[International research: a comment].
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Female↗
Biomedical subjects
B Norrild
Publications and source records attributed to B Norrild.
[Better screening of cervix cancer].
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Female↗
Herpes simplex virus type 1-infected human embryonic lung cells studied by optimized immunogold cryosection electron microscopy.
Herpes simplex virus type 1 (HSV-1) is a common human pathogen of skin and mucous membranes and is potentially dangerous when the infection is disseminated. Viral morphogenesis, especially the mechanism of viral envelopment and the exact pathway for processing and transport of HSV-1 glycoproteins, is still unclear. We report the results of optimized immunogold-labeled cryosection electron microscopy of HSV-1-infected cultured human fibroblasts (MRC-5). The simplified method presented has proved necessary to obtain reproducible results on cellular distribution of viral glycoproteins. It is now possible to demonstrate the viral glycoprotein gD-1, but not gC-1, in the nuclear membranes and to demonstrate gD-1- and gC-1-labeled viral particles in the perinuclear space, and to show the fate of the viral particles in the endoplasmic reticulum and Golgi area in infected cells.
Cells, Cultured↗
Development of cytomegalovirus (CMV) disease may be predicted in HIV-infected patients by CMV polymerase chain reaction and the antigenemia test.
OBJECTIVE: Cytomegalovirus (CMV) is a frequent opportunistic viral pathogen in patients with AIDS leading to retinitis and other serious manifestations. CMV disease may be successfully treated. Prophylactic antiviral therapy has been shown to reduce the risk of CMV disease if initiated early. We evaluated PCR and the antigenemia tests as methods for early detection of CMV disease. METHODS: Two-hundred HIV-seropositive subjects with CD4 T-cell counts below 100 x 10(6)/l were monitored with CMV polymerase chain reaction (PCR), the antigenemia test, blood cultures and CMV immunoglobulin (Ig) G and IgM titres every second month for 1 year. RESULTS: Thirty-eight patients (19%) developed CMV disease. The PCR test detected CMV DNA a median of 46 days before onset of disease. This was earlier than the median of 34 for the antigenemia test and a median of 1 day for CMV blood cultures. Univariate analysis showed that the CMV PCR, the antigenemia test and blood cultures all had significant predictive values for subsequent development of CMV disease with odds ratios (OR) of 30, 22 and 20. CMV serology had no predictive value. Multivariate analysis showed that the PCR method was superior to the other tests; OR: CMV PCR 10.0, antigenemia test 4.4 and CMV cultures 4.3. No clinical parameters had any significant predictive value in the stepwise multivariate model. CONCLUSIONS: The CMV PCR and the CMV antigenemia tests are both sensitive methods that may predict development of CMV disease up to several months prior to clinical disease. These methods make it possible to select patients at high risk for CMV disease and suitable for prophylactic therapy against CMV.
AIDS-Related Opportunistic Infections↗
Human papillomavirus in oral premalignant lesions.
The aetiology of oral premalignant lesions is generally accepted to be multifactorial. Tobacco and alcohol are established as important cofactors in malignant development in the oral cavity, but in addition microorganisms, such as human papillomavirus (HPV), have gained much interest over the past decade. For many years, HPV has been accepted as an important cofactor in the development of cervical cancer, originating from a mucous membrane with similarities to the oral mucosa. 49 patients with oral premalignant lesions and 20 control patients with normal oral mucosa and no history of HPV infection were examined for the presence of HPV by immune histochemical staining using the peroxidase anti-peroxidase technique (PAP), DNA-DNA in situ hybridisation (ISH), and polymerase chain reaction (PCR) analysed by Southern blot hybridisation with an HPV 16 specific probe. The investigations revealed that HPV was found in 62.5% of the verrucous leucoplakias, 50.0% of the erythroplakias, 45.5% of the homogeneous leucoplakias, 33.3% of erythroleucoplakias and in 12.5% of the nodular leucoplakias. An overall HPV detection rate in the examined premalignant lesions was 40.8% and no patients in the control sample were positive. Concerning oral cancer development, it seems likely that HPV may be a cofactor, as 100% of patients who developed oral cancers within 4-12 years were all positive for HPV, one being positive for HPV 16.
Adult↗
Analysis of human papillomavirus type 16 E5 oncogene expression in vitro and from bicistronic messenger RNAs.
During a human papillomavirus infection (HPV), a set of spliced messenger RNAs (mRNA) are produced from the coding strand of the 8-kilobase virus DNA genome. These mRNAs have been isolated from cervical intraepithelial neoplasia biopsies, tumor biopsies and HPV type 16 (HPV-16)-containing cell lines, and subsequent analyses have shown that the different mRNAs are obtained by often complex splicing events. In the present study we have investigated the coding capacity of the HPV-16 E5 open reading frame (ORF) in an in vitro translation system and by the expression of a HPV-16 E5-CAT fusion gene from bicistronic messenger RNAs derived from polycistronic mRNAs which have previously been identified by others. Our results show that the HPV-16 E5 protein synthesis can only be initiated from the authentic ATG codon in the presence of E2 ORF translation. The E5 gene is found to be expressed only from a bicistronic mRNA with an E2 ORF upstream of the E5 gene indicating that the E5 protein is synthesized coordinated with the E2 protein. The supposition that the E5 protein is synthesized early in infection is discussed.
3T3 Cells↗
Low-fading immunofluorescence with propidium iodide contrast compared with immunogold light microscopy in whole cells and semi-thin cryosections.
Conventional immunofluorescence produces excellent labeling but has drawbacks such as fading and the need for phase-contrast. Silver-enhanced colloidal gold probes allow counterstaining and permit permanent preparations with no fading if mounted correctly, but the most common limits of this technique are steric hindrance and the artifacts produced by silver enhancement. Our goal was to investigate Herpes simplex virus type 1 (HSV-1) morphogenesis by immunogold cryosection electron microscopy. We therefore needed a sensitive and reproducible immunocytochemical light microscopic method to confirm the immunofluorescence results in whole cells and to screen the cryopreparations before the time-consuming electron microscopic studies. We report data showing that the use of p-phenylenediamine to retard fading and propidium iodide to provide counter-staining results in brilliant fluorescence and contrast, minimal autofluorescence, and invisible fading at least for 5 min exposures, even in preparations with weak antigen presentation. Storage at -20 degrees C provides stable fluorescence. This method is superior to silver-enhanced colloidal gold light microscopy in our investigations.
Cells, Cultured↗
Detection of cytomegalovirus DNA in serum correlates with clinical cytomegalovirus retinitis in AIDS.
The high sensitivity of nested polymerase chain reaction (PCR) offers the possibility of rapid detection of cytomegalovirus (CMV) DNA in serum. Five consecutive serum samples were examined from 52 human immunodeficiency virus (HIV)-seropositive patients (19 of whom had clinically presumed diagnosis of CMV chorioretinitis). Presence of CMV DNA in serum was shown to precede development of clinical disease. Eleven patients who developed chorioretinitis were positive for CMV DNA in serum samples obtained 3 months before clinical disease, and 3 retinitis patients who initially were negative for CMV DNA became positive with the onset of clinical retinitis. In contrast, 29 of 33 HIV-seropositive subjects without clinical CMV chorioretinitis and matched with respect to age and CD4 T cell numbers were negative for CMV DNA in all 5 serum samples. Thus, the presence of CMV DNA in serum analyzed by PCR is a good predictive marker of CMV retinitis in HIV-seropositive subjects. A positive PCR results supports the clinical diagnosis and may be useful for monitoring response to antiviral treatment.
AIDS-Related Opportunistic Infections↗
The prevalence of human papillomavirus in cervical lesions with koilocytosis and/or CIN I.
Thirty-one patients with koilocytosis and/or concomitant CIN I were analysed for the presence of HPV types 11, 16 and 18 by in situ hybridization and Southern blot analysis. The prevalence of HPV was 48% and 55%, respectively, when measured by the two methods and among the HPV positive, HPV 11 and 16 were present in 47% and 60%, respectively, whereas HPV 18 was not found.
Blotting, Southern↗
CO2 laser vaporization in the treatment of cervical human papillomavirus infection in women with abnormal Papanicolaou smears.
In a randomized study, we have evaluated the treatment of cervical human papillomavirus (HPV) lesions by CO2 laser vaporization. Fifty patients with abnormal Papanicolaou smears and histological evidence of cervical HPV infection associated or not with cervical intraepithelial neoplasia (CIN) grade I were randomized to either a treatment or a control group. The cervical swabs were obtained every 3 months in both groups and examined for HPV type 16 DNA by the polymerase chain reaction. After a follow-up period of 12 months no significant differences were found between the laser treatment and the control groups in relation to the disappearance of the abnormal Papanicolaou smear. Two patients in the treatment group and 3 in the control group had a conization because of development of CIN I or aggravation of the concomitant CIN found at the initial visit. The percentage of women who demonstrated HPV in their cervical smears at 12 months' follow-up was identical in the two groups, supporting the hypothesis that HPV is a persistent infection during which the virus is widespread in the vaginal epithelium.
Biopsy↗
HPV 11, 16 and 18 DNA sequences in cervical swabs from women with cervical dysplasia: prevalence and associated risk of progression.
Cervical swabs obtained from 164 women with histologically proven preinvasive and invasive cervical neoplasia were analysed for HPV type 11, 16 and 18 DNA by filter in situ hybridisation. HPV 16 or 18 was detected in 8 of 24 swabs from patients with invasive squamous cell carcinoma (33%), in 59 of 100 patients with carcinoma in situ or severe dysplasia (59%) and in 16 of 40 patients with mild or moderate dysplasia (40%). HPV 6 or 11 was found in only 2% of all swabs. Thirty-eight of the patients participated in a prospective follow-up study and were monitored non-invasively by cytology and colposcopy for 8 to 36 months. 25 patients had persisting or progressive lesions, 13 of which harboured HPV 16 or 18. Of 13 patients who had complete resolution of the dysplasia, only 2 were HPV-positive. The study indicates a significantly higher risk of malignant progression when the cervical dysplasia is associated with HPV 16 or 18 infection.
Adolescent↗
Human papillomavirus type 16 in vulvar carcinoma, vulvar intraepithelial neoplasia, and associated cervical neoplasia.
Vulvar intraepithelial neoplasia (VIN) is becoming more widespread and the patients are becoming still younger. Although progression to invasive vulvar carcinoma is uncommon, local recurrences are frequent and about one-quarter of the patients have multicentric genital disease. The aim of the present study was to search for a possible significant association of human papillomavirus (HPV) infection with vulvar carcinoma, recurrences, and multicentric disease. We used the polymerase chain reaction to examine vulvar and cervical biopsies from 43 patients with vulvar neoplasia for HPV type 16, which is the subtype most often detected in genital malignant or premalignant lesions. HPV 16 DNA sequences were found in 14 of 24 (58%) vulvar squamous carcinomas and in 15 of 19 (79%) VIN lesions. Nine patients (21%) had associated cervical neoplasia and six of these harbored HPV 16 in both lesions. Patients with recurrent intraepithelial neoplasia had a significantly higher incidence of HPV 16-positive lesions. No association was found with regard to the occurrence of multicentric disease or risk of malignant progression.
Adult↗
The prevalence of human papillomavirus type 16 and 18 DNA in cervical cancer in different age groups: a study on the incidental cases of cervical cancer in Norway in 1983.
In a study carried out to estimate the prevalence of genital Human Papilloma Virus (HPV) infection among cervical cancer patients in Norway, the following values were recorded using the southern blot hybridization method: 54.1% HPV 16, 9.5% HPV 18, 0% HPV 11, and 1.4% HPV 16 and 18 coinfection, yielding a total of 62.2% HPV positive cases in the study. An attempt to include a normal biopsy as "internal control" from each patient failed, as macroscopically normal tissue was often histologically abnormal. HPV infection did not relate to stage of disease, age, or 5-year survival, whereas the classic relationship between stage and survival was observed. A very high percentage of HPV positive cases occurred among older women, a fact which does not support the existence of a new HPV epidemic. The studied population is unique, as it represents 75 cases which comprise a representative sample of the 372 incidental cases of cervical cancer in Norway in 1983. To our knowledge this is the first study of its kind.
Adult↗
Intracellular maturation and sorting of two herpes simplex virus type 1 glycoproteins. Immunogold staining of ultrathin cryosections.
Simultaneous immunocytochemical triple staining of ultrathin cryosections of herpes simplex virus type 1-infected cells was carried out using monoclonal antibodies specific for glycoprotein C, glycoprotein D and alpha + beta tubulin. The viral glycoproteins were identified in the cytoplasm, in the Golgi sacs, on the plasma membrane and on the surface of intra- and extracellular virus particles, but not on the nuclear membrane. The glycoproteins identified in the cytoplasm outside the Golgi region were not always confined to the membranes of vesicles, but were often located in close proximity to the tubulin-labelled structures. The glycoproteins C and D were usually codistributed in the cytoplasm, and both accumulated in the Golgi sacs in the same membrane domains. As the glycoproteins occur in close proximity to the microtubular structures, we speculate that these might be directly involved in the intracellular transport of viral glycoproteins.
Freezing↗
Cytokeratin intermediate filament pattern in uterine cervical biopsies.
Twenty-seven uterine cervical biopsies with histological diagnoses ranging from normal through dysplasia to invasive carcinoma were analysed for cytokeratin pattern in two-dimensional gel electrophoresis. No direct correlation between histological diagnosis and cytokeratin pattern was observed.
Adenocarcinoma↗
Cytokeratin intermediate filament pattern and human papillomavirus type in uterine cervical biopsies with different histological diagnosis.
The cytokeratin pattern and the presence of human papillomavirus (HPV) were analyzed in 53 uterine cervical biopsies. The biopsies were histologically characterized and the diagnosis ranged from normal through dysplasia to carcinoma. The cytokeratins were identified by their immunological reactivity with the monoclonal antibodies AE1 and AE3. The tissue was typed for the presence of HPV types 11, 16 and 18. We have previously shown that there was no correlation between the expression of cytokeratins No. 14, 15, 16 and 19 (K14, K15, K16 and K19) and the histological diagnosis of cervical biopsies. The present study shows that the cytokeratin pattern cannot be correlated to HPV infection of the cervical tissue either.
Autoradiography↗
[Herpes virus infection and cancer of the uterine cervix].
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Female↗