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

scholarly journals Cerebrospinal Fluid Flow Cytometry: Utility in Central Nervous System Lymphoma Diagnosis

2020 ◽  
Vol 47 (3) ◽  
pp. 382-388
Author(s):  
Ka Loong Kelvin Au ◽  
Sarah Latonas ◽  
Afshin Shameli ◽  
Iwona Auer ◽  
Christopher Hahn
Keyword(s):  
Central Nervous System ◽  
Flow Cytometry ◽  
Cerebrospinal Fluid ◽  
Nervous System ◽  
Hematological Malignancy ◽  
Previous History ◽  
Primary Cns Lymphoma ◽  
Cns Lymphoma ◽  
Csf Flow ◽  
Neurologic Symptoms

ABSTRACT:Background:Flow cytometry of the cerebrospinal fluid (CSF) is used in isolation or as an adjunct to cytology to increase the sensitivity of detecting central nervous system (CNS) lymphoma. We aimed to evaluate the sensitivity of CSF flow cytometry as a diagnostic screening tool for primary CNS lymphoma in patients presenting with undifferentiated neurologic symptoms.Methods:We retrospectively reviewed all CSF samples received by the Calgary Laboratory Services Flow Cytometry Laboratory from 2012 to 2015. Clinical data, laboratory investigations, radiologic imaging studies, and pathological data were analyzed. Clinical review extended to 2 years post-CSF flow cytometric testing.Results:Only 43/763 (5.6%) samples of CSF flow cytometry in 28/573 (4.9%) patients were found to be positive for a hematological malignancy in patients with undifferentiated neurologic symptoms. The overall sensitivity of the test was 13.8% with 25 patients with negative CSF flow cytometry later having a positive biopsy for CNS lymphoma. CSF flow cytometry was negative in all cases when at the time of CSF examination the patient did not have a previous hematological malignancy or findings of abnormal enhancement on MRI (n = 249).Conclusion:CSF flow cytometry has low utility in screening for primary CNS lymphoma in the absence of a previous history of hematologic malignancy or findings of abnormal enhancement on MRI.

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.

Primary intraocular lymphoma (PIOL): An International Primary CNS Lymphoma Collaborative group report

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 1534-1534
Author(s):  
S. Grimm ◽  
J. Pulido ◽  
K. Jahnke ◽  
D. Schiff ◽  
A. Hall ◽  
...  
Keyword(s):  
Performance Status ◽  
Primary Cns Lymphoma ◽  
Local Therapy ◽  
Initial Therapy ◽  
Collaborative Group ◽  
Intraocular Lymphoma ◽  
Cns Lymphoma ◽  
Treatment Type ◽  
Increased Risk ◽  
Sites Of Relapse

1534 Background: PIOL is a hemopoietic tumor that arises in the retina, vitreous or optic nerve head, and carries a high risk of ocular and CNS relapse. The natural history and optimal management are unknown. Methods: A retrospective series of 81 patients with PIOL was assembled from 15 centers in 7 countries. Only patients with isolated ocular lymphoma were included; none had brain, spinal cord, or systemic lymphoma at diagnosis. Results: The median age at diagnosis was 65 (24–85). 58% were women. The median ECOG performance status was 0, and only three had a score > 1. The median latency from symptom onset to diagnosis was 6 months (0– 36). Slit lamp exam was positive in 51, negative in 6, and not reported in 24. Vitrectomy was positive in 72 and negative in 2. 6 had a positive choroidal or retinal biopsy and 1 had no ocular surgery. CSF cytology was positive in 10 (17%), negative in 48, and unknown in 23. 21 received local therapy at diagnosis: 6 intra-ocular methotrexate (400 ug), 14 ocular radiation (median 3600 cGy), and 1 both modalities. 52 received more extensive therapy including systemic chemotherapy alone in 20 and a combination of chemotherapy and radiotherapy in 32. 5 received no treatment and details are unknown in 3. 47 patients (58%) relapsed a median of 19 months (0.5–180) after initial therapy. Sites of relapse included brain 47%, eyes 30%, brain and eyes 15%, and systemic 8%. Patients treated with ocular therapy alone did not have an increased risk of failing in the brain (p = 0.6). Progression free survival (PFS) and overall survival (OS) were 29.6 and 57 months respectively and were unaffected by the choice of therapy. CNS disease was the cause of death in 19/33 (58%). Conclusions: In this series, treatment type did not affect sites of relapse, PFS or OS in patients with PIOL. To minimize toxicity, the best initial therapy should be limited to intraocular chemotherapy or focal radiotherapy. Prospective clinical trials are needed to improve our understanding and treatment of this disease. No significant financial relationships to disclose.

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

Gut ◽  
2017 ◽  
Vol 67 (7) ◽  
pp. 1229-1238 ◽  
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.

scholarly journals NUCLEAR DNA CONTENT IN BLUEBERRY DETERMINED BY FLOW CYTOMETRY

HortScience ◽  
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.

scholarly journals Intraocular Lymphoma

1998 ◽  
Vol 5 (4) ◽  
pp. 317-325 ◽  
Author(s):  
David S. Bardenstein
Keyword(s):  
Primary Cns Lymphoma ◽  
Intraocular Lymphoma ◽  
Diagnostic Process ◽  
Cns Lymphoma ◽  
Ocular Involvement ◽  
Disease Treatment ◽  
Treatment Regimens ◽  
Ocular Lymphoma ◽  
Life Threatening ◽  
Distinct Subtype

Background Primary ocular lymphoma is a distinct subtype of intraocular lymphoma. Its clinical presentation can mimic benign conditions. Diagnosis is often based on obtaining an intraocular biopsy. Optimal management is not yet realized. Methods This report combines the experience of the author with a review of the current literature pertaining to intraocular lymphoma. Results Primary ocular lymphoma, a subtype of primary central nervous system (CNS) lymphoma, has a variable clinical course and frequently mimics benign inflammatory disease. Even when suspected, diagnosis can be elusive. Chemoradiation is the most effective treatment, but significant ocular and cerebral morbidity is associated with its use. Novel treatment regimens may reduce or eliminate side effects while preserving life, vision, and CNS function. Conclusions Primary CNS lymphoma with ocular involvement should be considered in patients with refractory uveitis, yellow-white choroidal masses, and CNS lymphoma. Aggressiveness in making the diagnosis should be tempered by the potential complications of the diagnostic process and advanced by the life-threatening nature of the disease. Treatment should attempt to maximize efficacy while incorporating considerations such as extent of disease and the patient's age, health, and mental state.

Results of Intensive Chemotherapy Followed by Hematopoietic Stem-Cell Rescue in 22 Patients With Refractory or Recurrent Primary CNS Lymphoma or Intraocular Lymphoma

2001 ◽  
Vol 19 (3) ◽  
pp. 742-749 ◽  
Author(s):  
Carole Soussain ◽  
Florence Suzan ◽  
Khê Hoang-Xuan ◽  
Nathalie Cassoux ◽  
Vincent Levy ◽  
...  
Keyword(s):  
Stem Cell ◽  
Primary Cns Lymphoma ◽  
Intraocular Lymphoma ◽  
Cns Lymphoma ◽  
High Dose ◽  
Refractory Disease ◽  
Intensive Chemotherapy ◽  
Hematopoietic Stem ◽  
Stem Cell Rescue ◽  
Entire Procedure

PURPOSE: To assess the feasibility and efficacy of intensive chemotherapy with hematopoietic stem-cell rescue (IC + HCR) in patients with refractory or recurrent primary CNS lymphoma (PCNSL) or intraocular lymphoma (IOL). PATIENTS AND METHODS: IC consisted of thiotepa 250 mg/m2/d days −9 through −7, busulfan 10 mg/kg (total dose) days −6 through −4, and cyclophosphamide 60 mg/kg/d days −3 and −2. Intravenous clonazepam 2 mg/d was given prophylactically from the day before initiation of busulfan therapy to the day after completion of busulfan therapy. Patients with refractory or recurrent PCNSL underwent IC + HCR only if they were chemosensitive to two cycles of salvage treatment with cytarabine (2 g/m2/d days 2 through 5 and 50 mg/m2/d days 1 through 5 in a 12-hour infusion) and etoposide (VP-16; 200 mg/m2/d days 2 through 5) (CYVE). Patients with IOL refractory to high-dose methotrexate (MTX) and cytarabine entered the IC + HCR program directly. RESULTS: Twenty-two patients (10 with relapses, 12 with refractory disease) were enrolled. Twenty patients entered the IC + HCR program: twelve entered after CYVE treatment, seven entered directly, and one had previously been retreated with high-dose MTX. Before IC, eight patients were in complete remission (CR), four were in partial remission (PR), one had stable disease, and seven had refractory disease. After IC + HCR, 16 patients entered CR, two remained in PR, one had stable disease, and one had disease progression. Fourteen patients remained alive (median follow-up time, 41.5 months). The overall probability of survival at 3 years was 63.7%. After IC, that probability was 60% and the 3-year probability of event-free survival was 53%. Seven patients had neurologic adverse events during the entire procedure. CONCLUSION: IC + HCR proved feasible and effective in patients with refractory or recurrent PCNSL or IOL. The entire procedure seemed to be most toxic in patients ≥ 60 years. A prospective multicenter study is ongoing.

Phase II trial of chemotherapy alone for primary CNS and intraocular lymphoma.

1998 ◽  
Vol 16 (9) ◽  
pp. 3000-3006 ◽  
Author(s):  
V Sandor ◽  
V Stark-Vancs ◽  
D Pearson ◽  
R Nussenblat ◽  
S M Whitcup ◽  
...  
Keyword(s):  
Radiation Therapy ◽  
Response Rate ◽  
Survival Rates ◽  
Primary Cns Lymphoma ◽  
Progression Free Survival ◽  
Complete Response ◽  
Intraocular Lymphoma ◽  
Cns Lymphoma ◽  
High Dose ◽  
Free Survival

PURPOSE Primary CNS lymphoma (PCNSL) and primary intraocular lymphoma (IOL) are usually treated with radiation therapy alone or in combination with chemotherapy. The neurotoxicity of these treatments can be substantial. This study attempts to define the toxicity and efficacy of the treatment of this disease with chemotherapy alone. PATIENTS AND METHODS Fourteen nonimmunocompromised patients were accrued to a chemotherapy regimen that incorporated a 24-hour infusion of high-dose methotrexate total dose of 8.4 g/m2 with leucovorin rescue; thiotepa 35 mg/m2; vincristine 1.4 mg/m2; dexamethasone; and intrathecal cytarabine (Ara-C) and methotrexate (MTV) administered in 21-day cycles. Seven patients were prospectively followed up with formal neuropsychologic assessments for evidence of CNS toxicity. RESULTS The response rate was 100% with 11 (79%) complete responses and three (21%) partial responses. Cumulative survival and progression-free survival rates at more than 4.5 years were 68.8% and 34.3%, respectively. Median survival has not been reached, and median progression-free survival was 16.5 months. Toxicity included severe leukoencephalopathy that was clearly attributable to chemotherapy (two patients), grade 3 or 4 neutropenia in 50% of the cycles administered, ileus (one patient), and seizures (two patients). Mucositis and renal and hepatic toxicity were mild and not therapy limiting. CONCLUSION The MTV regimen is generally well tolerated and produces a high complete response rate. Chemotherapy alone should be investigated further in this disease to assess the necessity of initial radiation therapy, either alone or in combined modality regimens, for the achievement of optimal response and survival.

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