Flow Cytometrically Determined DNA Content of Breast Carcinoma and Benign Lesions: Correlations with Histopathological Parameters

1986 ◽  
Vol 72 (2) ◽  
pp. 171-177 ◽  
Author(s):  
Raffaella Uccelli ◽  
Alberto Calugi ◽  
Donato Forte ◽  
Francesco Mauro ◽  
Paolo Polonio-Balbi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Cell Subpopulation ◽  
Aneuploid Cell ◽  
Breast Carcinomas ◽  
Benign Lesions ◽  
Dna Index ◽  
Tissue Samples ◽  
Normal Tissues ◽  
Relative Dna Content

The relative DNA content of cellular samples from 54 patients affected by breast carcinomas and 20 affected by benign breast lesions (including 11 fibroadenomas) was measured by flow cytometry. All normal tissue samples and 17/20 (85%) specimens from benign lesions exhibited a cytometrically diploid DNA distribution, 3/20 (15%) benign lesions an abnormal DNA content, and 35/54 (65%) carcinomas at least one aneuploid cell subpopulation. Furthermore, 9/54 (17%) tumors were characterized by the presence of more than one aneuploid cell subpopulation. The results also indicate that flow cytometry can be used to recognize lymph nodes infiltrated by aneuploid cells. Statistically significant correlations were evidenced between the occurrence of aneuploidy or the ploidy level measured as DNA index and the nodal infiltration status. The percentage of S cells can also be extracted from DNA content distribution histograms. Statistically significant differences (p < 0.01) were also observed for the percentage of S cells between normal tissues (6.2±3.2 SD) and benign lesions (11.1±6.6 SD), normal tissues (6.2 ± 3.2 SD) and aneuploid tumors (19.7 ± 10.3 SD), benign lesions (11.1 ± 6.6 SD) and aneuploid tumors (19.7 ± 10.3 SD), and diploid (7.9 ± 4.0 SD) and aneuploid tumors (19.7 ± 10.3 SD).

DNA content in human colon cancer and non-neoplastic adjacent mucosa

1995 ◽  
Vol 10 (1) ◽  
pp. 11-16 ◽  
Author(s):  
G. Saccani Jotti ◽  
M. Fontanesi ◽  
N. Orsi ◽  
L. Sarli ◽  
N. Pietra ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Human Colon ◽  
Human Colon Cancer ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Adjacent Mucosa ◽  
Aneuploid Tumor ◽  
The Relationship

DNA content was determined by flow cytometry in a series of 51 paired fresh tissue samples of primary colorectal carcinomas and the respective non-neoplastic adjacent mucosa in order to assess the relationship between DNA ploidy and the most commonly used prognostic factors. Aneuploidy was observed in 70.6% of the tumors and more than one aneuploid peak was present in 3.9%. Aneuploid tumor frequency was higher in left (93.3%) and right colon (64.7%) cancers than in rectal carcinomas (60.0%), and multiple aneuploid clones were detected more frequently in men than in women and in patients with advanced disease (Dukes stage D). Non-neoplastic mucosa adjacent to aneuploid tumors showed aneuploidy in 4 out of 51 samples (7.8%). The mucosa adjacent to diploid cancers had only diploid characteristics. Polidy did not correlate with histological abnormalities. These findings suggest that DNA content as determined by flow cytometry needs further study with adequate follow-up to evaluate possible correlations with relapse-free and overall survival. Furthermore the aneuploidy of non-neoplastic mucosa provides evidence for a field defect in mucosa adjacent to colorectal cancer and supports the concept that this alteration may be of influence on carcinogenesis.

scholarly journals Comparative DNA Flow Cytometric Study of Primary Intraocular and Central Nervous System Lymphomas

2020 ◽  
Vol 8 (2) ◽  
Author(s):  
Teresa Martinu ◽  
Claudia P. Correia ◽  
Andersson M. Figueiredo ◽  
Miguel N. Burnier Jr
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Primary Cns Lymphoma ◽  
Spatial Location ◽  
Intraocular Lymphoma ◽  
Dna Flow Cytometry ◽  
Cns Lymphoma ◽  
Primary Intraocular Lymphoma ◽  
Human Tonsil ◽  
Tissue Samples

Primary intraocular lymphoma is generally considered as a subset of primary CNS lymphoma. This study attempts to show that they may in fact represent distinct entities by comparing their respective proliferation rates using DNA flow cytometry. Four samples of primary intraocular lymphoma and seven samples of primary CNS lymphoma were analyzed, all from paraffin-embedded tissue. All tumors were of the large B-cell type. A normal human tonsil sample was used as a control. Tissue samples were analyzed by DNA flow cytometry, which is a precise and objective method to measure DNA content and cell proliferation of a tumor. S-phase fraction (SPF) and DNA content were measured for each sample. The average SPF for primary intraocular lymphoma was significantly higher than that of primary CNS lymphoma, 23.8 (range: 18.9 to 29.6) versus 15.1 (range: 1.1 to 25.1) respectively. Of the 11 tumors analyzed, 2 brain tumors were aneuploid and 1 eye tumor was peridiploid. All other tumors were diploid. Thus, no significant pattern was detected in the DNA content of the tumors. This lack of clinical significance of tumor aneuploidy is consistent with data reported in the literature. The results of this study indicate that primary intraocular lymphoma is more aggressive and of higher grade than primary CNS lymphoma. The different proliferation rates of intraocular and CNS lymphomas may be explained by either their different spatial location or a distinct genetic composition, the latter reinforcing the hypothesis that the two are fundamentally different entities

Flow Cytometric and Immunohistochemical Correlations in High Incidence Human Solid Tumors

1997 ◽  
Vol 83 (3) ◽  
pp. 689-697 ◽  
Author(s):  
Donatella Tirindelli Danesi ◽  
Marcello Spanò ◽  
Fabiana Antonini ◽  
Pierluigi AltaVista ◽  
Piera Catalano ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Rank Correlation ◽  
Breast Cancer Patients ◽  
Dna Index ◽  
Ploidy Levels ◽  
Spearman’S Rank Correlation ◽  
Flow Cytometric ◽  
Human Solid Tumors ◽  
Stem Cell Lines

475 patients with carcinoma at different sites (141 colon-rectum; 102 breast; 50 stomach; 48 kidney; 46 head and neck; 41 bladder; 47 other sites) submitted to surgery have been analyzed after histopathological staging and grading, by flow cytometry (monoparametric DNA content analysis) and immunohistochemistry (p53, c-erbB-2, and PCNA expression). In breast cancer patients the presence of receptors for estrogen (ER) and progesterone (PGR) has also been determined. Flow cytometry-derived parameters were DNA ploidy, fraction of cells in S-phase (SPF), and DNA content heterogeneity (multiclonal stem cell lines with different DNA index and/or more than one subpopulations with different ploidy levels in different samples from the same tumor). Correlations of the results obtained by the different techniques have been attempted by the non-parametric Spearman's rank correlation approach. Significant associations (P «0.05) were found between the histopathological, immunohistochemical and flow cytometric parameters considered in some anatomical regions, such as stomach (p53 vs DNA content aneuploidy and vs heterogeneity), colon-rectum (TNM vs p53 and vs heterogeneity), bladder (grading vs DNA content aneuploidy and vs heterogeneity). Tumor heterogeneity proved to be dependent on the number of tumor samples taken. The results of this preliminary assessment will subsequently be compared with the data obtained from a currently ongoing follow-up survey.

Intratumoral DNA content heterogeneity in breast carcinomas demonstrated by core punch tissue sampling and flow cytometry

2014 ◽  
Vol 67 (9) ◽  
pp. 821-824 ◽  
Author(s):  
Zhouwei Zhang ◽  
Donald L Weaver ◽  
Kavita Munjal ◽  
Mark F Evans
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Intratumoral Heterogeneity ◽  
Dna Index ◽  
Surgical Specimen ◽  
Flow Cytometric ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Relationship Of ◽  
Formalin Fixed

Further to advancements in instrumentation and fluorescent dye technologies, there has been a resurgence of interest in the flow cytometric assay of formalin-fixed, paraffin-embedded specimens. Here we present a novel, simple and effective alternative to whole block sectioning that allows selective multisampling of tissues within a specimen block and the investigation of intratumoral heterogeneity. Formalin-fixed, paraffin-embedded breast carcinoma specimens were core-punched using 1.0 mm diameter needles and assayed by flow cytometry using a modified Hedley method. Intratumoral heterogeneity for DNA index and per cent S-phase fraction was detected in 10 of 23 (44%) and 11 of 23 (47%) specimens respectively. Macro-level genomic heterogeneity is common in breast cancer even within a single surgical specimen block. Studies investigating the relationship of DNA content heterogeneity to other markers of genomic instability such as mutations, deletions, insertions and translocations are warranted.

scholarly journals DNA index and anatomical aspects of the micrografting of dragon fruit on different rootstocks

2021 ◽  
Vol 56 ◽  
Author(s):  
Renata Amato Moreira ◽  
Mariane Aparecida Rodrigues ◽  
Deniete Soares Magalhães ◽  
Leila Aparecida Salles Pio ◽  
Dalilhia Nazaré dos Santos ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Experimental Design ◽  
Dna Content ◽  
Dna Index ◽  
Dragon Fruit ◽  
Before And After ◽  
Hylocereus Undatus ◽  
Hylocereus Polyrhizus ◽  
Made In

Abstract The objective of this work was to evaluate the viability of the micrografting of yellow dragon fruit (Selenicereus megalanthus) on different rootstocks, based on DNA content and anatomical analyses. The used rootstocks were: yellow dragon fruit, white dragon fruit (Hylocereus undatus), Saborosa (Selenicereus setaceus) dragon fruit, and the Cebra and Orejona red dragon fruit (Hylocereus polyrhizus) varieties. The experimental design was completely randomized with five treatments and four replicates of five plants. After 30 days of cultivation, the following traits were evaluated: length and diameter of the micrografts and microrootstocks; and root length, percentage of setting, and fresh mass of the micrografts. Flow cytometry analyzes were performed before and after micrografting to verify genetic stability and the occurrence of endoreduplication. In addition, histological sections were made in the micrografting region to verify the connections of vessels and tissues between the graft and the rootstock. Endoreduplication was observed in all treatments. The amount of DNA in the yellow dragon fruit micrograft increased on the red Orejona variety. The presence of vessel connections was verified between the micrografts and microrootstocks. The yellow dragon fruit was also more vigorous when grafted on Orejona. Based on DNA content and anatomical analyses, in vitro yellow dragon fruit micrografting is feasible in all used rootstocks.

scholarly journals Estimation of the relative DNA content in species of the genus Spiraea, sections Chamaedryon and Glomerati by flow cytometry

2019 ◽  
Vol 9 (3) ◽  
pp. 142-149
Author(s):  
V. A. Kostikova ◽  
M. S. Voronkova ◽  
E. Yu. Mitrenina ◽  
A. A. Kuznetsov ◽  
A. S. Ernst ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Natural Populations ◽  
Internal Standard ◽  
Leaf Tissue ◽  
Cell Nuclei ◽  
Mountain Areas ◽  
Forest Zone ◽  
Standard Data ◽  
Relative Dna Content

The relative DNA content was studied in seven species of the genus Spiraea L., section Chamaedryon Ser., and in two species, section Glomerati Nakai, from 28 natural populations growing in Asian Russia. The cell nuclei were isolated from a leaf tissue. The relative intensity of fluorescence was measured using flow cytometry of propidium iodide-stained nuclei. The analysis was performed using a CyFlowSpace device (Germany, Sysmex Partec) with a laser radiation source of 532 nm. Fresh leaves of Solanum lycopersicum cv. ‘Stupice’ were used as an internal standard. Data on the relative DNA content are presented for the first time for S. flexuosa Fisch ex Cambess. (0.42–0.47 pg), S. ussuriensis Pojark. (0.49–0.52; 0.85 pg), S. alpina-Pall. (0.49–0.51 pg), S. media Schmidt. (0.45; 0.98–1.01 pg), S. trilobata L. (0.46 pg), S. hypericifolia L. (0.49–0.52 pg) and S. aquilegifolia Pall. (0.48–0.51 pg). Mesophytic species of the genus Spiraea growing in the forest zone (S. chamaedryfolia L. and S. media) exhibit a 2-fold higher relative DNA content (C-value) compared to xerophytic species (S. hypericifolia, S. crenata L., S. aquilegifolia and S. trilobata) and species confined only to mountain areas (S. alpinа, S. trilobata and S. aquilegifolia).

Aneuploid and Polyploid Cellular DNA Heterogeneity in Insect Cell Material of Diptera Species Analyzed by Flow Cytometry

1990 ◽  
Vol 45 (9-10) ◽  
pp. 1027-1030 ◽  
Author(s):  
Wolfgang Ulrich
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Insect Cell ◽  
Dna Analysis ◽  
Calliphora Vicina ◽  
Polyploid Cell ◽  
Cell Populations ◽  
Aneuploid Cell ◽  
Flow Cytometric ◽  
Cell Material

Abstract Flow cytometric 1-parameter DNA analysis and 2-parameter DNA /protein analysis have been performed with cell material of the diptera species Chironomus thummi, Drosophila melanogaster, Calliphora vicina and Musca domestica using an impulse cytophotometer with a new quartz objective, that was especially manufactured for cytofluorometric investigations. The occurrence of heterogenous cell populations with aneuploid and polyploid DNA content within the cell material of different developmental stages of diptera species have been determined, whereby in larvae polyploid cell populations and in imagos aneuploid cell populations predominate. Partially separation of 2C cells from other cell populations with higher DNA content can be done by Ficoll-Hypaque centrifugation as demonstrated with cell material from Chironomus larvae. For flow cytometric DNA analysis of insect cell material a simple and rapid cell preparation and staining technique is presented by using the DNA-specific fluorochrome DAPI in combination with the protein fluorochrome sulforhodamine 101. Employment of flow cytometry in diptera genetics might be a new tool for cytological and cytogenetic investigations as shown with the classical genetic objects Chironomus and Drosophila

scholarly journals ИССЛЕДОВАНИЕ РАЗМЕРА ГЕНОМА ПРЕДСТАВИТЕЛЕЙ РОДА POTENTILLA L. (ROSACEAE JUSS.)

2016 ◽  
Vol 6 (3) ◽  
pp. 229-233 ◽  
Author(s):  
A. A. Kechaykin ◽  
M. V. Skaptsov ◽  
S. V. Smirnov ◽  
M. G. Kutsev ◽  
A. I. Shmakov
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Dna Content ◽  
Pearson Correlation ◽  
Linear Regression Analysis ◽  
Genome Size Evolution ◽  
Size Evolution ◽  
Relative Dna Content ◽  
Pearson Correlation Analysis ◽  
First Time

<p>In our study, 15 representatives of the genus <em>Potentilla</em> L. were analyzed to obtain the relative DNA content and monoploid genome size in the seeds by flow cytometry method. For these <em>Potentilla</em> species, except <em>P. argentea </em>and<em> P. norvegica</em>, DNA content and genome size are revealed for the first time. As a result, the following data were obtaining: representatives of the older section <em>Desertae</em> are diploids with the large monoploid genome size. The genome of <em>P. asiatica</em> (Chrysanthae section) are decrease up to 0.35 pg, while the ploidy is increased to 4. Representatives of the <em>Multifidae</em> and <em>Supinae</em> section are presented with tetra- and hexaploid species with the 0.3 pg of the monoploid genome size. In section Tanacetifoliae are observed a further decrease of the genome size, representatives basically tetraploids. Thus, in the genome size evolution of a <em>Potentilla </em>genera are observed the increase of ploidy with significant reductions in monoploid genome size. This fact proves the general theory of genome evolution of angiosperms, directed towards its reduction. Linear regression analysis was showed a positive correlation between chromosome number and genome size of the representatives of the <em>Potentilla </em>genera. This suggests that between genome size and the number of chromosomes in <em>Potentilla </em>species are linear dependence. Pearson correlation analysis confirmed this result: the correlation coefficient is 0.97 at P&lt;0.05. </p>

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