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At least 73 records · Page 4Linked to original sources
PubMed1998

[Lecture of principles and applications of capillary electrophoresis (VI) Applications of CE in DNA analysis, carbohydrate analysis and capillary ion analysis].

A review with 87 references on the application and new development of CE in DNA analysis, carbohydrate analysis and capillary ion analysis is presented. The DNA analysis included determinations of nucleotide, nucleoside and bases by CZE and MECC. It also includes dsDNA, PCR products analysis, purity control of oligonucleotide and DNA sequencing by CGE. Sometimes LIF detector needs to be introduced into DNA analysis. The carbohydrate analysis includes the methods for chemical derivatization and direct determination of carbohydrates by different CE modes. The capillary ion analysis includes the principle and determination of anions and cations.

Animals↗
PubMed1989

[Generation of monoclonal anti-dermatan sulfate antibodies cross-reacting to calf thymus DNA. Analysis of "anti-DNA antibody" directly binding to dermatan sulfate].

Monoclonal anti-dermatan sulfate antibodies were produced by immunizing BALB/c mouse with dermatan sulfate (DS) derived from porcine skin. These antibodies were tested for cross-reactivity to double stranded calf thymus DNA(ds.DNA) and heat-denatured DNA (ss.DNA). Five clones (including 2H8 and 3C4 clones) had binding activities to DS, ds.DNA and ss.DNA in solid phase enzyme linked immunosorbent assay, but no clones binding to DS alone were produced. 2H8 and 3C4 clones could activate mouse complement when they bound to the DS and ss.DNA. The abilities of DS, ds.DNA and ss.DNA to inhibit binding of 2H8 and 3C4 clones to solid phased DS, ds.DNA and ss.DNA were examined. Their binding activities were inhibited by any of DS, ds.DNA and ss.DNA. DNase I treatment abolished the binding activities of 2H8 and 3C4 clones to ds.DNA and ss.DNA completely, but had no effect on the binding activities to Ds. On the other hand, chondroitinase ABC treatment of DS augmented the binding activities of 2H8 and 3C4 to DS. DS digested completely with chondroitinase ABC inhibited the binding activities to DS of 2H8 and 3C4 clones, but not any of delta Di-4S and delta Di-diSB produced from DS digested with this enzyme inhibited the binding activities. Therefore, it was suggested that the epitope recognized by 2H8 and 3C4 clones could not be in delta Di-4S or delta Di-diSB but in the linkage regions of DS.

Animals↗
PubMed2003

[DNA analysis in gestational trophoblastic disease].

OBJECTIVE: DNA analysis of different forms of gestational trophoblastic disease. DESIGN: Retrospective clinical study. SETTING: Slovak Center of Trophoblastic Disease, Bratislava, Slovak Republic. METHODS: In the period of September 1993 to April 2003, eighty-nine cases of gestational trophoblastic disease were analysed. There were 22 cases of partial hydatidiform moles, 58 cases of complete hydatidiform mole, 5 cases of invasive mole and 4 cases of gestational choriocarcinomas. Southern hybridization and polymerase chain reaction were used for DNA analysis. RESULTS: From 22 analyzed cases of partial hydatidiform moles 19 (86.4%) were triploid and 3 (13.6%) diploid ones. There were 58 cases of complete hydatidiform mole and out of them 29 (50%) were homozygous, 28 (48.3%) heterozygous, and in one case (1.7%) both paternal and maternal genome was detected. In 8 cases of heterozygous and in one case of homozygous complete hydatidiform mole occurred a malignant transformation to gestational choriocarcinoma. CONCLUSIONS: Molecular analysis can determine the nuclear DNA origin of complete hydatidiform mole and allow us to define the patients with higher risk of malignant transformation usually to gestational choriocarcinoma.

DNA, Neoplasm↗
PubMed1994

Comparative DNA analysis of breast cancer by flow cytometry and image analysis.

Measurement of DNA ploidy can be performed either with Flow Cytometry (FCM) and Image-Analysis (IA); both methods provide prognostic information in primary breast cancer. We compared the results of quantitative DNA analysis of formalin fixed, paraffin embedded tissue from 62 invasive ductal breast cancers. For FCM nuclear suspensions from disaggregated tumor were stained with Propidium Iodide and analyzed by means of Ortho Cytoron Absolute. For IA nuclear suspensions were stained by the Feulgen method and analyzed by means of Vidas system. We found a good correlation between flow cytometry DNA Index and Histogram Type, according to Auer classification (rs = 0.65, p < 0.001) and between DNA Index and Grading of Malignancy (MG) which had been measured by Bocking's algorithm (rs = 0.38, p < 0.05). Concerning to disease free survival (DFS), flow cytometric DNA Index showed a better correlation (rs = 0.56, p < 0.001). We concluded that the two methods provide comparable results, but offer individual advantages and are complementary for analyzing DNA ploidy in breast cancer.

Algorithms↗
PubMed1989

Prenatal diagnosis and heterozygote detection by DNA analysis in ornithine transcarbamylase deficiency.

This report summarizes our experience with DNA analysis using a complementary DNA probe for ornithine transcarbamylase in 24 individuals or families with deficiency of this enzyme. In four cases, including three reported elsewhere, a Taql restriction site alteration directly detected the mutation. In 10 additional cases, only an affected male was available, and results of DNA analysis using the Taql enzyme were normal. In 10 cases, family studies were performed with the use of restriction fragment length polymorphisms. Prenatal diagnostic studies were performed for three informative pregnancies, and two affected male fetuses were identified. Analysis of two restriction fragment length polymorphisms, Mspla and BamHl, was informative in 14 of 19 (74%) known carrier females and in 21 of 35 (60%) females (the total number studied). One female previously predicted to be a noncarrier by protein-loading test was determined to be a carrier by analysis of restriction fragment length polymorphisms. The frequency of Taql site alterations was 4 of 24 families (17%). These data illustrate the importance of DNA analysis, pedigree analysis, and biochemical testing in families with ornithine transcarbamylase deficiency to detect carriers and establish the diagnosis prenatally.

Blotting, Southern↗
PubMed1992

An image cytometric DNA-analysis in breast neoplasms. Parameters of DNA-aneuploidy and their relationship with conventional prognostic factors.

In this prospective study, an image cytometric DNA-analysis was performed in 86 women with breast neoplasms (72 primary invasive carcinomas and 14 benign lesions). Four DNA ploidy parameters were analysed: histogram type (according to AUER classification), DNA-index, tumor cells with DNA content above the 5n limit and DNA malignancy grade (DNA-MG, calculation according to Böcking). Their correlations with well established prognostic factors in breast carcinomas (tumor size, lymph node status, histologic grade, hormone receptor content) were studied. All but one benign lesions were diploid (13/14 cases), whereas the majority of the primary invasive breast carcinomas were aneuploid (58/72 cases). A predominance of carcinomas with a percentage of cells superior or equal to 1% with DNA content above the 5n limit was observed (54 cases out of 58). Most of the aneuploid tumors had a histogram type III or IV (53 cases) or a high DNA-index (50 cases). Of these 58 aneuploid cases, only 26 tumors had a DNA-MG superior to 1. Interestingly, 26 tumors had the 4 criteria of aneuploidy, 19 had 3 and 9 had 2 and only 4 tumors had one parameter. The DNA-MG was significantly related to hormonal receptors (p less than 0.001) and tumor size (p less than 0.01). The histogram types (Auer classification) and the DNA content above the 5n limit were correlated with histologic grade (SBR or SBRM) (p less than 0.02). Concerning the DNA-index no correlation was observed with well established prognostic factors. On the other hand no significant correlation was found between these new biologic variables and lymph node status.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1990

DNA analysis and prognosis of digestive tract cancers.

Cytophotometric DNA analysis was performed on esophageal and gastric carcinomas. In 35 cases of mucosal and submucosal carcinoma of the esophagus, patients with types I and II (relatively regular in DNA distribution) had an uneventful postoperative course and no recurrence, whereas 3 of 15 (20%), and 5 of 9 (55.6%) with type III and type IV, respectively (widely scattered DNA distribution, died following a recurrence. Cytophotometric DNA analysis using biopsy specimens from 75 patients with esophageal cancer in various stages also showed a close relationship between the DNA distribution pattern and prognosis. However, the growth mode and the DNA ploidy of mucosal gastric cancer correlated well in the investigation of 66 cases. Thus, data of this method closely reflected the outcome in patients with digestive tract cancers. These results suggested the potential usefulness of cytophotometric DNA analysis for assessing the prognosis, even in the early stage of cancers.

Adult↗
PubMed2003

Effects of environmental conditions to which nails are exposed on DNA analysis of them.

To investigate the feasibility of DNA analysis of nails taken from human remains after environmental exposure, we tested the quantity and quality of DNA that could be recovered from nails exposed to various conditions. We first investigated whether there is an inter- or intra-subject difference in the DNA content of nails using nails obtained from 333 volunteers. DNA content did not significantly differ between males and females, or between young and elderly groups. In addition, no statistically significant differences were observed among nail samples obtained from each finger of the same person. Then thumbnails of five volunteers were left in dried soil, wet soil, river water, sea water, distilled water or air for 3 months. Each nail sample was tested monthly for sex chromosome-specific repeats, ABO genotype, STRs (TH01, TPOX, CSF1PO, FES and vWA loci), and D1S80 locus. These markers were correctly detected from nails after 1 month, irrespective of environmental conditions. Thereafter, the detection rates were decreased to various degrees, except for nails left in air. The detection of longer DNA markers tended to be more difficult than the detection of shorter markers. Nails were more fragile in wet soil than in any other condition. However, nonspecific PCR products were not observed in any nail sample. Our results reconfirmed the usefulness of the nail as a specimen for forensic DNA analysis.

Adolescent↗
PubMed1991

[DNA analysis of breast cancer].

Several methods for DNA analysis including DNA histogram and proto-oncogene amplification in human breast cancer, and the results recently reported were reviewed. A large number of DNA histograms obtained by flow cytometry have provided a possible correlation between DNA ploidy pattern and clinical outcome of breast cancer patients. Poor clinicopathologic factors, however, are not always in association with DNA aneuploidy and/or S-phase fraction rate, suggesting that further investigations on DNA ploidy are required. Amplification and/or over expression of proto-oncogenes in human breast cancers have been reported. Among them c-erbB-2 amplification may be one of the candidates for prognostic indicators of breast cancer survival. To determine any reliable biological factors exist further analysis of tumor DNA will be required in breast cancer research.

Breast Neoplasms↗
PubMed2004

Image cytometry DNA-analysis of fine needle aspiration cytology to aid cytomorphology in the distinction of branchial cleft cyst from cystic metastasis of squamous cell carcinoma: a prospective study.

OBJECTIVE: Frequently, the distinction between branchial cleft cyst and cystic metastases from squamous cell carcinoma is difficult by cytomorphology. In a prospective study, we investigated the need for, and the value of, image cytometry DNA-analysis as a complement to cytologic evaluation of cystic lesions in the neck. STUDY DESIGN: Image cytometry DNA-analysis was performed on the fine needle aspiration cytology smears from 50 patients, referred to our department, with a solitary cystic lesion in the lateral region of the neck. METHODS: Smears from aspirates were Giemsa stained and cytologically evaluated. Ahrens image analysis was used for DNA analysis on smears stained with Schiff reagent, and lymphocytes were used as control cells. Epithelial cells with DNA values exceeding 5c were regarded as aneuploid, indicating malignancy. RESULTS: Nine lesions were diagnosed as squamous cell cancer metastases cytologically. DNA analysis showed aneuploidy in all of them except one. Three of these lesions had earlier been diagnosed as branchial cleft cyst at the referring hospital. Eight lesions were cytologically inconclusive and four of them were revealed as cystic metastasis at histopathologic analysis, and DNA analysis showed aneuploidy in all but one, which could not be analyzed. Two of these lesions were also diagnosed as branchial cleft cysts at the referring hospital. All benign lesions were diploid. Nine lesions were thyroid and salivary gland lesions. CONCLUSION: Image cytometry DNA-analysis was shown to help in the distinction between benign and malignant cystic lesions. Thus, when conventional cytomorphology does not suffice, DNA-analysis is clearly a valuable supplement.

Adolescent↗
PubMed1980

Clonal heterogeneity of small-cell anaplastic carcinoma of the lung demonstrated by flow-cytometric DNA analysis.

Flow-cytometric DNA analysis yields information on ploidy and proliferative characteristics of a cell population. The analysis was implemented on small-cell anaplastic carcinoma of the lung using a rapid detergent technique for the preparation of fine-needle aspirates for DNA determination and a newly developed procedure for storing aspirates at -80 degrees. Thirty-eight different metastases in 30 consecutive patients with small-cell anaplastic carcinoma of the lung were examined with a total of 273 fine-needle aspirations. The results on ploidy are reported in this paper. The degree of contamination of the aspirates with normal cells was determined by differential counts. The ratio of the peak channel numbers for the G1 phase of the tumor cells to that of the diploid standard (DNA index) was calculated and used for ploidy identification. Twenty-nine patients were evaluable with respect to DNA index determination. The coefficient of variation of the DNA index determinations was estimated as 0.039. In 23 (79%) patients, only one cell line could be detected. Evidence of the presence of 2 tumor cell clones with different ploidy was obtained in the remaining 6 (21%) patients. Of the 35 malignant clones thus demonstrated, 26 (74%) were significantly different from diploid (p less than or equal to 0.01). Four (11%) were hypodiploid, 3 (9%) were hypotriploid, and 19 (54%) were hypo- or near-tetraploid. Clonal heterogeneity in the tumors of 21% of the patients is a conservative estimate. Assessment of the detection limit set by the methodology used and the restricted number of samples studied in each patient indicate that the true occurrence of clonal heterogeneity in small-cell carcinoma of the lung may be much higher.

Aged↗
PubMed1998

Nuclear multiparameter flow cytometric DNA analysis of human brain tumors: correlation of DNA content with tumor histology and clinical behavior.

BACKGROUND: Using fresh surgical specimens of brain tumors to investigate cellular DNA content is uncommon. The aim of this study is to elucidate the relationship among cellular nuclear DNA content, tumor histology, and tumor clinical behavior of various tumors of the central nervous system. METHODS: Multiparameter flow cytometry (FACStar, Becton-Dickinson, San Jose, CA) was randomly applied to analyze 40 fresh surgical specimens from brain tumors. Histologically, there were 19 cases of benign tumors and 21 cases of malignant tumors. RESULTS: DNA distribution profiles in the benign tumors revealed 15 cases of diploidy and four cases of aneuploidy, while the malignant tumors had eight cases of diploidy and 13 cases of aneuploidy (p < 0.01). The mean DNA index (DI) in benign tumors was 1.13 +/- 0.27 and in malignant tumors 1.43 +/- 0.48 (p < 0.05). The mean proliferative index (PI) in benign tumors was 10.27 +/- 5.46% and 18.44 +/- 8.98% in malignant tumors (p < 0.05). Clinically, 10 patients had postoperative recurrence. The PI in recurrent and nonrecurrent tumors was 17.83 +/- 11.13% and 13.20 +/- 7.19%, respectively (p < 0.05). The 1-year cumulative survival rate in benign tumors was 78.9%, and 57.1% in malignant tumors (p < 0.05). CONCLUSIONS: Malignant tumors had a significantly higher incidence of aneuploid DNA histograms, a higher DNA index, and a higher proliferative index. Tumors with a high PI had a higher incidence of recurrence and a lower survival rate. Multiparameter DNA analysis by flow cytometry provides a valuable diagnostic aid for the histopathologic differentiation of human brain tumors.

Adolescent↗
PubMed2000

Specific detection of Leuconostoc mesenteroides subsp. mesenteroides with DNA primers identified by randomly amplified polymorphic DNA analysis.

Randomly amplified polymorphic DNA analysis using primer 239 (5' CTGAAGCGGA 3') was performed to characterize Leuconostoc sp. strains. All the strains of Leuconostoc mesenteroides subsp. mesenteroides (with the exception of two strains), two strains formerly identified as L. gelidum, and one strain of Leuconostoc showed a common band at about 1.1 kb. This DNA fragment was cloned and sequenced in order to verify its suitability for identifying L. mesenteroides subsp. mesenteroides strains.

DNA Primers↗
PubMed1991

[Flow cytometric DNA analysis of malignant potential in colorectal carcinoma--DNA ploidy pattern and liver metastasis].

On the purpose of elucidating the malignant potential, flow cytometric DNA analyses were carried out using paraffin-embedded materials of 82 subserosal, serosal and adventitial invasive colorectal carcinoma. The ratio of DNA diploidy against DNA aneuploidy was one to two. DNA ploidy pattern was not correlated with histological grade but with histopathological factors such as lymphatic permeation, lymph node metastasis and venous invasion. At the primary operation, the rate of the liver metastasis in DNA aneuploid cancer was 16.4% but in DNA diploid cancer the liver metastasis was not observed at all. On the liver metastasis, the 5-year disease free survival rates were lower in patients with DNA aneuploidy (72.8%) than those with DNA diploidy (95.2%). These results indicate that DNA ploidy pattern is related to liver metastasis. Furthermore, the 5-year survival rates in patients with absolute curative resection were lower significantly in DNA aneuploidy (62.5%) than DNA diploidy (92.9%). In conclusion flow cytometric DNA analysis is useful for evaluating the biological malignant potential and predicting the liver metastasis in colorectal carcinoma.

Adult↗
PubMed2002

The clinical value of image cytometry DNA analysis in distinguishing branchial cleft cysts from cystic metastases of head and neck cancer.

OBJECTIVES/HYPOTHESIS: A branchial cleft cyst presents as a lump in the neck that, generally, is easily cured by surgical excision. The preoperative diagnosis is based on clinical examination and, especially in the Scandinavian countries, fine-needle aspiration cytology. However, at times, the histopathological analysis of the excised cyst reveals a cystic metastasis of squamous cell carcinoma of the head and neck. If adequate diagnosis could be obtained preoperatively, patients would most likely fare better. The study was performed to investigate whether the diagnostic accuracy for these lesions could be improved preoperatively by image cytometry DNA analysis of the fine-needle aspiration cytology specimen. STUDY DESIGN: Image cytometry DNA analysis was performed on the preoperative fine-needle aspiration cytology specimen and the surgical specimens from 51 patients with solitary cysts in the lateral region of the neck. Thirty-six patients were selected because there was a discrepancy between findings on fine-needle aspiration cytology and the final histopathological diagnosis or an uncertain cytological diagnosis. There were 25 metastatic squamous cell carcinomas and 3 thyroid cancers, there was 1 lymphoma and 1 sialoadenitis, and there were 21 branchial cleft cysts. METHODS: The cytodiagnostic Giemsa-stained slides were destained in Methanol and then stained with Schiff's reagent. The paraffin-embedded material from excised cysts were cut and deparaffinized and then stained with Schiff's reagent. Ahrens image analysis was used for DNA analysis and lymphocytes were used as control cells. DNA valves exceeding 5c was regarded as aneuploid. RESULTS: Image cytometry DNA analysis of the preoperative cytological specimen was possible in 41 of 51 patients. We found that in 53% of the cases with cystic metastasis, image cytometry DNA analysis, when possible, revealed aneuploidy, thus indicating malignancy. DNA analysis showed diploidy in all benign cases. CONCLUSIONS: Aneuploidy is highly specific for malignancy. Image cytometry DNA analysis increases the diagnostic sensitivity for malignant cystic metastasis and therefore is a valuable supplement to conventional cytological study for these lesions.

Adult↗
PubMed2003

A functionalized THz sensor for marker-free DNA analysis.

We report on a novel resonant THz sensor for the label-free analysis of DNA molecules. The sensor allows the direct detection of DNA-probe molecules at functionalized electrodes via hybridization. Subsequent time resolved photoconductive sampling of the THz transmission identifies the binding state between probe and target DNA. Integrating neighbouring sensors on a chip, this technique can be extended to a parallel analysis of multiple DNA sequences. A clearly readable sensor response is obtained with less then 40 fmol of 20-mer single-stranded DNA molecules, indicating at least a sevenfold sensitivity increase compared to previous approaches.

Biosensing Techniques↗
PubMed1986

Prognostic significance of nuclear DNA analysis in histological sections in mammary carcinoma.

The nuclear DNA content in tumor cells from invasive ductal breast carcinomas was analyzed in 26 patients who survived more than 10 years and in 23 patients who died within 2 years after operation. The DNA content of individual tumor cells was measured in sections from the originally paraffin-embedded specimens. In short-term survivors, a large proportion of cells with very high DNA values was found in all tumors except one. Only two patients of the long-term survivors had tumors that exhibited such high DNA values. Prognostic information obtained by DNA analysis compared with histologic malignancy grading showed that the specificity using DNA analysis was distinctly higher. The data thus suggest that analysis of DNA content of tumor cells in breast adenocarcinomas can be a useful supplement to histologic malignancy grading to obtain prognostic guidance in individual patients.

Adenocarcinoma↗