DNA and nuclear protein measurement in isolated nuclei of human endometrium

Oud, Peter S.; Reubsaet‐Veldhuizen, José A. M.; Henderik, J. B. J.; Pahlplatz, M. M. M.; Hermkens, H. G.; Tas, Johan; James, J.; Vooijs, G. P.

doi:10.1002/cyto.990070405

Cited by 11 publications

(9 citation statements)

References 30 publications

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“…The clinically more advanced carcinomas do not show an increased proliferative activity compared to that of the other carcinomas. These results closely agree with those found in our previous study (11).…”

Section: Resultssupporting

confidence: 94%

“…Also, within the malignant population, no indication could be found that suggested a correlation with the histological grade or clinical stage. These results are somewhat different from those obtained in o w previous study (11). In that study, a complete overlap of the nuclear protein content of normal and malignant endometrium was also found.…”

Section: Iiiscussioncontrasting

confidence: 99%

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DNA and nuclear protein measurement in columnar epithelial cells of human endometrium

et al. 1986

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Propidium iodide DNA flow cytometry, Feulgen-DNA, and nuclear light green protein scanning cytometry were performed in columnar epithelial cells of normal, nonmalignant human endometrium and endometrial adenocarcinomas. Columnar cells were identified by immunohistochemical staining for cytokeratin 18, an intermediate filament protein specifically present in columnar cell epithelium. DNA measurements derived from flow and scanning cytometry showed comparable results. The DNA content of the GO/Gl fraction of the adenocarcinomas had a considerable overlap with that of normal endometrium, with that of the carcinomas shifted toward higher values. For the carcinomas, no correlation was found with the histological grade, with the exception of the adenosquamous carcinomas. Most of the clinical stage I tumors showed a DNA content in the normal diploid region. Three of the four carcinomas of clinical stage I1 and higher had an increased DNA content.For the carcinomas, the percentage of cells in the proliferative fraction, as determined In a preceding paper (111, we compared DNA and nuclear protein values in normal endometrium and endometrial adenocarcinomas to study the changes herein during neoplastic transformation. The results showed that the carcinoma cells had the same or a n increased DNA content compared with normal endometrium, a large variation in the percentage of cells in the proliferative fraction, and a comparable amount of nuclear protein. These measurements were performed in isolated nuclei obtained from homogenates of normal endometrium and endometrial adenocarcinomas. Normal endofrom scanning cytometric derived DNA histograms, was comparable to that of normal endometrium, or higher. No correlation was found with the histological grade. Tumors of clinical stage I1 and higher had intermediate values compared to carcinomas of lower stages.The nuclear protein/DNA ratio of malignant endometrium completely overlapped that of normal endometrium. Within the tumor poy;,ulation, no correlation was found with the hisitological grade, with the exception of the adenosquamous carcinomas, and clinical stage.Based on the aforementioned parameters, no discrimination could be obtained between normal and malignant endometrium. However, when the DNA content of the GO/G1 fraction was combined with the coefficient of variation of the nuclear protein/UNA ratio, a clear discrimination could be obtained with only two false-positive cases.Key terms: Adenocarcinomas, DNA cytometry, nuclear protein cytometry, cytokeratin metrium and endometrial adenocarcinomas are composed of several tissue types, however. Normal endometrium contains a varying number of glands, dependent. on the period of the menstrual cycle, which are lined up by columnar epithelial cells. The glands are Address reprint requests to Peter S. Oud,

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Section: Resultssupporting

confidence: 94%

Section: Iiiscussioncontrasting

confidence: 99%

DNA and nuclear protein measurement in columnar epithelial cells of human endometrium

et al. 1986

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“…In a previous study (14) using image cytometry, we found that with features deduced from the total amount of DNA (integrated optical density of the DNA dye) some differences among endometrial stages could be detected. However, a considerable overlap was still noted.…”

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confidence: 85%

“…Nuclear isolation and deposition of the nuclei on slides have been extensively described elsewhere (14). In brief, nuclei were prepared by homogenization of the defrosted tissue and fixed in suspension in a methanol:37% forma1dehyde:acetic acid mixture (85:10:5 by volume).…”

Section: Nuclear Isolation and Stainingmentioning

confidence: 99%

Evaluation of DNA and nuclear protein features for their use in describing normal and malignant endometrium

et al. 1987

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Different feature sets (geometric, densitometric, and textural) derived from DNA and nuclear protein staining were evaluated for their use in describing atrophic, secretory, and proliferative endometrium, and well-differentiated stage I and moderately differentiated stage I adenocarcinomas of the endometrium. It was found that the pattern of significant differences among these groups varied between feature sets, while remaining consistent within a set of features. The DNA density and run-length features were not very effective in describing group mean differences, whereas the co-occurrence features revealed significant differences among most groups. The protein run-length features were the only ones that consistently showed a difference between proliferative endometrium and well-differentiated adenocarcinomas. Analyses repeated on only cells in the GO/Gl DNA region improved the differentiation between moderately differentiated adenocarcinomas and the other groups. It was concluded that the use of DNA and nuclear protein texture features are effective in describing group differences that cannot be described by DNA content only.

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“…The use of tissue grinders with tight‐fitting pestles and cold sucrose solutions have a long history of use in the isolation of intact nuclei from rat tissues such as liver (Schneider, 1948; Wilbur & Anderson, 1951) and skeletal muscle (Edelman et al , 1965). Although not mentioned in protocols of sample preparation for FCM DNA analysis (Vindeløv et al , 1983; Petersen, 1985; Ormerod, 2000; Gomez et al , 2001; El‐Naggar & Vielh, 2004; Heinlein et al , 2010), the use of tissue grinders has been reported for DNA analysis of human tumours and cell lines in a few studies (Piwnicka et al , 1983; Oud et al , 1986; Remvikos et al , 1988). High‐density sucrose solution for tissue dissociation reportedly aids to split gap and tight junctions responsible for cell clusters while inclusion of sodium citrate aids to loosen intercellular bonds by sequestrating cations (Ca 2+ and Mg 2+ ) responsible for cell surface and intercellular structural matrix integrity (Pallavicini, 1987).…”

Section: Discussionmentioning

confidence: 99%

A robust flow‐cytometric protocol for assessing growth rate of hatchery‐reared barramundiLates calcariferlarvae

Domingos

Fromm

Smith-Keune

et al. 2012

Journal of Fish Biology

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In this study, a flow-cytometric cell cycle analysis method to assess instantaneous growth rate of whole larvae of the Australian barramundi Lates calcarifer was developed and validated. High-resolution DNA measurements of either fresh, frozen or RNAlater-preserved larvae (gap0-gap1, G(0) -G(1), coefficient of variation (c.v.) < 3, 4 and 5%, respectively) enabled the deconvolution of the DNA histogram and assignment of the proportion of nuclei into cell cycle compartments G(0) -G(1), S (DNA synthesis) and G(2) -M (Gap2-Mitosis). This technique can be also used for individual fish tissues such as brain, liver, fin and muscle. For the first time, the combined proportion of replicating nuclei (into S and G(2) -M phases) of whole fish larvae and absolute growth rate in length (mm day(-1)) has been correlated in commercial aquaculture conditions. Fast growing L. calcarifer larvae had an overall hyperplasia advantage as indicated by a greater proportion of cells in the S+G(2) -M phase compared with slow growing larvae, which might explain the increasing differences in size during culture. In a fasting trial, larvae ceased growth while maintaining the constant initial rates of cell division throughout a 6 day period. For a highly fed fast growing control group, cell division rates significantly increased after day 4. Flow-cytometric cell cycle analysis of whole fish larvae may provide fish biologists and aquaculturists with a better understanding of how cell division rates influence early growth in natural and artificial environments.

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DNA and nuclear protein measurement in isolated nuclei of human endometrium

Cited by 11 publications

References 30 publications

DNA and nuclear protein measurement in columnar epithelial cells of human endometrium

DNA and nuclear protein measurement in columnar epithelial cells of human endometrium

Evaluation of DNA and nuclear protein features for their use in describing normal and malignant endometrium

A robust flow‐cytometric protocol for assessing growth rate of hatchery‐reared barramundiLates calcariferlarvae

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