Human papillomavirus (HPV) related cervical cancer is still the most common gynaecological malignancy in developing countries and, as yet, there is no alternative to surgery for the treatment of HPV-associated pre-malignant lesions. HPV 'hijacks' the host-cell ubiquitin-proteasome system to degrade the p53 and Rb tumour suppressor proteins which in turn, leads to the development of cancer. Previous studies have shown that the HIV protease inhibitor lopinavir selectively inhibits the chymotryptic-like activity of the 26S proteasome which stabilises p53 and induces the apoptosis of HPV positive cervical carcinoma cells. Based on this it was hypothesised that lopinavir treatment of HPV positive cervical carcinoma cells would produce changes in the levels of a wide range of cellular proteins that are dis-regulated by HPV-related activation of the proteasome. In order to address this, antibody microarray screening was carried out on lopinavir treated and control untreated HPV positive SiHa cervical carcinoma cells. This showed lopinavir induced alterations in 51 proteins including the cellular antiviral defence protein RNase L. Lopinavir induced both a dose and time dependent increase in RNase L which was subsequently confirmed by western blotting. Transient siRNA silencing of RNase L expression reduced the lopinavir-dependent toxicity in SiHa cells, suggesting an important role for this protein in the toxicity of lopinavir in HPV infected cells. SiHa cells were much more sensitive to lopinavir than CaSKi cervical carcinoma cells which had much higher levels of the E6 protein and did not up regulate RNase L. Furthermore, lopinavir treated HPV16 E6/E7 immortalised keratinocytes were also shown to up regulate RNase L protein expression and these cells were much more sensitive to lopinavir induced apoptosis than mortal control keratinocytes. In addition, transient expression of RNase L in RNase L-deficient C33A cells and the same cells stably transfected with HPV16 E6 (C33AE6) demonstrated that E6 protected these cells from RNaseL-induced cell death. Surprisingly, analysis of RNase L protein levels in these cells demonstrated that E6 did not induce the degradation of the RNase L protein. Instead it was found that E6 stabilised the interaction between RNase L and its endogenous inhibitor protein, ABCE1, and that lopinavir de-stabilised this interaction. Given that C33A tumour cells, E6/E7 immortalised keratinocytes and hTert immortalised keratinocytes are all sensitive to lopinavir, this implies that this compound does not specifically target HPV immortalised cells but rather targets immortalised cells in general, regardless of how this was achieved. The optimum concentration of lopinavir for all these effects was 25 μM, which is 15-fold higher than is observed in cervico-vaginal secretions following oral dosing with the drug Kaletra. In conclusion these results have confirmed the potential of lopinavir to treat HPV related pre-cancerous cervical lesions and provided at least part of the mode-of-action. Indeed they strongly support the use of lopinavir as a low-cost, self-applied topical alternative to surgery for this disease which will be of particular benefit in low-resource countries. Finally, the ability of lopinavir to induce apoptosis of non-HPV related immortalised cells merits further investigation since this indicates this drug may be useful for the treatment of other non HPV related pre-malignant conditions.

Thesis (M. Sc. (Biochemistry)) -- University of Limpopo, 2019 / Cervical cancer is the fourth most common cancer in females, and the seventh of all cancer types in both genders, with an estimated 500,000 new cases each year. As with liver cancer, a large majority (around 85%) of the global burden occurs in the less developed regions, where it accounts for almost 12% of all female cancers. About 90% of cervical cases are associated with human papillomavirus (HPV) as a causative agent and this virus is frequently transmitted through sexual contact involving exchange of fluids (Walboomers et al., 1997). Due to the ineffectiveness, undesirable side effects and costly treatment for the disease the current study was aimed at determining the anti-proliferative effects of extracts of selected medicinal plants for their anticancer activity on HeLa cell line invitro. In order to accomplish the outcome of this research study, medicinal plants (Toona cilliata, Seriphium plumosum and Schkuhria pinnata) from Limpopo Province (South Africa) with history of traditional use on cervical cancer-associated patients were selected. The Toona cilliata plant leaves were collected from Tzaneen, area while Seriphium plumosum and Schkuhria pinnata leaves were collected from Mankweng area. The dried leaves were grounded into powder and extracted using acetone. Thereafter, extracted leaf materials of selected plants were subjected to fingerprint profiling using TLC silicon coated plates immersed in tanks with different mobile phases (TEA, CEF and EMW) of various increasing polarities since. The plates were sprayed with vanillin/H2SO4, dried and visualised under UV light. Scavenging ability of the plant extracts was determined through investigating the presence of antioxidant activities using 0.2% of the 2,2- diphenyl-1- picrylhydrazyl (DPPH) indicator. The quantitative presence of total phenolic and flavonoids contents was also determined using garlic and quercetin as standards, respectively. Quantitative antioxidant scavenging activities were also determined and ascorbic acid was used as a positive control. This was followed by quantitative determination of ferric reducing power and thereafter the EC50 values of the extracts were determined by linear regression. Cell proliferation or viability was determined using the 3[4, 5 dimethylthiazol-2-yl]-2-5 diphenyltetrazolium (MTT) assay with actinomycin as a xv positive control and untreated cells as the negative control. Apoptotic effects of the extracts were determined using the Annexin V Fluos staining kit. This was followed by determining whether apoptosis was calcium dependent or independent using a calorimetric assay. In comparing the acetone extract yield per 10 g leaves of plants, Toona cilliata leaves exhibited the highest yield followed by Seriphium plumosum and with the least yield from Schkuhria pinnata. The finger print profile showed the prominent separation and was achieved from all the plants when using the non-polar TEA solvent. All plants were shown to contain extracts with varying levels of antioxidant activity especially when using CEF and EMW mobile phases. When evaluating the total phenolic and flavonoids contents all plant extracts exhibited presence of phenolic compounds with high presence observed in Seriphium plumosum and Toona cilliata. Extracts from Seriphium plumosum and Toona cilliata showed to have higher concentrations of phytochemicals that may be of a benefit in antioxidant activities as compared to Schkuhria pinnata in relation to the positive control and a similar trend were observed in the ferric reducing power assay. Extracts from Seriphium plumosum were shown to have the best IC50 scavenging values followed by Toona cilliata and Schkuhria pinnata respectively. All the plants exhibited free radical scavenging abilities with Seriphium plumosum shown to possess higher activities in comparison with the positive control. All the plants exhibited a dose-dependent cytotoxicity activity against the HeLa cervical cell line. Evidence of induced apoptotic activity was observed in HeLa cells when using extracts from Seriphium plumosum and Toona cilliata. Induction of apoptosis by plant extracts was shown to be calcium dependent as there was a decrease in calcium concentration with a decrease in the number of viable cells. In conclusion, the leaf extracts from Toona cilliata, Seriphium plumosum and Schkuhria pinnata contain compounds of various polarities with freeradical, antioxidant and anti-cancerous activities that may be beneficial if further studies are conducted to identify chemical compounds that may inhibit anticervical cancer activities.

Thesis (MPH.) -- University of Limpopo, 2015 / Background Cervical cancer is the 2nd most common cancer in women globally representing 13% of female cancers and accounting for 11% of the total cancer deaths (Ahmedin et al.2011). Several studies demonstrated the association between HIV and HPV. In Zimbabwe the prevalence of HIV/AIDS is high and cervical cancer is the leading cause of cancer deaths among women of all age groups. The aim of the study was to determine the knowledge, attitude and practices of HIV infected women on cervical cancer and cervical cancer screening at Musiso Hospital, Masvingo Province, Zimbabwe. Methods 208 self administered questionnaires were used with a 100 per cent response rate. Quantitative data were analysed using STATA statistical package version 12 for descriptive and inferential statistics. Chi-squared tests were done for hypothesis testing at 5 per cent level of significance and 95 per cent confidence level. Multiple variable logistic regressions models were also used to assess association between outcomes of interest and socio-demographic characteristics. All open ended questions were analysed using qualitative methods. Results Out of the 208 participants, 45 (21.6 per cent) respondents claimed to know what cervical cancer is. About 55.3 per cent said cervical cancer is preventable. The majority (92.8 per cent) did not know any screening tests. Just above three quarters (77.3 per cent) of the respondents believed they were at risk of having cervical cancer. About 9 per cent (18) of all participants had screened for cervical cancer before and 95.8 per cent respondents reported would like to screen for cervical cancer in the future. Conclusion HIV infected women at Musiso mission hospital were found to be having inadequate knowledge, positive attitude and inadequate practices on cervical cancer and cervical cancer screening. There is need to equip these women with knowledge on cervical cancer and cervical cancer screening to increase cervical cancer screening uptake. Key Concepts: knowledge, attitude, practice, screening, cervical cancer

Though vaccine uptake and public support have risen since the release of the first HPV vaccines, the United States has far lower initiation and completion rates for the HPV vaccine series in comparison to other vaccines indicated for youth. Disparities are even greater in the Appalachian regions. Understanding factors contributing to these discrepancies is vital to improving raise vaccine rates in Appalachia. A comprehensive literature search identified all articles pertaining to HPV vaccination in children and adolescents living in Appalachia. The final 15 articles were included in a systematic review of the topic. Findings: HPV disease and HPV vaccine-related knowledge and communication were low in Appalachian communities, and vaccine uptake was lower in all areas of Appalachia as compared to non-Appalachian U.S. Moreover, large variations in uptake existed among Appalachian subregions. Many variables appear to contribute to this variation, including vaccine acceptance for younger adolescents, local and press-driven critical reports of the vaccine, physician communication, and views of the family matriarchs. Targeting the Appalachian subregions, specific campaigns or intervention may be more impactful than viewing the region as a homogenous whole.

Background: External Beam Radiotherapy is essential in the management of locally advanced cervical cancer (LACC). Generally, VMAT is thought to achieve higher conformity to the Planned Target Volume (PTV) and better sparing of organs at risk (OAR) when compared to 3D-CRT. This study focused on these principles as it applied to treatment and potential toxicity in the management of LACC. Aim: To compare dosimetric parameters between VMAT and 3D-CRT in the management of LACC. Setting: The study analysed patients treated at Groote Schuur Hospital between May and December 2017. Method: A non-randomized comparative retrospective study. EBRT plans for 3D-CRT and VMAT were generated and data on treatment parameters for PTV D50%, Dmax, Dmean, Conformity Index (CI), Homogeneity Index, Treated Volume (TV), Irradiated Volume (IV) and OAR constraints; femoral heads, bladder, bowel bag, rectum and bone marrow were collected. Results: Of the 45 patients assessed, VMAT showed significantly lower treatment parameter values for CI (1.09 vs 1.49; p< .001) whereas, 3D-CRT showed lower Dmax (48.1Gy vs 49.2Gy; p< .001) and rectum (88.5% vs 96%). A reduced 3D-CRT dose was noted for bladder Dmax (47.4Gy vs 48.3Gy; p< .001). Conclusion: VMAT offered a superior dosimetric option, with better OAR dose sparing and optimal tumour dosimetry.

Objective: To characterize the cervicovaginal microbiota of HPV-positive and HPV-negative asymptomatic Peruvian women, by identifying the presence of 13 representative bacteria genus. Results: A total of 100 HPV-positive and 100 HPV-negative women were matched by age for comparison of microbiota. The following bacteria were more frequently identified in HPV-positive patients compared to HPV-negative: Eubacterium (68 vs 32%), Actinobacteria (46 vs 33%), Fusobacterium (11 vs 6%) and Bacteroides (20 vs 13%). A comparison between high-risk and low-risk genotypes was performed and differences were found in the detection of Actinobacteria (50 vs 33.33%), Bifidobacterium (50 vs 20.83%) and Enterococcus (50 vs 29.17%). / Revisión por pares

The development of cervical cancer depends on high-risk human papillomavirus (HR-HPV) persistent infection in the cervix. The transformation process leading to invasive cancer can take many years even decades and provides ample opportunity to detect, prevent and cure true precursor lesions. Although cervical cancer is widely preventable, it is the fourth most common cancer among women throughout the world, being a real public health issue, especially in developing countries, as 85 % of deaths occur in low and middle-income countries. The situation in Bolivia is particularly alarming, as the cervical cancer incidence is around 38.5 per 100,000 women, which is estimated to be the highest incidence in Latin America. Prevention of cervical cancer in Bolivia is mainly based on the cytological examination of a Papanicolaou smear (Pap) and more recently on visual inspection after application of acetic acid (VIA). However, many economic, sociocultural, and geographic barriers impair this prevention program being successful, as reflected by the low coverage obtained with these screening tests.In order to reduce the cervical cancer incidence and mortality in the department of Cochabamba in Bolivia, we aimed to assess a low-cost HPV test applicable on self-samples. We believe that this strategy could improve the poor screening programs developed in our country. An evaluation was first made to knowledge of Bolivian women about human papillomavirus (HPV) and cervical cancer. As expected, Bolivian women, from rural, peri-urban and urban areas, knew little or nothing about those. Secondly, their degree of acceptability and confidence towards HPV self-sampling was assessed. Most of the women found self-sampling easier to perform (86.9 % to 93.2 %) and more comfortable (79.4 % to 83.3 %) compared to physician sampling. However, accuracy to detect cervical pre-cancer was higher in their point of view when it was performed on specimens taken by a physician (35.1% to 63.5%). Accordingly, the campaign of vaginal HPV self-sampling in a peri-urban area increased screening coverage, reaching in three months the annual rate average. Finally, the determination of accuracy to detect preneoplastic lesions was assessed for three screening methods, in 469 women, divided in two groups. The first group included 362 women that underwent three consecutively primary screening tests: self-collected sampling for HR-HPV detection, conventional cervical cytology and visual inspection under acetic acid (VIA). The second group included 107 women referred with a positive HR-HPV test that underwent were triage by conventional cervical cytology and VIA. The presence of high-grade intraepithelial neoplasia or invasive cancer (CIN 2+) was verified by colposcopy and biopsy.Among primary screening tests, the sensitivity of the HR-HPV test to detect CIN 2+ lesions was the highest (76 %). In HR-HPV positive women, the sensitivity of the VIA and cytology to detect CIN 2+ lesions were 100% and 81%, respectively.In conclusion, the knowledge about cervical cancer and HPV infection is poor in Bolivia. Despite greater acceptance of the vaginal self-sampling in all areas, women kept greater confidence in the screening performed by the gynecologist, although HPV self-sampling improved coverage rate. Finally, HPV testing on self- samples was the most sensitive screening test and VIA was the most sensitive method for the triage. / Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie) / info:eu-repo/semantics/nonPublished

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18894号 / 医博第4005号 / 新制||医||1009(附属図書館) / 31845 / 京都大学大学院医学研究科医学専攻 / (主査)教授 山田 泰広, 教授 戸井 雅和, 教授 小川 誠司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Cervical cancer is the second most common cancer affecting women in developing countries and accounted for 84% of the global incidence of cervical cancer in 2012. Nicaragua is one country illustrating this disparity, with an annual cervical cancer mortality six times the U.S. rate. This may be explained by lack and poor utilization of effective screening programs, especially the Papanicolaou, or Pap, smear. This study resulted from a partnership formed by faculty and students from two U.S. universities and a Nicaraguan nonprofit organization to conduct projects to benefit a community in Nicaragua. To promote a free Pap smear program provided by the local clinic, a community-wide survey regarding Pap smear utilization was conducted with local health promotoras (promoters). Of 1,117 women, 78.4% reported ever having a Pap smear, of whom 11.1% had not received their results, while results were reported as normal by 78.9%, and abnormal by 10%. The most common reasons for not having a Pap smear were refusal to test, fear, and pain. Proportions of women who ever had a Pap smear varied by etapa (stage/neighborhood, p < .001). Findings are useful for policy development to improve the clinic’s screening services and encourage full utilization of Pap smears.

Cervical cancer is the fourth most common cancer in women. It is caused by the hu- man papillomavirus (HPV). There are many different types of HPV, some of which are high-risk, highly associated with cancer, and low-risk. While HPV is very common— most sexually active individuals will contract some sexually transmitted HPV infec- tion in their lifetime—most infections are cleared without any complication. However, persistent infections may establish and develop into cancerous lesions. Two vaccines have been developed against the two most high-risk types, and have shown high lev- els of efficacy thus far. However, infections are still occurring and it is not clear why some individuals develop persistent infections while others do not. In this thesis, we develop a model to describe how the infection spreads within the host. We express the basic reproduction number R0, a threshold for the establishment of an infection. We solve for the diseased equilibrium, providing insight about whether an infection will persist or not. We develop a spatial model to examine how spatiality of the infec- tion process affects the establishment or clearance. Lastly, we develop a multi-type HPV model to examine whether competitive HPV types are able to coexist in the host for different levels of competition. Ultimately, this work provides groundwork for within-host modelling of HPV and can provide direction for future research. / Thesis / Master of Science (MSc) / The human papillomavirus (HPV) is a sexually transmitted infection that is known to cause cervical cancer in women along with other genital cancers. Cervical cancer is the fourth most common cancer in women, and thus researchers are looking to reduce the number of cervical cancer cases and the number of HPV infections. In order for HPV to cause cervical cancer, the infection must persist for a long time. Most individuals clear the infection without any complication; however, some individuals develop persistent infections. By using mathematical and computation models, we hope to understand why and how HPV infections spread in the host. We develop a criterion for when the infection may be able to establish in the host, and explore conditions that could lead to clearance. Understanding when and how infections will persist could inform treatment and monitoring of cervical cancer development.