bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differences between labeling index and DNA histograms in assessing s‐phase cells from a homogenous group of chronic phase CML patients
Authors: Raza Azra , Bhayana Ranjan , Ucar Kalust , Kirshner Jeffery , Preisler Harvey D. 
Published: August 2005

AbstractThe reliability of DNA histogram analysis in accurately estimating S‐phase cells from human tumors was tested by comparing the results to those of simultaneously obtained tritiated thymidine labeling index (LI) studies. Patients with chronic myelocytic leukemia (CML) during chronic phase were selected for study because the Philadelphia chromosome (Ph) was the only cytogenetic abnormality in each case and, since it is a balanced translocation, the frequently encountered problem of aneuploidy in human neoplastic cells was avoided. Unfortunately, when 30 CML patients were studied simultaneously by DNA histogram analysis and LI studies, the correlation coefficient between the two results was only r = 0.611. A comparison of three different mathematical programs for DNA histogram analysis showed that none was completely satisfactory. We conclude that DNA histogram analysis does not provide the same data as autoradiographically processed labeling index studies even in patients with Ph‐positive CML during the chronic phase when the situation is not complicated by additional aneuploidy.
Flow cytometric analysis of head and neck carcinoma DNA index and S‐fraction from paraffin‐embedded sections: Comparison with malignancy grading
Authors: Johnson Tod S. , Williamson Kenneth D. , Cramer Michael M. , Peters Lester J. 
Published: August 2005

AbstractArchival, paraffin‐embedded, pathology specimens representing pretreatment tissue biopsies from 73 patients with epidermoid carcinoma of the head and neck were analyzed for DNA Index and %S‐phase cells by flow cytometry and were scored for quantitative histomorphology. The DNA fluorescence/light scatter size patterns derived from paraffinembedded specimens were shown to be essentially the same as those from mechanically disaggregated, ethanol‐fixed cells obtained from the same tissue specimen. Patterns ranged from lymphocyte‐like to highly abnormal DNA Index cytokinetic patterns. The DNA Index values ranged from 0.70 to 3.50 (median 1.42), with an aneuploidy frequency of 63/73 (86%). DNA distribution %S ranged from 4% to 45% (mean 19), with the microscopic malignancy grading showing broad heterogeneity (mean 2.1, range 1.0–3.0, where 1.0–1.7 = well differentiated, 1.8–2.3 = moderately differentiated, 2.4–3.0 = poorly differentiated). Cross‐comparison of these data showed that (1) the tumor %S was dependent on DNA Index (higher %S at higher ploidy), (2) low to high malignancy tumors were randomly distributed between diploid/near diploid tumors and high‐degree DNA abnormality tumors, and (3) proliferative activity values broadly overlapped between low to high malignancy scored tumors. However, those carcinomas characterized by high DNA Index (≥ 1.50) and high %S‐phase fractions (≥ 20) had a five fold higher incidence of high‐degree malignancy, invasive tumors than diploid/near diploid (%S ⩽19) tumors.
Droplet Sorting of Large Particles
Authors: Jett James H. , Alexander Robert G. 
Published: August 2005

AbstractThe systematics of droplet formation conditions for orifices with diameters up to 200 μm are described. Sorting recovery experiments indicate that particles up to 44 μm in diameter can be recovered by charged droplet deflection of two drops with at least 75% recovery. By reducing the jet velocity, a deflection of greater than 1 cm was obtained for all droplet sizes.
Characterization of monoclonal antibodies to bromodeoxyuridine
Authors: Vanderlaan Martin , Thomas Cynthia B. 
Published: August 2005

AbstractThe characteristics of three mouse monoclonal antibodies to halogenated uridine derivatives are presented. Two, IU‐1 and IU‐2, are produced by hybridomas derived in our laboratory, and the third is the B‐44 hybridoma described by Gratzner (7) and obtained commercially from Becton‐Dickinson Monoclonal Center. Hybridomas IU‐1 and IU‐2 were derived from the fusion of spleen cells from a Biozzi High Responder mouse immunized with iododeoxyuridine (IdUrd) conjugated to bovine serum albumin and SP2/0 mouse myeloma cells. This paper presents methods and results for enzyme‐linked immunosorbent assays (ELISA) against whole cells labeled with bromodeoxyuridine (BrdUrd), ELISA against BrdUrd‐labeled DNA, and a competition ELISA for free BrdUrd. All three antibodies show similar binding affinities and specificities. The IU antibodies react with BrdUrd and IdUrd when the nucleosides are either free in solution or incorporated into single‐stranded DNA (ss‐DNA). The antibodies do not recognize either halogenated base in doublestranded DNA (ds‐DNA), nor do they react with uracil or bromocytidine. Weak binding to thymidine, 5‐fluorodeoxyuridine, and unsubstituted ss‐DNA occurs.
Cytochemistry for bromodeoxyuridine/DNA analysis: Stoichiometry and sensitivity
Authors: Dolbeare F. , Beisker W. , Pallavicini M. G. , Vanderlaan M. , Gray J. W. 
Published: August 2005

AbstractThis report describes an improved immunochemical procedure for staining cells in suspension for amount of incorporated bromodeoxyuridine (BrdUrd) and total DNA. In this procedure, cellular DNA is partially denatured by extracting the cells with 0. 1 M HCl and then heating them to 80°C in a 50% formamide solution. The cells are then immuno‐fluorescently stained using a monoclonal antibody against BrdUrd in single‐strand DNA (ssDNA) and counterstained for DNA content with propidium iodide (PI), a dye that fluoresces preferentially when bound to double‐strand DNA (dsDNA). We show that the relative amounts of immunofluorescently stained BrdUrd in ssDNA and PI in dsDNA can be altered reciprocally by changing the formamide concentration, denaturation time, and denaturation temperature. We show that this new immunochemical staining procedure allows more complete DNA denaturation so that fivefold lower levels of BrdUrd incorporation can be quantified. In addition, we show that the BrdUrd‐linked immunofluorescence achieved using the new denaturation procedure is more linearly related to cellular BrdUrd content than that achieved after acid DNA denaturation. However, cell loss is sufficiently severe with the thermal denaturation procedure that it may not be applicable to all cell types.
Cell cycle analysis using numerical simulation of bivariate DNA/bromodeoxyuridine distributions
Authors: Yanagisawa Masashi , Dolbeare Frank , Todoroki Takeshi , Gray Joe W. 
Published: August 2005

AbstractThis report describes a mathematical model of cell proliferation for simulation of bivariate DNA/bromodeoxyuridine (BrdUrd) distributions. The model formulates the change with time in the frequency of cells with any DNA content and in the amount of incorporated BrdUrd, according to given cytokinetic parameters, i.e., durations and dispersions of cell cycle phases and DNA synthesis rate during S‐phase. We have applied this model to sequential DNA/BrdUrd distributions measured for Chinese hamster ovary cells asynchronously grown in vitro, 1 for 30 min in 10 μM BrdUrd followed by growth in BrdUrd‐free medium for 0 to 24 h, or (2) during continuous incubation in 3 μM BrdUrd plus 30 μM thymidine for 2 to 24 h. The matches between the experimental and simulated distributions give the G1, S, G2M, and total cell cycle durations (and coefficients of variation) of 5.6h (0.08), 7.0h (0.07), 1.4h (0.16), and 14.0h (0.05), respectively. The model is shown to be useful for quantitative interpretation of the bivariate distributions.
Macrophage‐induced cytostasis: Kinetic analysis of bromodeoxyuridine‐pulsed cells
Authors: Stevenson Anita P. , Crissman Harry A. , Stewart Carleton C. 
Published: August 2005

AbstractThe effect of tumoricidal macrophages on the cell cycle progression of six different cell lines was studied using an anti‐bromodeoxyuridine (BrdUrd) monoclonal antibody to follow the traverse of BrdUrd‐labeled cells. Exponentially growing cultured mammalian cells, from six different cell lines, were prepulsed with BrdUrd before exposure to tumoricidal macrophages. The cultured cells were then analyzed as a function of time for DNA content (by propidium iodide staining) and for BrdUrd incorporation (using a fluoresceinisothiocyanate [FITC]‐conjugated anti‐BrdUrd monoclonal antibody). The position of the cells in cycle and the progression of the BrdUrd‐labeled cohort was followed using flow cytometry. The cell lines examined were: Colon 26, BALB/c‐3T3, ST3T3 (a spontaneously transformed, tumorigenic clone of 3T3), WCHE5 (a clone of whole Chinese hamster embryo cells), RIF (a radiation‐induced fibrosarcoma), and A101D (a human melanoma).The bivariate distributions showed that for all six cell lines the BrdUrd‐labeled cohort in the control cultures progressed around the cell cycle during the first 12h of culture, as the cells exponentially increased. In contrast, when each cell line was incubated with tumoricidal macrophages, the BrdUrd‐labeled cohort did not progress through cell cycle but remained in S phase throughout the 12‐h culture period. There was also no evidence for progression of cells out of G1. The data show that cells were arrested in every phase of cell cycle. This study suggests that cytostasis, as manifested by the termination of progression in all phases of the cell cycle, is a universal phenomenon induced by tumoricidal macrophages.
Phorbol ester enhances deoxynucleoside incorporation while inhibiting proliferation of K‐562 cells
Authors: Colamonici O. R. , Trepel J. B. , Neckers L. M. 
Published: August 2005

AbstractRecently a new technique has been developed to study bromodeoxyuridine incorporation into DNA as a function of the cell cycle phase distribution. We have made the observation that treatment of the hematopoietic cell line K‐562 results in a complete and immediate cessation of cell growth, althoughtritiated thymidine incorporation is markedly elevated for as long as 5 days. We have made use of that newly described technique to study this phenomenon.
Cell cycle kinetic measurements in an irradiated rat rhabdomyosarcoma using a monoclonal antibody to bromodeoxyuridine
Authors: Nüsse Michael , Afzal S. M. Javed , Carr Betsy , Kavanau Kris 
Published: August 2005

AbstractCell cycle kinetics after X‐irradiation were studied in a solid rat rhabdomosarcoma using a monoclonal antibody to bromodeoxyuricline (BrdUrd) In cells in which the DNA was labeled by BrdUrd. It could be shown that this tumor was composed of about 80% diploid host cells, and only 20% of the cells in the dissociated tumor were actually tetraploid tumor cells. When rats were injected intraperitoneally with BrdUrd to label S‐phase cells in the tumor, only a fraction of both types of cells became labeled with Brdtlrd during s‐phase, even 24 h after injection. The diploid BrdUrd‐labeled cells progressed rapidly into cycle; 4 h after injection of BrdUrd, labeled diploid s‐phase cells could be observed. Only 25% of the tetraploid S‐phase cells could be labeled by a single injection of BrdUrd (160 mg/kg body weight). These labeled tetraploid cells progressed through the cell cycle with similar velocities as did labeled diploid cells. Using a “Mini Osmotic Pump” containing bromodeoxycytidine (BrdCyd) at high concentration (0.3 mol/L) that released BrdCyd continuously into the organism where it was converted to BrdUrd, it could be shown that after 2 days about 60% of cells in S‐phase and 70% of cells in S‐phase were labeled. The fraction of labeled S‐phase cells in irradiated tumors (D = 10 and 20 Gy) was enhanced between 10 and 50 h after irradiation clue to a radiation‐induced G2 block in cycling tetraploid tumor cells. No effect of irradiation on incorporation of BrdUrd in S‐phase could be observed in the continuous labeling experiments in contrast to the pulse labeling with BrdUrd, where a small retardation of DNA synthesis was observed in the irradiated tumors during the first 15 h after irradiation. When dissociated cells of the tumor were plated in medium containing 20 μmol/L BrdUrd, only the tetraploid tumor cells were able to incorporate BrdUrd and to enter cycle.
A method to measure the duration of DNA syntheses and the potential doubling time from a single sample
Authors: Begg A. C. , McNally N. J. , Shrieve D. C. , Kärche H. 
Published: August 2005

AbstractA method is described whereby the DNA synthesis time, Ts, call be calculated using data of a single sample of cells taken several hours after labelling with bromodeoxyuridine (BrdUrd). The method involves a simple calculation using flow cytometry data of BrdUrd incorporation (green fluorescence, FITC‐labelled anti‐BrdUrd‐DNA antibody) and total DNA content (red fluorescence, propidium iodide). The movement of BrdUrd‐labelled cells through the S phase can be Ouantified by measuring their mean red fluorescence relative to that of G1 and G2 cells. Assuming the movement of the labelled cells toward G2 is linear with time, T., can be calculated by measuring their relative movement at any one time. The method was tested on cells in vitro and on bone marrow and tumor cells in vivo. Reasonable agreement was seen with published estimates of TS for these tissues.
Cell kinetic studies of in situ human brain tumors with bromodeoxyuridine
Authors: Hoshino Takao , Nagashima Tadashi , Murovic Judith , Levin Ellen M. , Levin Victor A. , Rupp Stephen M. 
Published: August 2005

AbstractAt the time of surgery, 18 patients with various brain tumors were given a 1‐h i.v. infusion of bromodeoxyuridine (BrdUrd), 170–200 mg/m2. At an infusion rate of 200 Mg/M2/h, serum BrdUrd levels of 8 μM were achieved. After the infusion, tumor tissue was obtained and divided into two portions. One portion was fixed in 70% ethanol, embedded in paraffin, and sectioned; the sections were deparaffinized, denatured with 2 N HCl, and reacted with monoclonal antibodies against BrdUrd (anti‐BrdUrd MAb). BrdUrd‐labeled nuclei were demonstrated satisfactorily by an indirect peroxidase method. The other portion was dissociated into single cells with a DNase enzyme cocktail and reacted with FITC‐conjugated anti‐BrdUrd MAb to determine the percentage of BrdUrd‐labeled cells or with chromomycin A3 for DNA analysis. The single‐cell suspensions were analyzed by flow cytometry. The fraction of S‐phase cells in the tissue sections was similar to both the percentage of BrdUrd‐labeled nuclei and the S‐phase fraction determined by flow cytometric analysis. The results obtained with BrdUrd‐labeled nuclei were similar to those obtained from previous autoradiographic studies of various brain tumors exposed to a pulse of 3H‐thymidine. Since BrdUUrd is not radioactive and is nontoxic at the dosage used, these techniques, together with the histopathological diagnosis, may help to predict the biological malignancy of individual tumors.
Double labeling and in vitro versus in vivo incorporation of bromodeoxyuridine in patients with acute nonlymphocytic leukemia
Authors: Raza Azra , Ucar Kalust , Preisler Harvey D. 
Published: August 2005

AbstractA monoclonal antibody against bromodeoxyuridine (BrdUrd) was produced, and a rapid slide technique (RPMB technique) was developed for the estimation of S‐phase cells in a population using this antibody. Bone marrow cells from patients with acute nonlymphocytic leukemia (ANIA) were studied by both the RPMB technique and tritiated thymidine (3HdThd) labeling index studies. The percentage of S‐phase cells obtained by each method was compared in 50 samples, and the correlation coefficient was r = 0.89.A “double label” method is also described in which cells were simultaneously incubated with either BrdUrd and 3HdThd or BrdUrd and tritiated cytosine arabinoside (3HAra‐C). The samples were first processed by the RPMB technique and then by autoradiography. Results showed only black grains overlying the nuclei of fluorescent cells in each group. An automated microphotometer was used to quantitate grains and fluorescence from each cell. This demonstrated an almost direct relationship between grains and fluorescence from BrdUrd + 3HdThd slides, whereas different patterns of relationship were noted from BrdUrd + 3HAra‐C slides of leukemic patients. Their implications are discussed in the text.Finally, intravenous infusions of BrdUrd was given to five leukemic patients. S‐phase cells were recognized distinctly within 5 min of starting the infusion. The percentage of Sphase cells was almost identical from in vivo and in vitro samples. Various possibilities of studying the biological behavior of acute leukemias and analyzing cell cycle characteristics are discussed.
Numerical evaluation of changes in the cytoplasmic microtubule complex of C3H mouse cells by optical diffractometry and of changes in cell shape by fourier analysis
Authors: Mareel Marc M. , De Bruyne Georges K. , Ostrowski Kazimierz , Rozycka Monika , Strojny Pawel , Dziedzic‐Goclawska Anna , Lenczowski Stanislaw , Grzesik Wojcech , Kieler Jorgen 
Published: August 2005

AbstractMo mouse cells in culture on glass were treated with taxol, or nocodazole, or incubated at 4°C to alter their cytoplasmic microtubule complex (CMTC). From each treated group and from an untreated group, 30 cells stained with an antiserum against tubulin, were photographed under the photomicroscope, and negatives were analysed by optical diffractometry. Differences between groups of cells were tested by variance analysis. Phasecontrast micrographs of the same cells were used for Fourier analysis of cell shape. Both types of analyses provided numerical objective data about changes in the CMTC and in cell shape that were typical for the kind of treatment. We conclude that optical diffratometry of immunostained cells and Fourier analysis of cell shape are complementary to photomicroscopy for the study of the CMTC in cell populations cultured on an artificial substrate.
On‐line three‐parameter data uptake, analysis, and display device for flow cytometry and other applications
Authors: Kachel V. , Schneider H. 
Published: August 2005

AbstractMultiparameter flow cytometric measurements are of growing interest in the study of complex features of biological cells. With state of the art instrumentation, three‐parameter (3‐P) data handling is relatively complicated and time consuming and the display methods are not satisfactory.As an alternative, an inteactive 3‐P analyzing module, Cytomic 123 is described, which displays 3‐P fields during and immediately after data uptake in the form of a cubic array of 32,768 channels. The fields can be randomly rotated by hardware and software. The event frequencies is the field are primarily visualized by brightness modulation of the display dots. Additionally, the display of the field may be confined to user selected ranges of event frequencies, which may also be superposed to mixed frequency displays. A set of preprogrammed functions is available for the following tasks: (a) uptake of 3‐P histograms combined with on‐line control of the transducer pulses, (b) automatic uptake of a series of 2‐P time correlated histograms in the cube, (c) generation and numerical evaluation of sections and projections of cube histograms, (d) interactive generation and evaluation of spatial subfields for integration, or as sorting matrix by successive erosion of section planes, or reprojection of projection windows, and (e) isometric display of sections and projections and exchange of data sets with other Cytomic modules or other data systems, especially the Cytomic 12 module, whose 2‐P capabilities can be used.The module is built with low cost Z80 microprocessor eurocards. A standard oscilloscope serves as a display unit.
Flow cytometric analysis of membrane permeability properties influencing intracellular accumulation and efflux of fluorescein
Authors: Prosperi E. , Croce A. C. , Bottiroli G. , Supino R. 
Published: August 2005

AbstractA flow cytometric investigation has been made on the membrane permeability properties that mediate intracellular turnover of fluorogenic substrates. The accumulation and efflux of fluorescein, consequent to the enzymatic turnover of fluorescein diacetate, were assessed in the presence of metabolic inhibitors and after treatment with membrane‐active compounds.The metabolic poisons KCN and rotenone greatly inhibited only the fluorescein efflux, reducing the rate constant to as little as one‐tenth in relation to control cells; in the presence of glucose such inhibition was partially removed.Glucose availability also affected fluorescein efflux: an increase of the rate constant was observed in cells treated with 20 mM glucose, and a decrease was measured in cells incubated for 1 hr in glucose‐free buffer.Membrane‐active compounds Triton X‐100 and hydrocortisone reduced fluorescein accumulation. Hydrocortisone strongly blocked also the efflux; the addition of glucose did not restore the rate significantly.The major evidence of these results is that fluorescein efflux is dependent on membrane integrity and on availability of metabolic energy. Fluorescein accumulation is only partially related to permeability properties regulating FDA uptake, due to the influence that treatments exhibit at the same time on FDA hydrolysis and/or fluorescein release.
Realtime acquisition, storage, and display of correlated three‐parameter flow cytometric data
Authors: Stewart Stephen S. , Price Gerald B. 
Published: August 2005

AbstractA microprocessor‐based system which performs realtime correlated acquisition, storage and display of multiparameter (3‐parameter) data from a flow cytometer (FACS‐III) is presented. List‐mode techniques are not employed.The 3‐parameter data is collected and correlated, then displayed along with cell‐frequency as a realtime 3‐parameter colour scattergram, while the experiment is in progress; in addition, correlated and uncorrelated higher‐resolution projectons of the 3‐parameter data are collected and stored. The data projections may also be displayed: as 1‐parameter histograms, or as 2‐parameter colour or grey‐scale scattergrams. Examples of 2‐and 3‐parameter colour scattergrams are presented.The speed and some characteristics of the realtime acquisition and display software are examined; methods to increase the realtime speed are discussed.
Fluorescence spectra of cells stained with a DNA‐specific dye, measured by flow cytometry
Authors: Steen Harald B. , Stokke Trond 
Published: August 2005

AbstractFluorescence spectra of ethanol‐fixed rat thymocytes stained with the DNA‐specific dye Hoechst 33258 have been measured in an arc lamp‐based flow cytometer including a grating monochromator in front of the fluorescence detector. Spectral resolution was 5–10 nm. Increasing dye concentration was found to yield an increasing shift of the fluorescence spectrum toward longer wavelengths, thus supporting previous work on soluble DNA that indicated several different binding modes of this dye. The results show that similar data may be obtained for all commonly used DNA‐specific dyes. It appears that this type of spectral information may be used to probe the structure of cell chromatin.