bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow‐Cytometric DNA analysis in primary breast carcinomas and clinicopthological correlations
Authors: Ewers Sven‐Börje , Baldetorp Eva Långström , Killander Dick 
Published: August 2005

AbstractFlow‐cytometric DNA analyses of single cell nuclei were performed on nuclear suspensions prepared from biopsies of primary breast carcinomas in 638 patients. Propidium‐iodide‐stained cell nuclei were analysed in an Ortho 50‐H Cytofluorograph. The patients were staged by the TMN classification. Sixty percent of all the patients had aneuploid primary breast carcinomas; of size T1, T2, T3, and T4 tumors, 51%, 63%, 67%, and 75% were aneuploid, respectively. The proportions of aneuploid tumors in each of the tumor stages SI, SIl, SII1, and SIV were 47%, 62%, 67%, and 69%, respectively. This trend to increasing aneuploidy proportions with more advanced disease was significant in contrast to the degree of aneuploidy found in relation to axillary nodal tumor involvement. Multiple aneuploid cell populations were found in 109 (17%) tumors.With a mean follow‐up time of 16 months, 92 patients have relapsed out of 540 completely staged patients with unilateral breast cancer with no distant metastases at the time of initial treatment. When the influence of various treatments and tumor stage are not considered, the recurrence rate was twice as high among patients with aneuploid primary tumors than among patients with euploid tumors. The differences in relapsing rates among patients with euploid and aneuploid primary tumors decreased with more advanced disease. Out of 170 patients with T1 tumors, 17 relapsed and 16 of these were aneuploid. No such difference in relapse rate in relation to ploidy was, however, found in patients with more advanced primary disease.
Cytophotometrical and biochemical characterization of nonpalpable, mammographically detected mammary adenocarcinomas
Authors: Fallenius Anders G. , Skoog Lambert K. , Svane Gunilla E. , Auer Gert U. 
Published: August 2005

AbstractFifty nonpalpable, mammographically detected, invasive breast carcinomas were analysed with respect to DNA distribution pattern, steroid receptor content, and histopathological criteria. No significant histomorphological differences were found as compared to palpable breast carcinomas. In contrast, DNA distributions of palpable and nonpalpable tumors differed. Ninety percent of these relatively small breast carcinomas were found to exhibit nuclear DNA amounts within the diploid and tetraploid regions of normal breast epithelium. In earlier findings in palpable breast carcinomas, 55% are of the diploid‐tetraploid type. The mean cellular content of the estrogen receptor was 1. 0 fmole/μg DNA in this group of mammographically detected carcinomas, which is significantly higher than in routinely detected, ie, larger, breast carcinomas. It is suggested that, despite the histomorphological findings, nonpalpable, mammographically detected breast carcinomas are dominated by biologically highly differentiated, slowly proliferating carcinomas with a favourable prognosis.
Morphologic changes in rat urothelial cells during carcinogenesis: II. Image cytometry
Authors: Young Ian T. , Vanderlaan Martin , Kromhout Lois , Jensen Ronald , Grover Abigail , King Eileen 
Published: August 2005

AbstractImproved early detection of neoplasia by screening of urothelial cells requires an understanding of the features distinguishing normal and neoplastic cell populations. We have begun a program of study based upon a rat model system for the controlled observation of early‐stage lesions produced by the carcinogen N‐butyl‐N‐(4‐hydroxybutyl)‐ nitrosamine. Cells dissociated directly from normal and malignant urothelium were characterized by conventional cytopathology techniques and by quantitative microscopy (for nuclear texture and nuclear and cytoplasmic size, shape, and stain content) to derive a comprehensive picture of bladder tumor development. By following the changes that occur in the dissociated urothelial cells we have found that the nuclear area, total nuclear stain, nuclear shape, and the nuclear chromatin change significantly over a 48‐wk interval as the lesions progress toward malignancy.
Classifying breast carcinomas with DNA measurements and morphometry
Authors: Boon Mathilde E. , Auer Gert U. , van Kaam Han , Schwinghammer Henriette 
Published: August 2005

AbstractThe relation between the DNA distribution pattern and the variables DNA content and nuclear area was investigated in 73 breast carcinomas. According to the DNA patterns, the tumors were subclassified into groups I, II, III, and IV (Auer's classification). The Auer's classes correlate with prognosis. In this investigation it was shown that Auer's classes strongly correlate with the standard deviation of DNA content and mean nuclear area. The carcinomas were subclassified using data of DNA content and nuclear area. For the establishment of this subclassification, multivariate analysis was applied. It was evident that Auer's group III was a hybrid group. Therefore a second classification rule was calculated without the data of Auer's group III. This second classification rule was even more successful in determining the Auer's group of each individual case. Our investigation shows that it is possible to subclassify breast carcinomas objectively using DNA content and nuclear area of the tumor cells.
A new disaggregation device for cytology specimens
Authors: Oud Peter S. , Zahniser David J. , Haag Ditty J. , van Boekel Maria C. G. , Hermkens Huub G. , Herman Chester J. , Vooijs G. Peter 
Published: August 2005

AbstractA new means of disaggregating cytology specimens in suspension using an immersible rotor device is described. The new rotor is compared to an automated syringing apparatus using cervical samples. Similar results using both devices are obtained for both normal and abnormal specimens.
Application of flow cytometry to studies on the distribution, persistence, and regeneration rate of Epstein‐Barr virus receptors
Authors: McCune Bryan , Volsky David J. 
Published: August 2005

AbstractFlow cytometry has been applied to study the persistence and regeneration rate of Epstein‐Barr virus (EBV) receptors and other membrane proteins in normal and tumor cells. EBV receptors were detected in a binding assay utilizing fluorescein isothiocyanate‐conjugated virions (FITC‐EBV). After stripping receptors from the cell surface with trypsin, binding of FITC‐EBV was rapidly regenerated in the lymphoma cell line Loukes, reaching 75% of the control levels by 3 h. Fresh human lymphocytes regenerated receptors at a much slower rate, reaching 50% of control levels by 24 h. The regenerating receptors did not reappear simultaneously on all the cells. Subpopulation of new receptor‐positive cells increased gradually during the incubation at 37°C. Reappearance of receptors was inhibited in the absence of protein synthesis. Conversely, receptors persisted in the nontrypsinized cells in the absence of protein synthesis. These results show that certain parameters of membrane receptor turnover, such as the change in receptor distribution within a cell population, receptor persistance, and regeneration rate, can be measured at the single‐cell level by flow cytometry.
Effects of temperature on the yeast cell cycle analyzed by flow cytometry
Authors: Vanoni Marco , Vai Marina , Frascotti Gianni 
Published: August 2005

AbstractThe effects of temperature (in the range 15–36°C) on growth and the nuclear and budding cycle have been studied in populations of the yeast Saccharomyces cerevisiae exponentially growing in batch on yeast nitrogen base (YNB) glucose medium. The maximal rate of exponential growth is achieved at 30°C, and a transition point is apparent at about 20°C. At all tested temperatures DNA replication begins when cells are still unbudded and both the budded period and the postreplicative period have the same temperature dependence. A temperature compensatory mechanism seems to operate in S phase, during which duration remains relatively constant, in the range 21–36°C, while duration of G2+M phases shows a much more pronounced temperature dependence. The results are discussed in terms of a cell‐cycle model for budding yeast.
Flow karyology of serially cultured chinese hamster cell lineages
Authors: Bartholdi Marty F. , Ray F. Andrew , Cram L. Scott , Kraemer Paul M. 
Published: August 2005

AbstractFlow karyology of serially cultured Chinese hamster cell lineages has been observed to be influenced by the degree of cellular heterogeneity in culture. The minimum coefficient of variation (CV) and debris fraction obtainable vary as a cell lineage evolves from a primary cell culture to an established cell line. The cell lineages pass through a stage of decreased cellular heterogeneity from which flow karyotypes can be obtained with lower CV and debris fraction. The influence of cellular heterogeneity on flow karyology is observed with constant preparative protocol and constant instrument performance, and can be an additional source of variability.
Flow cytometric analysis of immunogold cell surface label
Authors: Böhmer Ralph‐M. , King Nicholas J. C. 
Published: August 2005

AbstractGoat antimouse immunoglobulin antibodies conjugated to colloidal 40‐nm gold particles were used to label mouse spleen lymphocytes. The labeled cells were analysed with a flow cytometer, equipped with an argonion laser and a (0.5 mW) helium‐neon laser. The right‐angle (90°) light scatter signal of the red (632.8 nm) helium‐neon light was enhanced more than tenfold by the gold label. Dual labeling with gold and fluorescein isothiocyanate (FITC) showed no interference between the two labels. Thus immunogold provides a nonfluorescent cell surface label that can be combined with other cell labels for multiparametrical cell analysis.
A proposal for a flow cytometric data file standard
Authors: Murphy Robert F. , Chused Thomas M. 
Published: August 2005

AbstractThe increasing complexity of multiparameter data collection and analysis in flow cytometry and the development of relatively inexpensive arc‐lamp‐based flow cytometers, which increases the probability that laboratories or institutions may have more than one type of instrument, creates a need for sharable analysis programs and for the transport of flow cytometric data files within an installation or from one institution to another. To address this need, we propose a standard file format to be used for all flow cytometric data. The general principles of this proposal are: (1) The data file will contain a minimum of three segments, TEXT, DATA, and ANALYSIS; (2) The TEXT and ANALYSIS segments consist of KEYWORDS, which are the names of data fields, and their values; (3) All TEXT is encoded in ASCII; (4) KEYWORDS and their values may be of any length; (5) Certain KEYWORDS will be standard, i.e., having specified formats to be recognized by all programs. The structure of the DATA segment will be uniquely defined by the values of KEYWORDS in the TEXT area. It may be in any bit resolution, facilitating compatibility between machines with different word length and/or allowing bit compression of the data. The structured nature of the TEXT area should facilitate management of flow cytometric data using existing data base management systems. The proposed file format has been implemented on VAX, PDP‐11, and HP9920 based flow cytometry data acquisition systems.
Application of a high‐resolution TV‐microscope system to estimate the sequence of centromere separation in muntjak chromosomes
Authors: Gerlach B. , Solleder E. , Haucke M. , Harms H. , Schmid M. , Aus H. M. 
Published: August 2005

AbstractThe sequence of centromere separation in muntjak fibroblasts was studied using a high‐resolution TV‐microscope system. Computer programs that identify the chromosomes as well as measure the centromere separation are described. Important prerequisites for this study are the high sampling rate and the digital processing of the two images from the red and green channel of a color TV‐camera. The study demonstrates that the centromeres do not separate in a random sequence and that the separation sequence is not solely controlled by the amount of constitutive heterochromatin in the chromosomes.
Enumeration of cytoplasmic μ immunoglobulin positive acute lymphoblastic leukemia cells by flow cytometry: Comparison with fluorescence microscopy
Authors: Zipf T. F. , Bryant L. D. , Koskowich G. N. , MacGregor S. E. , Chin L. , Johnson H. 
Published: August 2005

AbstractThe pre‐B phenotype of childhood acute lymphoblastic leukemia, in which the μ immunoglobulin molecule is expressed in the cytoplasm of the blast cells, has been shown to have independent prognostic significance. Patients with this phenotype of the disease tend to have a poorer outcome on contemporary treatment regimens than do those with the other B‐precursor pheonotypes. For these clinical studies, the detection of the pre‐B phenotype has depended upon fluorescent‐microscope based techniques. A flow‐cytometry based technique for the detection of the cytoplasmic μ immunoglobulin molecule is presented here and the results compared in detail with those of fluorescent microscopy. The results show that the two techniques are equivalent. The flow cytometry method has the advantage of a standardized control, is less labor intensive, and is observer‐independent.
Proliferative characteristics of primary and metastatic human solid tumors by DNA flow cytometry
Authors: Frankfurt O. S. , Greco W. R. , Slocum H. K. , Arbuck S. G. , Gamarra M. , Pavelic Z. P. , Rustum Y. M. 
Published: August 2005

AbstractThe percentage of cells in S‐phase (S‐index) was calculated from DNA histograms of 453 primary and metastatic human solid tumors (predominantly bladder, breast, colorectal, renal, prostate, ovarian and lung carcinomas, melanomas, and sarcomas). S‐indices varied widely among both primary and metastatic tumors (1–48%); there was no significant difference in S‐indices between primary and metastatic tumors. The S‐indices for aneuploid tumors were signifycantly higher than for diploid tumors. When data for all aneuploid tumors were analyzed collectively, there was no significant relationship between S‐index and DNA ploidy index. However, for colorectal and ovarian carcinomas S‐indices increased, and for lung carcinomas S‐indices decreased with elevation in the degree of DNA ploidy. Lung carcinomas had the highest S‐indices. Comparison of flow cytometry (FCM) and cytology data indicated that for most diploid tumors S‐indices reflect the proportion of S‐phase cells among a mixed population of normal and tumor cells. For most aneuploid tumors, the proportion of tumor cells estimated cytologically was similar to the proportion of aneuploid cells estimated by FCM. For a small proportion of aneuploid tumors a comparison of cytology and FCM data indicated the presence of a predominant diploid tumor stemline and a minor stemline with aneuploid DNA content. There was a wide spread in the values of S‐indices within tumor groups defined by degree of differentiation and stage of disease at surgery.
An easy‐to‐build‐timer for kinetic measurements in flow cytometry
Authors: Beumer Tom , Lenssinck Hans , Pennings Arie , Haanen Clemens 
Published: August 2005

AbstractMeasuring kinetics of any time‐dependent intracellular process on a cell‐to‐cell basis is a powerful extension of flow cytometry (FCM) possibilities. A digital timer is described that is easy to build and that fits with all commercial or custom made multiparameter equipment. Of all possible applications of this timer (phagocytosis kinetics, staining of cell components, enzyme reactions, etc.), one example is given: enzymatic intracellular precipitation owing to myeloperoxidase in human peripheral blood cells.
Alignment and focusing unit for dual‐laser excitation in the fluorescence‐activated cell sorter
Authors: de Josselin de Jong J. E. , Hartwijk P. , Verkerk T. , Jongkind J. F. 
Published: August 2005

AbstractThe two laser beams in a dual‐laser fluorescence‐activated cell sorter FACS‐II can be aligned and focused independently on the sample stream with an additional unit, which can be fitted easily on the optical bench of the FACS. The unit consists of two spherical lenses, which have been mounted in separate holders and can be moved in three directions by way of micrometer gauges. The lenses, which have different focal lengths, have been cut off on one side so each laser beam only passes one lens. The setup has been tested using the flow analysis of a suspension of double‐stained chicken red blood cells. The histograms of both fluorescence signals showed normal distributions with a coefficient of variation of approximately 6%. After willful interference with the adjustments, the laser beams could be readily readjusted within five minutes.
Automatic cell identification and enrichment in lung cancer: V. Adenocarcinoma and large cell undifferentiated carcinoma
Authors: Tyrer Harry W. , Pressman Norman J. , Albright Craig D. , Frost John K. 
Published: August 2005

AbstractThe aims of this study were to develop a protocol for the identification and enrichment of cancer cells from sputum obtained from patients with adenocarcinoma of the lung (n = 6) and large‐cell undifferentiated carcinoma of the lung (n = 2), and to compare these findings with the results from our previous studies on other cell types from lung cancer. The hypotheses tested were: (1) Cancer cells in sputum can be preserved following flow sorting. (2) Enrichment for cancer cells from acridine orange (AO)‐stained specimens can be achieved. (3) Discrimination of cancer cells from noncancer cells is by AO green fluorescence and discrimination of lymphocytes from other cell types is by AO red fluorescence. (4) Cancer cells are consistently enriched in the AO high green and red fluorescence region, although, for a given cell type, maximal enrichment is patient‐dependent. (5) Finally, cancer cell enrichment and lymphocyte exclusion can be done simultaneously.Cells from sputum were initially fixed, stained with AO, sorted on a dual parameter flow sorter, and classified into six groups corresponding to two ranges of green and three ranges of red fluorescence intensities. Cells of each region were stained by the method of Papanicolaou and differential counts were performed to determine the relative frequencies (i.e., purities) of leukocytes, macrophages and unsorted (i.e., control) samples.The average purity of leukocytes (81%), macrophages (6%), squamous cells (11%), and cancer cells (2%) varied markedly from sample to sample. However, the largest enrichment values (i.e., ratio of purity of a cell type in a sorted sample to its purity in the unsorted control sample) achieved for cancer cells consistently occurred for each patient sample in the region corresponding to high green and high red fluorescence intensities. Experimentally, a cancer cell average enrichment of sixteen‐fold was obtained by this method. Additionally, fluorescence intensity ranges which increased the enrichment for macrophages by cell sorting typically excluded leukocytes and squamous cells, and vice versa. Finally, red fluorescence intensity was the primary discriminatory parameter for all cell types studied, although the additional use of green fluorescence intensity significantly increased cancer cell enrichment rates.In conclusion, sputum samples can be enriched for atypical and malignant cells, cells from these samples can be well preserved following machine processing, and sputum samples from patients with adenocarcinoma of the lung and large‐cell undifferentiated carcinoma have similar enrichment and preservation results to samples obtained from other types of lung cancer.
Cell‐cycle, protein content, and nuclear size in acute myeloid leukemia
Authors: Ffrench M. , Bryon P. A. , Fiere D. , Vu Van H. , Gentilhomme O. , Adeleine P. , Viala J. J. 
Published: August 2005

AbstractSimultaneous analysis of DNA and cellular proteins provides information on cell proliferation and metabolism. Cellular protein content coupled with nuclear geometric parameters can be used to evaluate cellular maturation and differentiation. In this study, leucoblasts from 50 cases of adult acute myeloid leukemia were analyzed by flow cytometry, and semiautomatic morphometry was performed on bone marrow smears. Ethanol‐fixed bone marrow blast cells were stained for DNA with propidium iodide (PI) and for proteins with fluorescein isothiocyanate (FITC). On the resulting FITC versus PI histograms we defined the cells with low protein content which are associated with a nonproliferating subpopulation (LPC fraction). Low protein content fraction and S‐phase are correlated (p < 0.01). The LPC fraction values are more dispersed than S‐phase values and thus should indicate more clearly eventual differences between cellular populations. This hypothesis has been tested with the prognostic significance of cell‐cycle variables: The LPC fraction was significantly higher in the complete remission group than in the other (p < 0.01), while S‐phase did not show any difference.The peak value of the protein content histograms is significantly lower in the granulocytic leukemias (M1, M2, M3) than in the leukemias with a monoblastic component (M4, M5). Furthermore, we showed that the differentiation and the maturation of the myeloid blast cells modify the nuclear size. The combination of these two parameters provides useful information for cytological classification.
A single‐parameter analog level discriminator: A cheap and powerful extension of FCM data acquisition
Authors: Beumer Tom , Lenssinck Hans , Pennings Arie , Haanen Clemens 
Published: August 2005

AbstractModern flow cytometers offer the possibility to measure more parameters of individual cells than the normally available data acquisition equipment can store. To enlarge the number of parameters that is taken into account while recording data, an analog level discriminator is described that is easy to build and easy to implement on any existing FCM system. An example of gated data acquisition in light scattering measurements on nucleated human bone marrow cells in given.The level discriminator may give a cheap and yet valuable extension of expensive data acquisition equipment.
Flow cytometry of leukocytes after intravenous fluorescein angiography
Authors: Felberg Norman T. , Haimowitz Anna J. 
Published: August 2005

AbstractBlood specimens were obtained for T lymphocyte analysis from patients undergoing intravenous fluorescein angiography (IVFA). Mononuclear cells were prepared by differential density centrifugation and analyzed by flow cytometry (FCM). Prior to IVFA, there was very little autofluorescence in the mononuclear cell population. After IVFA, there was an increase in the fluorescence of the mononuclear cells. Though barely visible by fluorescence microscopy, the fluorescence was uniformly distributed throughout the cytoplasm. By FCM, fluorescein was detected in 27% of the lymphocytes, 61% of the monocytes, and 75% of the granulocytes.Care must be taken when interpreting flow‐cytometric lymphocyte subset data because of the increased nonspecific fluorescence of mononuclear cells in patients who have undergone IVFA.
A comparison of different focus functions for use in autofocus algorithms
Authors: Groen Frans C. A. , Young Ian T. , Ligthart Guido 
Published: August 2005

AbstractA number of functions for the autofocusing of microscopes and other optical instruments are to be found in the literature. In this article we compare 11 of them to determine, in an objective manner, which functions are most suitable for implementation with real‐time video acquisition systems. Three different images, each representing a typical class of image, are used in the comparison. Among the best focus functions found in our study are the squared magnitude gradient, the squared Laplacian, and the normalized image standard deviation.