Exosomes: A Novel Zika Virus Vaccine Candidate

With the recent emergence of Zika virus (ZIKV) diseases, increasing global concern has driven the demand for a vaccine. One promising vaccine platform has presented itself in the form of exosomes: a subgroup of extracellular vesicles released by many human cell types that facilitate intercellular communication. The objective of this study is to engineer exosomes that incorporate ZIKV structural proteins into its phospholipid bilayer. Previous studies indicate that CD9 and CD63 proteins are highly enriched in exosomal membranes. From this, it was hypothesized that attaching ZIKV genes to CD9 or CD63 to produce a gene fusion may enable exosomes to act as antigen-presenting vesicles. These engineered exosomes may potentially stimulate T-cells to mount a strong immune response. The cDNA of the CD9, CD63, and the highly immunogenic ZIKV genes (envelope, precursor membrane, and NS1) were generated using RT-PCR. These products were used as a template for regular PCR, and cloned into pcDNA3.1/V5 vector. The chimeric gene fusion was assembled using the Gibson assembly kit, and transfected into human embryonic kidney epithelial (HEK293T) cells for expression. The exosomes were purified from the supernatant and subjected to immunoblotting and immunofluorescence assays to confirm the presence of ZIKV proteins. The results of this study are pending at the time of this abstract submission. A future study will be conducted using an in vitro activation assay to determine if the engineered exosomes induce T-cell activation. The potential candidates will be used in an animal study for immunity against ZIKV infection

Structure‐guided insights on the role of NS1 in flavivirus infection

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/111115/1/bies201400182.pd

Fekumi kihe potupotutatau ʻa e Tonga: Finding a balance of Tongan identity

RESEARCH QUESTION How can traditional Tongan narratives shape and inform cultural learning spaces? ABSTRACT Like most stories and traditions learned as children, it is through word of mouth. With every passing generation, slight changes to those practices are moulded into the perspectives of the time. A big misconception is that youth and young adults in Tonga are fully immersed and involved in the culture. This is simply not the case. The younger generation must learn these traditions before the ‘cultural library’ inside the generation above them is gone. The practices that will be explored in this project are the main three Tongan artforms, tufunga (material arts), nimamea’a (fifine arts), and faiva (performing arts). A fourth discipline will also be explored, lea (language), to help users navigate and utilise both Tongan and English. Providing a cultural learning space will help the youth and young adults who do not experience their heritage fully. By looking at the tala tupu’a e kava (Tongan creation myth of kava), an aspect of Tongan culture known and shared throughout its culture and tradition, a dialogue of connectivity and learning emerges. Kava encompasses multiple customs and connects all three artforms in traditions through protocols like the taumafa kava. Utilising kava’s inflfluence in these artforms can help embed them within the project. Breaking down the myth into core values and ideas, Balance and Honour, an architectural design process can be explored. Tevita Ka’ili’s tauhi vā theory will aid in understanding Tongan spatial relationships (vā ) and will be used to speculate how design might successfully incorporate these sociospatial dynamics. Using the tala tupu’a e kava as a catalyst, this project seeks to design a cultural learning space dedicated to the betterment of Tongan identity and the preservation of traditional Tongan artforms. OBJECTIVES • Design a space to bring back a traditional learning environment that locals feel comfortable in. • To provide an adjustable spatial design for the community for events and activities • To capture the essence of the myth in the design through the values it demonstrates and help establish a deeper connection to the architecture • Selecting a site that will achieve the greatest visibility, access, and success for the project • To provide an emphasis on traditional learning to allow youth and even adults, young and old, to reconnect on an identity level with their culture and each other • This project will also seek to address how architecture can help facilitate the relationships between people through socio spatial principles found in Tongan culture • Illustrate how architecture helps occupants reconnect to their Tongan cultures through learning traditional practices, using the four artforms tufunga, nimameaʻa, faiva, and lea Site: Neiāfu, Vavaʻu, Tong

Exploring the Dietary Experiences of Tongan Americans: Barriers and Facilitators to Healthy Dietary Behaviors

Ph.D

Illness beliefs and adherence in diabetes mellitus: a comparison between Tongan and European patients

Aims: The aim of this study was to determine whether there are cultural differences in the way in which Tongan and European people with Type 2 diabetes conceptualise their illness and treatment. The relationships between patients’ illness and treatment perceptions and their adherence to self-care regimes were also assessed.Methods: Participants completed either a Tongan or English version of a questionnaire, which included standardised measures of personal beliefs about diabetes and medication, and self-reported adherence. Information about the severity of patients’ diabetes was obtained from patients’ notes.Results: Comparisons of glycosylated haemoglobin levels showed that Tongan patients had significantly poorer control over their diabetes than did European patients. They were also significantly more likely than European patients to perceive their diabetes as acute and cyclical in nature, uncontrollable, and caused by factors such as God’s will, pollution in the environment, and poor medical care in the past. Tongan patients perceived less necessity for medication, and exhibited higher emotional distress related to their diabetes. The beliefs that characterised the Tongan patients tended to be associated with poorer adherence to diet and medication taking.Conclusions: This study highlights the need for assessment of patients’ personal and cultural beliefs about their illness. Understanding patients’ perceptions may provide an avenue for improving adherence to self-care regimens

Embedding digital technology into contemporary Māori and Pasifika architectural practise

The research paper will review how designers and architects can utilise digital technology to produce culturally respectful Māori and Pasifika architectural outputs within a contemporary landscape. The purpose of the research is to determine how digital fabrication technology can embody the same mana found within traditional design thinking and making process. The research will firstly aim to understand what social customs must be retained to ensure the design outcome are culturally appropriate. The second aim of the research is to determine what traditional building and artisan crafts must be employed in the fabrication process. The method will predominantly a review of various forms of literature, recorded interviews and case studies where possible. Three generations of Māori and Pasifika architectural practice having been selected for this review to ensure a large and diverse cultural representation are analysed. The research findings have indicated there are a large number of approaches to producing contemporary digital architecture. While some approaches ensure traditional craftsman are involved throughout the design and fabrication process, other designers opt to engage with modern craftsman with the blessing of the community. The value of this research is important, as it will serve as a mechanism to understand the conflicts between tradition and technological progress. Although it is essential to preserve cultural skill, expertise and craft, it is equally crucial to innovate technologically. The research goal is to enable digital architecture that can spiritually resonate mana and respect to ancestors of Māori and Pasifika culture

Potential Dual Role of West Nile Virus NS2B in Orchestrating NS3 Enzymatic Activity in Viral Replication

West Nile virus (WNV) nonstructural protein 3 (NS3) harbors the viral triphosphatase and helicase for viral RNA synthesis and, together with NS2B, constitutes the protease responsible for polyprotein processing. NS3 is a soluble protein, but it is localized to specialized compartments at the rough endoplasmic reticulum (RER), where its enzymatic functions are essential for virus replication. However, the mechanistic details behind the recruitment of NS3 from the cytoplasm to the RER have not yet been fully elucidated. In this study, we employed immunofluorescence and biochemical assays to demonstrate that NS3, when expressed individually and when cleaved from the viral polyprotein, is localized exclusively to the cytoplasm. Furthermore, NS3 appeared to be peripherally recruited to the RER and proteolytically active when NS2B was provided in trans. Thus, we provide evidence for a potential additional role for NS2B in not only serving as the cofactor for the NS3 protease, but also in recruiting NS3 from the cytoplasm to the RER for proper enzymatic activity. Results from our study suggest that targeting the interaction between NS2B and NS3 in disrupting the NS3 ER localization may be an attractive avenue for antiviral drug discovery

Potential Dual Role of West Nile Virus NS2B in Orchestrating NS3 Enzymatic Activity in Viral Replication

West Nile virus (WNV) nonstructural protein 3 (NS3) harbors the viral triphosphatase and helicase for viral RNA synthesis and, together with NS2B, constitutes the protease responsible for polyprotein processing. NS3 is a soluble protein, but it is localized to specialized compartments at the rough endoplasmic reticulum (RER), where its enzymatic functions are essential for virus replication. However, the mechanistic details behind the recruitment of NS3 from the cytoplasm to the RER have not yet been fully elucidated. In this study, we employed immunofluorescence and biochemical assays to demonstrate that NS3, when expressed individually and when cleaved from the viral polyprotein, is localized exclusively to the cytoplasm. Furthermore, NS3 appeared to be peripherally recruited to the RER and proteolytically active when NS2B was provided in trans. Thus, we provide evidence for a potential additional role for NS2B in not only serving as the cofactor for the NS3 protease, but also in recruiting NS3 from the cytoplasm to the RER for proper enzymatic activity. Results from our study suggest that targeting the interaction between NS2B and NS3 in disrupting the NS3 ER localization may be an attractive avenue for antiviral drug discovery

IMS localization and expression of WNV NS4B-GFP with or without the 2K-signal peptide in infected cells.

<p>(A) HEK293 cells were infected (b–d, f–h, j–l, and n–p) or mock-infected (a, e, i, and m) with WNV and after 2 hr transfected with C-4B (b–d), C-2K-4B (f–h), C-sig4B (j–l) or GFP (n–p). Cells were fixed at 24 hr after infection and immunostained with mouse anti-dsRNA antibody (red; c–d, g–h, k–l and o–p). The mock-infected cells were transfected and immunostained with mouse anti-dsRNA antibody (a, e, i, and m). Nuclear DNA was labeled with DAPI. The arrowheads indicate IMS. Confocal IF images were of optical slice thickness ∼1 µm. Scale bar, 10 µm. (B) Western blot analysis of WNV NS4B plasmids and stress response in transfected HEK293 cells. Fifty µg of cell lysate was loaded and electrophoresed on SDS-PAGE followed by immunoblotting with rabbit anti-GFP polyclonal antibody, an anti-GRP78 monoclonal antibody or anti-β-actin monoclonal antibody followed by a peroxidase-conjugated secondary antibody. ImageJ analysis of the western blot was conducted to determine GRP78 protein levels relative to β-actin.</p