Optimizing The Melanoma Tropism Of Mouse Parvovirus 1a For Use As A Viral Immunotherapy Vector

Melanoma is a prevalent, deadly disease with poor outcomes following metastasis. It is an immunogenic cancer with many unique tumor antigens, but the tumor microenvironment inhibits the immune system. Current monoclonal antibody treatment has been shown to be beneficial in metastatic melanoma by blocking inhibitory immune signals such as PD-1 or CTLA-4. The expression of immunostimulatory proteins such as B7.1 by tumor cells could provide additional immune targeting. Viral vectors based on the parvovirus genome could efficiently and selectively express B7.1 in melanoma cells, such that they become competent to directly activate cognate T cells. Once activated, T cells will become armed as cytotoxic effectors against cells expressing melanoma tumor antigens. To generate a viral vector, we first needed a parvovirus that infected melanoma. Parvoviruses are uniquely adapted to infect tumor cells as viral replication is dependent on the host cell advancing through S phase. However, most parvoviruses tested were unable to establish infection in the murine melanoma cell line B16F10. The most infectious parvovirus in B16F10 was mouse parvovirus 1a (MPV1a). B16F10 cells were infected with MPV1a, and the progeny virus was harvested and used to re-infect more B16F10 cells. After five serial passages of virus, the mutant polyclonal stock named MPV P5 was analyzed. At 24 hours following infection at a multiplicity of 5000 virions per cell, MPV1a infected 16% of cells, whereas MPV P5 infected 74%. Construction of a molecular clone from the polyclonal MPV P5 mixture yielded a construct consisting of the non-structural and virion polypeptide (NS and VP) genes that was able to infect 45% of cells under the same conditions. The mutations that were most responsible for the increase in B16F10 infectivity were isolated to the VP2 region, which encodes the major capsid protein. By incorporating the viral proteins from the clonal version of MPV P5 into a parvoviral vector system, we have generated a vector capable of transducing B16F10 cells and simultaneously expressing the Green Fluorescent Protein marker and B7.1. Further experiments are needed to determine if the B7.1 expression is sufficient at its current rate of infectivity to modulate an immune response, and to examine whether viral products within the tumor cells generate additional immunogenic signaling. Other aspects of the MPV P5 genome may be engineered into the vector to increase its efficiency

Towards a contextual approach to the place–homeless survival nexus: An exploratory case study of Los Angeles County

The characteristics of the immediate locale greatly affect the ability of homeless people to adapt to life on the street and in shelters, with different types of places nurturing different circumstances for survival. Current conceptualizations of the place–survival nexus are too narrow, relying on small-scale, intensive studies of particular places that are known to sustain homeless survival while ignoring more suburban and exurban locales, as well as failing to set these places of survival within the larger socio-economic spaces of the metropolitan area. Further, the literature is heavily qualitative, lacking any kind of ‘‘big picture” quantitative assessment of the nexus. In response, we contribute to the place–survival nexus literature by developing a typology of space for homeless survival and then use interview data to examine the variation in survival strategies across three types of urban space in Los Angeles County. Our results speak to how our innovative and exploratory approach enabled a broader, more extensive and variegated understanding of place–survival among homeless people than previous studie

Vínculos sociales y subsistencia en los «Templos de refugio» en Japón: Una exploración de las influencias en la situación de calle entre los jornaleros de los yoseba en Tokio

El trabajo por jornal se describe a menudo como no estructurado, lo que hace ambigua su capacidad de servir como trampa en la situación de calle o como paso hacia la economía formal. Exploramos esta cuestión describiendo los factores que influyen en la situación de calle entre los jornaleros. Analizamos datos de encuestas del mayor yoseba de Tokio, o enclave del trabajo por jornal, y hallamos que los jornaleros evitaron la situación de calle por más días trabajados, educación, permanencia en el cargo como jornalero, y conexiones interpersonales con los intermediarios. La atención a experiencias acumulativas y lazos sociales en los mercados de trabajo por jornal es crucial para entender su estructura, dinámica, funciones y efectos en las trayectorias a largo plazo de los trabajadores

Neighborhoods of Refuge: A PhotoVoice Project Proposal

Natalia Marques da Silva and Matthew Marr\u27s proposal for Neighborhoods of Refuge: A PhotoVoice Project introduces a collaboration between FIU, MDC, and Camillus House art therapy participants that follows the challenges, opportunities, stigma, and day-to-day realities related to homelessness through photograph

Assessing Agricultural Literacy in Urban and Suburban Mothers After Their Completion of the Illinois Farm Families Program

A true definition of what agricultural literacy actually means, has yet to be agreed upon by researchers. The agriculture industry, after all, is constantly evolving and changing. As time passes, much of society continues to become further and further removed from America’s agrarian heritage. The industry has been subject to much criticism in recent years, with allegations of mass malpractice. Due to the advent of various media platforms, people are able to publish falsified or inaccurate information for the world to see, and exploit the very industry that clothes and feeds them. What’s more, there are a variety of labels and marketing campaigns that have done nothing but mislead, and confuse consumers. However, a program by the name of Illinois Farm Families sought to combat these issues by providing urban and suburban mothers from Chicagoland with a yearlong learning experience in agriculture. Since 2012, IFF has taken cohorts of these mothers out to a variety of farming operations across the state of Illinois, with the intent to provide them with a firsthand experience in stepping foot on these farms, and conversing face to face with farmers. The researcher framed this study around two theories. The first being a centralized diffusion system, in which the Illinois Farm Families program fits as the formalized change agency. The second theory is Dewey’s experiential learning model, since this program is extremely experiential in nature. This census study of all mothers who are alumna of the Illinois Farm Families program assessed their levels of agricultural literacy. They were asked to complete the Food and Fiber Systems Literacy assessment, and answer six questions pertaining to their personal characteristics. The researcher compared their results based on personal characteristics such as: the year(s) they participated, level of completed education, number of children, ethnicity, age range, and proximity to Chicago proper. The findings revealed that these mothers are in fact agriculturally literate, based on a 60% benchmark. Although this was not a true pre/post-assessment, nor a true program evaluation, it is recommended that this study be replicated in these forms

Design Considerations for Embankment Protection During Road Overtopping Events

This report describes the research conducted by the University of Minnesota and project partners on roadway embankment overtopping by flood water. Roadway overtopping is a major safety concern for Minnesota transportation managers because of the potential for rapid soil erosion and mass wasting resulting in partial or complete failure of the roadway embankment. This multi-year research study focused on various aspects of the roadway embankment overtopping. A robust literature survey was performed to identify research, reports and other published knowledge that would inform the project. A field- based research campaign was developed with the goal of collecting data on the hydraulics associated with full-scale overtopping events. Finally, a series of laboratory experiments were conducted at the St. Anthony Falls Laboratory, University of Minnesota to study the hydraulic and erosional processes associated with embankment overtopping and in particular study of three slope protection techniques under overtopping flow. The largest component of the research project was the laboratory hydraulic testing, which focused on bare soil (base case) and three slope protection technologies. A full- scale laboratory facility was constructed to carry out the testing. Three erosion protection techniques were examined including 1) armored sod, 2) turf reinforcement mat, and 3) flexible concrete geogrid mat. Overtopping depths of up to 1-ft were used to determine the failure point of the protection technique and soil on both the 4h:1V and 6V:1H slopes. The full project report details the testing of each protection technique as well as observations and findings made during the testing

Dynamic clonal progression in xenografts of acute lymphoblastic leukemia with intrachromosomal amplification of chromosome 21

Intrachromosomal amplification of chromosome 21 is a heterogeneous chromosomal rearrangement occurring in 2% of childhood precursor B-cell acute lymphoblastic leukemia. There are no cell lines with iAMP21 and these abnormalities are too complex to faithfully engineer in animal models. As a resource for future functional and pre-clinical studies, we have created xenografts from intrachromosomal amplification of chromosome 21 leukemia patient blasts and characterised them by in-vivo and ex-vivo luminescent imaging, FLOW immunophenotyping, and histological and ultrastructural analysis of bone marrow and the central nervous system. Investigation of up to three generations of xenografts revealed phenotypic evolution, branching genomic architecture and, compared with other B-cell acute lymphoblastic leukemia genetic subtypes, greater clonal diversity of leukemia initiating cells. In support of intrachromosomal amplification of chromosome 21 as a primary genetic abnormality, it was always retained through generations of xenografts, although we also observed the first example of structural evolution of this rearrangement. Clonal segregation in xenografts revealed convergent evolution of different secondary genomic abnormalities implicating several known tumour suppressor genes and a region, containing the B-cell adaptor, PIK3AP1, and nuclear receptor co-repressor, LCOR, in the progression of B-ALL. Tracking of mutations in patients and derived xenografts provided evidence for co-operation between abnormalities activating the RAS pathway in B-ALL and for their aggressive clonal expansion in the xeno-environment. Bi-allelic loss of the CDKN2A/B locus was recurrently maintained or emergent in xenografts and also strongly selected as RNA sequencing demonstrated a complete absence of reads for genes associated with the deletions

Tale of two curricula: The performance of 2000 students in introductory electromagnetism

The performance of over 2000 students in introductory calculus-based electromagnetism (E&M) courses at four large research universities was measured using the Brief Electricity and Magnetism Assessment (BEMA). Two different curricula were used at these universities: a traditional E&M curriculum and the Matter & Interactions (M&I) curriculum. At each university, postinstruction BEMA test averages were significantly higher for the M&I curriculum than for the traditional curriculum. The differences in post-test averages cannot be explained by differences in variables such as preinstruction BEMA scores, grade point average, or SAT Reasoning Test (SAT) scores. BEMA performance on categories of items organized by subtopic was also compared at one of the universities; M&I averages were significantly higher in each topic. The results suggest that the M&I curriculum is more effective than the traditional curriculum at teaching E&M concepts to students, possibly because the learning progression in M&I reorganizes and augments the traditional sequence of topics, for example, by increasing early emphasis on the vector field concept and by emphasizing the effects of fields on matter at the microscopic level