[More effective screening prevents cervical cancer].
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Biomedical subjects
Publications and source records attributed to E Rylander.
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One hundred and forty-eight randomly chosen neutral-buffered formaldehyde-fixed cervical biopsies in which cervical intra-epithelial neoplasia (CIN) I-III had been diagnosed were tested for HPV (human papilloma virus) DNA by in situ hybridization (ISH) and polymerase chain reaction (PCR). For ISH, we utilized a biotinylated panprobe and type-specific, genomic probe sets. For PCR, we used the general primers GP5/GP6 and their recently described, elongated version GP5+/GP6+, and included the modification of hot-start PCR. Amplified DNA was detected by gel electrophoresis and slot blot hybridization. The positivity rate of ISH was 59% for all biopsies and 69%, 62% and 46% for CIN I, II and III, respectively. The sensitivity of GP5/GP6 was 74% with cold-start PCR and 78% with hot-start PCR. When GP5+/GP6+ was used, the sensitivity increased to 89% with cold-start PCR and to 95% with hot-start PCR. Based on the most sensitive PCR technique, HPV detection was 93%, 95% and 96% in CIN I, II and III, respectively. The number of HPV types decreased with the severity of the lesion, and HPV 16 was the predominant type. Multiple HPVs were rare and almost all HPV-positive cases could be typed. ISH and slot blot hybridization correlated well regarding HPV typing specificity. Our results confirm that distinct HPV types are present in a high proportion of cases of CIN. The sensitivity of ISH is lower than that of PCR. Furthermore, the modified general primers GP5+/GP6+ give a higher yield than GP5/GP6, while hot-start PCR increases sensitivity even further.
The polymerase chain reaction (PCR), used to detect human papillomavirus (HPV), is finding increasing applications in clinical laboratories. The standard method of analysis to detect amplified PCR products is ethidium bromide gel electrophoresis combined with labor intensive blot hybridization. In this study, we describe single-strand conformation polymorphism (SSCP) to detect and genotype simultaneously general primer GP5+/GP6+ amplified HPV DNA using semiautomated electrophoresis on polyacrylamide gels (PAGE) combined with sensitive silver staining. To establish a standard for the band patterns of the various HPV types, we used HPV plasmid DNA, which allowed us to distinguish HPV 6, 11, 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58, covering the most frequently recognized types. All the types tested are separated from each other, demonstrating diverse band patterns, HPV 16 being the most distinct. We also investigated PCR-SSCP for HPV detection and typing of 86 cervical biopsies diagnosed as cervical intraepithelial neoplasia (CIN) I-III and known to be HPV positive by PCR-slot blot hybridization and in situ hybridization. The correlation with SSCP was 91% for in situ hybridization and 98% for PCR-slot blot hybridization. SSCP is reproducible and specific. Its sensitivity is comparable to slot-blot hybridization. The interval to SSCP is approximately 2 h after PCR compared with several days' work when using conventional blot hybridization. We concluded that SSCP may be more advantageous than other PCR-based typing technologies.
Risk factors for severe wheezing bronchitis were studied in children aged 4 months to 4 years in need of hospitalization. The children included in the study consisted of all cases generated from a geographically defined population, the catchment area of St Göran's hospital in Stockholm. The incidence was 3/1000 children and year, during the two years of observation, with the highest rate in boys under the age of 18 months (4.7/1000). Symptoms of a preceding upper respiratory tract infection were reported in 90% of the cases, but a viral etiology could only be demonstrated with virus isolation in 26%. Respiratory syncytial virus was the most common finding in younger children. Rhinovirus was primarily seen in older children with a history of previous wheezing. Regardless of whether the cases had a positive or negative virus isolation they showed the same seasonal distribution. Furthermore, there was no difference in risk factors between children with a positive and negative virus isolation. Children older than 18 months with negative virus isolation had higher IgE levels than those with positive isolation, suggesting that atopy is of greater importance in this group.
BACKGROUND: A population-based case-control study was performed in Stockholm to assess the influence of air pollution on the occurrence of severe wheezing bronchitis in children. METHODS: The study included 197 children aged 4 months to 4 years, who were hospitalized because of breathing difficulties with wheezing, and 350 population controls. Information on potential risk factors for childhood wheezing and a residential history was obtained at home interview with parents. Outdoor nitrogen dioxide (NO2) concentrations at home addresses and day care centres from birth on were estimated from validated models, mainly using data on traffic intensity from municipal registers. RESULTS: The risk of wheezing bronchitis was related to time-weighted mean outdoor NO2 exposure in girls (P = 0.02), but not in boys. A gas stove in the home appeared to be a risk factor primarily for girls. All analyses controlled for parental asthma and maternal smoking, which were independent risk factors for wheezing bronchitis. CONCLUSIONS: The results suggest that exposure to combustion products containing NO2 may be of particular importance for the development of wheezing bronchitis in girls.
The prevalence of human papillomavirus (HPV) infection in cervical cell scrapes from a cohort of 276 young women was determined by a general two-step polymerase chain reaction. HPV infection fluctuated among young women during a 2-year interval. The total prevalence of HPV infection decreased from 21% to 8.3%. The most prevalent HPV types at enrollment were HPV-16 (3.3%) and HPV-6 (2.9%). At follow-up, the most common type was HPV-16 (2.9%), while no HPV-6 was detected. In 2 women only, the same HPV type persisted. Regression of HPV infection was found in 80% of the women. A new HPV type-specific infection was detected in 7.2% of the women and was independently associated with a new sex partner or an abnormal smear since enrollment.
We conducted a case-control study to assess the role of exposure to environmental tobacco smoke (ETS) in the development of wheezing bronchitis in children. The study included 199 children age 4 months to 4 years, who were treated in hospital for wheezing bronchitis, and 351 population controls of the same age group. We estimated exposure to ETS from urinary cotinine measurements as well as from questionnaires to parents. The median urinary cotinine concentration was 5.7 micrograms per liter in cases and 4.4 micrograms per liter in controls. Breast-feeding was related to urinary cotinine excretion in children with smoking mothers. The risk of wheezing bronchitis increased in relation to parental smoking and urinary cotinine concentration. This effect was most prominent in children up to 18 months of age, among whom the relative risk was 3.3 in those with a urinary cotinine level exceeding 20 micrograms per liter (95% confidence interval = 1.5-7.6). Our data confirm that ETS is an important risk factor for wheezing bronchitis in children and indicate that a single urinary cotinine measurement offers no major advantages to questionnaire data for assessment of long-term exposure to ETS.
OBJECTIVES: To assess the prevalence of lower genital tract symptoms and the association between reported symptoms and past and present signs of sexually transmitted diseases (STD) in young women. DESIGN: All women belonging to the 19-, 21-, 23- and 25-year age cohorts and living in the catchment area of the community health centre, were invited by mail to take part in a population-based study. The participants answered a structured questionnaire and a gynaecologic examination was performed. Samples for wet smear, cervical Pap smear, HPV DNA determination and Chlamydia trachomatis culture were taken at the gynaecologic examination. The presence of genital warts was noted. A blood sample was analysed for antibodies against C trachomatis and HSV-2. SETTING: The community health care centre was located in Umeå, a city in Northern Sweden. RESULTS: Of the 886 women who were eligible, 611 (70%) participated in the investigation. One out of four women reported symptoms from the lower genital tract. The most commonly reported symptoms were itching, followed by discharge, and soreness. The most commonly reported STD was C trachomatis (15%). The most prevalent present STD was HPV infection (20%) whereas C trachomatis infection could be isolated from 2.7% of the women. Antibodies against C trachomatis and HSV-2 were present among 22% and 6% of the women, respectively. There was a significant correlation between the women's complaint of vaginal discharge and previous C trachomatis infection, lack of lactobacilli and presence of leucocytosis in wet smear. CONCLUSIONS: We have in a population-based study of young healthy women found that one out of four women had some kind of lower genital tract complaint. Itching was the most commonly reported symptom and was associated with pseudohyphae and acetowhite patches. Reported vaginal discharge and soreness were associated with the history of a past C trachomatis infection and signs of a disturbed vaginal flora.
BACKGROUND AND OBJECTIVES: To assess the presence of human papillomavirus (HPV) DNA in vulvar papillomatosis, since some women with this complaint have symptoms associated with HPV infection, such as itching, burning, and dyspareunia. GOAL: To reassure the patients that they do not have a transmissible viral disease, by excluding a HPV origin of their condition. STUDY DESIGN: Vulvar biopsy specimens from 22 females with vulvar papillomatosis, from 10 females with prominent aceto-white vulvar lesions, and from 14 healthy controls were analyzed histologically for signs of HPV infection and by in situ hybridization and polymerase chain reaction for the presence of HPV DNA. RESULTS: Specimens from women with vulvar papillomatosis showed some histological signs associated with HPV infection, but no koilocytotic atypia or dysplasia and thus resembled normal-looking vulvar mucosa. Aceto-white lesions frequently displayed histological features suggestive of HPV infection, including dysplasia. HPV DNA was detected in 6 of 10 patients with aceto-white lesions, but only in 1 of 22 patients with papillomatosis and in no healthy controls. CONCLUSIONS: According to our results, HPV DNA, which is generally found in cervical lesions and subclinical infections, is not present in vulvar papillomatosis even though symptoms associated with HPV infection are frequent complaints.
In order to estimate the prevalence of HPV infection in male partners of women with squamous intraepithelial lesions and/or high risk HPV, we examined 25 men. In situ hybridisation or polymerase chain reaction was used to detect HPV DNA in biopsied tissue or cell samples from the genital epithelium. Twenty (80%) of the male consorts had clinical features suggestive of HPV infection. Of these, 18 (90%) had detectable HPV DNA, 11 (65%) of the high risk type. In 9 cases HPV DNA was detected by in situ hybridisation and in 9 by polymerase chain reaction. Concordance between female and male HPV type was found in 8 cases (32%), but regarding high-risk HPV carriage as such, 10 (40%) couples corresponded. A search for HPV infection in male partners of women known to be infected with high-risk HPV seems worthwhile.
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OBJECTIVE: To determine whether young women who have not experienced sexual intercourse may harbor genital human papillomavirus (HPV) infection in the vaginal-ectocervical mucosa. METHODS: We included 151 women, 10-25 years of age, attending two adolescent health care units (Stockholm and Uppsala) and one primary health care center (Umeå). The size of the hymenal orifice, use of tampons, and the habit of digital vaginal manipulation were registered. Samples of epithelial cells were collected from the vagina and analyzed for the presence of HPV using polymerase chain reaction. RESULTS: One hundred thirty of 154 samples contained an adequate number of cells. Two samples were HPV 6 DNA-positive. None were HPV 16 DNA-positive. None of the women had external genital warts. In 84%, the hymenal opening was 15 mm or less. Forty-eight percent of the women used tampons during periods. Fifty-four percent had inserted their own finger into the vagina and in 23%, a boyfriend's finger had penetrated the vagina. CONCLUSION: Human papillomavirus is rarely present vaginally in virginal women, even with the use of tampons or digital penetration.
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A population-based case-control study was performed to investigate etiologic factors for wheezing bronchitis and asthma in children up to four years of age. A total of 199 children hospitalized for the first time with these diagnoses at a major hospital in Stockholm in 1986-1988 constituted the cases, 351 children from the catchment area of the hospital were used as controls. Information on known and suspected risk factors was obtained through home interviews with a parent. Parental smoking was associated with a relative risk of 1.8 (95% confidence interval 1.3-2.6) corresponding to a population attributable proportion of 27%. The strongest association was seen for maternal smoking and children below 18 months of age. Other major risk factors included atopic heredity, recurrent upper respiratory tract infections and breast-feeding less than 3 months, which appeared to interact multiplicatively with parental smoking. The environmental factors had a stronger influence in the youngest age group, and the overall attributable proportion associated with parental smoking, short breast-feeding period and exposure to pets in the household was 43%. It is clear that successful primary prevention could dramatically reduce the incidence of wheezing bronchitis in children.
Cervicovaginal cell samples were analyzed for the presence of human papillomavirus type 16 deoxyribonucleic acid. All 99 women included in the study had normal Papanicolaou smear results, normal findings on wet smear, and no clinical signs of genital papillomavirus or any other genital infection. Deoxyribonucleic acid polymerase chain reaction was performed with three different human papillomavirus type 16-specific primer pairs from the early 6, early 7, and upstream regulatory regions. Human papillomavirus type 16 was detected in 21% of the 99 women. Seven of 69 women (10%) who were not taking any medication except for cyclic estrogen or progestin replacement therapy were carriers of human papillomavirus type 16. Seven of 19 women (37%) who used hormonal contraceptives or who sought early pregnancy termination were carriers of human papillomavirus type 16. All 7 women with diseases that required frequent hospital care were carriers of human papillomavirus type 16. The 4 women who had never had sexual intercourse were not carriers of human papillomavirus type 16. Our results indicate that the human papillomavirus type 16 prevalence in women may reflect to some extent the general health conditions of the patients.
AIMS: To compare two commercially available pan probes for the identification of human papillomavirus (HPV) DNA expression in histological sections and to type the HPV positive cases. METHODS: 97 formalin fixed, paraffin wax embedded biopsy specimens from the genital tract were tested for HPV positivity with in situ hybridisation using biotinylated cDNA pan probes--Probemix (Enzo) and OmniProbe (Digene). The HPV positive cases were further tested with HPV types 6/11, 16/18, and 31/33/35/51, and the HPV type was related to the histological diagnosis. Formalin fixed, HeLa cells (10-50 HPV 18 copies per cell) and SiHa cells (1-2 HPV 16 copies per cell) were used as reference cell lines. RESULTS: 32% of the specimens gave positive nucleic signals with both Probemix and OmniProbe. Of these, 84% could be further characterised with regard to HPV types 6/11, 16/18, and 31/33/35/51; 4% of all cases were positive with either Probemix or OmniProbe. The concordance of these probes was high, 96% altogether. HeLa cells stained positive but SiHa cells did not. CONCLUSION: There is no difference between Probemix and OmniProbe for the general detection of HPV. The mean detection limit of these probes is about 20 copies a cell.