Over the last decade, Scotland has witnessed a rising incidence in squamous cell carcinomas of the head and neck (HNSCC), a phenomenon thought to be linked to infection with high-risk Human Papillomavirus (HPV). HPV-associated HNSCC are a distinct disease presenting unique epidemiological, biological and clinical challenges. However, establishing HPV-related disease is impaired by non-standardised testing protocols and lack of a consensus on the efficacy of existing biomarkers such as p16. This is further complicated by the absence of additional biological markers and a dearth in our understanding of the molecular mechanisms underlying HPV-driven tumourigenesis. While HPV positivity is more commonly detected in the oropharynx, its prevalence and clinical impact in other head and neck subsites remains largely unexplored. The research presented in this thesis was undertaken to determine the prevalence of high-risk HPV in a heterogeneous cohort of 293 HNSCC patients from Tayside and to evaluate the validity of EBP50, a scaffolding protein involved in cell polarity which is targeted by high-risk HPV, as a potential marker for HPV-driven HNSCC. The p16 status of the patients in the cohort was already known and tissue specimens were genotyped for HPV using PCR. HPV infection, defined as p16 positivity and a positive HPV DNA status, was identified in 14% of the cohort. The majority (83%) of the HPV-positive tumours involved the oropharynx while the oral cavity, pharynx and the nasal cavity (17%) were involved to a much smaller extent. High-risk HPV type 16 was the most prevalent HPV type. Patients with HPV-positive tumours had significantly improved overall survival (OS) (2 year OS, 77% vs 57%) and recurrence free survival rates (RFS) (2 year RFS, 92% vs 77%) compared to patients with HPV-negative tumours. A positive tumour HPV status was found to be an independent prognostic indicator (HR 0.216; 95% CI 0.06 – 0.771) and so, given the high morbidity and debilitating physical and psychological problems associated with prevailing aggressive treatment regimens, it is imperative that this knowledge is harnessed to develop and improve treatment strategies. EBP50 expression was evaluated, by immunohistochemical analysis, first in normal oral mucosa and followed up in a smaller subset of 156 HNSCC patients from the main cohort. In the normal tissue EBP50 expression was predominantly membranous. In the tumour samples four distinct EBP50 expression patterns were observed and, of these, weak/ negligible cytoplasmic EBP50 expression showed a strong correlation, only marginally lower than p16 overexpression, with HPV DNA status and was observed largely in patients with tumours of the oropharynx and no history of smoking. Absence of EBP50 expression in the plasma membranes of tumour cells was a recurring pattern in a majority of the tumour samples. The scale of this study, comprising a Tayside cohort of unprecedented size, will undoubtedly contribute to the existing knowledge of HPV incidence in head and neck cancer in Scotland. Furthermore, this study presents compelling preliminary evidence for further researching weak/negligible cytoplasmic EBP50 expression as being a potential indicator of HPV-positivity in HNSCC.

Women infected with HIV are at an increased risk for infection of Human Papillomavirus (HPV), developing cervical lesions, and developing cervical cancer. Prior research has suggested disruptions in the immune system as well as circulating levels of stress and gonadal hormones as possible explanations for the increase of HPV infection in women with HIV. Additionally, psychosocial factors such as symptoms of depression and distress have also been associated with HPV infection, as well as disruptions in immune and endocrinologial systems, suggesting a psychoneuroimmunological pathway to disease progression. It was hypothesized that HIV+HPV+ women assigned to a Cognitive Behavioral Stress Management (CBSM) intervention will experience improvements in disease status, immune markers, circulating stress hormones, and reductions of depression and distress symptoms. An exploratory investigation of the effects of CBSM in levels circulating reproductive hormones was also tested. Follow-up hypotheses tested whether CBSM effects on immune variables were explained by reductions in symptoms of depression, distress, NE, cortisol, and increases of DHEA-S. Additionally, it was hypothesized that CBSM effects on stress hormones would be mediated by reductions in distress and depression symptoms. Finally, it was hypothesized that improvements in immune parameters would be correlated with decreases in risk of cervical dysplasia at a 9 month follow-up. Participants were 71 women co-infected with HIV and HPV that were mostly of African American, Haitian, Latina, and Caribbean descent. Hierarchical regression analyses were performed and showed a significant CBSM effect in decreases on BDI somatic depression subscale scores and increases in NK cell counts. Additionally, there was a marginally significant effect of CBSM on increases in CD4+ T-cells and decreases in urinary NE output. The bootstrapping method evidenced a mediation model, where the relationship between group assignment and CD4+ cell counts was explained by lower BDI somatic scores. More research is necessary to fully elucidate the psychobiological trajectories of disease as immunological changes in our sample did not explain the reduced odds of dysplasia in the women assigned to the CBSM group.

Syftet var att belysa kvinnliga sjuksköterskestuderandes kännedom om och inställning till HPV vaccination, samt att ta reda på hur de resonerade kring sitt eget val rörande HPV-vaccination. Metod: Kvalitativ studie med semistrukturerade intervjuer där åtta sjuksköterskestudenter deltog. Resultat: Studenterna hade en god kännedom gällande HPV-vaccination. Samtliga studenter var positivt inställda till HPV-vaccin. Däremot framkom en viss misstänksamhet mot HPV-vaccin då de uppfattade det som nytt och obeprövat. Studenterna ansåg att det är orättvist att inte pojkar erbjuds HPV-vaccin inom barnvaccinationsprogrammet. En del av studenterna menade även att det kan finnas risk för att HPV-vaccinerade känner en falsk säkerhet, vilket kan leda till att de inte använder kondom och går på cellprovskontroller i lika stor utsträckning som ovaccinerade. De studenter som inte hade vaccinerat sig mot HPV uppgav orsaker som otydlig information gällande kriterier för vaccination, att de hade en stadig partner, att de redan hade haft sex, var för gamla samt att vaccinet är för dyrt. De studenter som valt att vaccinera sig nämnde som främsta skäl, att de själva inte stått för kostnaden. Slutsats: Sjuksköterskestudenterna tycktes ha en god kännedom och huvudsakligen positiv inställning gällande HPV och HPV-vaccination. De visade även ett visst kritiskt tänkande gentemot vaccinet, då de menade att det kan vara svårt att veta långtidseffekterna, då det fortfarande är relativt nytt. Den ekonomiska aspekten, samt vilken information de fått, visade sig spela roll i studenternas beslut om vaccinering. Tydligare information från hälso- och sjukvård skulle kunna leda till att fler väljer att vaccinera sig.

<p>Human papillomavirus (HPV) infection of mucosal epithelium by `high-risk' HPV types has a prominent role in the development of anogenital intraepithelial neoplasias and carcinomas. Human epithelial cells transformed with the HPV E6 oncoprotein survive even under conditions that normally lead to cell apoptosis. This phenomenon has been attributed to HPV E6's ability to promote the degradation of the tumor suppressor protein p53. More recently, it has been demonstrated that HPV E6 contributes to activation of the NF-kB pathway. NF-kB is a transcription factor involved in the regulation of genes associated with cellular proliferation, apoptosis and inflammatory responses. In addition to p53 suppression, HPV E6 modulation of NF-kB activation presents another mechanism for HPV-driven tumorigenesis. However, it was not known how HPV E6 promotes NF-kB pathway activation. To address how HPV E6 leads to NF-kB activation, we identified an association between HPV E6 and the human cylindromatosis gene product (CYLD). CYLD is an endogenous inhibitor of canonical NF-kB activation. We showed that HPV E6 proteins could precipitate CYLD in vitro using a co-immunoprecipitation assay. Demonstrating that HPV E6 and CYLD proteins bind each other raised the possibility that this binding relationship would have a functional effect upon the NF-kB pathway by altering CYLD-mediated suppression of NF-kB activation. To identify HPV E6 functional relationship with CYLD and to determine how HPV E6 activates the NF-kB pathway, we transfected cells with either HPV E6 expression vectors containing the high-risk HPV type 16 E6 or the low-risk HPV type 11 E6 along with a CYLD expression vector. We showed HPV16 E6 expression in 293 cells blocked the ability of CYLD to inhibit CD40 ligand-stimulated NF-kB activation. Interestingly, HPV11 E6 was unable to inhibit CYLD mediated suppression of NF-kB in our system. CYLD had previously been shown to suppress NF-kB activation by removing stimulatory lysine 63-linked ubiquitin chains from TRAF2. We found CYLD expression in 293 cells leads to dose-dependent reduction in TRAF2 levels. This CYLD-mediated loss of TRAF2 is inhibited by co-expression of high-, but not low-risk E6 proteins. It was known that CYLD phosphorylation in vivo suppresses CYLD deubiquitination actions of canonical pathway proteins; we therefore tested the extent of CYLD phosphorylation when co-expressed with HPV E6, and discovered that CYLD phosphorylation was increased in the presence of HPV E6. This compilation of experiments suggests that with HPV16 E6 binding to CYLD, the E6 protein blocks CYLD-mediated TRAF2 loss and thereby TRAF2 is available to activate the canonical NF-κB pathway. Blocking HPV E6-mediated NF-kB activation may prove beneficial as a means for designing therapies that inhibit HPV-mediated tumorigenesis. The differences we detected between HPV11 E6 and HPV16 E6 are supported by other studies that showed E6 protein variations account for molecular and clinical differences among HPV infection outcomes. Similarly, there exist intratype E6 variations in HPV16. We obtained cervical specimens from patients with cytopathogenic changes consistent with the onset of cervical dysplasia infected with HPV16 E6 and the human immunodeficiency virus. We hypothesized the immunocompromised individual may harbor unique HPV16 E6 variants. Using PCR detection methods to amplify the HPV16 E6 DNA and sequencing technology, we identified that some of the samples indeed had nucleotide polymorphisms, resulting in amino acid sequence changes. However, the HPV E6 variants we detected were previously described, and fit know geographic HPV clades. Some of the HPV E6 variants we observed are suggested to be associated with progression to cervical cancer, but further evaluation is required.</p> / Dissertation

Syfte och frågeställningar: Syftet med studien är att undersöka på vilka sätt och vilka metoder två olika mediekanaler använder sig av för att övertyga och skapa trovärdighet om HPV-vaccinet.  Metod och material: Kritisk diskursanalys på ett avsnitt ur samhällsprogrammet TV4 Kalla Fakta och tidningsartiklar rörande HPV-vaccinet. Huvudresultat: Framställning av HPV-vaccinet skiljer sig väsentligt åt vid granskning av Kalla Fakta och tidningsartiklarna. Metoderna som används i texterna för att skapa trovärdighet är expertutlåtanden och skapa förtroende med tittarna och läsarna.

Bakgrund Humant papillomvirus (HPV) orsakar vårtor och är en vanligt förekommande könssjukdom världen över. Vaccination mot de vanligaste HPV-typerna som kan orsaka kondylom och leda till cancer ingår sedan 2012 i det allmänna vaccinationsprogrammet för flickor och unga kvinnor. Tidigare forskning visar att unga kvinnor trots låg kunskap om viruset, har en positiv inställning till vaccination. Syfte Att undersöka unga vaccinerade kvinnors kunskap om HPV samt deras kunskap om och inställning till HPV-vaccination. Metod En kvalitativ explorativ studie. The Health Belief Model användes som teoretisk modell. Individuella intervjuer med åtta unga kvinnor som vaccinerats mot HPV. Data analyserades med innehållsanalys. Resultat Totalt genomfördes åtta intervjuer med unga kvinnor födda 1993-1998. Tre kategorier skapades: 1) Bristande kunskap om HPV 2) Tillförlitligt skydd mot cancer samt 3) Vaccinet är tillgängligt. Kunskapen om HPV och HPV-vaccin var låg hos de unga kvinnorna. Den främsta anledningen till att de valde att vaccinera sig var rädsla för cancer, andras inflytande till vaccinering, främst från mödrar, en tilltro till hälso- och sjukvården och till vaccinet samt att vaccinet är tillgängligt. Slutsats Det är tydligt att kunskapen om HPV och vaccinet är låg bland de deltagande unga kvinnorna. Inför framtiden behövs anpassad information till unga kvinnor om viruset och vaccinet för att tillgodose behovet av information. Det är viktigt att unga kvinnor som vaccineras mot HPV har kunskap om vaccinet för att veta hur de skyddar sig mot HPV och att de även ska förstå vikten av att gå på gynekologisk cellprovskontroll som en del av prevention av HPV. / Background Human papillomavirus (HPV) cause warts and is a common sexually transmitted infection worldwide. Vaccination against the most common HPV types that can cause genital warts and cancer is implemented in the national vaccination programme for girls and young women since 2012. Previous research shows that young women, despite low knowledge about the virus, are in favour of the vaccine. Objective To explore young vaccinated women’s knowledge about HPV and knowledge and attitudes towards HPV-vaccination. Method An qualitative explorative study. The Health Belief Model was the  theoretical framework. Individual interviews were conducted with young women vaccinated against HPV. Data were analyzed with content analyses. Results In total eight interviews were undertaken with young women born in 1993-1998. Three categories were revealed through the interviews: 1) Lack of knowledge about HPV 2) Reliable protection against cancer and 3) The vaccine is available. The young women had low knowledge about HPV and HPV vaccine. The main reasons for vaccination were; fear of cancer, influence from others, especially the mothers, trust in the healthcare and the vaccine and the vaccine is available. Conclusion The knowledge of HPV and the vaccine was low among the included women. In the future the iformation about the virus and the vaccine needs to be adapted to the young women to provide the need of information. It is important that young women who are vaccinated against HPV have knowledge about the vaccine to be able to protect themselves against HPV and that they are aware of the importance of attending future cervical cancer screening controls as a part of the prevention against HPV.

The human papilloma virus (HPY) infection has a causal association with cervical intraepithelial neoplasia (CIN) and cervical cancer. However, pre-malignant or malignant transformation is not always observed with HPY infection. lILA molecules are important in the regulation of the immune response to foreign antigens. The role of genetic variation at the HLA class II loci (DR and DQ) in CIN was investigated in 176 British Caucasian patients and 420 controls (normal cervical cytology and negative for HPY 16, 18, 31 and 33). HLA DQB 1 *03 typing was performed by a novel polymerase chain reactionrestriction fragment length polymorphism method (A-RFLP). The technique uses PCR to mutate the first base of codon 40 (DQ alleles) from T to G to create an artificial restriction site for an enzyme, MluI, which distinguishes DQB 1 *03 from other alleles and is confirmed by digestion of amplified DNA with Mlul. Further HLA DR-DQ typing was performed by PCR DNA amplification and oligonucleotide probe typing. HPY types (16, 18, 31 & 33) were detected by using type-specific oligonucleotide primers and PCR. The alleles of the DQB 1 *03, DRB 1 *04 and DRB 1 * 11 groups were strongly associated with susceptibility to CIN. Specifically the haplotypes DRB 1 *040 I-DQB 1 *0301 and DRBl*1101-DQB1*0301 were significant and indicated susceptibility. The DQBl*03 locus was more contributory to this association than the DRB 1 loci. A weak protective effect was shown for the haplotype DRB 1 *0 10 I-DQB 1 *0501. Positive correlation was also observed for HPY-positive CIN, suggesting that specific HLA alleles may be important in determining the immune response to HPY antigens and the risk for CIN after HPY infection. Immunoaffinity purification of the susceptibility and protective HLA ~ molecules was performed and the naturally processed peptides were eluted and sequenced by Edman degradation. The data obtained was used for motif prediction of HPY 16 E6, E7, Ll and L2 sequences that may be capable of binding to these HLA molecules. Motif prediction as well as the binding affinity of predicted peptide motifs for HLA D RB 1 *0401 and DRB 1 *0 10 1 was accomplished using the published data' on the naturally bound peptide sequences bound to these HLA molecules. The results revealed significant differences in both the number and binding affinity of the HPV 16 derived peptides to the protective and susceptibility HLA molecules. These results should help in the rational design of vaccines against HPV.

Objective: Describe the prevalence of HPV genotypes via PCR and DNA sequencing in 397 women who attended to the gynecological outpatient center in the Hospital Regional Docente de Cajamarca from March to September 2017. Results: A positive PCR result for HPV was observed in 121 cervical samples. A high-risk genotype was found in 63.6% (77/121) of patients, a probably oncogenic type in 23.1% (28/121) and a low-risk type in 7.4%. Among the high-risk genotypes, HPV-31 was the most common one present in 20% (21/77), followed by HPV-16 in 11.4% (12/77). Coinfections between two or more genotypes were observed in 12 cases. / This work was supported by 4th research incentive of the Universidad Peruana de Ciencias Aplicadas (Grant: UPC‑EXP‑02‑2017). Lima, Peru. / Revisión por pares

Currently, the most common strategy employed to detect DNA sequences is PCR (Polymerase Chain Reaction). Nevertheless, in the last few years research on DNA biosensors has increased significantly. Such sensors represent an alternative to PCR in the detection of specific DNA sequences, once they exhibit fast response, low limits of detection, and require simpler sample preparation. The development of a biosensor for detection of DNA from Human Papillomavirus type 18 is reported. To immobilise DNA probe onto indium-tin oxide (ITO) electrodes, a silanisation was carried out using 3-Aminopropyltryethoxysilane (APTES). Silanisation was studied and optimised using ultra-violet absorption spectroscopy, atomic force microscopy, fluorescence microscopy, and cyclic voltammetry. After immobilisation, the hybridisation with target sequence is detected by changes in surface properties of ITO electrode by Cyclic Voltammetry and Electrochemical Impedance Spectroscopy, using the Ferri-Ferrocyante redox couple. The detection of synthetic target sequence was performed in the range of 12.5 to 100 nM, and 300nM for PCR products. The sensor did not show significative response for non-complementary sequence at 50 nM. This sensor can be applied for fast and low cost detection of HPV genetic material at nanomolar levels. / A estratégia mais empregada atualmente na detecção de sequência de DNA é a PCR (Reação em Cadeira da Polimerase). Contudo, nos últimos anos, a pesquisa em biossensores de DNA tem aumentado significativamente. Estes sensores representam uma alternativa a PCR na detecção de sequências específicas de DNA, uma vez que exibem resposta rápida, baixos limites de detecção e requerem preparação simples da amostra. Nesta dissertação descrito o desenvolvimento de um biossensor para a detecção do DNA do Papilomavirus Humano tipo 18. A fim de imobilizar a sequência de captura de DNA em eletrodos de óxido de estanho e índio (ITO), realizou-se uma silanização usando 3-Aminopropiltrietoxisilano (APTES). A reação de silanização foi estudada e otimizada através das técnicas de Espectroscopia de Absorção Ultravioleta, Microscopia de Força Atômica, Microscopia de Fluorescência e Voltametria Cíclica. Após a imobilização, a hibridização com a sequência alvo é detectada através de alterações nas propriedades de superfície do eletrodo através de Voltametria Cíclica e Espectroscopia de Impedância Eletroquímica, usando o par redox Ferri-ferrocianeto. A detecção da sequência alvo sintética foi realizada no intervalo de 12.5 a 100 nM, e para o produto de PCR, 300 nM. O sensor não demonstrou resposta significativa para sequência não complementar a 50 nM. Este sensor pode ser aplicado na detecção rápida e de baixo custo de material genético do HPV a níveis nanomolares.

Submitted by Ramon Santana (ramon.souza@ufpe.br) on 2015-03-10T17:05:17Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) TESE Albert Eduardo Silva Martins.pdf: 1520089 bytes, checksum: 6f85e6e88a6376e89af5aebd3d4d29fb (MD5) / Made available in DSpace on 2015-03-10T17:05:17Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) TESE Albert Eduardo Silva Martins.pdf: 1520089 bytes, checksum: 6f85e6e88a6376e89af5aebd3d4d29fb (MD5) Previous issue date: 2013-10-30 / Pacientes HIV-positivo possuem uma maior prevalência de co-infecção por HPV de alto risco oncogênico. A presença viral favorece a progressão de lesões escamosas intra-epiteliais e podem induzir ao câncer. O objetivo do presente estudo foi avaliar a prevalência, distribuição dos tipos virais e fatores de risco para a infecção pelo HPV em pacientes infectados por HIV. Amostras cervicais de 450 mulheres infectadas pelo HIV foram analisadas quanto à citologia oncótica, colposcopia, presença e tipagem de HPV através de PCR e sequenciamento utilizando os iniciadores MY09 e MY11. Os resultados foram analisados comparando dados demográficos, e outros relacionados com a infecção pelo HPV e HIV. A prevalência de HPV foi de 47,5%. Das amostras positivas para o HPV, 59% albergavam tipos de alto risco oncogênico. Análise multivariada confirmou a associação da infecção pelo HPV com a presença de alterações à citologia (p=0,003), idade maior ou igual a 35 anos (p=0,002); número de parceiros maior que três (p=0,002); contagem de linfócitos T CD4+ < 200/mm3 (p=0,041), e etilismo (p=0,004). Apesar da presença de HPV de alto risco na maioria das lesões estudadas, a baixa freqüência de HPV 16 (3,3%) e o estado imunológico preservado na maioria das pacientes HIV-positivas, são fatores que podem explicar a baixa ocorrência de lesões cervicais pré-cancerosas nessa população. A persistência da infecção cervical por tipos de alto risco oncogênicos de papiloma vírus humano (HPV) pode levar a neoplasisas intra-epitliais cervicais (NIC). O objetivo do presente estudo foi o de avaliar, em mulheres infectadas pelo HIV, se a presença de aneuploidia em amostras de células cervicais está associda à presença e à evolução de NIC. O presente estudo constou de 2 etapas. Na primeira etapa, correspondendo a um corte transversal , analisou-se a associação entre a presença de aneuloidia por citometria de fluxo e características sócio-demográficas, hábitos e características ligadas à infecção pelo HPV e HIV. Na segunda etapa, correspondendo a uma coorte, verificou-se se a aneuploidia era preditiva da evolução da NIC. Não observou-se associação entre a presença de aneuploidia e a infecção por HPV, nem com alterações à citologia oncótica. Por outro lado, a aneuploidia esteve associada à presença de NIC (p=0,03?) no exame histológico e ao não uso de TARV (p=0,001). A maioria das mulheres infectadas por HIV (234/272) apresentaram contagem de linfócito T CD4+ normal (acima de 350células /mm3) e mostraram uma maior taxa de regressão (77,5%) da aneuploidia comparadas à taxa de progressão (23,9%) em até dois anos de seguimento. Embora tenha sido encontrada uma associação entre a presença da lesão tecidual cervical e o índice de DNA, este não foi preditivo da evolução da lesão cervical, sugerindo que a progressão da lesão cervical para o câncer em mulheres HIV-positivas também pode ser alterada pela melhoria do estado imunológico propiciado pelo uso de terapia anti-retrovirial (TARV).