DNA flow cytometry of acinic cell carcinomas of major salivary glands

AbstractFifteen acinic cell carcinomas from an equal number of patients were analysed for their DNA content and proliferative (S-phase) index by flow cytometry from archival tissues. Seven of the carcinomas manifested a diploid DNA content. None of the patients with diploid acinic cell carcinomas died of their carcinomas and none developed metastases in follow-up periods extending for 10 or more years. Four of eight patients with aneuploid acinic cell carcinomas have died because of their malignancies within a 10 year period after the first surgical removal of the carcinoma. Five of the eight patients exhibited metastases. Although the number of cases does not permit strong correlations between histopathological features, abnormalities in DNA content and outcome of patients, it was noted that carcinomas with prominent necrosis, tubulo ductal differentiation and ‘dedifferentiated’ areas displayed more aggressive biological courses.

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Persistent or recurrent Barrett’s neoplasia after an endoscopic therapy session is associated with DNA content abnormality and can be detected by DNA flow cytometric analysis of paraffin-embedded tissue

Modern Pathology ◽

10.1038/s41379-021-00832-8 ◽

2021 ◽

Author(s):

Christopher J. Bowman ◽

Ruth Zhang ◽

Dana Balitzer ◽

Dongliang Wang ◽

Peter S. Rabinovitch ◽

...

Keyword(s):

Risk Factors ◽

Flow Cytometry ◽

Dna Content ◽

Endoscopic Therapy ◽

Significant Risk ◽

Dna Flow Cytometry ◽

Low Grade ◽

Complete Eradication ◽

Increased Risk

AbstractEndoscopic therapy is currently the standard of care for the treatment of high-grade dysplasia (HGD) or intramucosal adenocarcinoma (IMC) in patients with Barrett’s esophagus (BE). Visible lesions are treated with endoscopic mucosal resection (EMR), which is often coupled with radiofrequency ablation (RFA). However, endoscopic therapy may require multiple sessions (one session every 2-3 months) and does not always assure complete eradication of neoplasia. Furthermore, despite complete eradication, recurrences are not uncommon. This study assesses which potential risk factors can predict a poor response after endoscopic sessions. Forty-five BE patients who underwent at least one endoscopic session (EMR alone or ablation with or without preceding EMR) for the treatment of HGD/IMC, low-grade dysplasia (LGD), or indefinite for dysplasia (IND) were analyzed. DNA flow cytometry was performed on 82 formalin-fixed paraffin-embedded samples from the 45 patients, including 78 HGD/IMC, 2 LGD, and 2 IND. Eight non-dysplastic BE samples were used as controls. Three to four 60-micron thick sections were cut from each tissue block, and the area of HGD/IMC, LGD, or IND was manually dissected. Potential associations between clinicopathologic risk factors and persistent/recurrent HGD/IMC following each endoscopic session were examined using univariate and multivariate Cox models with frailty terms. Sixty (73%) of the 82 specimens showed abnormal DNA content (aneuploidy or elevated 4N fraction). These were all specimens with HGD/IMC (representing 77% of that group). Of these 60 HGD/IMC samples with abnormal DNA content, 42 (70%) were associated with subsequent development of persistent/recurrent HGD/IMC (n = 41) or esophageal adenocarcinoma (EAC; n = 1) within a mean follow-up time of 16 months (range: 1 month to 9.4 years). In contrast, only 6 (27%, all HGD/IMC) of the 22 remaining samples (all with normal DNA content) were associated with persistent/recurrent HGD/IMC. For outcome analysis per patient, 11 (24%) of the 45 patients developed persistent/recurrent HGD/IMC or EAC, despite multiple endoscopic sessions (mean: 3.6, range: 1–11). In a univariate Cox model, the presence of abnormal DNA content (hazard ratio [HR] = 3.8, p = 0.007), long BE segment ≥ 3 cm (HR = 3.4, p = 0.002), endoscopic nodularity (HR = 2.5, p = 0.042), and treatment with EMR alone (HR = 2.9, p = 0.006) were significantly associated with an increased risk for persistent/recurrent HGD/IMC or EAC. However, only abnormal DNA content (HR = 6.0, p = 0.003) and treatment with EMR alone (HR = 2.7, p = 0.047) remained as significant risk factors in a multivariate analysis. Age ≥ 60 years, gender, ethnicity, body mass index (BMI) ≥ 30 kg/m2, presence of hiatal hernia, and positive EMR lateral margin for neoplasia were not significant risk factors for persistent/recurrent HGD/IMC or EAC (p > 0.05). Three-month, 6-month, 1-year, 3-year, and 6-year adjusted probabilities of persistent/recurrent HGD/IMC or EAC in the setting of abnormal DNA content were 31%, 56%, 67%, 79%, and 83%, respectively. The corresponding probabilities in the setting of normal DNA content were 10%, 21%, 28%, 38%, and 43%, respectively. In conclusion, in BE patients with baseline HGD/IMC, both DNA content abnormality and treatment with EMR alone were significantly associated with persistent/recurrent HGD/IMC or EAC following each endoscopic session. DNA content abnormality as detected by DNA flow cytometry identifies HGD/IMC patients at highest risk for persistent/recurrent HGD/IMC or EAC, and it also serves as a diagnostic marker of HGD/IMC with an estimated sensitivity of 77%. The diagnosis of HGD/IMC in the setting of abnormal DNA content may warrant alternative treatment strategies as well as long-term follow-up with shorter surveillance intervals.

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Flow Cytometry in Bladder Tumours: Our Experience

Urologia Journal ◽

10.1177/039156039306000206 ◽

1993 ◽

Vol 60 (2) ◽

pp. 158-161 ◽

Cited By ~ 1

Author(s):

M. De Siati ◽

D. Grassi ◽

N. Franzolin ◽

F. Marchioretto ◽

L.S. Azzolina

Keyword(s):

Flow Cytometry ◽

Initial Data ◽

Patient Survival ◽

Tumour Progression ◽

Tumour Stage ◽

Number Of Patients ◽

Bladder Tumours

From January 1992 to January 1993, specimens from bladder tumours of 75 patients were analyzed by cytometry (FCM). Our data show a good correlation between tumour stage and grade, and tumour ploidy. Furthermore, aneuploid tumours showed progression more often than diploid tumours. A larger number of patients admitted to the study and a more prolonged period of follow-up will be necessary to confirm our initial data and to investigate whether a correlation exists between FCM and tumour progression and patient survival.

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Flow cytometric DNA analysis in conservatively treated renal tumours

Urologia Journal ◽

10.1177/039156039506201s39 ◽

1995 ◽

Vol 62 (1_suppl) ◽

pp. 141-143

Author(s):

P. Beltrami ◽

M. Lazzarotto ◽

G. Giusti ◽

C. Tallarigo ◽

G. Malossini ◽

...

Keyword(s):

Flow Cytometry ◽

Dna Analysis ◽

Tumour Size ◽

Prognostic Significance ◽

Dna Flow Cytometry ◽

Additional Parameter ◽

Flow Cytometric ◽

Renal Tumours ◽

Dna Histograms

— The DNA histograms of 21 conservatively resected renal tumours were studied using DNA flow cytometry. Five patients had an imperative and sixteen an elective indication for conservative resection of the renal tumour. On the basis of DNA histograms twelve aneuploid tumours were pointed out. A mean follow-up of 34.2 months was considered to see whether the ploidy would provide criteria with a prognostic significance, to be useful as an additional parameter. None of the twenty-one patients had local recurrence or distant metastasis: in our series the DNA analysis had no influence on the prognosis of this group of patients. The tumour size seems to be the only selective parameter for choosing renal-conserving surgery.

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

The International Journal of Biological Markers ◽

10.1177/172460089501000103 ◽

1995 ◽

Vol 10 (1) ◽

pp. 11-16 ◽

Cited By ~ 10

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.

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Diagnosis and risk stratification of Barrett’s dysplasia by flow cytometric DNA analysis of paraffin-embedded tissue

Gut ◽

10.1136/gutjnl-2017-313815 ◽

2017 ◽

Vol 67 (7) ◽

pp. 1229-1238 ◽

Cited By ~ 14

Author(s):

Won-Tak Choi ◽

Jia-Huei Tsai ◽

Peter S Rabinovitch ◽

Thomas Small ◽

Danning Huang ◽

...

Keyword(s):

Flow Cytometry ◽

Risk Stratification ◽

Dna Content ◽

Dna Analysis ◽

Diagnostic Marker ◽

Oesophageal Adenocarcinoma ◽

Dna Flow Cytometry ◽

Low Grade ◽

Area Of Interest ◽

Formalin Fixed Paraffin Embedded

ObjectiveThe diagnosis of dysplasia in Barrett’s oesophagus (BO) can be challenging, and reliable ancillary techniques are not available. This study examines if DNA content abnormality detected by flow cytometry can serve as a diagnostic marker of dysplasia and facilitate risk stratification of low-grade dysplasia (LGD) and indefinite for dysplasia (IND) patients using formalin-fixed paraffin-embedded (FFPE) BO samples with varying degrees of dysplasia.DesignDNA flow cytometry was performed on 80 FFPE BO samples with high-grade dysplasia (HGD), 38 LGD, 21 IND and 14 negative for dysplasia (ND). Three to four 60-micron thick sections were cut from each tissue block, and the area of interest was manually dissected.ResultsDNA content abnormality was identified in 76 HGD (95%), 8 LGD (21.1%), 2 IND (9.5%) and 0 ND samples. As a diagnostic marker of HGD, the estimated sensitivity and specificity of DNA content abnormality were 95% and 85%, respectively. For patients with DNA content abnormality detected at baseline LGD or IND, the univariate HRs for subsequent detection of HGD or oesophageal adenocarcinoma (OAC) were 7.0 and 20.0, respectively (p =<0.001).ConclusionsThis study demonstrates the promise of DNA flow cytometry using FFPE tissue in the diagnosis and risk stratification of dysplasia in BO. The presence of DNA content abnormality correlates with increasing levels of dysplasia, as 95% of HGD samples showed DNA content abnormality. DNA flow cytometry also identifies a subset of patients with LGD and IND who are at higher risk for subsequent detection of HGD or OAC.

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NUCLEAR DNA CONTENT IN BLUEBERRY DETERMINED BY FLOW CYTOMETRY

HortScience ◽

10.21273/hortsci.27.6.580e ◽

1992 ◽

Vol 27 (6) ◽

pp. 580e-580

Author(s):

Rodomiro Ortiz ◽

D.E. Costich ◽

T.P. Meagher ◽

N. Vorsa

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Haploid Genome ◽

Dna Flow Cytometry ◽

Dna Amount ◽

Ploidy Levels ◽

Polyploid Evolution ◽

Haploid Genome Size

DNA flow cytometry was used to determine nuclear DNA content in diploid blueberry species, and 3x, 4x, 5x, and 6x ploidy levels. Relative fluorescence intensity of stained nuclei measured by flow cytometry was a function of the number of chromosome sets (X): Y = 3.7X – 2.3 (r2 = 95.1%). DNA flow cytometry should be useful for ploidy level determination in the seedling stage. A significant linear relationship was established between nuclear DNA content and number of chromosomes (x); DNA (pg) = 0.52 x1 (r2 = 99.8%). Based on this equation the haploid genome DNA amount (1C) was calculated as 0.62 ± 0.08 pg, with an approximate haploid genome size of 602 Mbp/1C. The results indicate that conventional polyploid evolution occured in the section Cyanococcus, genus Vaccinium: the increase in DNA was concurrent with increase in chromosome number. DNA content differences among 2x species were correlated with Nei's genetic distance estimates based on 20 isozyme markers. Most of the variation was among species (49%), with 26% between populations within species, and 25% within populations.

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

McGill Journal of Medicine ◽

10.26443/mjm.v8i2.674 ◽

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

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Prevalence of Large Granular Lymphocyte Proliferation in Chronic Myeloid Leukemia (CML) Patients Treated with Dasatinib.

Blood ◽

10.1182/blood.v114.22.1111.1111 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1111-1111

Author(s):

Jason Valent ◽

Charles A. Schiffer

Keyword(s):

Flow Cytometry ◽

Side Effects ◽

Peripheral Blood ◽

Chronic Phase ◽

Cell Transplant ◽

Pleural Effusions ◽

Blast Phase ◽

Number Of Patients ◽

Dasatinib Treatment

Abstract Abstract 1111 Poster Board I-133 Dasatinib is a potent inhibitor of the BCR-ABL tyrosine kinase which is effective in the treatment of imatinib refractory CML. While hematologic toxicities of neutropenia and thrombocytopenia are well known, large granular lymphocytosis has been reported in only a small number of patients without prior allogeneic stem cell transplant treated with dasatinib for CML. During routine follow up of leukocyte counts in 15 consecutive patients (age range 27-77 years) treated with dasatinib, 4 patients (2 chronic-phase, 1 accelerated phase with clonal cytogenetic progression, 1 blast-phase) developed a lymphocytosis (> 3800/mm3). Peripheral blood smear and peripheral blood flow cytometry revealed a population of large granular lymphocytes (LGLs) expressing CD3, CD8, CD57, and variable expression of CD56. Lymphocytosis was first noted between 1 and 9 months after initiation of dasatinib and has persisted in 3 of the patients with a median follow up of 33 months from the onset of lymphocytosis. Peak absolute lymphocyte count ranged from 5000/mm3 to 6900/mm3 and approximately 40 to 60% of the lymphocytes were LGLs by flow cytometry with the remainder being predominantly T lymphocytes. These 4 patients with LGL lymphocytosis have all have major molecular responses and the patient with blast-phase has remained in a complete cytogenetic remission with a major molecular response 44 months after initiation of dasatinib. The 11 other patients (6 chronic-phase, 2 accelerated-phase, 3 blast-phase) treated with dasatinib for CML have not developed lymphocytosis. These patients have been followed for a median of 25 months (range 3-50 months) although some were treated with dasatinib for a relatively short period of time because of poor response of their advanced CML. Review of the peripheral blood smears from 3 of the 6 chronic phase patients without lymphocytosis who remain on dasatinib treatment did not reveal any LGLs. All of these 6 patients have had complete, sustained cytogenetic responses. A persistent pleural effusion developed in the blast phase patient with lymphocytosis approximately 12 months after lymphocytosis developed; no significant side effects were noted in the other 3 patients although one remains thrombocytopenic. Pleural effusions developed in 2 of the 6 patients without lymphocytosis who remain on dasatinib treatment. Previous reports have suggested an increased incidence of “inflammatory” type side effects such as pleural effusions and pneumonitis in patients with dasatinib related LGL proliferation, although the small number of patients in this series precludes analysis of this association. In summary, LGL proliferation was detected in a minimum of 27% of dasatinib recipients and may be associated with a beneficial response. While the mechanism of LGL proliferation has not been fully explained, it has been suggested that dasatinib mediated inhibition of immunoregulatory kinases such as Src is permissive of LGL proliferation. Further evaluation of the frequency and clinical impact of this phenomenon in the large clinical trials of patients treated with dasatinib is warranted. Disclosures Schiffer: Bristol Myers: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding.

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Flow Cytometry Analysis in Voided Urine of Patients with Bladder Cancer

Urologia Journal ◽

10.1177/039156039306000207 ◽

1993 ◽

Vol 60 (2) ◽

pp. 162-166

Author(s):

A. Lotto ◽

G. Carluccio ◽

A. Calisti ◽

A. Disperati ◽

E. Capuzzo ◽

...

Keyword(s):

Flow Cytometry ◽

Bladder Cancer ◽

Quantitative Evaluation ◽

Dna Content ◽

Urine Cytology ◽

Urinary Cytology ◽

Flow Cytometry Analysis ◽

Excellent Correlation ◽

First Diagnosis

Flow cytometry is known to be able to give a quantitative evaluation of the DNA of cellular populations (grade of ploidy), as well as to estimate the percentages of phases (S + G2M) providing useful information about the pathology in question and its aggressivity. This method has been applied in diagnosing patients with bladder cancer, using their voided urine and comparing with urine cytology. Our data, from 59 patients, indicate flow cytometry utility in diagnosing bladder cancer; in fact there is an excellent correlation between the urinary cytology and the DNA content in cytometry which increases in higher grade bladder cancer. The sensitivity of CFM is in the range of 92% to 94%, and is superior to that of conventional voided urine cytology (range 64% to 84%). It is felt that cytofluorometric analysis permits a reliable evaluation of voided urine, not only at first diagnosis, but especially during follow-up.

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