scholarly journals Oral Cancer Genesis and Progression: DNA Near-Diploid Aneuploidization and Endoreduplication by High Resolution Flow Cytometry

2010 ◽  
Vol 32 (5-6) ◽  
pp. 373-383
Author(s):  
Alessandra Donadini ◽  
Massimo Maffei ◽  
Antonio Cavallero ◽  
Monica Pentenero ◽  
Davide Malacarne ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Resolution ◽  
Human Lymphocytes ◽  
Mitotic Division ◽  
Dna Index ◽  
Normal Oral Mucosa ◽  
Oral Carcinogenesis ◽  
Clinically Normal ◽  
Risk Of Progression ◽  
Potentially Malignant

Oral potentially malignant lesions (OPMLs) with dysplasia and aneuploidy are thought to have a high risk of progression into oral squamous cell carcinomas (OSCCs). Non-dysplastic “oral distant fields” (ODFs), characterized by clinically normal appearing mucosa sited at a distance from co-existing OPMLs, and non-dysplastic OPMLs may also represent an early pre-cancerous state. ODFs, OPMLs without and with dysplasia and OSCCs were investigated by high resolution DNA content flow cytometry (FCM). ODFs and OPMLs without dysplasia were DNA aneuploid respectively in 7/82 (8.5%) and 25/109 (23%) cases. “True normal oral mucosa” and human lymphocytes from healthy donors were DNA diploid in all cases and were used as sex specific DNA diploid controls. Dysplastic OPMLs and OSCCs were DNA aneuploid in 12/26 (46%) and 12/13 (92%) cases. The DNA aneuploid sublines were characterized by the DNA Index (DI ≠ 1). Aneuploid sublines in ODFs and in non-dysplastic and dysplastic OPMLs were near-diploid (DI < 1.4) respectively in all, 2/3 and 1/3 of the cases. DNA aneuploid OSCCs, instead, were characterized prevalently by multiple aneuploid sublines (67%), which were commonly (57%) high-aneuploid (DI ≥ 1.4). DNA near-diploid aneuploid sublines in ODFs and OPMLs appear as early events of the oral carcinogenesis in agreement with the concept of field effect. Near-diploid aneuploidization is likely to reflect mechanisms of loss of symmetry in the chromosome mitotic division. High DNA aneuploid and multiple sublines in OPMLs with dysplasia and OSCCs suggest, instead, mechanisms of “endoreduplication” of diploid and near-diploid aneuploid cells and chromosomal loss. High resolution DNA FCM seems to enable the separation of subsequent progression steps of the oral carcinogenesis.

New Criteria To Identify Risk of Progression in Monoclonal Gammopathy of Uncertain Significance: Multiparametric Flow Cytometry Analysis of Bone Marrow Plasma Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3517-3517
Author(s):  
Ernesto Perez-Persona ◽  
María-Belén Vidriales ◽  
Gema Mateo ◽  
Ramón Garcia Sanz ◽  
Marivi Mateos ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Monoclonal Gammopathy ◽  
Plasma Cells ◽  
Flow Cytometry Analysis ◽  
Dna Index ◽  
Multiparametric Flow Cytometry ◽  
Uncertain Significance ◽  
Risk Of Progression ◽  
Bone Marrow Plasma

Abstract Monoclonal Gammopathy of Uncertain Significance (MGUS) is a monoclonal disorder defined by the presence of a serum monoclonal protein <3g/dL, bone marrow plasma cells < 10% and absence of end-organ damage. The risk of progression to multiple myeloma (MM) is about 1% per year, and therefore these patients require long follow-up. Accordingly, the definition of new parameters that could be used for the identification of patients at risk of progression could be of great value. The aim of the present study is to evaluate the utility of multiparameter flow cytometry analysis of bone marrow (BM) plasma cells (PC) for predicting the risk of progression of MGUS patients. From January 1996 to September 2004, bone marrow aspirate samples from 350 patients, who fulfil the criteria of MGUS according to the International Myeloma Working Group criteria, were analysed by multiparametric flow cytometry. A specific gate on PC was performed based on CD138/CD38 expression and FSC/SSC characteristics and PC were immunophenotypically classified as normal (polyclonal) or aberrant (clonal) according to the expression of CD138, CD38, CD45, CD19 and CD56 antigens. Twenty seven patients (8 %) progressed to MM, with a median time to progression (TTP) of 46 months (range 9 to 109 months). Interestingly, the percentage of aberrant PC within the total BM PC compartment (aPC/BMPCc) clearly identify patients at different risk of progression. Thus, TTP in patients with ≥ 95% aPC/BMPCc was 85 months vs not reached cases with <95% aPC/BMPCc (p=0.0000). Other parameters with a significant influence on progression in the univariate analysis were: paraprotein level (higher vs lower of 2 mg/dl; p= 0.0004), the presence of immunoparesis (no paresis vs. decreased levels in one or two Ig. p= 0.0005), Bence-Jones proteinuria (p= 0.0003), PC BM infiltration assessed both by morphology and flow cytometry (p=0.0074; and p= 0.001, respectively), and DNA index assessed by flow cytometry (diploid vs aneuploid; p=0.0064). Moreover, the cut off level of 95% aPC/BMPCc, also allows the discrimination of two risk categories upon considering only patients at low risk of progression, based on a low paraprotein level or absence of inmunoparesis (p= 0.0000 and p= 0.0000, respectively). On multivariate analysis only the percentage of aPC/BMPCc (≥95%) (p=0.000), the DNA index (p=0.007), and the Bence-Jones proteinuria (p=0.000) showed independent prognostic value. In summary, our results show that multiparameter FC evaluation of BMPC at diagnosis is a simple, cost-effective and valuable tool for predicting the risk of progression of MGUS patients.

scholarly journals Diagnostic Value of Cytokeratin 17 during Oral Carcinogenesis: An Immunohistochemical Study

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Sirima Sanguansin ◽  
Theerachai Kosanwat ◽  
Rachai Juengsomjit ◽  
Sopee Poomsawat
Keyword(s):  
Diagnostic Marker ◽  
Diagnostic Value ◽  
Premalignant Lesions ◽  
Oral Potentially Malignant Disorders ◽  
Normal Oral Mucosa ◽  
Cytokeratin 17 ◽  
Oral Carcinogenesis ◽  
Clinicopathologic Factors ◽  
Potentially Malignant

Background. Little is known about the role of cytokeratin 17 (CK17) during oral carcinogenesis. CK17 expression in oral leukoplakia (OL), the most encountered oral potentially malignant disorders and oral squamous cell carcinoma (OSCC), remains very limited. To determine the role of CK17 during oral carcinogenesis and its potential diagnostic marker in oral premalignant and malignant lesions, this study evaluated CK17 expression in OL without dysplasia, OL with dysplasia, and OSCC. CK17 expression in these tissues was compared with those of normal oral mucosa (NOM). Additionally, the relationship between CK17 expression and clinicopathologic factors of OSCC was investigated. Methods. CK17 expression was evaluated in 186 samples consisting of 12 NOM, 33 OL without dysplasia, 58 OL with dysplasia, and 83 OSCC using immunohistochemistry. The proportion of positively immunostained cells was evaluated and scored. Results. CK17 was expressed in 8.3%, 54.5%, 74.1%, and 90.4% of NOM, OL without dysplasia, OL with dysplasia, and OSCC, respectively. NOM had a significantly lower CK17 score than OL with dysplasia ( p = 0.0003 ) and OSCC ( p < 0.0001 ). A significant association between CK17 expression and histopathologic differentiation of OSCC was found. Tumors with well differentiation had high CK17 expression compared with those of moderate and poor differentiation. Conclusion. CK17 was overexpressed in OL with dysplasia and OSCC, suggesting that CK17 plays a pivotal role in the development of premalignant lesions and OSCC. Of clinical significance, CK17 may be a good diagnostic marker for oral premalignant lesions and OSCC. Additionally, CK17 could be used as an objective tool to classify histopathologic grade in OSCC. The findings that CK17 expression is high in OSCC but low in NOM imply that CK17 may serve as a potential therapeutic target for OSCC.

scholarly journals Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles

Biomedicines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 124
Author(s):  
Jaco Botha ◽  
Haley R. Pugsley ◽  
Aase Handberg
Keyword(s):  
Flow Cytometry ◽  
High Resolution ◽  
Extracellular Vesicles ◽  
Single Particles ◽  
Multiple Parameters ◽  
Imaging Flow Cytometry ◽  
Highly Sensitive ◽  
Individual Project ◽  
Benchmarking Performance ◽  
High Throughput Manner

Flow cytometry remains a commonly used methodology due to its ability to characterise multiple parameters on single particles in a high-throughput manner. In order to address limitations with lacking sensitivity of conventional flow cytometry to characterise extracellular vesicles (EVs), novel, highly sensitive platforms, such as high-resolution and imaging flow cytometers, have been developed. We provided comparative benchmarks of a conventional FACS Aria III, a high-resolution Apogee A60 Micro-PLUS and the ImageStream X Mk II imaging flow cytometry platform. Nanospheres were used to systematically characterise the abilities of each platform to detect and quantify populations with different sizes, refractive indices and fluorescence properties, and the repeatability in concentration determinations was reported for each population. We evaluated the ability of the three platforms to detect different EV phenotypes in blood plasma and the intra-day, inter-day and global variabilities in determining EV concentrations. By applying this or similar methodology to characterise methods, researchers would be able to make informed decisions on choice of platforms and thereby be able to match suitable flow cytometry platforms with projects based on the needs of each individual project. This would greatly contribute to improving the robustness and reproducibility of EV studies.

scholarly journals Imaging flow cytometry as a sensitive tool to detect low-dose-induced DNA damage by analyzing 53BP1 and γH2AX foci in human lymphocytes

2015 ◽  
Vol 87 (12) ◽  
pp. 1070-1078 ◽  
Author(s):  
Matus Durdik ◽  
Pavol Kosik ◽  
Jan Gursky ◽  
Lenka Vokalova ◽  
Eva Markova ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Damage ◽  
Low Dose ◽  
Human Lymphocytes ◽  
Imaging Flow Cytometry ◽  
Γh2ax Foci ◽  
Sensitive Tool

High resolution DNA flow cytometry of boar sperm cells in identification of boars carrying cytogenetic aberrations

2004 ◽  
Vol 62 (3-4) ◽  
pp. 501-511
Author(s):  
Jacob Larsen ◽  
Knud Christensen ◽  
Jørgen K Larsen ◽  
Peter Østrup Jensen ◽  
Ingemar Gustavsson ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Resolution ◽  
Dna Flow Cytometry ◽  
Sperm Cells ◽  
Cytogenetic Aberrations ◽  
Boar Sperm

scholarly journals Prerequisites for the analysis and sorting of extracellular vesicle subpopulations by high-resolution flow cytometry

2015 ◽  
Vol 89 (2) ◽  
pp. 135-147 ◽  
Author(s):  
Tom Groot Kormelink ◽  
Ger J. A. Arkesteijn ◽  
Frans A. Nauwelaers ◽  
Ger van den Engh ◽  
Esther N. M. Nolte-'t Hoen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Resolution ◽  
Extracellular Vesicle

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).

Assessment of Persisting Chromosome Aberrations by Flow Karyotyping of Cloned Chinese Hamster Cells

1988 ◽  
Vol 43 (11-12) ◽  
pp. 948-954 ◽  
Author(s):  
Friedrich J. Otto
Keyword(s):  
Flow Cytometry ◽  
High Resolution ◽  
Cell Lines ◽  
Chromosomal Aberrations ◽  
Chromosome Aberrations ◽  
Chinese Hamster ◽  
Chinese Hamster Cells ◽  
Cell Clones ◽  
Permanent Cell

Abstract A preparation, staining and measuring protocol for high resolution flow cytometry of chromosomes was developed. This method allows us to identify all chromosome types and is suited for characterization of permanent cell lines and cell clones by establishing their flow karyotypes. In cell clones this procedure can be used for the detection of chromosomal aberrations which appear spontaneously or are induced by mutagen treatment and persist in the cell population.

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