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At least 469 records · Page 26Linked to original sources
PubMed2001

Mother-offspring bonding in farmed red deer: accuracy of visual observation verified by DNA analysis.

The accuracy of a maternity assessment based on visual observation was tested during the post parturient phase in farmed red deer. The mother of the calf was determined using visual observation of the hind's peri-parturient and early maternal behaviour during the first week of the calf's life. This assessment was compared with genetic analysis based on DNA microsatellite polymorphism. Data for 48 new-born calves and 50 hinds were compared. The visual assessment of maternity was correct in 43 of the 48 cases (89.58%). In the five remaining cases the mother was identified by DNA testing whereas adoptive mother was determined by visual observation. Altogether 14 cases of adoption were seen in this study and in 9 cases the biological mother was correctly determined by visual observation. Detailed observation of the hind-calf interactions during calving and the first week after the parturition allowed an accurate prediction of maternity. However, such observation is technically demanding.

Journal Article↗
PubMed1993

Diagnostic radioimmunoassay and DNA-analysis in Swedish and Japanese patients with familial amyloidotic polyneuropathy. Homozygosity for the TTR met30 gene.

Eighteen Swedish patients with familial amyloidotic polyneuropathy were tested for the met30 mutation of the transthyretin (TTR) (prealbumin) gene by RFLP analysis of genomic DNA using the restriction enzyme NsiI. The results confirmed previous findings that the Swedish variant of familial polyneuropathy has the same valine by methionine substitution at position 30, as seen in patients with FAP from Japan, Portugal or patients of Swedish descent from USA. However, two of the patients were homozygous, totally lacking the wild type allele. Measurable serum values for the variant transthyretin (TTR) was detected with a RIA-method in all the 18 Swedish FAP patients studied with a mean concentration of 12.1 +/- 5.1 (SD) mg/100 ml, (11.0 +/- 4.1 when the two homozygotes were excluded). In 45 Japanese patients the mean was 9.2 +/- 2.7 mg/100 ml. The variant TTR was not detected in the healthy controls. The value for the variant TTR was nearly twice that high in the two homozygous patients, 21.14 and 21.16 mg/100 ml, respectively. There was no correlation between the serum levels of variant TTR and the duration of disease or levels of serum albumin in the FAP-patients.

Adolescent↗
PubMed1999

[Hematologic improvement of Pearson's syndrome confirmed by mitochondrial DNA analysis].

We report on an 8-month-old girl with Pearson's syndrome who presented with transfusion-dependent pancytopenia, exocrine pancreatic dysfunction, and lactic acidosis. Bone marrow findings were consistent with sideroblastic anemia and marked vacuolization of myeloid and erythroid precursors. Southern blot analysis of mitochondrial DNA (mtDNA) revealed a 4.5 kb deletion in peripheral blood cells. Gradual hematologic improvement was observed thereafter, and the patient was relieved of the need for blood transfusions. We were able to confirm a decrease of the mtDNA deletion in lymphocytes as well as in lymphoblastoid cell lines cultured from peripheral lymphocytes as the patient made steady hematologic progress.

DNA, Mitochondrial↗
PubMed1989

Sources and nature of variation in DNA analysis of follicular thyroid adenoma.

We investigated the sources and nature of variation that may occur in the DNA analysis of thyroid adenomas from cytological and histological samples. Imprints and smears gave identical results. However, the nuclear area was higher in smears where the optical density of the nuclei was lower. In measuring imprints, the interactive selection of nuclei was preferred to the automatic, because the risk of measuring nuclear fragments or undesired objects was thus avoided. The reproducibility and the variation of the DNA measurements depended on the degree of observer training in quantitative pathology, the method of field selection, and the type of instrumentation. Biological variation in the spatial distribution of nuclei with different ploidy values in some adenomas seemed to hide the influence of section thickness on measurements. Our data seem to suggest that it is best to apply a constant section thickness and 5 micron sections seem acceptable.

Adenoma↗
PubMed1984

A DNA analysis program designed for computer novices working in an industrial-research environment.

A program designed for DNA sequence analysis by people with no computer background or experience is described. The criteria used in the program design were (i) user friendliness, (ii) ability to handle large DNA files (greater than 10 kilobases), (iii) low storage requirements so that it could be put on a personal computer (IBM), (iv) written in a widely used language (FORTRAN 77) and (v) ability to handle all of the needed tasks for DNA analysis. The program consists of an executive that controls a set of modular subprograms. A file management system using 'SEQ' and 'SITE' files is also in place. SEQ files contain long DNA nucleotide sequences (e.g. restriction enzyme sites). Related SEQ and SITE files are grouped into categories. A SITE category can include related sequences a user wishes to search for in a longer SEQ sequence from a designated category. For example, all of the restriction enzyme recognition sites can group into a single category called 'Enzymes' and can be searched for in a SEQ sequence which has been placed in the SEQ file category 'Plasmids'. Colour graphics is also used as a visual aid.

Base Sequence↗
PubMed1987

Quantitative DNA analysis of low grade cervical intraepithelial neoplasia and human papillomavirus infection by static and flow cytometry.

Quantitative deoxyribonucleic acid (DNA) analysis of cervical biopsy specimens from 26 women with cytological, colposcopic, and histological evidence of mild cervical atypia consistent with cervical intraepithelial neoplasia grade I, reactive atypia, or human papillomavirus infection alone or in combination was performed in a comparative evaluation of Feulgen microspectrophotometry, the fast interval processor image analysis system, and flow cytometry. The fast interval processor image analysis system showed a distinct advantage over the other methods, being faster and allowing the operator to see the cells that were selected for measurement. The three methods of measurement together showed that the DNA content of at least 2% of the cells measured exceeded 5C (C being the haploid amount of DNA in a normal cell and 2C representing the diploid complement of a normal cell) in all cases of cervical intraepithelial neoplasia grade I and reactive atypia and in 87% of those reported as showing human papillomavirus infection alone. In contrast, the DNA content of cervical biopsy specimens from the transformation zone of 11 normal controls did not exceed 4C. This study shows the value of using a DNA threshold--that is, the "5C exceeding rate"--to distinguish between normal and neoplastic appearances of the cervix. These results support the view that cervical infection by human papillomavirus is a true precursor of neoplasia.

Aneuploidy↗
PubMed1994

Comparative flow cytometric DNA analysis on fresh and paraffin-embedded tumor tissue in different neoplasms.

The results of flow cytometric DNA-analysis on fresh and paraffin-embedded tissue were compared in four various groups of tumors. The best correlation of the results was observed in the group of non Hodgkin's lymphomas. Comparative analysis of the ploidy on fresh and paraffin embedded tissue demonstrated discordance in particular in mammary carcinoma and malignant melanoma groups. The values of proliferative fraction assessed on fresh and paraffin embedded tissues were interrelated.

Breast Neoplasms↗
PubMed1989

Bone marrow transplantation monitoring by DNA analysis.

Variable Number of Tandem Repeats (VNTR) DNA polymorphisms analysis was used in bone marrow transplantation (BMT) follow up. Three Acute Myeloid Leukemia (AML) transplants were investigated with YNH 24/MspI and with EFD 64.2/Rsa or HinfI highly polymorphic VNTRs. Absence of mosaicism and complete engraftment of donor cells was observed in two cases, while mixed hematopoietic chimerism was present in a case in which T cell depleted marrow was transplanted. VNTR systems represent accurate and sensitive individual specific markers for monitoring the clinical course of patients undergoing BMT, and for detecting the biological origin of relapses.

Bone Marrow Transplantation↗
PubMed1986

Malignant potentiality of squamous cell carcinoma of the esophagus predictable by DNA analysis.

Cell nuclear deoxyribonucleic acid (DNA) content was microspectrophotometrically determined in biopsy specimens from 75 patients with squamous cell carcinoma of the esophagus. The relationships among DNA distribution patterns, pathohistologic features, and prognosis were investigated. In patients with a high hyperploid DNA content (type IV), there was a high frequency of lymph node metastasis and lymphatic invasion, as compared with those exhibiting the near diploid DNA content (type II). In the type IV group, 22 of 23 patients died within 2 years, whereas patients with type II had a good clinical course and the 5-year survival rate was 60%. The DNA distribution pattern, determined by a multivariate Cox model analysis, was one of independent and significant prognostic variables. As these findings suggest that the DNA distribution pattern reflects the malignant potentiality of the tumor, the preoperative determinations of the DNA pattern in biopsy specimens of the esophageal carcinoma should provide a valuable parameter for predicting the prognosis.

Adult↗
PubMed1995

Characterization of Paracoccidioides brasiliensis isolates by random amplified polymorphic DNA analysis.

We initially used 25 different random primers in order to test their ability to generate random amplified polymorphic DNA fragments from the dimorphic human pathogenic fungus Paracoccidioides brasiliensis. From the tested primers we chose five to distinguish between seven isolates of this microorganism. The DNA amplification patterns allowed clear differentiation of the seven isolates into two distinct groups with only 35% genomic identity. One of these groups contained two subgroups with 81% genetic similarity. The random amplified polymorphic DNA analysis method proved to be a good tool for analyzing and comparing different genomes of P. brasiliensis isolates.

Base Sequence↗
PubMed1995

Expression of p53 and flow cytometric DNA analysis of isolated neoplastic glands of the stomach: an application of the gland isolation method.

The expression of p53 was studied immunohistochemically in combination with the DNA ploidy pattern by gland isolation in 97 alcohol-fixed gastric lesions. A polyclonal antibody, CM-1, was applied to the paraffin-embedded sections in this study. Overexpression of the p53 protein was found in 73.2% of 41 well or moderately differentiated gastric carcinomas and 52.2% of 23 cases with poor differentiation (P < 0.05). Immunoreactivity of p53 was also detected in isolated cancerous glands. No p53 immunoreactivity was detected in benign gastric lesions including adenomas, hyperplastic polyps and regions of intestinal metaplasia. In addition, flow cytometric DNA analysis was performed on isolated glandular epithelium adjacent to the portions used for immunostaining. DNA aneuploidy (DA) was detected in 85.7% of the well or moderately differentiated carcinomas and 42.9% of those with poor differentiation (P < 0.05). There was a positive correlation between DA, p53 positivity and the presence of regional lymph node metastasis, but not with other clinicopathological variables. In spite of the limited applicability of this method to poorly differentiated gastric cancer, we found that immunostaining and flow cytometry in combination with the gland isolation method facilitates analysis of gastric carcinogenesis.

DNA, Neoplasm↗
PubMed2001

DNA analysis and the Freedom of Information Act: information or invasion?

This note explores the possibility of release of an individual's DNA analysis to any person who requests it through the Freedom of Information Act (FOIA), after an individual's post-aircraft accident DNA profile has been developed by the Federal Aviation Administration's (FAA) Civil Aerospace Medical Institute (CAMI). It analyzes whether the request would fall under the FOIA's 552(b)(6) exemption, which weighs a person's privacy interest against any public interest in such information, or if the release would constitute a "clearly unwarranted invasion of personal privacy."

Access to Information↗
PubMed1994

[Feasibility of DNA analysis for prognostic evaluations of colorectal cancer using touch preparations and image cytometry].

The purpose of this study is to demonstrate the applicability of DNA ploidy analysis by using touch preparations obtained from fresh and frozen tissues on the prognostic evaluations of colorectal cancer, especially the development of hepatic metastases. The specimens obtained from surgically resected colorectal carcinomas (32), and hepatic metastases (4), were lightly touched with the slide glasses. Among the 32 patients with colorectal cancers, 7 had synchronous liver metastases, and 5 metastases developed metachronously. After formalin-fixation and Feulgen staining, the DNA analysis was performed by image cytometer (ICM, CAS200R). The data were expressed as the ploidy patterns and DNA indices (p-DNAI: peak DNA index, m-DNAI: mean DNA index), and were correlated very well with those obtained from cytophotometry. The DNA ploidy pattern among the 32 colorectal cancer specimens were Aneuploid(A) in 16, Polyploid (P) in 7, Diploid (D) in 4 and D+P in 5. The cases with synchronous or metachronous liver metastases had a higher m-DNAI (1.74 +/- 0.28) and were Aneuploid in 8, P in 3, and D+P in 1 case. Aneuploidy or Polyploidy predominated among the cases with metachronous liver metastases (4 in 5 cases), with m-DNAI higher than 1.5. Thus, "m-DNA > 1.5" could be a useful prognostic indicator of the high risk group of liver metastases. The liver metastases specimens had Aneuploid and Polyploid patterns in 2 cases each, with m-DNAI ranging from 1.88 to 2.31. DNA ploidy pattern analysis using touch preparations with image cytometry may be useful in determining the prognosis for colorectal cancer.

Adult↗
PubMed1988

Cross-contamination of human esophageal squamous carcinoma cell lines detected by DNA fingerprint analysis.

DNA "fingerprint" analysis has recently become known as a valuable technique for positive identification of any given individual. The chances for mistaken identity have been estimated to be 10(-6) for close siblings or as little as 10(-23) for randomly selected individuals. This methodology thus represents a significant improvement over previously established identification tests using protein or enzyme analysis techniques and has already found application in forensic medicine. One of the chief problems in tissue culture studies is the question of the unequivocal identity of the cultured cells used and the very real possibility of their being contaminated by cells of a similar morphological appearance. We report here the application of the DNA "fingerprint" technique to the genotypic analysis of cultured human squamous carcinoma cells. The results show that a number of lines, designation HCu, have become cross-contaminated. Lines SNO, HCu 10, and HCu 13 are genetically distinct, however lines HCu 10, 18, 33, 37, and 39 are genetically identical and are in fact subcultures of the same cells. In addition, a myocardial line known as Girardi is shown to be identical to HeLa cells. The introduction of this technique to tissue culture laboratories could therefore prevent contaminated cultures from being disseminated or used in research studies.

Carcinoma, Squamous Cell↗
PubMed1988

A rapid procedure for flow cytometric DNA analysis in cultures of normal and transformed epidermal cells.

A simple, rapid, and highly reproducible procedure for flow cytometric DNA analysis has been adapted for studying cell cycle kinetics in epidermal cell cultures. The preparation of cell nuclei and their staining with the fluorescent dye propidium iodide were performed directly on the culture dish, without prior suspension and fixation of the cells. Singly dispersed nuclei were produced by mild trypsinization of cells in the presence of the nonionic detergent Nonidet P-40 and spermine. The culture dishes could be kept frozen for prolonged periods of time before trypsinization and staining, without affecting either the recovery of nuclei or the cell cycle distribution profiles. This remarkable stability of cell nuclei greatly simplified the analysis of multiple samples in cell cycle kinetic studies. This method was used to analyze the cell cycle distribution in cultures of normal and transformed mouse epidermal cells, human colon carcinoma cells, primary bovine aortic endothelial cells, and fibroblastic and myogenic cell lines. This procedure should be very useful in studying growth kinetics, differentiation, and transformation of epidermal as well as other adherent cell types.

Animals↗
PubMed1997

[Identification of 5 variants of the hemoglobin beta chain using DNA analysis (amplification, sequencing and restriction enzymes)].

PURPOSE: To confirm the conventional techniques for studying structural haemoglobinopathies and to show off the simplicity and efficacy of new methods based on the study of DNA. PATIENTS AND METHODS: Peripheral blood samples of 17 patients with 5 haemoglobin variants detected by conventional and shown off by means of sequencing according to Sanger's method, plus PCR-RFLP, were studied. RESULTS: Five structural haemoglobin variants were found, which distributed as follows: 7 cases of Hb Complutense (beta 127 Gln-->Glu), 1 Hb D-Punjab (beta 121 Glu-->Gln), 3 Hb Hofu (beta 126 Val-->Glu), 3 Hb J-Baltimore (beta 16 Gly-->Asp) and 3 Hb San Diego (beta 109 Val-->Met). CONCLUSIONS: These results allow us to stress the simplicity and usefulness of DNA analysis (sequencing , amplification and enzymatic digestion) to identify haemoglobin variants as opposed to laborious analysis of the primary structure by means of HPLC peptide separation.

DNA Mutational Analysis↗
PubMed1993

DNA analysis as a predictor of the outcome of induction chemotherapy in advanced head and neck carcinomas.

We investigated whether flow cytometric DNA index and/or ploidy status are predictors of response to chemotherapy and survival. Fifty consecutive patients with previously untreated locally advanced squamous cell carcinomas of the head and neck received induction chemotherapy consisting of three courses of cisplatin (100 mg/m2) and a subsequent 120-hour infusion of fluorouracil (1000 mg/m2 per 24 hours) repeated every 3 weeks. Chemotherapy was followed by radiotherapy to a median target dose of 65 Gy and subsequent surgery for residual tumor. The median observation time was 27 months (range, 24 to 57 months). Flow cytometric DNA analysis was based on formalin-fixed and paraffin-embedded tissue from pretreatment tumor biopsy specimens. Complete response after induction chemotherapy was achieved in only 12% (2/17) of patients with diploid tumors compared with 39% (13/33) of those with nondiploid tumors. Among patients with nondiploid tumors, DNA index was higher for those responding to chemotherapy compared with the nonresponders. Complete response to chemotherapy was apparently a prerequisite for survival in the nondiploid group. Of the patients not responding to chemotherapy but responding to subsequent radiotherapy, survival was better among those with diploid tumors than among those with nondiploid tumors.

Adult↗