Michael Sindos

Differential regulation of different human papilloma virus variants by the POU family transcription factor Brn-3a

The Brn-3a POU family transcription factor is overexpressed in human cervical carcinoma biopsies and is able to activate expression of the human papilloma virus type 16 (HPV-16) upstream regulatory region (URR), which drives the expression of the E6 and E7 oncoproteins. Inhibition of Brn-3a expression in human cervical cancer cells inhibits HPV gene expression and reduces cellular growth and anchorage independence in vitro as well as the ability to form tumours in vivo. Here, we show that Brn-3a differentially regulates different HPV-16 variants that have previously been shown to be associated with different risks of progression to cervical carcinoma. In human cervical material, Brn-3a levels correlate directly with HPV E6 levels in individuals infected with a high risk variant of HPV-16, whereas this is not the case for a low-risk variant. Moreover, the URRs of high- and intermediate-risk variants are activated by Brn-3a in transfection assays, whereas the URR of a low-risk variant is not. The change of one or two bases in a low-risk variant URR to their equivalent in a higher-risk URR can render the URR responsive to Brn-3a and vice versa. These results help explain why the specific interplay between viral and cellular factors necessary for the progression to cervical carcinoma only occurs in a minority of those infected with HPV-16.

Triage by HPV-DNA testing: is it useful in women with persistent minor smear abnormalities?

Background.  This study was carried out to evaluate the efficacy of HPV‐DNA (Human Papilloma Virus) testing as a triage strategy for persistent borderline and mild cytological abnormalities.

Prenatal diagnosis of Wolf–Hirschhorn syndrome: ultrasonography and genetics

A 36-year-old primigravida attended our Fetal Medicine Unit for the routine second trimester anomaly scan at 19 weeks of her pregnancy. Her first trimester screening for chromosomal abnormalities (nuchal translucency, nasal bone, ductus venosus combined with b-hCG and PAPP-A) had given a low risk of 1/2320 for trisomy 21. The ultrasound examination of the fetus revealed increased nuchal fold (NF) thickness, with an abnormal flat facial profile with hypoplastic nasal bone, hypertelorism and possible hypospadias (Figure 1a–d). The rest of the fetal anatomy including fetal echocardiography was unremarkable. After genetic counseling the patient was advised to undergo invasive testing to check the fetal karyotype. An amniocentesis was performed and the karyotype showed that the short arm of chromosome 4 was missing some genetic material. In particular R-banded chromosome analysis was performed at the 550 band level using standard procedures [1]. The karyotype was 46, XY, del (4) (p16.3) de novo. Multiplex ligation-dependent probe amplification specific molecular probe detected a 19-Mb deletion of the gene LETM1 1.81 Mb from the telomere. A further molecular cytogenetic analysis with fluorescent in situ hybridization (FISH) using specific probe locus-specific identification (LSI) Wolf–Hirschhorn/CEP4 (4p16.3) was performed and confirmed the initial karyotype diagnosis compatible with Wolf–Hirschhorn syndrome (WHS; Figure 2). Totally 100 nuclei were studied, one copy of the region 4p16.3 was observed which defines deletion in this region. Both parental karyotypes were normal. The parents after genetic and pediatric counseling about the prognosis of WHS elected to terminate the pregnancy. Post-mortem examination of a 350 g fetus confirmed the ultrasound findings.

The cellular transcription factor Brn-3a and the smoking-related substance nicotine interact to regulate the activity of the HPV URR in the cervix

The cellular transcription factor Brn-3a differentially regulates different human papilloma virus (HPV)-16 variants that are associated with different risks of progression to cervical carcinoma in infected humans. The upstream regulatory regions (URRs) of high- and intermediate-risk HPV-16 variants are activated by the cellular transcription factor Brn-3a, whereas the URR of a low-risk HPV-16 variant is not. In this study, we show in transfection assays that Brn-3a and the smoking-related substance nicotine produce stronger responsiveness of the URR of the low- and high-risk variants than with either factor alone, but not the intermediate-risk variant. We determined that this synergistic activity of Brn-3a/nicotine is due to two nucleotide differences in the URR, crucial for oncogenic E6/E7 transactivation. Mutant constructs in which the nucleotide residues were substituted alter Brn-3a/nicotine responsiveness. Importantly, women smokers with high levels of Brn-3a infected with low- or high-risk HPV-16 variants have augmented E6 levels, and were more frequently diagnosed with higher grades of cervical intraepithelial neoplasia (CIN) and cancer, as compared with non-smokers who were infected with similar variants and expressed similar levels of Brn-3a. Therefore, this study defines the specific interplay between the cellular transactivator Brn-3a, the environmental smoking-related substance nicotine and specific HPV variants in cervical carcinogenesis, and thus helps to explain why some women are susceptible to rapid CIN progression and cancer and others are not.

Trends in twin pregnancies and mode of delivery during the last 30 years: inconsistency between guidelines and clinical practice.

Abstract Aim: To investigate the characteristics of twin pregnancies and their mode of delivery over a 30-year period, in order to define the current trends in clinical practice regarding twin deliveries and the factors that influence these practices. Materials and methods: A retrospective study of twin deliveries at ≥24 weeks of gestation between January 1st, 1981 and December 31, 2010. Data concerning number of twin deliveries, mode of delivery, maternal age, use of ART techniques, parity, gestational age at delivery, birth weight and Apgar scores were collected from labor ward records. Results: Over the years there has been a statistically significant increase in the number of twin deliveries, twins after IVF, cesarean section rate, maternal age, nulliparity and a statistically significant reduction in term deliveries, mean birth weight and assisted vaginal deliveries. Maternal age, gestational age, parity and conception after IVF are factors related to the risk of performing a cesarean section. Conclusions: Despite the lack of adequate scientific evidence concerning the optimal route of delivery in twin pregnancies it seems that over time cesarean section has become the standard of care in most of twin pregnancies. Changes in the demographic characteristics contribute among other factors to this practice.

Bilateral hematosalpinx in a case of ectopic pregnancy: a clinical dilemma

A 31-year-old woman with a positive pregnancy test and a transvaginal ultrasound scan result that was suggestive of a right tubal ectopic pregnancy underwent a laparoscopy, which showed bilateral hematosalpinx. In the presence of active bleeding and deteriorating hemodynamic status of the patient, a minilaparotomy was performed that revealed a right-sided hematosalpinx and a left-sided ectopic gestation.

Risk for fetal loss and prematurity after 12,413 second trimester amniocenteses in a single center.

Abstract Objectives: The purpose of this retrospective controlled study is to estimate the risk for fetal loss and preterm delivery attributed to second trimester amniocentesis from a single tertiary center. Methods: The study group consists of 12,413 singleton pregnancies with consecutive amniocenteses, performed in a single tertiary center during a 15-year period (1996–2010) with known pregnancy outcome. The control group consisted of 6993 pregnancies with negative second trimester screening for aneuploidies during the same period who did not have any invasive test. The two groups were compared in terms of fetal loss rate up to 24 weeks and premature deliveries. Results: Total fetal loss up to 24 weeks in the study group, excluding terminations of pregnancy, was estimated at 1.25% (1.05%–1.45%, confidence interval [CI]: 95%). In the control group the loss rate was 0.65% giving a procedure related fetal loss rate of 0.6% which was not found to be a statistically significant difference. Delivery before the 28th, 32nd, 34th, and 37th week in the study group was reported in 0.2%, 0.8%, 1.2% and 8.1% respectively, and it was not statistically different from controls. Conclusion: The present study has shown that the risk of miscarriage that can be attributed to amniocentesis in our institution is 0.6%, and this is not statistically significant when compared with cases without any invasive procedure during pregnancy. Similarly, the risk for preterm labor was not statistically significant when compared with controls.

How significant is a cervical smear showing glandular dyskaryosis

OBJECTIVE To evaluate the incidence, outcome and predictive value of cytology showing glandular dyskaryosis. PARTICIPANTS Fifty-seven women with a smear diagnosis of glandular dyskaryosis registered between January 1997 and December 2001. SETTING Colposcopy and cytopathology units in a large district general hospital. RESULTS Sixty smears in 57 women showing glandular dyskaryosis were identified from a cohort of 135,120 smears, giving an incidence of 0.05%. Hospital records were available for 50 women. Final diagnosis included 13 cases of cervical glandular intraepithelial neoplasia (CGIN), 4 microinvasive cervical adenocarcinomas, 2 undifferentiated tumours, 1 microinvasive squamous carcinoma, 21 cases of CIN and 13 cases of endometrial pathology (8 endometrial cancers). Twelve women had coexistent squamous and glandular disease. Forty-five out of 50 women had significant pathology (positive predictive value 90%). Colposcopy was seen to be of limited value in assessment of smears showing glandular dyskaryosis. Only 1 out of 13 glandular lesions was diagnosed by colposcopy. CONCLUSION Smears showing glandular dyskaryosis are associated with significant pathology in 90% of cases and malignancy in 32% of cases. Hence, women with a smear showing glandular dyskaryosis should be referred urgently to a colposcopy clinic and flagged up as suspected cancer. Glandular dyskaryosis should be included in the national referral criteria for suspected gynaecological cancer.

Asymptomatic vasculitis of the uterine cervix in presence of cervical intraepithelial neoplasia grade III

Case reportA 34-year-old woman was diagnosed to have a high-grade cervical intraepithelial neoplasia and was treated by large loop excision of the transformation zone. Histology of the excised cone confirmed the diagnosis but also showed evidence of vasculitis of medium-sized vessels of the cervix. The woman was referred to a physician to rule out underlying systemic disease. Extensive laboratory and clinical screening was negative.DiscussionThe clinical significance and management of asymptomatic isolated vasculitis of the uterine cervix are discussed.

Cervical cancer diagnosed after simple hysterectomy for persistent abnormal cervical smears: a clinical dilemma.

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