Orientador: Regina Helena Garcia Martins / Resumo: Mercuri G. Incidência e Prevalência da Papilomatose Laríngea no Estado de São Paulo [dissertação]. Botucatu, SP: Faculdade de Medicina de Botucatu, Universidade Estadual Paulista; 2019. Introdução: A papilomatose respiratória recorrente ou papilomatose laríngea recorrente é a neoplasia benigna da laringe causada pelo Papiloma Vírus Humano, caracterizando-se pela presença de lesões proliferativas exofíticas recorrentes. A ausência de tratamento curativo, os custos relacionados ao diagnóstico, prevenção e tratamento das doenças causadas pelo HPV apresentam importante impacto econômico em todo mundo. No entanto, a falta de dados epidemiológicos nacionais, quanto à incidência e prevalência da papilomatose laríngea no Brasil não nos permite analisar o comportamento da doença em nossa população, bem como os benefícios da vacinação. Objetivo: Estimar a incidência e a prevalência da Papilomatose Laríngea no Estado de São Paulo. Métodos: Estudo populacional que objetivou levantar todos os casos diagnosticados de Papilomatose Laríngea do Estado de São Paulo, considerando os principais centros de diagnóstico desta enfermidade. Foram mapeados e selecionados os principais serviços de residência e especialização em Otorrinolaringologia do Estado de São Paulo, a partir dos dados fornecidos pela Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial (ABORL-CCF). Através do preenchimento de um questionário obtivemos os dados para estimar a prevalência e a incidência mínimas p... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Mercuri G. Incidence and Prevalence of Laryngeal Papillomatosis in São Paulo State [thesis]. Botucatu, SP: Faculty of Medicine of Botucatu, Universidade Estadual Paulista; 2019. Introduction: Recurrent respiratory papillomatosis or recurrent laryngeal papillomatosis is a benign laryngeal neoplasm caused by Human Papilloma Virus, characterized by the presence of recurrent exophytic proliferative lesions. Without curative treatment, costs related to the diagnosis, prevention, and treatment of HPV-related diseases have a significant economic impact worldwide. However, the lack of national epidemiological data on the incidence and prevalence of laryngeal papillomatosis in Brazil does not allow us to analyze the behavior of the disease in our population, as well as the benefits of vaccination. Objective: To estimate the incidence and prevalence of Laryngeal Papillomatosis in the State of São Paulo. Methods: A population study was conducted aiming to raise all the diagnosed cases of Laryngeal Papillomatosis of the State of São Paulo, considering the main centers of diagnosis of this disease. The main services of residence and specialization in Otorhinolaryngology of the State of São Paulo were mapped and selected, based on the data provided by the Brazilian Association of Otorhinolaryngology and Cervical-Facial Surgery (ABORL-CCF). By completing a questionnaire we obtained the data to estimate the minimum prevalence and incidence for the State in 2017. Results: The questionnaire wa... (Complete abstract click electronic access below) / Mestre

<p><strong><em>Uvod:</em></strong>&nbsp; U novije vreme sve veći broj studija sugeri&scaron;e da se genotipske varijante humanih papiloma virusa (HPV),&nbsp; uprkos filogentskoj srodnosti, mogu razlikovati u patogenosti i&nbsp; različito doprineti razvoju cervikalnih neoplazija.&nbsp; Cilj ovog rada je bio da se defini&scaron;e zastupljenost različitih onkogenih tipova HPV kod ispitanica sa područja Južnobačkog okruga (AP&nbsp; Vojvodina) i&nbsp; odredi genotipska varijabilnost najprevalentnijih genotipova HPV, analizom DNK sekvenci i RFLP metodom.</p><p><strong><em>Materijal i metode:</em></strong>&nbsp; Istraživanje je sprovedeno u periodu od januara 2014. do novembra 2015.&nbsp;godine. Studija je obuhvatila 564 osobe ženskog pola, starosti od 18 do 69 godina. Genotipizacija 12 visokorizičnih tipova HPV izvr&scaron;ena je upotrebom komercijalnog kita&nbsp; HPV High Risk Typing Real-TM (Sacace Biotechnologies, Italy, CE, IVD). Automatsko&nbsp;sekvenciranje amplifikovanih fragmenata L1 gena HPV rađeno je na automatskom sekvenatoru ABI Prism BigDye 3.1 (PE Applied Biosystems, Foster, CA, SAD).&nbsp; Bioinformatičkom analizom utvrđene su genetičke distance i filogenetski odnosi HPV izolata dobijenih u AP Vojvodini u odnosu na izolate iz drugih geografskih područja za tipove: 16, 18, 31 i 33.&nbsp; PCR-RFLP metodom ispitivana je heterogenost unutar HPV tipa 51, na nivou E1, L1 i L2 gena 11 odabranih HPV DNK pozitivnih izolata, primenom restrikcionih enima DraI, TaqI i PstI.</p><p><em><strong>Rezultati</strong></em>:&nbsp; Prevalencija HPV infekcije kod&nbsp; testiranih žena je iznosila 51,8%. Učestalost različitih tipova vema je varirala, pri čemu su&nbsp; najprevalentniji bili: HPV16 (34,5%); HPV 31 (20,5%); HPV 51 (13%); HPV33 (11,6%); HPV52 (11%) i HPV18&nbsp; (11%). HPV tipovi&nbsp; 16&nbsp;&nbsp;&nbsp; i 33 su&nbsp; najče&scaron;će dijagnostikovani u starosnoj grupi od 31-40 godina&nbsp; (25,5%&nbsp; i&nbsp; 6,21%), dok su&nbsp; HPV18 (8,16%), HPV31 (7,04%&nbsp; )&nbsp; i HPV51 (12,3%) &nbsp;najče&scaron;će dokazani kod žena mlađih od 30 godina.&nbsp; Određene&nbsp; su&nbsp; genetičke distance&nbsp; i filogenetski&nbsp; odnosi&nbsp; u&nbsp; okviru&nbsp; populacije&nbsp; izolata&nbsp; iz Južnobačkog&nbsp; okruga,&nbsp; AP Vojvodine,&nbsp; u&nbsp; odnosu&nbsp; na druge&nbsp; izolate&nbsp; čije&nbsp; su&nbsp; genomske&nbsp; sekvence dostupne&nbsp;u&nbsp; GenBank&nbsp; bazi&nbsp; podataka.&nbsp; Filogenetska analiza nukleotidnih sekvenci HPV tipa 16 je potvrdila da se najveći broj izolata iz područja AP Vojvodine grupi&scaron;e u liniju A, (podlinije A3 i A4 ) dok se jedan izolat izdvojio u liniju D (podlinija D1). Tri izolata HPV tipa 18 su grupisana&nbsp; u liniju A, a dva izolata kao posebna grupa. Utvrđeno&nbsp; je&nbsp; da genotipske varijante&nbsp; HPV 31 iz na&scaron;eg područja&nbsp;&nbsp;&nbsp; pripadaju linijama A, B i C. Izolati HPV tipa 33 pripadali su liniji A (podlinije&nbsp; A1 i A2). Dobijene nukleotidne distance su pokazale da je razlika između analiziranih i referentnih sekvenci manja od 2%, &scaron;to&nbsp;potvrđuje sličnost na nivou varijante.&nbsp; Na osnovu PCR-RFLP analize u odabranim&nbsp; DNK pozitivnim na HPV tip 51, restrikcionom analizom PCR produkata na nivou&nbsp; L1, L2 i E1 gena, utvrđeno je da su prisutni samo genotipovi &nbsp;koji odgovaraju liniji A sa podlinijom A1.</p><p><strong><em>Zaključak</em></strong>:&nbsp; Rezultati studije predstavljaju prve dostupne&nbsp; podatke o rasprostranjenosti 12 visokorizičnih tipova HPV, kao i podatke o genomskoj&nbsp; arijabilnosti i filogenetskoj srodnosti najprevalentnijih HPV tipova kod žena sa&nbsp; područja Južnobačkog okruga.&nbsp; Najprevalentniji&nbsp; HPV&nbsp; tipovi na&scaron;eg regiona su&nbsp; pokazali usklađenost sa evropskim izolatima, ali su nađene i ne-evropske varijante.&nbsp;Nizak procenat genetičkih distanci u okviru istog HPV tipa je u skladu sa niskom stopom mutacija kod ovih virusa.&nbsp; Iako se u dana&scaron;nje vreme RFLP metoda primenjuje u manjoj meri zbog uspona tehnologija sekvenciranja DNK, ova tehnika se može koristiti kao brza i jeftinija metoda&nbsp; za određivanje pojedinih linija ovih virusa.&nbsp; S obzirom da je&nbsp; u&nbsp; većini kliničkih laboratorija sekvenciranje DNK nedostupno,razvijena metoda se može koristiti za diferenciranje dve glavne linije HPV tipa 51, koje imaju različit onkogeni potencijal. Rezultati predstavljaju izuzetan doprinos epidemiolo&scaron;koj proceni realnih potreba uvođenja rutinske&nbsp; imunizacije protiv &nbsp;visokoprevalentnih genotipova prisutnih u prekanceroznim promena na grliću materice inficiranih žena sa na&scaron;eg područja. Tipizacija&nbsp; HPV virusa primenjenim metodama može biti od velike koristi u ranom otkrivanju maligne transformacije&nbsp;inficiranih ćelija i prevenciji karcinoma grlića materice.</p> / <p><strong><em>Introduction:</em></strong>&nbsp;In recent times increasing number of researches suggests that&nbsp; genotypic variants of human papillomavirus (HPV), in spite of phylogenic relations, could differ in virulence and contribute the development of cervical neoplasia. The aim of this work was to define the partition of different oncogenic HPV types in examined women from the area of Juznobacka&nbsp; district (AP Vojvodina) and to determine genotypic variability of the most prevalent HPV genotypes, by DNA sequences&rsquo; analysis and RFLP method.</p><p><strong><em>Material and methods:</em></strong>&nbsp;The research was&nbsp;conducted in period from January 2014 till November 2015. Study included 564 female persons, age from 18 to 69. The genotypization of 12 high-risk HPV types was performed with use of commercial kit HPV High Risk Typing Real-TM (Sacace Biotechnologies, Italy, CE, IVD). The automatic sequencing of amplified fragments of L1 HPV gene was performed on automatic sequencer ABI Prism BigDye 3.1&nbsp; (PE Applied Biosystems,Foster, CA, USA). By bioinformatic analysis genetic distances, as well as phylogenic relations of HPV isolates in AP&nbsp; Vojvodina were determined in relation with isolates from other regions for types 16,18, 31 and 33. By PCR-RFLP, the&nbsp; heterogeneity within HPV type 51, at the level of E1, L1 and L2 genes 11 selected HPV DNA positive isolates, by use of restriction enzymes DraI, TaqI and PstI.</p><p><strong><em>Results:</em></strong>&nbsp;The prevalence of HPV infection in tested women was 51,8%. The&nbsp; frequency of different types varied considerably, where the most prevalent types were: HPV16 (34,5%); HPV 31 (20,5%); HPV 51 (13%); HPV33 (11,6%); HPV52 (11%) and&nbsp; HPV18 (11%). The HPV types 16 and 33 are diagnosed the most in age group from 31-40 (25,5% and 6,21%), while HPV 18 (8,16%), HPV31 (7,04% ) and HPV51 (12,3%) are the most diagnosed in women younger than 30. The genetic distances and phylogenic relations within samples from population of Južnobacka region, AP Vojvodina,&nbsp; are determined in relation to other samples whose genomic sequences are available in&nbsp; GenBank&nbsp; data base. Phylogenetic analysis of nucleotide sequences&nbsp; of HPV 16 confirmed that the most of isolates from the area of AP Vojvodina are grouped in line A (sublines A3 and A4), while one isolate separated to the line D (subline D1). Three isolates of HPV type 18 are grouped in line A, and two&nbsp; isolates grouped as separate&nbsp; group. It was confirmed that genotypic variants of HPV&nbsp; 31 from our region belong to lines A, B and C. Isolates of HPV type 33 belonged to line A&nbsp; (sublines A1 and A2). Gained nucleotide distances showed that difference between analyzed and referent sequences is lower than 2%, which confirme the similarity at the level of variant.&nbsp; Based on PCR-RFLP analysis in selected DNA of persons positive on HPV type 51, by restriction analysis of PCR products on the level of L1, L2&nbsp; and E1 genes, it is confirmed that only genotypes that correspond to line A, subline&nbsp; A1 are present .</p><p><strong><em>Conclusions:</em></strong>&nbsp; The results of study present the first available data on abundance of 12 &nbsp;high-risk types, as well as data on genomic variability and phylogenic relations of the most prevalent HPV types in females from the Juznobacka region.&nbsp; The most prevalent HPV types of our region showed concordance with European isolates, but non-european variants were also found. Low percentage of genetic distances within the same HPV type is in concordance with low mutation rate of these viruses.&nbsp; Although today RFLP method is applied at lower scale because of rise of DNA&nbsp; technologies,this technique could be used as fast and efficient method for determination of particular lines of these viruses. Regarding that DNA sequencing is unavailable&nbsp; to the most of clinical laboratories,developed technique could be used for discrimination of two main lines of HPV type 51 that have different oncogenic &nbsp;potential. The results present considerable contribution to the epidemiological assessment of real needs for implementation of routine immunization against the high prevalent genotypes, present in precancerous alterations on the cervix of infected women from our area. Typization of HPV viruses by applied methods could be of great&nbsp; benefit in early detection&nbsp; of malignant transformations of infected cells and prevention of cancer of cervix.</p>

<p>The study aimed to describe the knowledge and awareness of HPV infection and vaccine of female university students and to determine the predictors of vaccine acceptability.&nbsp / The study found that 70% of the participants were sexually active. Awareness and knowledge on HPV/vaccine were poor / with only 22% being aware of HPV and that a HPV vaccine was available in South Africa. A greater proportion (80%) reported willingness to be vaccinated. Being aware of the existence of a pap smear, higher knowledge about HPV, higher perceived vaccine effectiveness and higher perceived severity of HPV infection were significantly associated with increased willingness to be vaccinated.</p>

Bakgrund    HPV, humant papillomvirus, är den vanligast sexuellt överförbara sjukdomen i världen. HPV kan orsaka kondylom, vilket innebär könsvårtor, men det kan även ge gynekologiska cellförändringar som kan leda till livmoderhalscancer. Syfte            Syftet var att undersöka vilken kunskap och inställning gymnasieelever, som inte ingår i vaccinationsåldern, hade till HPV och HPV-vaccination samt om det fanns några könsskillnader. Metod   En kvantitativ enkätstudie genomfördes på en gymnasieskola i Uppsala, Sverige. Enkätdistribueringen skedde vid två tillfällen och 58 (98,3 %) ifyllda enkäter kunde inhämtas, varav 38 var flickor och 20 var pojkar. Resultat    Flickorna på skolan hade en generellt högre andel rätt svar på enkäten, och totalt var det 21 (55,2 %) av flickorna som var vaccinerade medan endast 1 (5 %) av pojkarna. Av samtliga elever var det 22 (37,9 %) som kunde besvara frågan rätt angående hur många typer av HPV-virus det finns, dock kunde 51 (87,9 %) besvara frågan rätt om hur man skyddar sig mot HPV-viruset. Signifikant könsskillnad hittades i kunskapsfrågan gällande om HPV kan orsaka livmoderhalscancer (p&lt;0,045) där flickorna hade flest rätt svar på frågan. Det var fler vaccinerade flickor jämfört med pojkar (p&lt;0,001) och signifikant fler pojkar som kunde tänka sig att vaccineras (p&lt;0,019). Majoriteten, 68 %, av eleverna var positiva till den befintliga HPV-vaccinationen. Slutsats     Eleverna hade generellt sett en låg kunskapsnivå gällande HPV men trots det var det ändå mer än hälften av flickorna som var vaccinerade. Behov av ytterligare kunskap om HPV och HPV-vaccination till befolkningen behövs. / Background HPV, human papilloma virus, is the most common sexually transmitted disease in the world. HPV can cause genital warts and gynaecological dysplasia, which can lead to cervical cancer. Objective The objective was to describe the amount of knowledge the students had regarding HPV and the HPV-vaccination. Another objective was to describe how many of the students who were vaccinated and to study the existing attitudes towards HPV and the HPV-vaccination. The last objective was to compare whether there was any gender differences. Methods  A quantitative survey study was conduced at an upper secondary school in Uppsala, Sweden. The distribution of the surveys occurred on two occasions and 58 (98,3 %) completed surveys were collected. Results The girls had a generally higher percentage of correct answers on the survey. About 55 % (n=21) of the girls were vaccinated but only 5 %(n=1) of the boys. Regarding how many types of HPV there are 22(37,9 %) of the students answered correctly, although 51 (87,9 %) knew how to protect themselves against a possible infection. Significant differences in gender were found in the question whether HPV can cause cervical cancer or not (p=0,045) and whether the students would be willing to take the vaccine or not (p=0,019). The majority of the students, 68 %, were in favour of the existing HPV vaccination. Conclusion Although the students generally had poor knowledge regarding HPV the majority of the girls had taken the vaccine. The need of further knowledge is vast.

<p>The study aimed to describe the knowledge and awareness of HPV infection and vaccine of female university students and to determine the predictors of vaccine acceptability.&nbsp / The study found that 70% of the participants were sexually active. Awareness and knowledge on HPV/vaccine were poor / with only 22% being aware of HPV and that a HPV vaccine was available in South Africa. A greater proportion (80%) reported willingness to be vaccinated. Being aware of the existence of a pap smear, higher knowledge about HPV, higher perceived vaccine effectiveness and higher perceived severity of HPV infection were significantly associated with increased willingness to be vaccinated.</p>

Introduction: Human papillomavirus (HPV) is changing the prevalence, survival and treatment paradigms in oropharyngeal squamous cell carcinoma (OPSCC). Improved survival of patients with HPV positive compared to HPV negative OPSCC has led to trials of treatment de-escalation. Current HPV detection methods are imprecise, therefore standardised assessment of transcriptionally active HPV in OPSCC is required. Furthermore, the differences in immune characteristics and/or the hypoxia response/effects could explain observed differences in prognosis between HPV positive and negative OPSCC. Rigorous HPV detection and subsequent biomarker evaluation should provide additional information required before introduction of treatment de-escalation in broad patient groupings. Methods: The study cohort was 218 patients with OPSCC who received radiotherapy with curative intent. HPV status was determined on pre-treatment, formalin-fixed paraffin-embedded blocks using: 1) polymerase chain reaction (PCR); 2) in-situ hybridisation (ISH) and 3) immuno-histochemistry (IHC). QuantiGene multiplex assay was designed to detect mRNA of reference sequences of the common high-risk HPV types (16, 18, 33, 35, 45, 52 and 58). HPV detection methods were compared with mRNA quantification. Multimarker IHC of immune cell markers using chromogenic and fluorescent staining was performed, analysed and compared with single marker IHC using automated multispectral image analysis. A validated multiplex IHC method was used for a) chromogenic (CD3, CD4, CD8, and FoxP3) and b) fluorescent (CD8, CD68 and PD1/PD-L1) evaluation in tumour and stroma compartments. Single marker IHC was used to investigate tumour hypoxia markers (HIF-1α and CA-IX) in HPV positive and negative OPSCC. Results: p16 IHC and ISH were the most sensitive and specific, respectively, for classifying HPV status. The combination of the three tests had the highest positive/negative predictive values compared with QuantiGene mRNA detection. Multiplex validation showed that, for serial sections up to 6 μm apart, there were highly significant correlations (P<0.0001) between single and multiplex counts for both chromogenic and fluorescent IHC. Overall there was less variation in cell counts with fluorescent staining when compared to chromogenic staining. Multiplex IHC of TILs in HPV positive and negative OPSCC showed higher infiltration in both tumour and stromal areas of CD3+CD4+ and CD3+CD8+ T cells but not CD4+FoxP3 Tregs in HPV positive compared with HPV negative OPSCC. Only CD3+CD8+ stromal and not tumour area infiltration was associated with increased survival (P=0.02). PD-L1 expression was higher in HPV negative OPSCC and this was related to macrophage (CD68) expression of PD-L1. In HPV negative tumours infiltration with CD68+PD-L1 was associated with a good prognosis. HPV negative patients had higher expression of HIF-1α but not CA-IX. High expression of both markers was associated with a poor prognosis irrespective of HPV status. Conclusions: There are other prognostic factors operating in the larger subdivision of HPV positive and negative OPSCC. Precise HPV detection and inclusion of other prognostic factors is required before treatment de-escalation is used. Expression of immune inhibitory factors (PD1/PD-L1) alone without contextualisation with immune cell density is insufficient for patient prognostication and potential selection for therapy using immune checkpoint inhibitors. Hypoxia modification of radiotherapy should be explored in both HPV positive and negative OPSCC.

Denna studie undersökte kvinnors upplevelser när de fick ett HPV-positivt screeningresultat brevledes. Den svenska screeningen för förebyggande av livmoderhalscancer meddelar resultatet i ett kort, standardiserat brev. Efter att ha inkluderat humant papillomvirus (HPV) i analysen år 2014 verkade kvinnor bli mer oroliga för resultatet som kunde meddela en sexuellt överförd infektion (STI).En intervjustudie genomfördes med åtta kvinnor och empiriska data analyserades med receptionsteori. Resultaten visade vilka känslor kvinnor kämpade med, om de upplevde en negativ inverkan på deras sexualitet och om de var nöjda med ett brev som kommunikationssätt.Okunnighet om HPV och rädsla för cancer var centrala resultat. Kvinnor beskrev till och med en förändring i attityd till andra sexuella praktiker än vaginalt samlag. Resultatbrevet ansågs vara för kort och inte så informativt som nödvändigt. Detta fick samtliga kvinnor att söka efter mer information främst på internet för att hitta svar på deras frågor.Sammanfattningsvis finns det ett behov av grundlig och strukturerad information relaterad till HPV och påverkan på livmoderhalsen. Personal inom hälso- och sjukvårdssystemet bör vara lyhörda för kvinnors oro över deras screeningresultat. / This study explores women´s experiences when receiving their HPV-positive pap smear result as a letter. The Swedish screening programme for the prevention of cervical cancer notifies of the result in a short, standardised letter. After including human papilloma virus (HPV) in the analysis in 2014 women appeared to become more worried about the result showing a sexually transmitted infection (STI). An interview study was performed with eight women and the empirical data was analysed with reception theory. The study-results show which feelings women struggled with, whether they experienced a negative impact on their sexuality, and whether they were content with a letter as communication method for their results. Ignorance of HPV and fear of cancer were central findings. Women even described a change in attitude towards sexual practices other than vaginal intercourse. The result letter was considered to be too short and not as informative as needed. This caused all the women to look for more information foremost on the internet in order to find answers to their questions.In conclusion there is a need for thorough and structured information related to HPV and impact on the cervix. Healthcare system personnel should have a sensitive ear for women´s concerns about their pap smear results.

La implicación del papiloma virus humano (HPV) en carcinomas de cérvix está totalmente demostrada, pero esta infección se puede producir en otras regiones anatómicas con similares consecuencias patológicas. El estudio presenta la detección de HPV en carcinomas de pene y vulva mediante PCR, analizando la sensibilidad de diferentes sistemas, primers GP5+/6* y MY09/11. También se lleva a cabo la detección inmunohistoquímica de proteínas celulares como posibles marcadores diagnóstico de infección por HPV, las proteínas P53 y P16 su expresión supuestamente se altera por la infección de HPV en carcinomas de pene (77,5%), mientras que en los carcinomas de vulva la tasa de detección está dentro de los rangos esperados según trabajos previos publicados (29, 7). El estudio de la relación entre la presencia de HPV y la expresión de los marcadores celulares muestra su no asociación en estas patologías. / The infection of human papilloma virus (HPV) is clearly related with cervical carcinomas, but this infection can be found in other regions whit PCR, analyzing the sensitivity of HPV in penile and vulvar carcinomas whit PCR, analyzing the sensitivity of two differents systems, MY09/11 and GP 5+/6+. Also the immunohistochemical detection of cell proteines as diagnostic markers for the HPV in penile carcinomas in the study populations (77,5%), whereas in vulgar carcinomas . The detection rates is similar to the previously published data ( 29, 7%). The study of the relatioship between HPV and the expression of the cell markers shows no asocciation with the pahologies.

Cervical cancer represents an enormous health, psychological and social stress for every woman. The most important risk factor in the development of cervical carcinoma, which the second most common malignant cancer in women, is infection with a high-risk strain of human papillomavirus - a very frequent sexually transmitted disease. More than 100 types of HPV are acknowledged to exist, with HPV 16 and 18 being classified as high-risk types in particular. Worldwide, 500,000 new cases of cervical cancer are diagnosed every year. In the Czech Republic, there are 1,000 new cases of cervical cancer each year, out of which up to 400 women die. It is estimated that there will be up to 1,000,000 new cases of cervical cancer by 2050 unless the prevention is improved. Every woman is at risk of developing cervical cancer. HPV is sexually transmitted, however not only by sexual intercourse but also by skin-to-skin-contact with infected areas. Other risk factors in the development of the disease are: first sexual intercourse at early age, the number of sexual partners, smoking, other sexually transmitted diseases and a long term use of hormonal contraception. Use of condoms, which protects against sexually transmitted diseases, reduces the transmission of HPV by up to 70%. Having regular gynaecological check-ups with Pap smears is crucial for cervical cancer screening, as the screening suggests the presence of cytological abnormalities and pre-cancer. However, it cannot detect all types of premalignant changes and early stages of the carcinoma. Two vaccines have recently been developed, effective against the most frequent oncogenic strains of HPV (16 and 18), which currently cause about 70% of cervical cancer cases. Active immunisation against human papillomavirus is the first vaccination against carcinoma. Together with screening, it represents the best prevention method against cervical carcinoma. Based on the research of technical literature, the first part of the dissertation gives an overall view of the issue of cervical carcinoma. The second part of the dissertation deals with the research, eliciting the knowledge and attitude of women from Southern Bohemian towns in the field of cervical carcinoma prevention in the period of December 2008 - March 2009 and comparing it to technical literature.

L’intégration du génome du virus papilloma humain (VPH) a été reconnu jusqu’`a récemment comme étant un événnement fréquent mais pourtant tardif dans la progression de la maladie du col de l’utérus. La perspective temporelle vient, pourtant, d’être mise au défi par la détection de formes intégrées de VPH dans les tissus normaux et dans les lésions prénéoplasiques. Notre objectif était de déterminer la charge virale de VPH-16 et son état physique dans une série de 220 échantillons provenant de cols uterins normaux et avec des lésions de bas-grade. La technique quantitative de PCR en temps réel, méthode Taqman, nous a permis de quantifier le nombre de copies des gènes E6, E2, et de la B-globine, permettant ainsi l’évaluation de la charge virale et le ratio de E6/E2 pour chaque spécimen. Le ratio E6/E2 de 1.2 ou plus était suggestif d’intégration. Par la suite, le site d’intégration du VPH dans le génome humain a été déterminé par la téchnique de RS-PCR. La charge virale moyenne était de 57.5±324.6 copies d'ADN par cellule et le ratio E6/E2 a évalué neuf échantillons avec des formes d’HPV intégrées. Ces intégrants ont été amplifiés par RS-PCR, suivi de séquençage, et l’homologie des amplicons a été déterminée par le programme BLAST de NCBI afin d’identifier les jonctions virales-humaines. On a réussi `a identifier les jonctions humaines-virales pour le contrôle positif, c'est-à-dire les cellules SiHa, pourtant nous n’avons pas detecté d’intégration par la technique de RS-PCR dans les échantillons de cellules cervicales exfoliées provenant de tissus normaux et de lésions de bas-grade. Le VPH-16 est rarement intégré dans les spécimens de jeunes patientes. / Integration of human papillomavirus (HPV) has, until recently, been a frequent but late event in cervical carcinogenesis. The temporal view has, however, been challenged lately as integrated forms of HPV have been detected even in normal and preneoplastic lesions. Our objective was to describe HPV 16 load and physical state in a series of 220 normal and low grade cervical samples. We used quantitative real-time PCR, Taqman method, targeting E6, E2 and B-globin to calculate the HPV 16 load and the E6/E2 ratio in each sample. An E6/E2 ratio of 1.2 was used as a surrogate marker of integration. The site of integration was determined by restriction site PCR. Results show that the average viral load was 57.5±324.6 copies of DNA per cell, while E6/E2 ratio identified 9 samples with integrants. These integrants underwent amplification by restriction site PCR, followed by sequencing and nucleotide blast to identify the human-viral junctions. In conclusion, although it was possible to identify viral-host junctions with the integration positive control, that is, the SiHa cell line, the exfoliated cells of normal and low grade cervical lesions were negative for integration site by RS-PCR. HPV-16 is seldom integrated in specimens from young patients.