Tissue-specific markers in flow cytometry of urological cancers. III. Comparing chromosomal and flow cytometric dna analysis of bladder tumors

1987 ◽  
Vol 39 (3) ◽  
pp. 304-310 ◽  
Author(s):  
A. W. G. B. Smeets ◽  
L. Laarakkers ◽  
R. P. E. Pauwels ◽  
J. L. M. Beck ◽  
G. P. Vooijs ◽  
...  
1995 ◽  
Vol 62 (1_suppl) ◽  
pp. 141-143
Author(s):  
P. Beltrami ◽  
M. Lazzarotto ◽  
G. Giusti ◽  
C. Tallarigo ◽  
G. Malossini ◽  
...  

— 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.


1986 ◽  
Vol 37 (2) ◽  
pp. 201-207 ◽  
Author(s):  
W. F. J. Feitz ◽  
H. F. M. Karthaus ◽  
H. L. M. Beck ◽  
C. Romijn ◽  
A. P. M. van der Meyden ◽  
...  

1985 ◽  
Vol 36 (3) ◽  
pp. 349-356 ◽  
Author(s):  
W.F.J. FEITZ ◽  
H.L.M. BECK ◽  
A.W.G.B. SMEETS ◽  
F.M.J. DEBRUYNE ◽  
G.P. VOOIJS ◽  
...  

2000 ◽  
Vol 46 (2) ◽  
pp. 224-233 ◽  
Author(s):  
Bent Raungaard ◽  
Finn Heath ◽  
Peter Steen Hansen ◽  
Jens Uffe Brorholt-Petersen ◽  
Henrik Kjærulf Jensen ◽  
...  

Abstract Background: Familial defective apolipoprotein (apo) B-100 (FDB) is caused by a mutation in the apoB gene and characterized by decreased binding of LDL to LDL receptors because of reduced function of the apoB-100 ligand. FDB may be associated with severe hypercholesterolemia and cannot always be distinguished from familial hypercholesterolemia phenotypically. Methods: We used a fluorescence flow cytometry assay with Epstein-Barr virus-transformed lymphocytes to detect reduced LDL ligand function by competitive binding with fluorescently conjugated LDL (DiI-LDL). The assay was tested and validated using LDL from patients heterozygous for the Arg3500-Gln mutation and their first-degree relatives. Knowing the actual apoB genotype of patients and relatives allowed us to assess the ability of the assay to predict the results of DNA analysis. The results were compared to measurements of LDL ligand function in unrelated healthy control subjects to characterize functionally the Arg3500-Gln mutation. Results: Fluorescence was significantly increased in cells incubated with DiI-LDL in competition with unlabeled LDL from FDBR3500Q heterozygotes compared with cells incubated with DiI-LDL in competition with unlabeled LDL from relatives or unrelated healthy control subjects. Thus, patients heterozygous for the Arg3500-Gln mutation had significantly reduced LDL ligand function. The binding affinity of LDL from FDBR3500Q heterozygotes was 32% of that in non-FDB relatives and healthy controls. The assay had a diagnostic sensitivity of 0.95 and diagnostic specificity of 0.89. Conclusions: The diagnostic accuracy of the assay was too low to allow reliable diagnosis of individual cases of heterozygous FDBR3500Q. However, fluorescence flow cytometry may supplement genetic identification of FDB and functionally characterize gene mutations associated with major reductions in LDL ligand function.


1987 ◽  
Vol 137 (6) ◽  
Author(s):  
Masaaki Tachibana ◽  
Seido Jitsukawa ◽  
Tomohiko Iigaya ◽  
Shiro Baba ◽  
Makoto Hata ◽  
...  

1990 ◽  
Vol 45 (9-10) ◽  
pp. 1027-1030 ◽  
Author(s):  
Wolfgang Ulrich

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


1988 ◽  
Vol 69 (4) ◽  
pp. 588-592 ◽  
Author(s):  
Kyung G. Cho ◽  
Tadashi Nagashima ◽  
Stanley Barnwell ◽  
Takao Hoshino

✓ Paraffin-embedded specimens of brain tumors from 256 patients who had received an intravenous infusion of bromodeoxyuridine (BUdR) at the time of craniotomy were analyzed retrospectively by flow cytometry to determine the modal deoxyribonucleic acid (DNA) population. A single G1 peak was considered to represent a unimodal DNA population; two or more G1 peaks indicated a multimodal population. Most of the pituitary tumors and moderately anaplastic astrocytomas had unimodal DNA populations, whereas a higher percentage of other slow-growing tumors, such as meningiomas, ependymomas, and neurilemomas, had multimodal populations (46.2%, 50.0%, and 60.0%, respectively). A relatively high percentage of the rapidly growing or highly malignant brain tumors, including highly anaplastic astrocytomas, glioblastomas multiforme, metastatic tumors, and medulloblastomas, also had multimodal populations (52.9%, 48.7%, 57.1%, and 66.7%, respectively). In most tumor groups, however, the percentage of tumors with a multimodal DNA population did not correlate with the BUdR labeling index or with the percentage of BUdR-labeled S-phase cells. Thus, modal DNA analysis by flow cytometry may provide information about the degree of heterogeneity and the biological behavior of individual brain tumors, but the results do not necessarily correlate with the rate of tumor growth or the prognosis in individual patients.


2021 ◽  
pp. 1-10
Author(s):  
Viktória Temesfői ◽  
Kinga Molnár ◽  
Péter Kaltenecker ◽  
Barbara Réger ◽  
Árpád Szomor ◽  
...  

BACKGROUND: Recent studies proved that metabolic changes in malignant disorders have an impact on protein glycosylation, however, only a few attempts have been made so far to use O-GlcNAc analysis as a prognostic tool. Glucose metabolism is reported to be altered in hematological malignancies thus, we hypothesized that monitoring intracellular O-GlcNAc levels in Rai stage 0-I (Binet A) CLL patients could give deeper insights regarding subtle metabolic changes of progression which are not completely detected by the routine follow-up procedures. OBJECTIVE: In this proof of concept study we established a flow cytometric detection method for the assessment of O-GlcNAcylation as a possible prognostic marker in CLL malignancy which was supported by fluorescence microscopy. METHODS: Healthy volunteers and CLL patients were recruited for this study. Lymphocytes were isolated, fixed and permeabilised by various methods to find the optimal experimental condition for O-GlcNAc detection by flow cytometry. O-GlcNAc levels were measured and compared to lymphocyte count and various blood parameters including plasma glucose level. RESULTS: The protocol we developed includes red blood cell lysis, formalin fixation, 0.1% Tween 20 permeabilisation and employs standardized cell number per sample and unstained controls. We have found significant correlation between O-GlcNAc levels and WBC (R2= 0.8535, p< 0.0029) and lymphocyte count (R2= 0.9225, p< 0.0006) in CLL patients. Interestingly, there was no such correlation in healthy individuals (R2= 0.05664 for O-GlcNAc vs WBC and R2= 0.04379 for O-GlcNAc vs lymphocytes). CONCLUSION: Analyzing O-GlcNAc changes in malignant disorders, specifically in malignant hematologic diseases such as CLL, could be a useful tool to monitor the progression of the disease.


Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1320
Author(s):  
Kristýna Pekárková ◽  
Jakub Soukup ◽  
Marie Kostelanská ◽  
Jan Širc ◽  
Zbyněk Straňák ◽  
...  

Extracellular vesicles (EVs) from liquid biopsies are extensively analyzed by flow cytometry, a technology that is continuously evolving. Thresholding utilizing a violet 405 nm laser side scatter (VSSC) has recently been implemented. Here, we collected set of large EV (lEV) samples from cord blood, which we analyzed using a standard flow cytometer improved via a 405 nm laser side scatter. Samples were analyzed using two distinct thresholding methods—one based on VSSC, and one based on VSSC combined with fluorescence thresholding on stained phosphatidylserine. Through these thresholding methods, we compared lEVs from pre-term births and control cord blood. Double-labeled lEVs with platelet CD36+/CD41+, activated platelet CD41+/CD62P+ and endothelial CD31+/CD105+ antibodies were used. Apart from comparing the two groups together, we also correlated measured lEVs with the thresholding methods. We also correlated the results of this study with data analyzed in our previous study in which we used a conventional 488 nm laser SSC. We did not find any difference between the two cord blood groups. However, we found highly concurrent data via our correlation of the thresholding methods, with correlation coefficients ranging from 0.80 to 0.96 even though the numbers of detected lEVs differed between thresholding methods. In conclusion, our approaches to thresholding provided concurrent data and it seems that improving the cytometer with the use of a VSSC increases its sensitivity, despite not being particularly critical to the validity of flow cytometric studies that compare pathological and physiological conditions in liquid biopsies.


2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Omri Nahor ◽  
Cristina F. Morales-Reyes ◽  
Gianmaria Califano ◽  
Thomas Wichard ◽  
Alexander Golberg ◽  
...  

Abstract Controlling the life cycle of the green macroalga Ulva (Chlorophyta) is essential to maintain its efficient aquaculture. A fundamental shift in cultivation occurs by transforming the thallus cells into gametangia and sporangia (sporulation), with the subsequent release of gametes and zoids. Sporulation occurrence depends on algal age and abiotic stimuli and is controlled by sporulation inhibitors. Thus, quantification of sporulation intensity is critical for identifying the biotic and abiotic factors that influence the transition to reproductive growth. Here, we propose to determine the sporulation index by measuring the number of released gametes using flow cytometry, in proportion to the total number of thallus cells present before the occurrence of the sporulation event. The flow cytometric measurements were validated by manually counting the number of released gametes. We observed a variation in the autofluorescence levels of the gametes which were released from the gametangia. High autofluorescence level correlated to phototactically active behaviour of the gametes. As autofluorescence levels varied between different groups of gametes related to their mobility, flow cytometry can also determine the physiological status of the gametes used as feedstock in seaweed cultivation.


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