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At least 127 records · Page 7Linked to original sources

Flow cytometric DNA analysis of benign hyperfunctioning parathyroid glands: significant difference in the S phase fraction and proliferative index between adenomas and hyperplasias.

AIMS: Flow cytometric DNA analysis was performed to measure the DNA content of benign parathyroid tumours in patients with primary hyperparathyroidism. METHODS: DNA analysis of paraffin-embedded parathyroid samples was performed on 51 parathyroid glands from 29 patients after parathyroidectomy. Histopathology showed parathyroid adenoma in 25 cases and hyperplasia in four patients. DNA ploidy status, DNA index (DI), percentage of cells in S phase and proliferative index (PI) were determined. RESULTS: Normal cells from normal glands were all diploid. DNA cytometry showed 12 aneuploid and 13 diploid adenomas. There were 12 diploid and four aneuploid hyperplastic glands. Incidence of aneuploid DNA histograms did not show a statistically significant difference between adenomas and hyperplasias (P=0.216). Mean S phase fraction was 3.45% in adenomas and 1.53% in hyperplasias (P= 0.015). Mean PI was 6.48% in adenomas and 2.78% in hyperplastic parathyroid glands. This difference was statistically significant (P=0.006). Diploid cases had a mean PI of 4.78% and aneuploid glands a mean PI of 7.7% (P=0.08). Aneuploid DNA content did not reveal statistically significant correlation with age, gender, pre-operative Ca, alkaline phosphatase, i-PTH levels, and tumour size. The mean S phase fraction and PI were 2.25% and 4.78% in diploid glands, and 4.5% and 7.7% in aneuploid cases. CONCLUSION: Aneuploid DNA content may be present in benign parathyroid diseases, but not in normal parathyroid glands. Aneuploid DNA histograms and higher PI occur more often in adenomas compared with hyperplasias, but the nuclear DNA analysis is unable to make a distinction between adenomas and hyperplasias.

Adenoma↗

[Flow cytometry DNA analysis of a gastric cancer before and after intra-arterial anticancer chemotherapy--report of a case].

Flow cytometric DNA analysis has been performed on tumor tissue obtained by endoscopic biopsy before and after intraarterial infusion of ADM and 5Fu. The patient was a 74-year-old woman with a gastric cancer with an involvement of a para-aortic and cervical lymph nodes. Histological examination revealed a poorly differentiated adenocarcinoma, and flow cytometric DNA analysis disclosed that the DNA ploidy (DNA index) did not differ before and after intraarterial infusion of ADM and 5Fu. The fraction of the tumor cells in the S + G2.M phase of the cell cycle however, decreased from 56% to 44% with anticancer chemotherapy. A flow cytometric DNA analysis, using biopsied samples of tumor tissue, was found to be a useful tool for monitoring the effect of anti-cancer chemotherapy for solid tumors.

Adenocarcinoma↗

Prenatal diagnosis of zygosity by fetal DNA analysis, a contribution to the management of multiple pregnancies. A series of 31 cases.

OBJECTIVE: To evaluate the contribution of prenatal diagnosis of zygosity by fetal DNA analysis to the management of multiple pregnancies. METHODS: Between March 1999 and March 2000, 31 same-sex multiple pregnancies, were referred to our Institute during their second trimester to have amniocentesis done. Fetal DNA variants were studied in addition to karyotype. The main indications for amniocentesis were fetal growth discordance, fetal malformations or selective pregnancy termination. Zygosity results were compared to the first-trimester ultrasound diagnosis. RESULTS: 21 out of the 31 multiple pregnancies were dizygous, and 10 out of the 31 pregnancies were monozygous. First-trimester ultrasound provided information on chorionicity only in 24 out of the 31 cases (77%). Of these 24 reports, 21 proved to be correct (87.5%). Moreover, in 2 cases among 14 pregnancies with growth discordance, DNA analysis ruled out a twin-to-twin transfusion syndrome, while ultrasound could not yield a conclusion. In 2 twin pregnancies with fetal malformations affecting one of the fetuses, DNA analysis made it possible to assess the risk for the other twin. In the 5 cases leading to selective termination, prenatal diagnosis of zygosity had a decisive value for the surviving fetus. CONCLUSION: Fetal DNA analysis is a simple and highly useful test for prenatal diagnosis of zygosity in the management of complicated multiple pregnancies.

Amniocentesis↗

Miniaturized pyrosequencer for DNA analysis with capillaries to deliver deoxynucleotides.

As the human genome project proceeds, various types of DNA analysis tools are required for life sciences and medical sciences including DNA diagnostics. For example, a small DNA sequencer for sequencing a short DNA is required for bed-side DNA testing as well as DNA analysis in a small laboratory. Here, a new handy DNA sequencing system (pyrosequencer) based on the detection of inorganic pyrophosphate (PPi) released by polymerase incorporation is demonstrated. The system uses the bioluminescence detection system. The key point for the miniaturized DNA sequencer is to make a deoxynucleotide triphosphate (dNTP) delivery system small and inexpensive. It has been realized by using narrow capillaries to connect a reaction chamber and four dNTP reservoirs. Each dNTP is introduced into the reaction chamber by applying a pressure to the reservoir. Compared with other microdispensers, it is much cheaper and easier. By optimizing the conditions, an excellent sequencing ability is achieved while it is a simple and inexpensive system. In most cases, more than 40 bases can be successfully sequenced. A homopolymeric region, which can not be easily sequenced by a conventional gel-based DNA sequencer, is readily sequenced with this system. The new system is successfully applied to sequence a GC rich region or a region close to a priming region where misreading frequently occurs. A rapid analysis for a short DNA was easily achieved with this small instrument.

Base Sequence↗

Characterization of taeniid cestodes by DNA analysis.

High molecular weight DNA samples free of contaminating proteins and RNA obtained from one isolate (of Guangzhou origin) of Cysticercus tenuicollis and five isolates (of Tianjin, Harbin, Lanzhou, Shenyang and Zhengzhou origin) of Cysticercus cellulosae were subjected to thermal denaturation, restriction endonuclease digestion, Southern blotting and hybridization analysis. C. cellulosae DNA showed a melting temperature (Tm) of 82 degrees C corresponding to a 31% GC content whereas C. tenuicollis DNA melted at 85 degrees C suggesting 38.3% GC content. Visual inspection of ethidium bromide-stained gel showed differences not only between the DNAs of the two species of Taeniid cestodes, but also among the five isolates of C. cellulosae. Furthermore, we used two fragments (1.9kb and 5.5kb) of HindIII-derived restriction fragments of C. cellulosae DNA (Harbin origin) and pTS10 as probes to hybridize the DNAs of the Taeniid cestodes from six origins to detect inter- and intra-species genetic variation, the restriction fragment length polymorphisms (RFLPs) were identified.

Animals↗

Flow cytometric DNA analysis in gynecological oncology.

The relevance of flow cytometric DNA analysis in neoplasia of the female genital tract is reviewed. The virtues and limitations of the technique are discussed. There is good evidence, mainly from retrospective studies, that DNA ploidy and/or the tumor S-phase fraction are valuable prognostic indicators in patients with carcinoma of the ovary and endometrium. Further prospective studies are needed, however, to establish the precise value of flow cytometric DNA analysis before it can be used safely for stratification of therapeutic regimes.

Journal Article↗

Technical and statistical improvements for flow cytometric DNA analysis of paraffin-embedded tissue.

Flow cytometric DNA analysis of paraffin-embedded solid tumors has permitted review of large series of archival tissue in attempts to relate abnormal DNA content to prognosis. Limitations of the technique include: 1) a laborious, time-consuming procedure; 2) variation in technique between laboratories; and 3) lack of an objective method of computing DNA indices. Critical evaluation of our technique has shortened the time involved in dewaxing and rehydration, selectively utilized patient's own normal tissue as the internal standard, proved reproducibility of stored specimens, standardized DNA index computation, and developed a statistical analysis to confirm aneuploidy. These technical improvements and the development of a statistical analysis provide a way to shorten the procedure time and standardize the data generated from flow cytometric DNA analysis so as to improve the quality of retrospective reviews of paraffin-embedded tumors and accelerate the definition of flow cytometry's role as a prognostic indicator.

Breast Neoplasms↗

Value of DNA analysis in addition to cytological testing in the diagnosis of malignant pleural effusions.

BACKGROUND: Aneuploidy appears to be a highly specific marker for cancer, and measurement of cellular DNA content by flow cytometry is rapid and reliable. This study was undertaken to determine if the addition of DNA analysis improved the sensitivity of cytological diagnosis of malignancy in pleural fluid. METHODS: Pleural effusions from 92 patients were studied by cytological examination and flow cytometry. RESULTS: In 41 patients the final diagnosis was malignancy, there were 40 cases of benign effusions including 22 with pleural tuberculosis, and in the remaining 11 patients with biopsy proven cancer the presence of malignant cells was not found by cytological and histological means in the pleural fluid. Aneuploidy and cytological malignancy were found in 14 samples. There were seven cases with abnormal flow cytometry and negative cytological results. In 12 patients the cytological test results were positive but DNA analysis was normal. Thirty six samples of fluid were both diploid and cytologically negative. Of the 22 tuberculous effusions seven contained aneuploid cells. The sensitivity of DNA and cytological analysis was 51.2% and 63.4%, respectively. The specificity of DNA analysis was 74.5%. CONCLUSIONS: DNA analysis of cells in malignant pleural effusions is both less sensitive and specific than the cytological diagnosis. Flow cytometric analysis is not recommended for routine use in the diagnosis of pleural effusions.

Adolescent↗

Flow cytometric DNA analysis combined with fine needle aspiration biopsy in the diagnosis of palpable metastases.

To determine the diagnostic value of flow cytometric (FCM) DNA analysis in combination with fine needle aspiration (FNA) biopsy, the nuclear DNA content was analyzed in FNA specimens from 155 superficial metastases and 60 benign lymph nodes. The results of FCM DNA analysis were considered to be positive for malignancy if DNA aneuploidy was found or if more than 12% S+G2M cells were present in the aspirate in diploid cases. The diagnostic accuracy of FCM DNA analysis alone in detecting malignancy was 92%, with a sensitivity of 91% and a specificity of 95%. FCM suggested the correct diagnosis in 5 of the 7 cytologically false-negative cases and in 9 of the 12 cytologically indeterminate or suspicious cases.

Aneuploidy↗

DNA analysis of ornithine transcarbamylase deficiency.

By analysing the restriction fragment length polymorphism (RFLP) detected by an ornithinetranscarbamylase (OTC) gene specific DNA probe, we followed the segregation of the defective gene in two families with OTC deficiency (X-linked disease). We were able to exclude some female family members as carriers. In one case a doubtful result obtained in a biochemical carrier detection test (by examining the renal orotic acid excretion after a protein load) could be clarified by DNA analysis. In every family with OTC deficiency, carrier detection should be biochemical with additional DNA analysis. Previous results of the biochemical carrier test should be controlled by DNA analysis, especially when "normal" results were obtained.

Amino Acid Metabolism, Inborn Errors↗

Clear cell sarcoma of the kidney--immunohistochemical study and flow cytometric DNA analysis of 7 cases.

Immunohistochemical study and flow cytometric DNA analysis were done on seven cases of clear cell sarcoma of the kidney (CCSK) to speculate its histogenesis and to access the diagnostic usefulness of these methods in the differential diagnosis of Wilm's tumor. Clinically, CCSK is a rare malignant renal tumor of children with a propensity to metastasize to bone. Arborizing vascular pattern surrounding the tumor cells which have clear cytoplasm is characteristic histologic finding. Immunohistochemically, only vimentin was diffusely demonstrated in the tumor cell membrane and cytoplasm. In flow cytometric DNA analysis, four cases showed diploidy and two cases near diploidy. CCSK is a separate disease entity with characteristic clinicopathologic, immunohistochemical and flow cytometric findings in distinction from Wilms' tumor. Considering the histologic and immunohistochemical findings, the possible histogenetic mechanism of CCSK seems to be in common with congenital mesoblastic nephroma (CMN), that is primitive mesenchymal cells which committed early stromagenic activity.

Child, Preschool↗

Prediction of the resistance of human tumors to adriamycin by chemosensitivity tests and DNA analysis of the multidrug resistance gene.

In order to predict natural resistance to Adriamycin (ADM), the amplification of multidrug resistance gene 1 (MDR1) was investigated in 50 human cancer specimens using Southern blot analysis. Genomic DNA was extracted from both human solid tumors and adjacent normal tissues for the analysis. MDR1 gene amplification was not observed in any of the patients tested, including 5 patients in whom ADM was not clinically effective. On the other hand, chemosensitivity tests performed on the tumor cells of these 5 patients indicated resistance to ADM. Our results therefore indicate that MDR1 gene amplification is rarely seen among clinical samples and that conventional chemosensitivity tests might be more useful for the prediction of ADM resistance in cancer patients than the analysis of MDR1 gene amplification.

Blotting, Southern↗

DNA analysis in the diagnosis of infection and in the speciation of nematode parasites.

DNA analysis is playing an increasingly important role in characterising and classifying nematode parasites. Though less emphasis has been placed on utilising DNA elements to study nematodes of veterinary importance, correct diagnosis is, nevertheless, critical to proper treatment and control. The genus Trichinella presents a particularly interesting problem since the level of classification within this genus remains unclear. Herein we discuss the application of DNA analysis to the diagnosis and speciation of the parasitic nematode Trichinella spiralis, where the study of cloned repetitive DNA elements and ribosomal RNA genes has led to significant advances in the understanding of the species-level systematics of this nematode. Despite advances in the recognition of multiple gene pools within the genus Trichinella, their taxonomic level is still uncertain because of insufficient knowledge regarding intraspecific variation within this genus. The utility of mitochondrial DNA and ribosomal DNA sequence analysis in studying the phylogeny and evolutionary history of parasites within this genus is also discussed.

Animals↗

Flow-cytometric DNA analysis of intracranial tumors in children.

The objective of this study was to investigate flow-cytometric DNA values of pediatric intracranial tumors, and to establish DNA analysis as a potential prognostic parameter. Twenty-nine brain tumor specimens from 26 pediatric patients were cryo-preserved within a 3-year period. The DNA content was measured by flow cytometry. Six of the tumor specimens had aneuploid DNA patterns. The median of the proliferation index was lower in the survivor group compared with the non-survivor group (36.4% and 47.5%, respectively). Ten of the 26 patients are still alive, eight were lost to follow up, and eight died. Flow-cytometric DNA analysis may be a helpful tool for examining brain tumors in children. The small size of this study could not establish flow cytometry as a definite prognostic factor, but further prospective multicenter studies will evaluate the prognostic significance of flow-cytometric DNA analysis.

Adolescent↗

Identification of Campylobacter pyloridis isolates by restriction endonuclease DNA analysis.

Campylobacter pyloridis isolates recovered from gastric biopsy specimens of 16 patients were examined by restriction endonuclease DNA analysis with HindIII. For 8 of these 16 patients two different isolates were compared to study the persistence of the colonizing strains and the stability of their DNA digest patterns during a period of 2 years (two patients), the identity or nonidentity of different colony types within one culture (two patients), and the nature of the relapses after apparently successful antibacterial therapy (four patients). The isolates from the 16 patients all produced different DNA digest patterns. Comparison of the two different isolates recovered from the same patients showed that these isolates were identical in all eight cases. Laboratory subculturing of a C. pyloridis strain (10 times) did not change its DNA digest pattern. These results indicate the stability of the DNA digest patterns and a marked variability of these patterns among isolates from different patients. Using restriction endonuclease DNA analysis, we found the persistence in the stomach of the same C. pyloridis strain during a period of 2 years and the identity of different colony types within one culture. The relapses after apparently successful antibacterial treatment could be attributed to recrudescence rather than reinfection. Restriction endonuclease DNA analysis is a sensitive and useful method for identifying C. pyloridis isolates.

Campylobacter↗

[The diagnostic value of flow cytometric DNA analysis of bronchial washing and lavage fluids in lung cancer].

OBJECTIVE: To evaluate the diagnostic value of flow cytometric DNA analysis of bronchial washing and lavage fluids in lung cancer. METHODS: The cellular DNA contents of bronchial washing and lavage fluids from 68 patients (43 with lung cancer and 25 with benign pulmonary disorder) were analyzed by flow cytometry, and the diagnostic value of this method in lung cancer was compared with that of fiberoptic bronchoscopic biopsy and brushing. RESULTS: The presence of aneuploidy was used as a diagnostic criterion, the sensitivity of DNA analysis was 81%, and the specificity was 92%. The positive rate of aneuploidy in central lung cancer (29) was 83%, and there was no statistical difference when compared with biopsy (90%) and brushing (69%). The positive rate of aneuploidy in peripheral lung cancer (14) was 79%, significantly higher than biopsy (29%) (P < 0.025) and brushing (29%) (P < 0.025). In a few patients from both central and peripheral lung cancer groups, biopsy and brushing appeared negative, but aneuploidy was obtained from cytometry. CONCLUSION: These results suggest that flow cytometric DNA analysis of bronchial washing and lavage fluids may be a valuable adjunctive method in the diagnosis of lung cancer, particularly in peripheral lung cancer.

Adenocarcinoma↗

[Flow cytometric DNA analysis of gynecologic malignant tumors using paraffin-embedded tissue].

We have studied flow cytometric DNA analysis of gynecologic malignant tumors using paraffin-embedded tissue for histopathology. A close correlation was observed between the DNA indices obtained from fresh unfixed tissue and from paraffin-embedded tissue (r = 0.993). The coefficient of variation (CV) for the diploid G0G1 peak of the paraffin-embedded tissue was one and half greater than that obtained from fresh tissue on the average. In the DNA analysis of 64 endometrial carcinomas, DNA aneuploidy was detected in 3% of nuclear grade 1, in 25% of nuclear grade 2 and in 63% of nuclear grade 3. That is, the more abnormal the nuclear findings, the higher the percentage of DNA aneuploidy. In the analysis of 48 ovarian tumors, DNA aneuploidy was detected in 79% of serous adenocarcinoma, in no of mucinous adenocarcinoma, in 17% of endometrioid carcinoma, in all of clear cell carcinoma and dysgerminoma, that is, its incidence was different among histological subtypes.

Aneuploidy↗

Central giant cell granulomas of the jaws. Nuclear DNA analysis using image cytometry.

Giant cell nuclear DNA, in 30 giant cell lesions of the jaws, was quantified by computer-assisted image analysis. DNA content was then used to predict clinical behavior and outcome. 4 nuclei in each of 25 giant cells (total = 100 nuclei) were randomly selected and the DNA content was quantified by the Leitz Texture-Analysis-System-Plus. DNA in nuclei of normal appearing stromal fibroblasts (n = 20) was similarly measured. The DNA index was calculated as the mean nuclear DNA content of giant cells divided by the mean DNA content of control fibroblasts. The mean DNA-index of aggressive lesions (1.09, SD = 0.12) was not significantly different from that of non-aggressive lesions (1.18, SD = 0.15) (p = 0.093). The results indicate that the nuclear DNA content of giant cells is not useful as a predictor of the clinical behavior of giant cell lesions of the jaws.

Adolescent↗