Antonio Frega

Sensitivity, Specificity, and Clinical Value of Human Papillomavirus (HPV) E6/E7 mRNA Assay as a Triage Test for Cervical Cytology and HPV DNA Test

ABSTRACT There is evidence that testing for human papillomavirus (HPV) E6/E7 mRNA is more specific than testing for HPV DNA. A retrospective study was carried out to evaluate the performance of the PreTect HPV-Proofer E6/E7 mRNA assay (Norchip) as a triage test for cytology and HPV DNA testing. This study analyzed 1,201 women, 688 of whom had a colposcopy follow-up and 195 of whom had histology-confirmed high-grade intraepithelial neoplasia or worse (CIN2+). The proportion of positive results and the sensitivity and specificity for CIN2+ were determined for HPV mRNA in comparison to HPV DNA and cytology. All data were adjusted for follow-up completeness. Stratified by cytological grades, the HPV mRNA sensitivity was 83% (95% confidence interval [CI] = 63 to 94%) in ASC-US (atypical squamous cells of undetermined significance), 62% (95% CI = 47 to 75%) in L-SIL (low-grade squamous intraepithelial lesion), and 67% (95% CI = 57 to 76%) in H-SIL (high-grade squamous intraepithelial lesion). The corresponding figures were 99, 91, and 96%, respectively, for HPV DNA. The specificities were 82, 76, and 45%, respectively, for HPV mRNA and 29, 13, and 4%, respectively, for HPV DNA. Used as a triage test for ASC-US and L-SIL, mRNA reduced colposcopies by 79% (95% CI = 74 to 83%) and 69% (95% CI = 65 to 74%), respectively, while HPV DNA reduced colposcopies by 38% (95% CI = 32 to 44%) and by 15% (95% CI = 12 to 19%), respectively. As a HPV DNA positivity triage test, mRNA reduced colposcopies by 63% (95% CI = 60 to 66%), having 68% sensitivity (95% CI = 61 to 75%), whereas cytology at the ASC-US+ threshold reduced colposcopies by 23% (95% CI = 20 to 26%), showing 92% sensitivity (95% CI = 87 to 95%). In conclusion, PreTect HPV-Proofer mRNA can serve as a better triage test than HPV DNA to reduce colposcopy referral in both ASC-US and L-SIL. It is also more efficient than cytology for the triage of HPV DNA-positive women. Nevertheless, its low sensitivity demands a strict follow-up of HPV DNA positive-mRNA negative cases.

Imiquimod treatment of vulvitis circumscripta plasmacellularis

Vulvitis circumscripta plasmacellularis (VCP) is a rare but well-described condition affecting women of any age [1]. A biopsy is mandatory for VCP diagnosis because its clinical aspect is similar to that of several other conditions [2,3]. The present report describes 2 cases of VCP successfully treated with topical imiquimod (Aldara; 3-M Pharmaceuticals, Saint Paul, MN, USA). In March 1997, a 40-year-old woman underwent carbon dioxide laser vaporization of a vulvar wart caused by human papillomavirus (HPV). The results of an HPV test, performed by polymerase chain reaction, were positive for virus types 6 and 11. Follow-up tests results were negative. In November 2002 the patient complained that she had been experiencing vaginal discharge, vulvar erythema, pruritus, burning, and dyspareunia over

Human papillomavirus in virgins and behaviour at risk

Human papillomavirus (HPV) infection is one of the most common sexual transmitted diseases (STDs). We compared two groups of virgins with genital HPV lesions to evaluate the behaviour at risk in the transmission of HPV infection. Partners were also examined. HPV lesions were detected in 88 virgins, who have never had sexual intercourse. This can be due to vertical transmission, fomities and skin-to-skin contact. Many other hypothesis can be proposed to explain HPV genital infection, however, further studies are required.

Urinary hMG (Meropur) versus recombinant FSH plus recombinant LH (Pergoveris) in IVF: a multicenter, prospective, randomized controlled trial

To compare IVF outcome in ovarian stimulation protocols with recombinant FSH plus recombinant LH versus hMG, 122 patients were randomized into two study groups: group A, patients treated with urinary hMG, and group B, patients treated with rFSH plus rLH. The two groups proved to be comparable to the main IVF outcome (pregnancy rate, implantation rate, oocytes, and embryos quality), with an increasing risk of ovarian hyperstimulation in the Pergoveris group.

Pregnancy outcome after loop electrosurgical excision procedure for cervical intraepithelial neoplasia

To determine pregnancy outcomes among women who underwent loop electrosurgical excision procedure (LEEP).

Expression of HPV16 E5 down-modulates the TGFbeta signaling pathway

BackgroundInfection with high-risk human papillomavirus (HR-HPV) genotypes, mainly HPV16 and HPV18, is a major risk factor for cervical cancer and responsible for its progression. While the transforming role of the HPV E6 and E7 proteins is more characterized, the molecular mechanisms of the oncogenic activity of the E5 product are still only partially understood, but appear to involve deregulation of growth factor receptor expression. Since the signaling of the transforming growth factor beta (TGFbeta) is known to play crucial roles in the epithelial carcinogenesis, aim of this study was to investigate if HPV16 E5 would modulate the TGF-BRII expression and TGFbeta/Smad signaling.FindingsThe HPV16 E5 mRNA expression pattern was variable in low-grade squamous intraepithelial lesions (LSIL), while homogeneously reduced in high-grade lesions (HSIL). Parallel analysis of TGFBRII mRNA showed that the receptor transcript levels were also variable in LSILs and inversely related to those of the viral protein. In vitro quantitation of the TGFBRII mRNA and protein in human keratinocytes expressing 16E5 in a dose-dependent and time-dependent manner showed a progressive down-modulation of the receptor. Phosphorylation of Smad2 and nuclear translocation of Smad4 were also decreased in E5-expressing cells stimulated with TGFbeta1.ConclusionsTaken together our results indicate that HPV16 E5 expression is able to attenuate the TGFbeta1/Smad signaling and propose that this loss of signal transduction, leading to destabilization of the epithelial homeostasis at very early stages of viral infection, may represent a crucial mechanism of promotion of the HPV-mediated cervical carcinogenesis.

Prognostic Value of HPV E6/E7 mRNA Assay in Women with Negative Colposcopy or CIN1 Histology Result: A Follow-Up Study

Pap test, and especially HPV DNA test, identify a large group of women who do not have any clinically relevant lesions, i.e., CIN2+ (Cervical Intraepithelial Neoplasia grade 2 or worse), but who are at greater risk of getting lesions in the future. The follow up of these women needs new biomarkers with prognostic value. The objective of this study is to evaluate the prognostic value of E6/E7 mRNA over-expression assay (PreTect HPV-Proofer, Norchip) for 5 HR-HPV types (16, 18, 31, 33, and 45) for progression to CIN2+ after a negative colposcopy. This prospective study, conducted at four Italian centres, enrolled 673 women with either a negative colposcopy or a negative or CIN1 histology. The clinical end-point was histological confirmation of CIN2+. Women were classified at baseline according to mRNA results and managed according to local colposcopy protocols. At least one conclusive follow-up test was obtained for 347 women (25 months average lapse since recruitment, range 5–74). Only seven CIN2+ were detected during follow up, three among the 82 women positive for mRNA at baseline, two among the 250 negative (Fisher exact test, p = 0.02), and two among the 12 with an invalid test. Absolute CIN2+ risk was 6.7/1,000 person/years in the whole cohort. The absolute CIN2+ risk was 18.4/1,000 person/years and 3.6/1,000 person/years in mRNA-positive and mRNA-negative women, respectively. In conclusion, E6/E7 mRNA over-expression appears to be a good candidate as a prognostic biomarker to manage HR-HPV DNA-positive women with negative colposcopy or histology, particularly in order to decrease follow-up intensity in those who are negative.

Obstetric and neonatal outcomes of women with FGM I and II in San Camillo Hospital, Burkina Faso

PurposeFemale genital mutilation (FGM) is still performed in the world. Women who underwent FGM have marked psychological, gynecological and obstetric consequences. This article contributes to the spread of knowledge about obstetric and neonatal outcomes in women with FGM I and II.MethodsOur observational study compared the obstetric outcomes of 85 women with FGM I and II (case group) and 95 women without it (control group). We evaluated age, need of oxytocin during labor, duration of the expulsion phase, need of episiotomy, weight of the newborn, Apgar score at birth, resuscitation of the newborn, stillbirth. We observed the rate of cesarean sections and their main indications. We compared the rate of cesarean sections among the cases and the controls.ResultsControls were younger than women who underwent FGM. Intravenous oxytocin injection was higher in cases. The expulsion phase was longer in women with FGM than in the controls. FGM is related to a higher risk of episiotomy. Apgar score 9/10 was more frequently assigned to babies from mothers without FGM. There were more resuscitated babies and more stillbirth in the group of cases. Ten percent of all women underwent cesarean section. FGM is related to a higher incidence of cesarean section.ConclusionFGM is associated with a higher risk of gynecological and obstetrical consequences, acting on women’s health and also on the economy of resource limited countries. Because of migration, health professionals could interface with women who underwent FGM and have to know their related complications.

In vivo HPV 16 E5 mRNA: Expression pattern in patients with squamous intra-epithelial lesions of the cervix

BACKGROUND Human Papillomavirus (HPV) type 16 E5 is a small protein, which is reported to display transforming activity in vitro and in animal studies. The E5 transcriptional activity, however, has been rarely reported in vivo in literature. OBJECTIVES (a) To detect the E5 transcripts in vivo in a population of HPV 16 positive patients with abnormal cytology and (b) to correlate the level of expression to the degree of the cytological lesion. STUDY DESIGN AND METHODS 250 cytological samples of HPV positive women were obtained and tested for the E6/E7 mRNA expression. Patients were selected if HPV 16 only mRNA positive and with a cytology consistent with low-grade/high-grade squamous intra-epithelial (LSIL/HSIL) lesions. Selected patients were tested for the E5 transcripts by reverse RT PCR, comparing the expression level in vivo with a transfected HPV 16 E5 HaCaT cell line. RESULTS 27 HPV 16 E6/E7 mRNA positive LSIL/HSIL patients were selected. 13 out of 17 LSIL patients were tested positive for the E5 mRNA, showing an ample range of positivity. In the HSIL group 7 out of 10 patients were tested positive, displaying lower and more homogeneous levels of expression if compared with the transfected cells. CONCLUSION The HPV 16 E5 transcripts levels showed a broad distribution in vivo; the discrepancy was wider in LSIL patients, with HSIL patients displaying a more homogeneous profile. However, because of the limited number of patients, we could not draw a firm conclusion about the correlation between the E5 expression and the disease progression.

Impact of human papillomavirus infection on the neovaginal and vulval tissues of women who underwent surgical treatment for Mayer-Rokitansky-Kuster-Hauser syndrome

OBJECTIVE To evaluate the impact of human papillomavirus (HPV) infections on the neovaginal and vulval tissues of women who underwent surgical treatment for Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome. DESIGN Multicenter observational study. SETTING University and community hospitals. PATIENT(S) Thirty-three women who had previously undergone neovagina reconstruction due to MRKH and who were referred to our centers for the evaluation and treatment of HPV neovaginal/vulval-related lesions. INTERVENTION(S) HPV infections were confirmed by polymerase chain reaction analysis or hybrid capture 2 tests; the patients underwent vaginoscopy, pap smear, and biopsy of the lesion and were treated by laser vaporization. Follow-up was conducted for 5 years. MAIN OUTCOME MEASURE(S) HPV-related neovaginal/vulval lesions, HPV testing, follow-up, recurrence rate. RESULT(S) Seventeen patients showed vulval lesions, and 16 patients neovaginal lesions. HPV testing results were positive for low-risk HPVs in 27 patients and high-risk HPVs in six patients. All the vulval lesions were condylomata, whereas 10 neovaginal lesions were condylomata, three were vaginal intraepithelial neoplasia (VAIN) degree 1, two were VAIN degree 2, and one was an adenocarcinoma. Eight patients were lost to follow-up. Twenty patients tested positive for an HPV infection, and seven patients (28%) had a recurrence of the lesion in the follow-up time. CONCLUSION(S) Patients who underwent neovagina reconstruction have sexual relationships and are HPV exposed. These patients should be evaluated after surgery for HPV infections to prevent HPV-related diseases and cancers.