmental change...
2017: Frontiers in Systems Neuroscience
https://www.readbyqxmd.com/read/28712009/analysis-of-molecular-subtypes-for-the-increased-her2-equivocal-cases-caused-by-application-of-the-updated-2013-asco-cap-her2-testing-guidelines-in-breast-cancer
#19
Lei Guo, Pei Yuan, Jing Zhang, Yun Ling, Wenbin Li, Bohui Zhao, Jianming Ying, Lixue Xuan
PURPOSE: Accurate testing of the status of human epidermal growth factor receptor type 2 (HER2) is a prerequisite for HER2-directed therapy. The American Society of Clinical Oncology (ASCO) and the College of American Pathologists (CAP) published joint guideline recommendations for HER2 testing in breast cancer in 2007 and it was updated in 2013. We compared the HER2 gene amplification status based on these two guidelines and analyzed the molecular characteristics of the equivocal cases...
July 15, 2017: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/28711302/potential-selection-of-genetically-balanced-spermatozoa-based-on-the-hypo-osmotic-swelling-test-in-chromosomal-rearrangement-carriers
#20
Alexandre Rouen, Léa Carlier, Solveig Heide, Matthieu Egloff, Pauline Marzin, Flavie Ader, Mathias Schwartz, Eli Rogers, Nicole Joyé, Richard Balet, Nathalie Lédée, Laura Prat-Ellenberg, Nino Guy Cassuto, Jean-Pierre Siffroi
Chromosomal translocations and other balanced rearrangements, although usually associated with a normal phenotype, can lead to the transmission of an abnormal unbalanced genome to the offspring. Balanced and unbalanced spermatozoa, being indistinguishable, cannot be selected or deselected for prior to IVF and pre-implantation genetic diagnosis. Spermatozoa from 16 chromosomal rearrangement carriers were studied. After incubation in a hypo-osmotic solution (hypo-osmotic swelling test, or HOST), spermatozoa were fixed on microscope slides...
June 27, 2017: Reproductive Biomedicine Online
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