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D V Coleman

Publications and source records attributed to D V Coleman.

At least 37 records · Page 2Linked to original sources
PubMed1994

DNA adduct assay in cervical epithelium.

Numerous epidemiological studies have shown that there is an association between smoking and cervical cancer. However, the essential evidence to show whether this relationship is casual or causal is lacking. The demonstration of DNA modification by tobacco components in the cervical epithelium would provide biochemical evidence to support a causal role. In this study, DNA from 39 cervical biopsies was analysed for the presence of DNA adducts using the 32P-postlabeling technique. A questionnaire on smoking habit and a urinary cotinine assay were used to identify smokers and nonsmokers. DNA samples from smokers [identified from questionnaire] were found to have significantly higher adduct levels than nonsmokers (Mann-Whitney one-tailed U-test, 95% CI > 0.339, P = 0.024). Exclusion of the women whose urinary cotinine levels did not confirm their self-reported smoking status (smoker or nonsmoker) increased this significance (95% CI > 0.508, P = 0.01). Women who had abnormal cervical smears had significantly higher DNA adduct levels than those with normal smears (95% CI > 0.439, P = 0.015). Monitoring of women with high DNA adduct levels may be a way of identifying women at risk of cervical cancer. These findings demonstrate that tobacco smoking by women leads to elevated levels of DNA adducts in cervical epithelium and provides the biochemical evidence to support the concept that smoking is a cause of cervical cancer.

Adult↗
PubMed1994

Maternal contamination of amniotic fluid demonstrated by DNA analysis.

DNA from 16 sets of samples comprising DNA from uncultured amniotic fluid cells, cultured amniotic fluid cells, fetal tissue, and maternal blood was analysed by the polymerase chain reaction (PCR) with AC-repeat primers. The analysis was performed to investigate the presence of contaminating maternal cells in amniotic fluid which would affect the reliability of DNA studies for prenatal diagnosis. In three sets, maternal contamination of uncultured amniotic fluid cells was detected. In one of the three sets, maternal contamination was present in both uncultured and cultured amniotic fluid cells. The use of amniotic fluid cells as a source of DNA for prenatal diagnosis should be limited to cases where the purity of the DNA can be demonstrated prior to the diagnostic test being performed. This limitation in the use of amniotic fluid DNA also extends to other forms of diagnosis relying on the purity of amniotic fluid samples, particularly the new in situ hybridization methods currently being developed.

Amniotic Fluid↗
PubMed1994

Quality assurance in cervical cancer screening: results of a proficiency testing scheme for cytology laboratories in the north west Thames region.

OBJECTIVES: To examine the effectiveness of introducing External Quality Assessment (EQA) into all laboratories which undertake gynaecological cytopathology. To assess pathologists and cytotechnologists regularly for their competence to screen cervical smears, regardless of their standing in the laboratory hierarchy or their experience of gynaecological cytopathology. METHODS: Each participant was asked to screen and report on 10 slides during a 2 h period. The assessment was carried out by a facilitator under the direction of a specially appointed EQA Committee. A maximum score of 20 points was awarded for a completely correct set of answers. A minus score was awarded for a missed abnormal smear. Seventeen pathology laboratories in North West Thames Regional Health Authority participated; 146 cytologists were assessed. RESULTS: A pilot and four rounds of EQA have been completed and a total 5350 smears examined. Out of 2568 dyskaryotic (abnormal) smears screened, 0.7% were not identified correctly. Of the 146 cytologists taking part in the assessment, 95% achieved a score of 17 or more. Three participants were identified who did not reach an acceptable level of competence and appropriate remedial action was taken. CONCLUSION: The EQA scheme detected unacceptable levels of performance which can be quickly rectified. Participation of 100% has been maintained on a voluntary basis, and 4 years experience of the scheme confirms that a very high standard of screening prevails in the Region. The study illustrates that voluntary self-regulation is acceptable in the NHS, and the introduction of similar EQA schemes on a national scale will go a long way to establishing confidence in the cervical cancer screening programme.

Female↗
PubMed1994

Evaluation of the sensitivity, specificity, and predictive value of monoclonal antibody 3F6 for the detection of Pneumocystis carinii pneumonia in bronchoalveolar lavage specimens and induced sputum.

The sensitivity and specificity of different staining procedures for the detection of Pneumocystis carinii organisms were compared. Three conventional stains (Papanicolaou, Giemsa and Grocott's) and one immunocytochemical stain using 3F6 antibody were used on smears prepared from the same specimen. Bronchoalveolar lavage (BAL) and induced sputum (IS) specimens were used for this purpose. One hundred and sixty-five episodes from 142 patients were investigated by the four different staining techniques. Cysts of P. carinii were detected in 64 episodes from 63 patients. Immunocytochemical staining with 3F6 was found to be slightly more sensitive at detecting the cysts than Grocott's, Giemsa, or Papanicolaou stain.

AIDS-Related Opportunistic Infections↗
PubMed1994

Pneumocystis carinii pneumonia: relationship between cyst size and response to treatment.

Pneumocystis carinii pneumonia is a common cause of death in patients with AIDS. Diagnosis is based on cytological examination of smears prepared from induced sputum samples and bronchoalveolar lavage (BAL) specimens which have been stained with methenamine silver. We have examined 46 BAL/induced sputum specimens from patients who had clinical symptoms and signs suggestive of P. carinii pneumonia and measured the diameter of a minimum 100 cysts in each specimen. We found that cyst size correlated with response to treatment with co-trimoxazole. This observation has implications for the therapeutic management of patients with this infection.

AIDS-Related Opportunistic Infections↗
PubMed1994

EB9, a new antibody for the detection of trophozoites of Pneumocystis carinii in bronchoalveolar lavage specimens in AIDS.

AIM: To prepare a monoclonal antibody (EB9) against the trophozoite form of Pneumocystis carinii and to test its efficacy for detecting infection with this organism. METHOD: The sensitivity and specificity of the EB9 antibody were assessed by comparing it with other conventional stains (Papanicolaou, Giemsa and Grocott) and 3F6 antibody in 33 bronchoalveolar lavage specimens from HIV positive patients suspected of having P carinii pneumonia. RESULTS: P carinii infection was detected in 15 of 33 patients. In 14 cases the organism was detected by two or more of the staining methods used; however, EB9 failed to detect infection in two cases which were positive by other staining techniques. In one case P carinii infection was detected by EB9 only. CONCLUSION: The results of this study suggest that P carinii infection in the lung may occur in two forms: the cyst form and the trophozoite form, which may explain the observed variation in response to treatment.

Acquired Immunodeficiency Syndrome↗
PubMed1993

Damage to DNA in cervical epithelium related to smoking tobacco.

OBJECTIVE: To determine whether tobacco smoking causes increased DNA modification (adducts) in human cervical epithelium. DESIGN: Comparison of DNA adducts measured by the technique of postlabelling with phosphorus-32 in normal ectocervical epithelium of smokers and non-smokers. A questionnaire on smoking habit and a urinary cotinine assay were used to identify smokers and non-smokers. SETTING: Cytology unit in large teaching hospital. SUBJECTS: 39 women (11 current smokers, seven former smokers, and 21 who had never smoked) undergoing gynaecological treatment (colposcopy or hysterectomy). Nineteen members of staff who did not smoke as controls. INTERVENTIONS: Biopsy of normal ectocervical epithelium. Urine sample. MAIN OUTCOME MEASURES: Measurement of DNA adducts in cervical epithelial tissue of smokers and non-smokers. Smoking habit derived from results of questionnaire and urinary cotinine:creatinine ratio. Proportion of adducts in women with abnormal and normal results of cervical smear test. RESULTS: DNA samples from smokers (identified from questionnaire) had significantly higher median proportions of DNA adducts that non-smokers (4.62 (95% confidence interval 4.04 to 7.74) v 3.47 (2.84 to 4.78) adducts/10(8) nucleotides; p = 0.048). Exclusion of women whose urinary cotinine:creatinine ratio did not confirm their self reported smoking habit (smoker or non-smoker) increased this difference (4.7 (3.85 to 8.08) v 3.52 (2.32 to 4.95) adducts/10(8) nucleotides; p = 0.03). Women who had abnormal results of cervical smear tests had significantly higher proportions of adducts than those with normal results (4.7 (3.90 to 8.13) v 3.47 (3.06 to 5.36) adducts/10(8) nucleotides; p = 0.03). CONCLUSIONS: Tobacco smoking by women leads to increased modification of DNA in cervical epithelium, suggesting biochemical evidence consistent with smoking as a cause of cervical cancer.

Adult↗
PubMed1993

Use of fine needle aspiration cytology for investigating lymphadenopathy in HIV positive patients.

The cytological diagnoses of 27 lymph node aspirates were compared with the histological diagnoses or clinical outcome in 23 HIV positive patients. There was agreement between the cytological and histological diagnoses in 14 of the 16 surgically biopsied cases. The clinical outcome in the remaining 11 cases was consistent with the cytodiagnosis. Fine needle aspiration (FNA) is a reliable, minimally traumatic, cost effective method with high specificity. It is suitable for an initial rapid diagnosis in HIV positive patients with lymphadenopathy.

Adult↗
PubMed1993

Phase contrast microscopic examination of urinary erythrocytes to localise source of bleeding: an overlooked technique?

AIMS: To localise the source of bleeding in the urinary tract in patients presenting with haematuria. METHODS: Urine samples were obtained from 109 patients with symptoms referable to the urinary tract. The sample was examined for the presence of red blood cells by phase contrast microscopy (PCM) and the proportion of dysmorphic and isomorphic red blood cells was determined. If more than 20% of the red blood cells were dysmorphic a glomerular origin for the site of bleeding was suspected; if less than 20% of the red blood cells were isomorphic a non-glomerular origin was suspected. Phase contrast microscopy and clinical findings were correlated. RESULTS: The correct bleeding site was shown in 27 of 30 (90%) patients with glomerulopathy and in all 17 patients with bleeding from the lower urinary tract, indicating that this method of analysis has a sensitivity of 90% and specificity of 100% for detecting the glomerular source of bleeding. CONCLUSIONS: The examination of urine for dysmorphic and isomorphic red blood cells by phase contrast microscopy is strongly recommended in routine clinical practice for the detection of glomerular and non-glomerular lesions. This technique may avoid unnecessary investigations for the diagnosis of the site of bleeding in patients with haematuria.

Adult↗
PubMed1992

Prevalence of human papillomavirus (HPV) infection in Basque Country women using slot-blot hybridization: a survey of women at low risk of developing cervical cancer.

Cervical smears from 1,178 women with cytologically normal cervices and 67 women with cervical intra-epithelial neoplasia (CIN) or cervical carcinoma were analyzed for the presence of HPV 6, 11, 16 and 18 DNA by slot-blot hybridization. HPV DNA was detected in 17% (95% CI: 14%-19%) of the women with a normal smear; 11% of infected women harboured HPV 16 and 18 (95% CI: 9%-13%, each). HPV DNA was detected in 54% (95% CI: 41%-66%) of patients with abnormal smear; the most prevalent virus type in this group was HPV 16 (45%; 95% CI: 32%-38%). In order to verify the slot-blot results, a proportion of the samples was also investigated by PCR. There was 88% correlation between the 2 tests. The high prevalence of HPV 16 and 18 infection demonstrated in our low-cervical-cancer-risk area further support the role of as yet unidentified co-factors as determinants of the different geographic rates of cervical-cancer incidence.

DNA, Viral↗
PubMed1991

Cytogenetic studies of amniotic fluid taken before the 15th week of pregnancy for earlier prenatal diagnosis: a report of 114 consecutive cases.

One hundred and fourteen samples of amniotic fluid taken before 15 weeks of gestation were cultured for cytogenetic studies. The results of culturing these early amniotic fluid (EAF) samples were compared with the results of culturing 114 standard amniotic fluid (SAF) samples taken after 15 weeks of gestation matched for maternal age and received in the laboratory within the same week. Cell culture was successful in all 114 of the EAF samples and in 111 SAF samples. There was no significant difference in the days to harvesting and days to reporting in the two groups. Three samples of SAF failed to grow and two EAF samples produced tetraploid karyotypes, so that in these five cases amniocentesis had to be repeated. These problems were attributed to toxicity of a fungicide used in the culture medium. Pseudo-mosaicism was noted in two EAF samples and one SAF sample; and maternal cell contamination was noted in one EAF and one SAF sample. Thus, culturing and karyotyping cells harvested from EAF and SAF are similar, indicating that EAF samples from 12-14-week pregnancies could be used for prenatal diagnosis.

Adult↗
PubMed1991

Extraction of DNA from amniotic fluid cells for the early prenatal diagnosis of genetic disease.

Ten-ml samples of amniotic fluid were taken from pregnancies being terminated at 8-14 weeks' gestation. DNA was extracted from the amniotic cells by sequential centrifugation and analysed using the polymerase chain reaction (PCR). Fifteen samples were analysed for evidence of maternal contamination using Mfd5 oligonucleotide primers for repeat polymorphisms. Ten amniotic fluid samples were tested for the Delta-F508 deletion characteristic of cystic fibrosis to demonstrate a diagnostic application for the technique. In each case, DNA extracted from fetal tissue from the same pregnancy was included in the controls. In 14 of the 15 cases tested with the Mfd5 primers, both the amniotic fluid DNA and the fetal DNA showed no evidence of contaminating DNA. In one case, neither the amniotic fluid cells nor the fetal cells yielded results. In nine of the ten cases tested with the Delta-F508 primers, the amniotic fluid cell DNA provided accurate information about the genetic status of the fetus; in the tenth, the fetal DNA failed to amplify. The results indicate that adequate DNA can be extracted from amniotic fluid from 8 weeks' gestation onward and these samples are suitable for prenatal diagnosis using PCR.

Adult↗
PubMed1991

Early amniocentesis at 11-14 weeks' gestation for the diagnosis of fetal chromosomal abnormality--a clinical evaluation.

Early amniocentesis between 11 and 14 weeks' gestation was offered to 110 women at risk of a chromosomally abnormal fetus due to maternal age. Four were found to be unsuitable for the procedure, and 106 early amniocenteses were performed. In 102 cases, clear amniotic fluid was obtained with a single tap. There were two dry taps and two bloodstained taps; sampling was repeated in three of these cases before 15 weeks. In the fourth case, placental biopsy was performed at 16 weeks. Thus, we were able to obtain a satisfactory sample in all but three cases (2.8 per cent). Karyotyping of cells harvested from the early amniotic fluid samples was successful in all the 105 cases. Cell culture from the initial samples revealed a normal karyotype in 99 cases, two balanced translocations, two tetraploid karyotypes, and two cases of pseudomosaicism. Of the 105 pregnancies successfully sampled, there have been two losses to date (1.8 per cent). Two further patients presented with premature rupture of membranes, both pregnancies having successful outcomes. Sixty-two babies have delivered to date, with four congenital anomalies. There were no respiratory problems. Twenty-nine pregnancies are continuing without known complications, and details are not yet available on the remaining 12. The results indicate that early amniocentesis may replace the traditional test at 15-17 weeks.

Adult↗
PubMed1991

Evaluation of immunocytochemical staining as a method of improving diagnostic accuracy in a routine cytopathology laboratory.

Immunocytochemical stains in a routine cytopathology laboratory can be used to distinguish between benign and malignant cells, and to identify tumour type. In our laboratory 30 problematic cases were selected for immunocytochemical stains and the results analysed in this paper. The following markers were used: cytokeratin (CAM5.2), carcinoembryonic antigen (CEA), kappa and lambda light chains, leucocytic common antigen (LCA), chorionic gonadotrophin (hCG), prostate specific antigen (PSA), L26, UCHL1, S100-protein and vimentin. Twelve FNA (four lymph nodes, one parotid swelling, two from lungs, two from pleura and chest wall, one from lumbar region, two from soft tissue masses), and 18 effusions (12 pleural effusions, five ascitic fluids, one pericardial effusion) were investigated. We found immunocytochemical stains of value in formulating the cytological diagnosis in 11/12 of FNA and 15/18 of effusions.

Cytodiagnosis↗
PubMed1991

C-myc expression in exfoliated cells in serous effusions.

In an attempt to improve the discrimination between benign and malignant cells in pleural and peritoneal fluids, 32 effusions were investigated for c-myc expression. Smears were prepared from cells harvested from the fluids for immunocytochemical staining to identify the presence of c-myc protein. Recombinant DNA technology (Northern blotting, slot blotting and in situ hybridization) was used to detect c-myc mRNA. No significant difference in expression of c-myc was noted in benign or malignant effusions. Although the results are inconclusive, the recombinant DNA technology developed for this research could be used to investigate the expression of other oncogenes in cytological material.

Adult↗