Recent Trends in HPV Infection and Type Distribution in Greece.

BACKGROUND/AIM: This study gives an insight into recent trends for Human papillomavirus (HPV)-specific infection and its fluctuation over the years 2011-2016. MATERIALS AND METHODS: A total of 2,417 Caucasian women between the age of 18 and 71 years underwent their annual gynaecologic examination at the Outpatient Gynaecological Clinic in the study period. RESULTS: Overall HPV prevalence was 43.9%, and high-risk HPV accounted for 31.3%. HPV16 was the most common high-risk type followed by HPV51 and HPV31. HPV positivity was higher in those with low- and high-grade squamous intraepithelial lesions (HSILs) than in women with a normal cytological test. HPV infection was highest in 2011.The prevalence of single infections remained higher than multiple infections over the entire study period. HPV16 prevalence was very high in the first years of the study and HPV18 exhibited highest prevalence in 2011. Younger women exhibited a significant increase in HPV infection from 2014. Overall HPV infection decreased over the study period. CONCLUSION: These data suggest that HPV vaccination might reduce the frequency of HSILs and cervical cancer and are useful for the development of a national screening programme.

Oropharyngeal and laryngeal but not oral cancers are strongly associated with high-risk human papillomavirus in 172 Greek patients.

A strong and consistent association has been reported between human papillomavirus (HPV) infection and oropharyngeal cancer, whereas a similar link has not yet been clarified in oral and laryngeal cancer. The purpose of this study was to investigate the association between HPV infection and head and neck squamous cell carcinoma (HNSCC) in Greek patients. Cytological or tissue specimens from 172 cases patients with HNSCC and cytological specimens from 91 control subjects were analyzed for HPV DNA detection and genotyping using a microarray-based assay. Multivariate logistic regression was used to estimate odds ratios (ORs) for the association between the presence of HPV infection and HNSCC for each of the tumor site, after adjustment for potential confounders. The adjusted ORs for positivity to high-risk HPV infection for oropharyngeal and laryngeal cancer were 20.3 (95% CI: 1.7-250.1) and 22.8 (95% CI: 2.5-206.2), respectively. High-risk HPV infection was not significantly associated with oral cancer. HPV infection was independently associated with poorly differentiated tumors (OR = 2.8; 95% CI: 1.1-7.5). Our results suggest a strong association of high-risk HPV infection with oropharyngeal and laryngeal cancer. J. Med. Virol. 89:170-176, 2017. © 2016 Wiley Periodicals, Inc.

Bax and Bak expression in cervical smears of women with low-and high-risk HPV types: A study of 120 cases.

BACKGROUND: Human papillomavirus (HPV) is known to be involved in the carcinogenesis of squamous cells in uterine cervix cancer, mostly by binding and inactivating the p53 and pRb tumor suppressor genes. Lately, evidence has emerged suggesting that HPV oncoproteins may interact with proteins involved in cellular apoptosis as well. AIM: This study aimed to investigate the expression of proapoptotic proteins Bax and Bak in women with low-risk and high-risk HPV types as opposed to HPV-negative women, and in women with normal pap smear compared to women with abnormal Papanicolau test (Pap) smear. MATERIALS AND METHODS: A total of 120 liquid-based cervical samples were subtyped for HPV types with microarray hybridization and then stained and evaluated immunocytochemically for Bax and Bak expression. Statistical analysis was performed on the Bax and Bak scores (percentage of positive cells × staining intensity), the overall percentage of positive cells, and the most prevalent staining intensity group found in each sample. RESULTS: A weak association between negative Bax staining and cytologically normal Pap smears was discovered, whereas cytologically abnormal samples tended to stain weakly or moderately positive. No other statistically significant difference was found in the other analyzed parameters. CONCLUSION: Cytologically normal pap smears seem to have a slight tendency to stain negative for Bax as opposed to cytologically abnormal pap smears. Although the association is weak, it is an indication that there might be a connection between the expression of Bax and the development of cervical intraepithelial dysplasia, which warrants further investigation in larger-scale studies.

Prevalence of human papillomavirus infection in Greek patients with squamous cell carcinoma of the larynx.

BACKGROUND/AIM: The causal relation between human papillomavirus (HPV) infection and squamous cell carcinoma (SCC) of the larynx has not been yet clarified. The aim of the present study was to investigate HPV infection in 54 SCC of the larynx and correlate it with patients' epidemiological and clinicopathological data. MATERIALS AND METHODS: Fifty-four biopsies were collected from patients with laryngeal SCC and tested for HPV DNA. Local recurrence analysis was performed at the 2- year follow-up. RESULTS: HPV DNA was detected in 18.5% (10/54) of laryngeal SCC; infection from high risk (hr) HPV and low risk (lr) HPV types was found in 16.7% (9/54) and 1.8% (1/54) of the samples, respectively. HPV 16 was the commonest type detected in 7.5% (4/54). The presence of HPV DNA was significantly associated with the absence of tobacco use (p=0.001) and poorly differentiated tumors (p=0.003). CONCLUSION: This study confirms the prevalence of HPV infection among patients with SCC of the larynx.

Prevalence and expression of human papillomavirus in 53 patients with oral tongue squamous cell carcinoma.

BACKGROUND/AIM: Human papillomavirus (HPV) infection has been considered a potential risk factor for the development of oral tongue squamous cell carcinoma (SCC). The purpose of the present study was to investigate HPV infection and high-risk HPV E6/E7 mRNA expression in SCC of the oral tongue in 53 Greek patients. MATERIALS AND METHODS: Fifty-three biopsies were collected from patients with SCC of the oral tongue and tested for HPV DNA and E6/E7 mRNA expression. RESULTS: The overall prevalence of HPV DNA was 11.3% (6/53), while high-risk HPV DNA was found in 7.5% (4/53) of SCC of the oral tongue. E6/E7 mRNA expression was observed in 9.4% (5/53) samples examined. HPV 16 was the commonest genotype identified by both DNA and RNA assays. A total of 28.3% (15/53) of the patients were non-smokers and non-drinkers. HPV infection was strongly associated with abstinence from tobacco and alcohol (p=0.013). CONCLUSION: The presence of high-risk HPV E6/E7 mRNA expression suggests that HPV may be implicated in the pathogenesis of SCC of the oral tongue.

E6/E7 mRNA expression of high-risk HPV types in 849 Greek women.

AIM: The goal of this study was to investigate human papillomavirus (HPV) geno typing and high-risk HPV E6/E7 mRNA expression in 849 women. PATIENTS AND METHODS: 849 Cervical samples were obtained from patients attending an outpatient clinic to have their annual gynaecological check-up. All patients underwent a conventional Pap test. The patients were also offered HPV test with the knowledge that it is not part of the screening. RESULTS: Overall prevalence of HPV DNA was 41.3%, while E6/E7 mRNA expression was found in 20.7% of the patients. HPV DNA and E6/E7 mRNA expression were detected in 21.1% and 9.1% of normal cytological samples, respectively, 38.1% and 23.8% of atypical squamous cells of unknown significance, 84.8% and 40.7% of low-grade squamous intraepithelial lesions, and 96.4% and 89.3% of high-grade squamous intraepithelial lesions, respectively. HPV 16 was the most frequent genotype identified. CONCLUSION: E6/E7mRNA detection might be useful as a screening marker for the early prediction of active infections and subsequent progression to severe dysplasia.

Prevalence of human papillomavirus in 45 greek patients with oral cancer.

The relation between HPV and head and neck cancer has recently and extensively been investigated. The purpose of this study was to indentify HPV genotypes, as well as E6/E7 mRNA expression of high-risk HPVs (16, 18, 31, 33 and 45) in oral squamous cell carcinomas (OSCCs) from 45 Greek patients. The overall prevalence of HPV DNA positive OSCCs was 11.1% (5/45), while high-risk HPV DNA was found in 6.7% (3/45) of OSCCs. E6/E7 mRNA expression was detected in 8.9% (4/45) of the oral cavity samples. Our data indicated that HPV 16 was the commonest genotype identified in HPV-positive OSCCs by both DNA and RNA tests. This study confirms the prevalence of HPV infections among patients with OSCCs. Future analysis and followup of more OSCCs will enable us to correlate HPV detection and clinical outcome.

A cross sectional study of HPV type prevalence according to age and cytology.

BACKGROUND: A cross sectional study to investigate HPV prevalence according to age and cytology. METHODS: Women presenting to a gynaecological outpatient clinic for a Pap smear test were included in the study (n=3177). All women had cervical cytology and HPV testing. RESULTS: Overall prevalence of any 24 HPV type analysed was 33.1% (95% CI 31.5% to 34.7%) and HPV 16 and HPV 42 were the most frequent (6.7% (95% CI 5.8% to 7.6%), 6.8% (95% CI 5.9% to 7.6%)), in total samples. Multiple HPV infection rate was 12.9% (95% CI 11.8% to 14.1%). High risk HPV (hrHPV) types were present in 27.4% (95% CI 25.8% to 28.9%) of the samples.HPV prevalence was highest among 14 to 19 y.o (46.6% (95% CI 40.7%-52.4%)) and second highest among 30-34 y.o. (39.7%, 95% CI 35.4%-44%). HPV 16 was highest among 20-24 (9.0% (95% CI 6.4%-11.6%)) and second highest among 50 to 54 y.o. (6.3% (95% CI 2.9% to 9.8%).In Low-grade Squamous Intraepithelial Lesions (LgSIL) cytology samples, the most frequently detected hrHPV types were: 16 (14.5% (95% CI 12.1% to 16.9%)), 51 (13.0% (95% CI 10.7% to 15.3%)) and 53 (9.1% (95% CI 7.2% to 11.1%)) and in High-grade Squamous Intraepithelial Lesions (HgSIL) were: HPV 16 (37.2% (95% CI 26.5% to 47.9%)), HPV 51 (17.9% (95% CI 9.4% to 26.5%)) and HPV 18 (12.8% (95% CI 5.4% to 20.2%)). CONCLUSIONS: In the population studied, HPV 16 and 51 were the most frequent detected hrHPV types. HPV positivity, hrHPV and multiple HPV types infections were higher in young women, while HPV prevalence declined with increasing age and presented two peaks a higher (14-19 y.o.) and a lower one (30-34 y.o.) These results may contribute to the creation of a national screening programme.

Human Papilloma Virus infection in sexually active adolescent girls.

PURPOSE: Adolescents are a vulnerable group with regard to sexually transmitted infections, including Human Papilloma Virus (HPV). This is thought to be both because of their more liberal sexual behavior and also the relative immaturity of their genital tract. The aim of the study was to examine trends in HPV infection among sexually active adolescents attending for a sexual health screen. METHODS: Sexually active adolescents were offered cervical screening, HPV typing and conventional genital cultures as part of a sexual health prevention protocol. Participating adolescents also completed a sexual health questionnaire. RESULTS: Between January 2008 and July 2011, 149 sexually active girls were examined for reproductive health issues (mean age 17.3 years). 62 (42%) tested positive for HPV DNA, 30 were infected by multiple types and 54 had at least one high risk type identified. The most commonly identified HPV DNA was for low risk type 42 (15 cases, 7.8%), followed by high risk types 51 (14 cases, 7.3%) and 59 (11 cases, 5.7%). E6-E7 mRNA expression was detected in 16 girls (11%), of which half were for HPV 16. Girls who tested positive for HPV DNA and mRNA had similar age at sexual debut with those that tested negative (15.1 years in all groups) but had on average more sexual partners (4.1 vs 1.9 p=0.007). Only 9 girls in this cohort had been vaccinated against HPV. Approximately half of the girls reported using inadequate or no contraception. CONCLUSIONS: HPV infection rate was high in our sample. Furthermore a high percentage was infected with high risk types and a proportion of girls demonstrated mRNA HPV expression. As age at first sexual intercourse drops, vaccination against HPV and sexual education should be instituted in early adolescence, in order to reduce long term reproductive sequelae of unsafe sexual practices.

Human papillomavirus genotyping and e6/e7 mRNA expression in greek women with intraepithelial neoplasia and squamous cell carcinoma of the vagina and vulva.

A large proportion of vaginal and vulvar squamous cell carcinomas (SCCs) and intraepithelial neoplasias (VAIN and VIN) are associated with HPV infection, mainly type 16. The purpose of this study was to identify HPV genotypes, as well as E6/E7 mRNA expression of high-risk HPVs (16, 18, 31, 33, and 45) in 56 histology samples of VAIN, VIN, vaginal, and vulvar SCCs. HPV was identified in 56% of VAIN and 50% of vaginal SCCs, 71.4% of VIN and 50% of vulvar SCCs. E6/E7 mRNA expression was found in one-third of VAIN and in all vaginal SCCs, 42.9% of VIN and 83.3% of vulvar SCCs. Our data indicated that HPV 16 was the commonest genotype identified in VAIN and VIN and the only genotype found in SCCs of the vagina and vulva. These findings may suggest, in accordance with other studies, that mRNA assay might be useful in triaging lesions with increased risk of progression to cancer.

Isomorph expression of BAG-1 gene, ER and PR in endometrial cancer.

BACKGROUND: BAG-1 isomorphs are regulating proteins with antiapoptotic action in endometrium. ERa and PRA isomorphs seem to have an important role in endometrial cancer. PATIENTS AND METHODS: We investigated the expression of BAG-1, ERa and PRA isomorphs in endometrioid adenocarcinoma and we correlated them with clinicopathological findings of the tumor. Fresh endometrial tissues were obtained from 33 patients with endometrial carcinoma and 191 paraffin-embedded tissues were analyzed by real-time PCR and immunochemistry for BAG-1, ER and PR. RESULTS: BAG-1 protein is expressed in both nucleus and cytoplasm. Grade 3 tumors were considered to have the highest intensity. Only 4 out of 79 samples showed intense expression of ERa, while 37 samples out of 72 samples strongly expressed PRA. CONCLUSION: BAG-1 nuclear isomorph appeared more frequently in grade 2 tumors than in grade 1 and 3 tumors, and the cytoplasmatic isomorph was expressed more strongly than the nuclear one.

BCL-2, BAX and P53 expression profiles in endometrial carcinoma as studied by real-time PCR and immunohistochemistry.

BACKGROUND: Apoptotic genes regulate apoptosis by the action of their pro- and antiapoptotic products. Among the most important proteins are p53 and Bcl-x family proteins. PATIENTS AND METHODS: The differential expression of these apoptotic genes were analyzed in relation to clinicopathological criteria in women with endometrial carcinoma. Thirty-three fresh tissues and 191 paraffin-embedded tissues were analyzed by real-time PCR for bcl-2/bax ratio and immunohistochemistry for p53, bcl-2 and bax proteins. RESULTS: Bcl-2/bax ratio tended to increase in grade 3 samples compared to grade 1 tumors. Mutated p53 was frequently observed in serous-papillary endometrial carcinomas (p=0.018). Low (<10%) and moderate (10-50%) expression of mutated p53 was observed in tumors with high expression of bax protein (>0.7). CONCLUSION: The Bcl-2/bax ratio is increased in grade 3 tumors. Bax protein shows a strong tendency for expression in the third group of clinical staging (stage IIb, III and IV). Poorly differentiated tumors highly expressed mutated p53.

Prevalence of human papillomavirus types 6, 11, 16, 18, 31, and 33 in a cohort of Greek women.

To study HPV prevalence and HPV types 6, 11, 16, 18, 31, and 33 distribution in cervical smears in a cohort of Greek women. One thousand six hundred thirty-six samples were cytologically evaluated and molecularly analyzed, by PCR based assay. Abnormal cytology was identified in 997 women and 75.4% of them were HPV DNA positive, while 639 had normal cytology and 24.6% were HPV DNA positive. HPV was detected in 62.9% of 256 ASCUS smears, 89.3% of 516 LSIL, 86.7% of 60 HSIL and 47.3% of 165 with cervical carcinoma. Overall, HPV 11 was the most common type (13.4%), followed by 18 (10.3%), 6 (7.2%), 16 (6.4%), 31 (3.4%) and 33 (3.4%). Multiple infections with two (11.3%) or more types, primarily 11 and 18 (4.8%), were also identified. Low-risk types 11 and 6 were common in ASCUS (36.6% and 26.4%, respectively), and high-risk types 16 and 18 in HSIL (42.3% and 30.8%, respectively) and in cancer (51.3% and 41%, respectively). Multiple infections were detected in 2.2% of normal and 31.7% of HSIL. HPV prevalence was 75.4% in abnormal and 24.6% in normal cervical smears. HPV 16 and 18 were the most common types in cancer. Single infection with type 11 and multiple infections with 11 and 18 were more frequent.

Herpes simplex virus type 1 in peptic ulcer disease: an inverse association with Helicobacter pylori.

AIM: To assess the frequency of herpes simplex virus type I in upper gastrointestinal tract ulcers and normal mucosa with the modern and better assays and also with a larger number of well characterized patients and controls and its relationship to Helicobacter pylori(H pylori). METHODS: Biopsy specimens from 90 patients (34 with gastric ulcer of the prepyloric area and 56 with duodenal ulcer) were evaluated. Biopsies from 50 patients with endoscopically healthy mucosa were considered as the control group. The method used to identify herpes simplex virus-1 (HSV-1) was polymerase chain reaction. H pylori was detected by the CLO-test and by histological method. RESULTS: Herpes simplex virus-1 was detected in 28 of 90 patients with peptic ulcer (31%) [11 of 34 patients with gastric ulcer (32.4%) and 17 of 56 with duodenal ulcer (30.4%)] exclusively close to the ulcerous lesion. All control group samples were negative for HSV-1. The likelihood of H pylori negativity among peptic ulcer patients was significantly higher in HSV-1 positive cases than in HSV-1 negative cases (P = 0.009). Gastric ulcer patients with HSV-1 positivity were strongly associated with an increased possibility of Helicobacter pylori negativity compared to duodenal ulcer patients (P = 0.010). CONCLUSION: HSV-1 is frequent in upper gastro-intestinal tract ulcers but not in normal gastric and duodenal mucosa. There is an inverse association between HSV-1 and H pylori infection.