Recurrence of vulval intraepithelial neoplasia following treatment with cidofovir or imiquimod: results from a multicentre, randomised, phase II trial (RT3VIN).

OBJECTIVE: To compare the recurrence rates after complete response to topical treatment with either cidofovir or imiquimod for vulval intraepithelial neoplasia (VIN) 3. DESIGN: A prospective, open, randomised multicentre trial. SETTING: 32 general hospitals located in Wales and England. POPULATION OR SAMPLE: 180 patients were randomised consecutively between 21 October 2009 and 11 January 2013, 89 to cidofoovir (of whom 41 completely responded to treatment) and 91 to imiquimod (of whom 42 completely responded to treatment). METHODS: After 24 weeks of treatment, complete responders were followed up at 6-monthly intervals for 24 months. At each visit, the Common Terminology Criteria for Adverse Events (CTCAE) v3.0 was assessed and any new lesions were biopsied for histology. MAIN OUTCOME MEASURES: Time to histologically confirmed disease recurrence (any grade of VIN). RESULTS: The median length of follow up was 18.4 months. At 18 months, more participants were VIN-free in the cidofovir arm: 94% (95% CI 78.2-98.5) versus 71.6% (95% CI 52.0-84.3) [univariable hazard ratio (HR) 3.46, 95% CI 0.95-12.60, P = 0.059; multivariable HR 3.53, 95% CI 0.96-12.98, P = 0.057). The number of grade 2+ events was similar between treatment arms (imiquimod: 24/42 (57%) versus cidofovir: 27/41 (66%), χ2 = 0.665, P = 0.415), with no grade 4+. CONCLUSIONS: Long-term data indicates a trend towards response being maintained for longer following treatment with cidofovir than with imiquimod, with similar low rates of adverse events for each drug. Adverse event rates indicated acceptable safety of both drugs TWEETABLE ABSTRACT: Long-term follow up in the RT3VIN trial suggests cidofovir may maintain response for longer than imiquimod.

Effect of diindolylmethane supplementation on low-grade cervical cytological abnormalities: double-blind, randomised, controlled trial.

BACKGROUND: Cervical screening identifies many women with low-grade abnormalities. In vitro and in vivo studies have shown that diindolylmethane (DIM) could potentially halt (cervical) carcinogenesis. We report on a randomised controlled trial of the effect of DIM in women with low-grade cervical cytological abnormalities. METHODS: We conducted a pragmatic double-blind, randomised controlled trial of 150 mg DIM (from BioResponse DIM) or placebo daily for 6 months in women with newly diagnosed, low-grade cytological abnormalities. Randomisation was in the ratio 2 (DIM) to 1 (placebo). All women were invited for colposcopy at 6 months with biopsy of any abnormality. RESULTS: Of the 551 randomised women available for analysis, 9% on DIM and 12% on placebo had cervical intraepithelial neoplasia-2 (CIN2) or worse after 6-month supplementation (risk ratio (RR) 0.7 (95% confidence interval (CI): 0.4-1.2)), whereas 4.6% and 5.1%, respectively, had CIN3 or worse (RR 0.9 (95% CI: 0.4-2.0)). A total of 27.3% of women on DIM and 34.3% on placebo had no sign of disease (negative cytology, colposcopy and human papilloma virus (HPV) tests) at 6 months (RR 0.8 (95% CI: 0.6-1.0)). Of those HPV-positive at baseline, 69% (114 out of 166) of the DIM group were positive at 6 months compared with 61% (43 out of 71) of the placebo group: RR 1.1 (95% CI: 0.9-1.4). Diindolylmethane supplementation was well tolerated. CONCLUSION: The results suggest that short-term DIM supplementation (150 mg day(-1)) is well tolerated, but is unlikely to have an effect on cytology or HPV infection. Uncertainty remains regarding its effect on CIN2+.

Development of optimal liquid based cytology sample processing methods for HPV testing: minimising the 'inadequate' test result.

Incorporation of HPV testing into cervical screening is anticipated and robust methods for DNA extraction from liquid based cytology (LBC) samples are required. This study compared QIAamp extraction with Proteinase K digestion and developed methods to address DNA extraction failure (ß-globin PCR negative) from clinical specimens. Proteinase K and QIAamp extraction methods in paired LBC samples were comparable with adequate DNA retrieved from 93.3% of clinical specimens. An HPV prevalence cohort (n=10,000) found 7% (n=676) LBC samples tested negative for ß-globin, and were classified as inadequate. This 'failure' rate is unsuitable for population screening, particularly as the sampling method is intrusive. 379/676 samples were assessed to determine the cause of test failure. Re-testing confirmed adequate DNA in 21.6% of the original extracts; re-extraction from stored material identified 56.2% samples contained adequate material; dilution to overcome sample inhibition (1:10) resolved 51.7% cases in original extracts and 28% in new extracts. A standardised approach to HPV testing with an optimal DNA concentration input rather than standard volume input is recommended. Samples failing initial DNA extraction should be repeat extracted and assessed for sample inhibition to reduce the 7% of HPV tests being reported as inadequate and reduce the need for retesting of those women to <1%.

The prevention of cervical cancer in Africa.

In this article we aim to draw attention to the burden of cervical cancer in Africa for reproductive health and review strategies for prevention, including appropriate noncytology-based cervical screening and prophylactic human papillomavirus vaccination. We consider the heavy burden of disease attributable to human papillomavirus infection borne by developing countries, particularly in Africa. Following identification of the human papillomavirus as the infectious etiological agent and elucidation of the long natural history of cervical neoplasia, cervical cancer is now one of the most preventable of all cancers. Opportunities for primary prevention by prophylactic vaccination and secondary prevention by appropriate cervical screening are discussed, together with the importance of population coverage. Qualitative work on attitudes towards cervical cancer prevention, education needs, the creation of an environment for informed choice and uptake are essential aspects of effective prevention programs. Cervical cancer poses a huge health burden in Africa. It is a disease that is eminently preventable given political will, the availability of affordable vaccines, appropriate cervical screening and access to cheap, point-of-care human papillomavirus testing. There are a number of unanswered questions for the prevention of cervical cancer and a need for demonstration projects to address these and further develop prevention strategies.

Prophylactic HPV vaccination: a major breakthrough in the fight against cervical cancer?

Cervical cancer is the second most common female cancer with 500000 new cases and 290000 deaths occurring worldwide per annum. Organized cervical screening programs have reduced the incidence and mortality of cervical cancer. However, in developing countries scarce resources, poverty, lack of infrastructure and disenfranchisement of women have been major hurdles in the effective implementation of routine screening programmes. As a result, 83% of cervical cancers still occur in the developing countries and account for 15% of all female cancers. Epidemiological studies have established a causative role of Human Papillomavi-rus (HPV) infection in the development of cervical cancer. The development and implementation of a prophylactic HPV vaccine will have a major impact on preventing this global disease. However, long-term surveillance of the HPV vaccination program will be required to confirm the expected reduction in cervical cancer incidence. This article reviews the role of HPV in the development of cancer and the burden of HPV related cancers; types and pharmacokinetics of HPV vaccines; challenges and issues in implementing vaccination programmes; screening in the developing and developed countries and screening options in the post-vaccination era.

Comparison of the PapilloCheck DNA micro-array Human Papillomavirus detection assay with Hybrid Capture II and PCR-enzyme immunoassay using the GP5/6+ primer set.

BACKGROUND: Cervical screening detects precancerous cells and routine screening could be improved by testing for Human Papillomavirus (HPV), the virus that causes cervical cancer. HPV infection is common and the benefit of HPV testing would be identification of women who are HPV negative and at low risk of developing cancer. STUDY DESIGN: The aim of this study was to evaluate the Greiner Bio-one PapilloCheck micro-array assay (PapilloCheck) for detection of HPV in comparison with Hybrid Capture II (hc2) and PCR-enzyme immunoassay (PCR-EIA) using the GP5/6+ primers. RESULTS: Samples from a cytologically defined population (n=878) were analysed and 187 samples also had histology information. Overall, 674 out of 878 samples gave a consistent result (76.8%; 95% CI 73.83-79.52%) on all three platforms. The genotype results obtained by PapilloCheck and PCR-EIA were compared and 94% were consistent (95% CI 92.1-96.4%). The main difference was the poor Kappa agreement for detection of high risk (HR) type 35 (Kappa=0.190) with all inconsistent results being HR positive by PCR-EIA assay but negative on the PapilloCheck platform. There was no statistically significant difference between the performance of each assay when HR HPV positive samples were linked with clinical result (cytology and histology grade). PapilloCheck detected the highest number of HR HPV infections in samples with histology confirmed as CIN1, CIN2 and CIN 3 (76.6%, 85% and 91.7%, respectively). CONCLUSIONS: Overall, PapilloCheck proved to be a sensitive, reproducible, robust molecular assay for HPV genotyping with the potential for high throughput of specimens in a clinical setting.

Human Papillomavirus negative but dyskaryotic cervical cytology: re-analysis of molecular testing.

BACKGROUND: Evaluation of molecular Human Papillomavirus (HPV) testing into UK Cervical Screening Programmes is underway. In South Wales the current HPV prevalence in women attending routine screening is 13.5% with 76.3% HR HPV positive in cases with reported dyskaryotic cervical cytology [Hibbitts S, Jones J, Powell N, Dallimore N, McRea J, Beer H, et al. Human papillomavirus prevalence in women attending routine cervical screening in South Wales, UK: a cross-sectional study. Br J Cancer 2008;99(December (11)):1929-33]. OBJECTIVES: The aim of this study was to re-analyse the 23.7% cases with reported dyskaryotic cytology that were HR HPV negative (n=52 out of 219 in a population of 10,000). STUDY DESIGN: Three procedures were performed: (i) GP5+/GP6+ PCR-EIA repeat on original DNA extracts; (ii) DNA extraction and GP5+/GP6+ HPV PCR-EIA; (iii) DNA extraction and HPV typing using Greiner Bio-One PapilloCheck DNA microarray. RESULTS: 51 out of 52 samples were re-analysed. Direct repeat HPV PCR-EIA identified 24% (n=12/51) of samples positive for HR HPV. Re-extracted DNA and PCR-EIA increased detection to 41.2% (n=21/51) and PapilloCheck detected 78.4% (n=40/51). HR HPV detection by PapilloCheck was significantly higher compared with the other methods of re-analysis. Eleven samples were persistently HR HPV negative but 4 tested positive for low risk HPV. CONCLUSIONS: This study identifies that up to 78% of samples with dyskaryotic cervical cytology that test negative for HPV can be found to be HPV positive on re-analysis. The reliance on a single negative HPV test result could lead to missed HPV related disease in a subset of patients, the number dependant on which HPV test is performed. The clinical significance of a false negative HPV result depends on the screening interval and how HPV testing is incorporated into screening.

Prophylactic HPV vaccination for women over 18 years of age.

Cervical screening has resulted in a major reduction in the incidence and mortality of cervical cancer in the UK and other developed countries. Nevertheless approximately 2700 women present with cervical cancer in the UK each year with mortality in excess of 1000 cases. Prophylactic HPV vaccination against HPV 16 and 18 has been shown to be highly effective in preventing HPV related malignancy in clinical trials. Newly introduced HPV vaccination programmes in the UK and elsewhere are ultimately likely to result in a further significant reduction in the incidence and mortality of cervical cancer. These vaccination programmes will be most effective in early adolescence when prevalence of HPV infection is low. Consequently, vaccination programmes in the UK have been initially targeted at 12 to 13-year olds. In Australia favourable estimates of cost effectiveness have supported funding of a 'catch-up' programme to 26 years. In the UK the catch up programme has for the present been restricted to 18 years for cost effectiveness reasons. In addition the value of HPV vaccination beyond 26 years has not yet been fully clarified. Nevertheless women up to 45 years of age have been shown to exhibit strong immune responses to the bivalent HPV vaccine which might be expected to reduce the risk of HPV re-infection and address the second peak of HPV related malignancy in later life, evident over 45 years of age. Early data from randomised trials testing the quadrivalent HPV vaccine in women over 25 years has suggested high vaccine efficacy comparable to younger women. This paper will explore the evidence supporting HPV vaccination in HPV naïve and HPV exposed sexually active women up to 26 years and beyond this age group.

Human papillomavirus prevalence in women attending routine cervical screening in South Wales, UK: a cross-sectional study.

In this cross-sectional population-based study we determine human papillomavirus (HPV) prevalence in South Wales to provide comprehensive baseline data for future assessment of the impact of prophylactic HPV vaccination and to help inform future screening strategies. Liquid-based cytology samples from women attending routine cervical screening were collected (n=10 000: mean age 38 years, 93% cytology negative, and 64.8% from the 50% least deprived LSOA according to social deprivation score (SDS)). High-Risk (HR) and Low-Risk HPV screening was performed using HPV PCR-EIA with genotyping of HR positives and data correlated with age, SDS and cytology. Overall HPV prevalence was 13.5% (9.3% age standardised) and the most frequent HR types were HPV 16, 31, 18 and 58. In HR HPV-positive cases 42.4% had a single HR type and they were predominant in women with severe cytological abnormalities. Here, 66% of all HR HPV cases were in women aged 30 years of age or less and SDS had no significant effect on HPV status. HPV prevalence increased significantly with degree of dyskaryosis from 7% in cytology negative samples to 80% in samples with severe cytological abnormalities (P-value <0.0001). Overall, 46% of HR HPV cases were positive for the two HR types targeted by the prophylactic vaccines (HPV 16 and HPV 18). The data presented represents the largest type-specific investigation of HPV prevalence in an unselected UK population.

Human papillomavirus infection in Barrett's oesophagus in the UK: an infrequent event.

BACKGROUND: Human papillomavirus (HPV) infection has been reported in squamous cell carcinomas of the oesophagus and has been recently described in Barrett's oesophagus, a premalignant condition which may give rise to oesophageal adenocarcinoma. OBJECTIVES: To investigate HPV infection in Barrett's oesophagus in a UK population. STUDY DESIGN: DNA was extracted from 73 Barrett's oesophagus biopsies and examined for the presence of DNA for 14 high risk (HR) and 6 low risk (LR) HPV types. RESULTS: HPV DNA was present in only 1 of 73 samples; genotyping indicated this was a high risk type 51 infection. CONCLUSIONS: HPV infection appears unlikely to be a significant factor in the aetiology of Barrett's oesophagus in the UK.

Contribution of demographic, psychological and disease-related factors to quality of life in women with high-grade vulval intraepithelial neoplasia.

OBJECTIVES: To quantify the effect of demographic, psychological and disease-related factors on Quality of life (QoL) outcomes in women with high-grade vulval intraepithelial neoplasia (VIN2-3). To obtain qualitative data on the effect of disease and treatment in these women and their partners. To assess the participants' perception of their risk of developing of vulval cancer and its relation to QoL outcomes. METHODS: A questionnaire was constructed using existing instruments to measure the effect of demographic, psychological and disease-related factors on QoL outcomes. Free text space was provided for qualitative data. The questionnaire was mailed to women attending two specialist VIN clinics. RESULTS: One hundred and fifty women were invited for the study. Eighty-two responded (54.6%) of which forty-four (53.6%) were sexually active. Demographic factors (age and or living situation) had a significant effect on emotional health and body image. Psychological factors (anxiety and depression) had a significant effect on all aspects of QoL. Disease-related factors did not have a measurable effect on QoL outcomes, although the qualitative data revealed that various aspects of VIN had affected the lives of these women and their partners. There was a significant positive association between a perceived risk of developing vulval cancer with worsening general and emotional health. CONCLUSION: Psychological co-morbidity and various demographic factors should be considered while managing women with VIN. Accurate information regarding the development of vulval cancer should be given. The findings of this preliminary study will assist the construction of VIN-specific QoL instruments in the future.

Will vaccination against human papillomavirus prevent eye disease? A review of the evidence.

The role of human papillomavirus (HPV) infection in eye disease is controversial. However, a recent case illustrates the possible role of HPV in conjunctival squamous carcinoma and the potentially devastating effects of this disease. The development of two vaccines to prevent infection with HPV types most commonly associated with anogenital cancers has led to debate about the pros and cons of a national immunisation programme to prevent cervical cancer. The introduction of such a vaccination programme may have an additional beneficial effect on the occurrence of some head and neck, including ocular, cancers. This review discusses the nature of papillomaviruses, mechanisms of infection and carcinogenesis, the possible role of HPV in eye disease, and finally the likely impact of the new prophylactic vaccines.

Variation in human papillomavirus type-16 viral load within different histological grades of cervical neoplasia.

The objective of this study was to investigate variation in human papillomavirus (HPV) type-16 load within histologically defined grades of cervical intraepithelial neoplasia. Two hundred and thirty-seven liquid based cytology samples were collected from women attending colposcopy clinics, DNA was extracted, and presence of virus determined by PCR-enzyme immunoassay. Quantitative real-time PCR was used to determine viral load for 70 HPV-16 positive single infections. Viral load was expressed as the ratio of copies of the viral L1 gene to copies of the human beta-globin gene. Measurements varied from 0.019 to 4,194 HPV genomes per cell. Our data demonstrate that in cervical neoplasia, HPV load tends to correlate with disease severity, but that the number of viral genomes/cell varies considerably within histological grades. This variation within disease grades currently limits the clinical utility of viral load measurement.

Human papilloma virus (HPV) prophylactic vaccination: challenges for public health and implications for screening.

Prophylactic vaccination against high risk human papilloma virus (HPV) 16 and 18 represents an exciting means of protection against HPV related malignancy. However, this strategy alone, even if there is a level of cross protection against other oncogenic viruses, cannot completely prevent cervical cancer. In some developed countries cervical screening programmes have reduced the incidence of invasive cervical cancer by up to 80% although this decline has now reached a plateau with current cancers occurring in patients who have failed to attend for screening or where the sensitivity of the tests have proved inadequate. Cervical screening is inevitably associated with significant anxiety for the many women who require investigation and treatment following abnormal cervical cytology. However, it is vitally important to stress the need for continued cervical screening to complement vaccination in order to optimise prevention in vaccinees and prevent cervical cancer in older women where the value of vaccination is currently unclear. It is likely that vaccination will ultimately change the natural history of HPV disease by reducing the influence of the highly oncogenic types HPV 16 and 18. In the long term this is likely to lead to an increase in recommended screening intervals. HPV vaccination may also reduce the positive predictive value of cervical cytology by reducing the number of truly positive abnormal smears. Careful consideration is required to ensure vaccination occurs at an age when the vaccine is most effective immunologically and when uptake is likely to be high. Antibody titres following vaccination in girls 12-16 years have been shown to be significantly higher than in older women, favouring vaccination in early adolescence prior contact with the virus. Highest prevalence rates for HPV infection are seen following the onset of sexual activity and therefore vaccination would need to be given prior to sexual debut. Since 20% of adolescents are sexually active at the age of 14 years, vaccination has been suggested at 10-12 years. However, parental concerns over the sexual implications of HPV vaccination may reduce uptake of vaccination thereby reducing the efficacy of an HPV vaccination programme. Concerns have already been raised over the acceptability of a vaccine preventing a sexually transmitted infection in young adolescents, particularly amongst parents or communities who consider their children to be at low risk of infection. This may be a particularly sensitive issue for ethnic minority groups. This paper considers the factors which will influence the efficacy of a public HPV vaccination programme and its impact on cervical screening.

Survival in ovarian cancer in Wales: Prior to introduction of all Wales guidelines.

The objective of the study was to review referral practice, overall management, and survival in women with suspected ovarian cancer in Wales. This study was done prior to introduction of cancer management guidelines in the region. A confidential study questionnaire was sent to 20 participating hospitals. Data on 287 consecutive women with suspected ovarian cancer were collected, of which 250 women underwent primary laparotomy. Information was obtained on referral pattern, preoperative investigations, place of primary surgery, specialty of the primary surgeon, surgical parameters recorded at the time of operation, a final overall stage, adjuvant treatment, and survival outcome. There was a wide variation in referral practice and management of ovarian cancer in Wales. Stage of the disease, attempt at optimal debulking, residual disease, management by a cancer centre multidisciplinary team, and platinum-based chemotherapy were associated with improved overall survival and progression-free survival. More women were alive if managed in the cancer centre at 1 and 3 year after diagnosis (P = 0.022). This study has highlighted the acute issue of the standards of clinical care in the area of ovarian cancer management and will emphasize the implementation of better care pathways for ovarian cancers.

Human papillomavirus infection: an anonymous prevalence study in South Wales, UK.

The objective of this study was to describe human papillomavirus (HPV) prevalence in South Wales in relation to age, cytology and social deprivation. This was an unlinked, prospective, anonymous, population-based study. DNA was purified from 1911 liquid-based cytology samples (mean age 37.7 years, cytology 93.2% negative, social deprivation average score 17.9) using quality assured techniques and the presence of virus determined by PCR-Enzyme Immuno Assay (PCR-EIA). 209 (10.9%) samples contained high-risk (HR) HPV infection of which 36.4% had multiple HR-HPV types. The most frequent HR types were HPV 16 (19.6%), HPV 35 (9.5%), HPV 66 (9.2%), HPV 59 (8.5%) and HPV 56 (7.6%). There was a strong association between HPV infection and cytological abnormality. Significantly more HR-HPV infections were detected in women under the age of 30 years (68.9% of all HR-HPV infections Fisher's exact test P=0.0001) compared to 30 years and above. There was no difference in HPV prevalence between different socioeconomic groups. The data presented suggest a different HPV type distribution in South Wales in comparison to that reported for other populations.

Cervical screening in 20-24-year olds.

BACKGROUND: In 2003 the NHS Cervical Screening Programme in England changed the age at which women are first invited for cervical screening from 20 to 25 years. The aim of this review was to assess the evidence for benefit and harm of undertaking cervical screening in Wales for women aged 20-24 years. METHOD: A literature review looking for evidence of the effectiveness and potential harmful effects of cervical screening was undertaken. Welsh data for the number of cases of high-grade cervical intraepithelial neoplasia (CIN3) were examined and rates of invasive cervical cancer (1981-2003) for young women calculated. The medical notes of women less than 25 years old diagnosed with cervical cancer were reviewed and a cost analysis was performed. RESULTS: The literature review failed to identify any randomized controlled studies of the effectiveness of cervical screening in young women, but demonstrated that organized screening programmes result in a decrease in the incidence of cervical carcinoma. Following the introduction of the Welsh organized call/recall cervical screening programme in 1988, cervical cancer has been reduced by 58% in women aged 20-24 years and 45% in women aged 25-29 years (mean age-specific rate per 100,000 women aged 20-24 years: 4.2 in 1981-88 compared with 2.2 in 1989-2003). If these changes can be attributed to the screening of women aged 20-24, then the costs of at least pound sterling 82,500 are estimated to prevent one cervical cancer in this age group. CONCLUSIONS: The incidence of cervical cancer in young women has halved since the introduction of the Welsh-organized call-recall cervical screening programme. In Wales we recommend that women continue to be invited for cervical screening from 20 years of age. This will provide the information required to compare the incidence and stage at diagnosis of cervical cancer in young women invited for first time cervical screening at different ages across the UK.