1Objectives. Infection with oncogenic human papillomaviruses (HPV) is a prerequisite 2 for the development of cervical cancer. In many cases of cervical cancer and all cervical 3 cancer derived cell lines oncogenic HPV DNA is found to be integrated, indicating the 4 importance of integration in disease development. In this study, 176 HPV 16 positive 5 precancerous cervical lesions were analyzed for the physical state of viral genome to 6 determine the sites of integration into a host cell DNA and to evaluate the incidence of the 7 integration in different stages of cervical lesions. 8Methods. The detection of integrated papillomavirus sequences (DIPS) method in 9 combination with the amplification by polymerase chain reaction (PCR) of E1/E2 region was 10 used to identify the physical state of HPV 16 genome. The site of integration within a host 11 cell genome was determined by sequencing of DIPS unusual sized amplicons. 12

Objective: To demonstrate the clinical course in a young female with gonadotroph adenoma causing ovarian stimulation. Patient and methods: Our patient was a 23-year-old woman with a history of oligomenorrhea who had previously undergone bilateral ovarian wedge resection owing to the clinical appearance of polycystic ovaries. Two years later, she sought treatment for headache, galactorrhea, history of spotting and lower abdominal distension. FSH, LH, b-LH, inhibin A and B, estradiol, prolactin (PRL), and bchorionic gonadotrophin (b-CG) were measured, and the responses of FSH, LH and b-LH to thyrotrophin-releasing hormone (TRH) were documented. Immunohistochemical analysis of the tumor tissue was performed after surgery. Five years after the trans-sphenoidal surgery, the patient again became oligomenorrheic. A large recurrent adenoma was diagnosed on CT one year later. Transvaginal ultrasound showed ovaries of normal size with multiple small cystic formations simulating a polycystic pattern, While the patient was awaiting surgery, a pituitary apoplexy occurred. Emergency decompressive surgery was performed and the patient fully recovered. Results: Enlarged ovaries were found on ultrasound examination simulating a hyperstimulationlike pattern. At that time, elevated levels of FSH (13.4 IU/l) and marginally elevated levels of b-LH (1.43 ng/ml) were found, whereas the level of LH (0.5 IU/l) was subnormal. Plasma estradiol was markedly supranormal (6150 pmol/l). Levels of inhibin A and B were elevated (326 pg/ml and 588 pg/ml respectively). The prolactin level (70 ng/ml) was increased, whereas b-chorionic gonadotrophin (b-CG) was normal. Signi®cantly increased FSH, LH, and b-LH responses to TRH stimulation were documented. Pituitary macroadenoma was found on MRI scan and removed by trans-sphenoidal surgery. Immunohistochemical examination showed high positivity for b-CG and LH, and slight positivity for FSH. Five years after the surgery, estradiol was elevated (1160 pmol/l), whereas basal levels of LH (4.65 IU/l) and FSH (3.98 IU/l) were not suppressed. After the second operation, immunostaining of the adenoma tissue con®rmed the previous ®ndings. Conclusions: Measurement of gonadotrophins in our case did not prove to be a method for identifying a large recurrent gonadotroph pituitary adenoma. The sonographic ovarian imaging varied from a polycystic-to an ovarian hyperstimulation-like pattern during the evolution of the tumour.

The main etiological factor of precancerous lesion and invasive cervical cancer are oncogenic human papillomaviruses types (HPVs). The objective of this study was to establish the distribution of the most common HPVs in different cervical lesions and cancer prior to the implementation of organized population-based cervical screening and HPV vaccination in Croatia. In this study, 4,432 cervical specimens, collected through a 16-year period, were tested for the presence of HPV-DNA by polymerase chain reaction (PCR) with three sets of broad-spectrum primers and type-specific primers for most common low-risk (LR) types (HPV-6, 11) and the most common high-risk (HR) types (HPV-16, 18, 31, 33, 45, 52, 58). Additional 35 archival formalin-fixed, paraffin embedded tissue of cervical cancer specimens were analyzed using LiPA25 assay. The highest age-specific HPV-prevalence was in the group 18–24 years, which decreased continuously with age (P<0.0001) regardless of the cytological diagnosis. The prevalence of HR-HPV types significantly increased (P<0.0001) with the severity of cervical lesions. HPV-16 was the most common type found with a prevalence (with or without another HPV-type) of 6.9% in normal cytology, 15.5% in atypical squamous cells of undetermined significance, 14.4% in low-grade squamous intraepithelial lesions, 33.3% in high-grade squamous intraepithelial lesions, and 60.9% in cervical cancer specimens (P<0.0001). This study provides comprehensive and extensive data on the distribution of the most common HPV types among Croatian women, which will enable to predict and to monitor the impact of HPV-vaccination and to design effective screening strategies in Croatia.

Purpose We explored the value of transvaginal sonography as a screening technique for endometrial abnormalities. Methods Transvaginal sonography was used to evaluate the likelihood that an endometrial abnormality was present in a group of 103 women with postmenopausal bleeding. Results Sonographic findings showed a mean endometrial thickness of 6.2 mm in patients with endometrial atrophy, 12.4 mm in patients with simple hyperplasia, 13.4 mm in patients with complex hyperplasia, and 14.1 mm in patients with endometrial carcinoma. There was no statistically significant difference in endometrial thickness measurements between the hyperplasia and endometrial carcinoma groups. However, the difference between these groups and the endometrial atrophy group was statistically significant. The mean age was significantly higher for patients with endometrial carcinoma (62 years) than for the patients in the other groups. Conclusions Our study shows that transvaginal sonography is a useful method of screening for endometrial abnormalities. © 1997 John Wiley & Sons, Inc. J Clin Ultrasound 25: 431–435, 1997.

Change in the host and/or human papillomavirus (HPV) DNA methylation profile is probably one of the main factors responsible for the malignant progression of cervical lesions to cancer. To investigate those changes we studied 173 cervical samples with different grades of cervical lesion, from normal to cervical cancer. The methylation status of nine cellular gene promoters, CCNA1, CDH1, C13ORF18, DAPK1, HIC1, RARβ2, hTERT1, hTERT2 and TWIST1, was investigated by Methylation Specific Polymerase Chain Reaction (MSP). The methylation of HPV18 L1-gene was also investigated by MSP, while the methylated cytosines within four regions, L1, 5’LCR, enhancer, and promoter of the HPV16 genome covering 19 CpG sites were evaluated by bisulfite sequencing. Statistically significant methylation biomarkers distinguishing between cervical precursor lesions from normal cervix were primarily C13ORF18 and secondly CCNA1, and those distinguishing cervical cancer from normal or cervical precursor lesions were CCNA1, C13ORF18, hTERT1, hTERT2 and TWIST1. In addition, the methylation analysis of individual CpG sites of the HPV16 genome in different sample groups, notably the 7455 and 7694 sites, proved to be more important than the overall methylation frequency. The majority of HPV18 positive samples contained both methylated and unmethylated L1 gene, and samples with L1-gene methylated forms alone had better prognosis when correlated with the host cell gene promoters’ methylation profiles. In conclusion, both cellular and viral methylation biomarkers should be used for monitoring cervical lesion progression to prevent invasive cervical cancer.