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Biomedical subjects

D V Coleman

Publications and source records attributed to D V Coleman.

At least 73 records · Page 4Linked to original sources

Use of hybridot assay to screen for BK and JC polyomaviruses in non-immunosuppressed patients.

Urine samples from 50 patients attending a genitourinary outpatient clinic and from 13 renal allograft recipients were investigated for evidence of infection with human BK and JC polyomaviruses using cytology and a new DNA hybridot assay. Forty four per cent of samples from the renal allograft recipients were positive by cytology and 75% by DNA hybridisation, indicating that hybridot assay is more sensitive than cytological screening. BK and JC viral DNA was found in 20% of the patients attending the genitourinary clinic, showing infection with BK virus and JC virus in a group of patients with clinical conditions not normally associated with immunological deficiency-a finding that has not been reported before.

Adult↗

DNA hybridization of cervical tissues.

The increasing frequency of cervical neoplasia among younger women and the increased invasiveness of these tumors has led to a considerable growth in research into this disease. Conventional methods (epidemiology, cytology, and immunology), while being extremely useful, also have significant limitations. Recent advances in techniques for the manipulation of DNA now make it possible to analyze tissues for the presence of viral genomes. This review introduces these techniques and describes their application to the search for herpes simplex virus and human papillomavirus sequences in cervical tissue. The significance of the findings both for the mechanism of transmission of the disease, and also the consequences for early detection and hence more successful treatment, are also discussed.

Cloning, Molecular↗

Association of human papillomavirus with squamous carcinoma of the uterine cervix.

The frequent association of human papillomavirus infection with preinvasive and invasive cancers of the cervix has led to speculation that the viruses may be involved in the neoplastic process. We have carried out some epidemiological studies of the prevalence of human papillomavirus infection in the female genital tract to evaluate more fully the oncogenic potential of these viruses in the cervix. A non-invasive method of detecting the virus has been developed for this purpose using DNA hybridization of cervical scrapings. The technique has been used to carry out prospective studies of patients with cervical intraepithelial neoplasia (CIN) and to investigate infection in women treated by laser therapy for CIN. We have also studied women with normal cervices to determine the prevalence of human papillomaviruses in the normal population. Further studies have involved the investigation of the effect of interferon on virus replication and epithelial proliferation in women with CIN III.

Carcinoma in Situ↗

The effect of interferon on human papillomaviruses associated with cervical intraepithelial neoplasia.

A double-blind, placebo-controlled, trial of leucocyte interferon showed that, contrary to previous reports, interferon had no significant effect on cervical intraepithelial neoplasia (CIN) when applied topically in a gel. DNA hybridization of cervical scrapes was used to monitor the effect of interferon on the human papillomaviruses (HPV) associated with CIN. There was, however, no significant difference in the expression of HPV 6 or 16 in the cervical epithelium of patients treated with interferon compared with those given a placebo. By using superficial cells scraped from the surface of the cervical epithelium as a source of DNA for viral studies, we were able to investigate the relation between HPV and CIN without interfering with the natural history of the disease. HPV 16 was detected in lesions which persisted while HPV 6 only was detected in one lesion that regressed. Regression was clearly associated with reduction in the number of copies of viral DNA per cell in this case. Dual infection with HPV types 6 and 16 were recorded in two patients with persistent lesions. In one patient, hybridization studies indicated that infection with HPV 16 could have occurred after infection with type 6 was established, and it is postulated that this may have changed the nature of the cervical lesion.

Adolescent↗

Screening for wart virus infection in normal and abnormal cervices by DNA hybridisation of cervical scrapes.

Cervical scrapings from 78 women attending a sexually transmitted diseases (STD) clinic, family planning clinic, and colposcopy clinic were assayed for human papillomavirus type 6 (HPV-6) by DNA hybridisation. Results of hybridisation were compared with clinical, colposcopic, and cytological findings. Scrapings from 2 of the 4 women with genital warts gave positive results with the HPV probe. No HPV-6 DNA was detected in scrapings from 18 women attending an FPC whereas scrapings from 2 of 19 STD clinic patients with normal colposcopic and cytological examination were positive-this means that DNA hybridisation detected wart virus infection where previously none was suspected. HPV-6 DNA was detected in about 10% of cervical scrapings from women with cervical intraepithelial neoplasia (CIN) both before and after treatment. Indeed in 2 patients in whom viral DNA persisted after laser therapy, it was associated with local recurrence of neoplasia. This non-invasive technique is suitable for use as a screening test for HPV infection. In this study it identified women who have a high risk of developing CIN and in whom close cytological and colposcopic surveillance is indicated.

Animals↗

A practical assessment of ultrasound-guided transcervical aspiration of chorionic villi and subsequent chromosomal analysis.

Aspiration biopsy of trophoblastic villi was performed on 56 patients immediately preceding suction termination of pregnancy, using a malleable metal cannula and ultrasound guidance. Villi were obtained from 47 patients (84%), with sampling success rising to 93% after experience. Immediate complications were noted in 14% of patients and correlated with placental positions situated furthest from the cervical canal. Karyotyping from cultured and/or direct preparations was attempted on the villus samples and was successful in 42. Clarity of the karyotypes obtained from direct preparations in this series was not found to be adequate for diagnostic purposes. A number of practical suggestions which facilitate chorion villus sampling are described.

Biopsy, Needle↗

Monoclonal antibodies for the histopathological diagnosis of cervical neoplasia.

Five monoclonal antibodies (Ca1, HMFG 1 and 2, 8.30.3 and 77.1) were used to study the distribution of antibody binding sites in cervical tissue with a view to identifying a marker which would distinguish between benign and malignant cervical epithelium. Both benign tissue (mature and immature metaplastic squamous epithelium, congenital transformation zone and glandular epithelium) and neoplastic tissue (cervical intraepithelial neoplasia, 1, 2 and 3 and invasive squamous cell carcinoma) were stained by these antibodies. Although immature metaplastic epithelium stained strongly with all the antibodies, the intensity and distribution of staining in general did not distinguish between benign and neoplastic conditions. All five antibodies, raised against three different antigens, stained cervical tissue in a similar way and thus were unsuitable for use as specific tumour markers in equivocal cases. Further studies on other tumour markers are indicated in view of the potential value of this approach.

Antibodies, Monoclonal↗

Epithelial membrane antigen in cells from the uterine cervix: immunocytochemical staining of cervical smears.

Smears made from cervical scrapes have been stained immunocytochemically for epithelial membrane antigen using a polyclonal antiserum and two monoclonal antibodies. With the polyclonal antiserum malignant cells and those showing dysplasia consistently expressed the antigen. Normal cells were generally negative, with the exception of some metaplastic cells. The monoclonal antibodies, although they stained the abnormal cells less consistently, gave the same pattern of staining. All three antibodies showed considerable heterogeneity in the intensity of stain. This appears to be a general feature of the expression of this type of epitope in epithelial cells. While the results confirm that an immunohistochemical stain might have potential application for improved diagnostic methods, the staining of metaplastic cells with the presently available antibodies limits the usefulness of an antiserum to epithelial membrane antigen.

Antibodies, Monoclonal↗

Clinical significance of human papillomavirus infection of the uterine cervix in the development of cervical intraepithelial neoplasia.

A group of 25 controls and 30 women with lesions of the cervix associated with papillomavirus were studied prospectively by colposcopy and cytology for up to two years to investigate the relation between human papillomavirus (HPV) infection of the cervix and resolution or progression of cervical intraepithelial neoplasia (CIN). We found that the viral changes were transient and that active infection, as judged by repeated colposcopic and cytological examination, resolved in 83% of the patients. Resolution or persistence of the viral infection did not appear to affect the development of the CIN lesion. The proportion of CIN lesions that resolved and persisted were the same for the study group and the controls. Possible reactivation of latent papillomavirus was noted in three control group patients. Our findings indicate that changes in the cervix associated with papillomavirus should not influence the clinical management or follow up of patients with CIN.

Adolescent↗

Immunocytochemical staining of cervical smears for the diagnosis of cervical intraepithelial neoplasia.

A total of 233 cervical smears were stained by immunocytochemical methods for epithelial membrane antigen (EMA); the findings were compared with those from Papanicolaou-stained smears from the same women. Squamous epithelial cells from normal cervices did not stain, but cells shed from cervices with cervical intraepithelial neoplasia (CIN) did express the EMA marker. Metaplastic cells from normal and abnormal cervices also frequently stained. The results confirm that this marker detects cervical intraepithelial neoplasia in vitro, but its potential use in an automated screening program may be limited by the staining of the metaplastic cells.

Antigens, Neoplasm↗

Presence of human papillomavirus DNA sequences in cervical intraepithelial neoplasia.

Twenty two patients referred to a district colposcopy clinic because of an abnormal cervical cytology report or a suspicious cervix and found to have a cervical epithelial abnormality were studied. The techniques of cytology, histology, immunohistochemistry, and DNA-DNA hybridisation were used to detect infection by human papillomavirus. Using an indirect immunoalkaline phosphatase technique human papillomavirus antigen was found in biopsy specimens from six of the 22 patients and DNA of papillomavirus type 6 in biopsy specimens from 13 of these women, including four out of six whose histological diagnosis was cervical intraepithelial neoplasia grade 3. In eight cases where cytological, colposcopical, and histological investigations all indicated the presence of wart virus infection, papillomavirus type 6 DNA was found in seven. Papillomavirus type 6 DNA was found in more than half of the proved cases of cervical intraepithelial neoplasia. The presence of this viral DNA in women with no cervical abnormality is to be studied.

Adolescent↗

Absence of maternal contamination of chorionic villi used for fetal-gene analysis.

Chorionic villi can be obtained by direct transcervical aspiration at 9 to 10 weeks' gestation and used for analysis of fetal DNA. However, for the method to be reliable, there must be no detectable contamination by maternal DNA. To investigate the question of contamination, we compared the DNA of chorionic villi from five fetuses with that obtained from maternal lymphocytes, using the restriction endonuclease Taql and specific DNA probes for a pair of alleles on the X chromosome. The alleles yield fragments of different lengths when digested with Taql (length polymorphism), which can be demonstrated by electrophoresis and hybridization with the radioactive DNA probes. If the pattern obtained with the chorionic DNA is different from that obtained with the maternal DNA, contamination is not present. In two cases the fetal DNA of the chorionic villi was shown to be uncontaminated by maternal tissue. In one of these cases the mother was heterozygous and the fetus was homozygous; in the other the mother was homozygous and the fetus was heterozygous. In three other cases no definitive conclusions could be drawn, because the genotypes of the fetus and mother were identical. We conclude that chorionic villi at 9 to 10 weeks' gestation are a source of fetal DNA that can be used for gene analysis, with no detectable contamination by maternal DNA.

Alleles↗

Colposcopy in the diagnosis of papillomavirus infection of the uterine cervix.

Morphological criteria are described for the diagnosis of papillomavirus lesions at colposcopy. Using these criteria, evidence of cervical papillomavirus infection was found in 58 (29.0%) of 200 women referred for colposcopy because of an abnormal cytology report. Directed biopsies were obtained from 152 of these patients and histological changes suggestive of papillomavirus infection were found in 47 (30.9%); additional biopsy material from 139 was studied using an immunohistochemical technique and human papillomavirus antigen was demonstrated in 28 (20.1%). Comparison between the colposcopic results and those obtained by histology suggests that whereas it is not possible to make a distinction at colposcopy between lesions due to papillomavirus and those of cervical intraepithelial neoplasia, it is possible to identify those epithelial abnormalities that are most likely to be associated with a papillomavirus infection.

Animals↗

Assessment of a transcervical aspiration technique for chorionic villus biopsy in the first trimester of pregnancy.

A cannula attached to a syringe was passed through the cervical canal into the uterine cavity in an attempt to aspirate chorionic villi just before termination of a first trimester pregnancy in 82 patients. Chorionic villi were obtained from 40% of these patients. In view of the value of this technique for the first trimester diagnosis of genetic disorder, development of the technique and further trials of transcervical aspiration of chorionic villi would seem warranted.

Biopsy, Needle↗

Prognostic significance of herpes simplex virus antibody status in women with cervical intraepithelial neoplasia (CIN).

A total of 107 women with abnormal cervical smears showing cytological changes consistent with cervical intraepithelial neoplasia (CIN) 1 or CIN 2 were kept under regular cytological, colposcopic, virological and serological surveillance for an average of 18 months (range 9 months-3 years). Regression of the cervical lesion was noted in 31 (29%) and progression to CIN 3 in nine women (8.4%). We found a positive correlation between the presence of type 2 antibody and progression of CIN 1 and 2 to CIN 3 and a negative association with the presence of type 1 antibody and suggest the antibody status of a woman with CIN 1 or CIN 2 may provide a useful basis for follow-up. We found no association between the outcome of the cervical lesion and active infection with herpes simplex or cytomegalovirus or any other infectious agent or sex-related factors.

Adolescent↗