Matching Items (12)

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Multiplexed, In-Solution Protein Array (MISPA) for Identification of Novel Protein Interactions and Early Detection of Pathogen Induced Cancers

Description

Disturbances in the protein interactome often play a large role in cancer progression. Investigation of protein-protein interactions (PPI) can increase our understanding of cancer pathways and will disclose unknown targets

Disturbances in the protein interactome often play a large role in cancer progression. Investigation of protein-protein interactions (PPI) can increase our understanding of cancer pathways and will disclose unknown targets involved in cancer disease biology. Although numerous methods are available to study protein interactions, most platforms suffer from drawbacks including high false positive rates, low throughput, and lack of quantification. Moreover, most methods are not compatible for use in a clinical setting. To address these limitations, we have developed a multiplexed, in-solution protein microarray (MISPA) platform with broad applications in proteomics. MISPA can be used to quantitatively profile PPIs and as a robust technology for early detection of cancers. This method utilizes unique DNA barcoding of individual proteins coupled with next generation sequencing to quantitatively assess interactions via barcode enrichment. We have tested the feasibility of this technology in the detection of patient immune responses to oropharyngeal carcinomas and in the discovery of novel PPIs in the B-cell receptor (BCR) pathway. To achieve this goal, 96 human papillomavirus (HPV) antigen genes were cloned into pJFT7-cHalo (99% success) and pJFT7-n3xFlag-Halo (100% success) expression vectors. These libraries were expressed via a cell-free in vitro transcription-translation system with 93% and 96% success, respectively. A small-scale study of patient serum interactions with barcoded HPV16 antigens was performed and a HPV proteome-wide study will follow using additional patient samples. In addition, 15 query proteins were cloned into pJFT7_nGST expression vectors, expressed, and purified with 93% success to probe a library of 100 BCR pathway proteins and detect novel PPIs.

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Date Created
  • 2016-12

Developing and Pilot Testing Digital Storytelling Interventions to Promote HPV Vaccinations among Vietnamese American Adolescents

Description

Significance Background: Human papilloma virus (HPV) is the most common sexually transmitted infection, affecting 79 million Americans today and an additional 14 million Americans becoming infected with HPV each year.

Significance Background: Human papilloma virus (HPV) is the most common sexually transmitted infection, affecting 79 million Americans today and an additional 14 million Americans becoming infected with HPV each year. HPV infection may lead to the development of genital warts and several types of cancers including both cervical and oropharyngeal cancers. The promotion of currently available HPV vaccines is important to prevent HPV transmission and reduce the prevalence of the comorbidities associated with infection. Promotion to Vietnamese-Americans in particular is important because of the increased rates of cervical cancers seen in this population. As Vietnamese-American mothers often act as the primary healthcare decision maker for their children, they were chosen as the target population for this intervention. Purpose: This study aims to (1) develop personal digital stories about HPV and HPV vaccination among Vietnamese women with adolescent children who are vaccinated against HPV; and (2) share these stories with a group of Vietnamese American mothers and assess the effect of the stories in changing the attitudes, beliefs, and intention to vaccinate for HPV. Methods: This study used a two-step process to design, implement, and evaluate digital stories to improve Vietnamese mothers' attitudes, beliefs, and intention to vaccinate their adolescent children against HPV. The first step was a formative research design to develop the digital stories. The second step was quasi-experimental with a pre and posttest design to evaluate the effect of the stories. Results: The first phase has produced two digital stories which will be screened recruitment has been completed for phase two. Content analysis showed the importance of community resources, the desire to protect children, a history of familial and/or personal cancer, concerns about side effects, and the influence of healthcare providers as themes in both stories. Recruitment efforts are underway to recruit eligible Vietnamese mothers to assess the effect of these stories. Data collection is ongoing. Conclusions and lessons learned: The project has yielded two digital stories and recruitment for phase two is underway. This project has been successful in obtaining IRB approval, recruiting phase one participants, holding a digital storytelling workshop, designing the phase two survey, and beginning data collection efforts. The phase two recruitment has been challenging and will necessitate a change in strategy to find participants.

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Date Created
  • 2017-05

HPV Vaccine Administration in Community Pharmacies

Description

The human papillomavirus (HPV) vaccine can help prevent numerous cancers and genital warts. Traditionally, pediatricians and family medicine providers administer the vaccine. However, pharmacists can also vaccinate against HPV. The

The human papillomavirus (HPV) vaccine can help prevent numerous cancers and genital warts. Traditionally, pediatricians and family medicine providers administer the vaccine. However, pharmacists can also vaccinate against HPV. The objective for this study is to assess Arizona pharmacists’ behaviors and influences in relation to administering the HPV vaccine. We administered a survey to Arizona pharmacists at a statewide virtual conference. The key points that are assessed: pharmacists’ behaviors, intentions, attitudes, subjective norms, and perceived behavioral in relation to the human papillomavirus vaccination. Looking at the measures, the leading outcomes of the study involved the HPV vaccination behavior and intentions to administer the vaccine. Secondary outcomes related more to the Theory of Planned Behavior constructs, which ended up being stronger with predictions of HPV vaccine administration intentions and behavior. Our results show that most of pharmacists held very positive attitudes (on a Likert Scale of 1-5)towards the HPV vaccine. It looked like attitude, perceived behavioral control, and subjective norm combined had a big influence on HPV vaccination intentions; however, the strongest predictor came down to the subjective norms, in administering the vaccine. Pharmacists believed strongly with implementing the HPV vaccine, and want to do so in the near future. In conclusion, the overall point of the study is that there should be a need in increasing pharmacy professionals’ subjective norms to vaccinate against the HPV in order to accelerate pharmacy-based HPV immunizations. Implementing human papillomavirus vaccine promotions in the near future could help engage leadership in pharmacy, and further encourage pharmacists’ awareness to administer the vaccine. Additionally, raising pharmacists’ awareness to administer the vaccine among adolescents could add facilitation in increasing human papillomavirus rates.

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Created

Date Created
  • 2020-12

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College Students’ HPV Vaccine Perceptions: What they need to know to get the vaccine

Description

Aims: The aim of this research is to identify factors that would lead to increased utilization of the HPV vaccine among college students.
Methods: We conducted 11 focus groups with

Aims: The aim of this research is to identify factors that would lead to increased utilization of the HPV vaccine among college students.
Methods: We conducted 11 focus groups with a total of 28 students, averaging 3 per group. Using an inductive approach, we hand-coded focus group transcripts, developed a coding structure, and discussed themes as they emerged from the data.
Results: Although more than half of the students had never heard of the HPV vaccine, students generally held positive views about vaccines. Barriers to receiving the HPV vaccine included lack of awareness and knowledge about the HPV vaccine, as well as lack of perceived need for the vaccine. When asked about the most important information that they needed in order to make an informed decision about whether or not to get the vaccine, participants mentioned the following: 1) prevalence of HPV, 2) HPV-related diseases, 3) what the HPV vaccine protects against, 4) HPV vaccine safety, 5) HPV vaccine efficacy, and 6) how they can access the vaccine. Participants differed in their preferences for types of health education, from an in-person informative lecture, to YouTube videos, to posters placed in dorms or in bathrooms. They preferred the conveyor of this information to be a trustworthy source; they identified healthcare providers, professors/researchers, and other students who have received the vaccine as credible sources. In terms of message appeal, many students described wanting the facts, statistics about HPV prevalence and the vaccine as well as narratives from students who have been diagnosed with HPV and those who have experience receiving the vaccine.
Conclusions: Although this cancer prevention resource has been commercially available since 2006, college students still a lack of awareness and perceived need for the HPV vaccine. Future health education efforts should utilize participants’ recommendations to increase students’ understanding of HPV and the HPV vaccine and, therefore, impact their perceived susceptibility to HPV, the benefits of the vaccine to their health, and therefore increase utilization of this resource. Strategies to increase vaccination should include health education and vaccine implementation strategies, as well as strategies to reduce the cost of the vaccine for college students, thereby increasing the accessibility of the vaccine for this population.

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Created

Date Created
  • 2020-05

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College Students’ HPV Vaccine Perceptions: What they need to know to get the vaccine

Description

Aims: The aim of this research is to identify factors that would lead to increased utilization of the HPV vaccine among college students.

Methods: We conducted 11 focus groups with a

Aims: The aim of this research is to identify factors that would lead to increased utilization of the HPV vaccine among college students.

Methods: We conducted 11 focus groups with a total of 28 students, averaging 3 per group. Using an inductive approach, we hand-coded focus group transcripts, developed a coding structure, and discussed themes as they emerged from the data.

Results: Although more than half of the students had never heard of the HPV vaccine, students generally held positive views about vaccines. Barriers to receiving the HPV vaccine included lack of awareness and knowledge about the HPV vaccine, as well as lack of perceived need for the vaccine. When asked about the most important information that they needed in order to make an informed decision about whether or not to get the vaccine, participants mentioned the following: 1) prevalence of HPV, 2) HPV-related diseases, 3) what the HPV vaccine protects against, 4) HPV vaccine safety, 5) HPV vaccine efficacy, and 6) how they can access the vaccine. Participants differed in their preferences for types of health education, from an in-person informative lecture, to YouTube videos, to posters placed in dorms or in bathrooms. They preferred the conveyor of this information to be a trustworthy source; they identified healthcare providers, professors/researchers, and other students who have received the vaccine as credible sources. In terms of message appeal, many students described wanting the facts, statistics about HPV prevalence and the vaccine as well as narratives from students who have been diagnosed with HPV and those who have experience receiving the vaccine.

Conclusions: Although this cancer prevention resource has been commercially available since 2006, college students still a lack of awareness and perceived need for the HPV vaccine. Future health education efforts should utilize participants’ recommendations to increase students’ understanding of HPV and the HPV vaccine and, therefore, impact their perceived susceptibility to HPV, the benefits of the vaccine to their health, and therefore increase utilization of this resource. Strategies to increase vaccination should include health education and vaccine implementation strategies, as well as strategies to reduce the cost of the vaccine for college students, thereby increasing the accessibility of the vaccine for this population.

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Created

Date Created
  • 2020-05

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HPV Vaccination and Perceptions Among College Males

Description

In the US, individuals between ages 18-26 years old have the highest incidence of new HPV infection, the most common sexually transmitted infection. HPV infection can lead to the development

In the US, individuals between ages 18-26 years old have the highest incidence of new HPV infection, the most common sexually transmitted infection. HPV infection can lead to the development of non-cancerous genital warts and persistent infection with cancerous strains can cause various cancers. An HPV vaccine which offers protection against seven cancerous strains and the two non-cancerous strains which cause genital warts has recommended for use by the Centers for Disease Control and Prevention for both men and women beginning in 2011. However, young adult men, including college-aged men, are lagging in HPV vaccine uptake and completion. This cross-sectional study, guided by the Health Belief Model and Theory of Planned Behavior, seeks to explore college men’s behaviors and intentions to getting the HPV vaccine (primary outcomes). It also seeks to explore their perceived susceptibility and severity of HPV infection; attitudes about the HPV vaccine, self-efficacy and social norms to vaccinate, and stigma related to HPV infection (secondary outcomes). Study results showed that only 33% of college men reported knowing they had received at least one dose of the HPV vaccine. The majority of men reported they had never had a conversation about the HPV vaccine with their primary care provider. Overall, men had positive attitudes about the HPV vaccine, high self-efficacy to communicate and access the vaccine, but low intentions and low social norms to vaccinate against this virus. Outcomes of this research suggest the need for future intervention work to focus on increasing college males’ social norms to vaccinate against HPV. Influencing parents, peers, and other influential individuals to encourage college males to vaccinate against HPV may, in fact, increase their intentions and behaviors to utilize this cancer prevention resource.

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Created

Date Created
  • 2020-05

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Expression of 12 High and Low Risk HPV Type Proteomes for the Development of a Protein Microarray

Description

Introduction: Human papillomavirus (HPV) infection is seen in up to 90% of cases of cervical cancer, the third leading cancer cause of death in women. Current HPV screening focuses on

Introduction: Human papillomavirus (HPV) infection is seen in up to 90% of cases of cervical cancer, the third leading cancer cause of death in women. Current HPV screening focuses on only two HPV types and covers roughly 75% of HPV-associated cervical cancers. A protein based assay to test for antibody biomarkers against 98 HPV antigens from both high and low risk types could provide an inexpensive and reliable method to screen for patients at risk of developing invasive cervical cancer. Methods: 98 codon optimized, commercially produced HPV genes were cloned into the pANT7_cGST vector, amplified in a bacterial host, and purified for mammalian expression using in vitro transcription/translation (IVTT) in a luminescence-based RAPID ELISA (RELISA) assay. Monoclonal antibodies were used to determine immune cross-reactivity between phylogenetically similar antigens. Lastly, several protein characteristics were examined to determine if they correlated with protein expression. Results: All genes were successfully moved into the destination vector and 86 of the 98 genes (88%) expressed protein at an adequate level. A difference was noted in expression by gene across HPV types but no correlation was found between protein size, pI, or aliphatic index and expression. Discussion: Further testing is needed to express the remaining 12 HPV genes. Once all genes have been successfully expressed and purified at high concentrations, DNA will be printed on microscope slides to create a protein microarray. This microarray will be used to screen HPV-positive patient sera for antibody biomarkers that may be indicative of cervical cancer and precancerous cervical neoplasias.

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Date Created
  • 2015-05

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Detection of antibodies to HPV16-associated oropharyngeal cancer using custom bead arrays

Description

Oropharyngeal cancer (OPC) is the world's sixth most common cancer and in many cases is associated with infection with human papillomavirus (HPV) type 16. Antibodies (Abs) to HPV16 viral antigens

Oropharyngeal cancer (OPC) is the world's sixth most common cancer and in many cases is associated with infection with human papillomavirus (HPV) type 16. Antibodies (Abs) to HPV16 viral antigens are potential diagnostic biomarkers of HPV-associated OPC (HPV OPC). A custom multiplexed bead array assay was used to detect Abs to HPV16 antigens E1, CE2, NE2, E4, E5, E6, E7, L1, and L2. Following extensive optimization of the assay, these genes were expressed as GST-fusion proteins and captured onto anti-GST magnetic beads. Serum was obtained from 256 OPC patients at the time of diagnosis and from 78 healthy controls. The median fluorescent intensity (MFI) was determined for each antigen and ratios of MFI to control GST-fusion protein were determined for each serum sample. Cutoff values were set as the mean + 3 SD of the MFIs of healthy controls and p-values were calculated using Wilcoxon unpaired and Fisher's exact test. Results of this experiment showed that HPV16 E1, CE2, NE2, E4, E6, and E7 Ab levels were elevated in OPC patients compared to controls (p<0.001), as were Ab levels to L1 (p = 0.013) and L2 (p = 0.023), per Fischer's exact test. Abs to CE2, NE2, E6, and E7 were identified as a potential biomarker panel for early detection of HPV OPC. For the 111 patients with known HPV+ tumors as measured by tumor PCR of E6 and/or E7, this assay had a sensitivity of 90% and specificity of 87% (AUC = 0.96). From these results, we conclude that custom bead array assays can be used to detect HPV16 Abs in patient sera, and we have identified a 4-Ab biomarker panel for the early detection of HPV OPC.

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Date Created
  • 2013-05

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Creating an artificial antigen presenting cell system for HPV16 proteins

Description

Background: High risk types of human papillomavirus (HPV) are known to cause cancer, including cervical (99%) and oropharyngeal cancer (70%). HPV type 16 is the most common subtype. Three antigens

Background: High risk types of human papillomavirus (HPV) are known to cause cancer, including cervical (99%) and oropharyngeal cancer (70%). HPV type 16 is the most common subtype. Three antigens that are critical for integration or tumor progression are E2, E6 and E7. In this study, we developed a systematic approach to identify naturally-processed HPV16-derived HLA class I epitopes for immunotherapy development. Methods: K562 cells, which lack HLA expression, were transduced with each HPV16 antigen using lentivirus and supertransfected with HLA-A2 by nucleofection. Stable cell lines expressing each antigen were selected for and maintained throughout the investigation. In order to establish a Gateway-compatible vector for robust transient gene expression, a Gateway recombination expression cloning cassette was inserted into the commercial Lonza pMAX GFP backbone, which has been experimentally shown to display high transfection expression efficiency. GFP was cloned into the vector and plain K562 cells were transfected with the plasmid by nucleofection. Results: Expression of K562-A2 was tested at various time points by flow cytometry and A2 expression was confirmed. Protein expression was shown for the transduced K562 E7 by Western blot analysis. High transfection efficiency of the pMAX_GFP_Dest vector (up to 97% GFP+ cells) was obtained 48 hours post transfection, comparable to the commercial GFP-plasmid. Conclusion: We have established a rapid system for target viral antigen co-expression with single HLA molecules for analysis of antigen presentation. Using HPV as a model system, our goal is to identify specific antigenic peptide sequences to develop immunotherapeutic treatments for HPV-associated cancers.

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Created

Date Created
  • 2016-05

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Describing Virus Vaccines and Controversies: Comparison of Three Vaccines in Medical and Social Contexts

Description

In the United States, a dispute has arisen over the safety and need for vaccination, particularly in regard to compulsory vaccination laws. New outlets and social media sites publish countless

In the United States, a dispute has arisen over the safety and need for vaccination, particularly in regard to compulsory vaccination laws. New outlets and social media sites publish countless reports about the dangers of vaccines or of known adverse reactions as well as imagined or unproven worries. Individuals' rights to choose to get vaccinated or allow their children to be vaccinated comes to direct conflict with measures needed to protect communities from preventable viral diseases. The controversy surrounding vaccines is not new, nor necessarily are the fundamental reasons for skepticism. Looking back through the history of vaccines as a medical tool, the evolution of the controversy can be observed taking place with each new historical context, scientific development, and social conditions. Despite scientific research and assurances of vaccine safety, opposition and unease about vaccination appear to take Looking individually at the development and distribution of the smallpox (variola virus), polio (poliovirus) and human papilloma virus(HPV) vaccines, concerns regarding the violation of personal rights, safety of vaccines themselves, and social stigmas and connotations surrounding vaccines can be seen to evolve and change. Due to the way doubt can manifest in different ways over time, it may be impossible to fully end the vaccine debate. However, nderstanding the sociological factors behind anti-vaccine sentiment may allow healthcare professionals to work with concerned people with a particular care to address these visceral and sometimes irrational fears surrounding vaccination.

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Date Created
  • 2014-05