Grants

Josephine County

A Street Building Petroleum Cleanup Project

NIAID - National Institute of Allergy and Infectious Diseases

Post-TB lung disease (PTLD) contributes substantially to the overall morbidity and mortality associated with TB infection. On a histopathological level, fibrosis is a significant feature of PTLD and likely underpins many associated symptoms and clinical findings. Adjunctive therapies that could be used with traditional anti-TB antibiotics to modulate tissue destruction and pathologic remodeling could improve post-treatment lung health, quality of life, and longevity for TB survivors. To date, our insight into the pathological processes driving TB- associated fibrosis is limited, consequently limiting the development of such adjunctive treatments. In the proposed work, we will begin to address that knowledge gap by creating a baseline understanding of the molecules and cells associated with fibrogenesis during TB infection. Our work will make use of unique resources in the Steyn group, including a Human Tissue Biobank of lung specimens that represent a range of TB- associated fibrotic pathologies, and the unique resources of the Barczak laboratory, who are applying a mouse model to study pathogenic mechanisms of TB-associated fibrosis. In Aim 1, we will use our mouse model and a combination of histopathology, immunohistochemistry, and immunofluorescence to identify spatial correlates of fibrogenesis. We will first test our hypothesis that defined macrophage subsets are spatially associated with fibrogenesis (1A). The idiopathic pulmonary fibrosis (IPF) mouse model is the canonical model for fibrogenesis in lung; we will next test whether molecular and cellular factors identified in the IPF model are associated with TB-associated fibrogenesis (1B). Complementing our hypothesis-driven work, we will use spatial transcriptomics to identify novel candidate molecules, cells, and pathways (1C). In Aim 2, we will use the Steyn lab Human Tissue Bank to benchmark findings from the mouse model and identify clinical drivers of TB-associated fibrosis. We will use metadata to identify clinical correlates of four intermediate-stage fibrosis morphologies (2A). We will then test associations between fibrosis and macrophage subsets (2B), additional molecular and cellular factors relevant in the IPF model (2C), and novel cellular and molecular factors identified in Aim 1C (2D). We will explicitly compare results between mouse and human specimens. We will then test associations between clinical factors and cellular and pathway correlates of fibrosis (2E). In Aim 3, we will use µCT and artificial intelligence approaches to develop algorithms that characterize and comprehensively quantify fibrosis in human lung sections and whole mouse lungs to identify candidate microanatomic contributors to fibrogenesis during TB infection. In addition to enabling a full comparison between mouse and human fibrosis, results of Aim 3 will enable the development of new hypotheses around microenvironmental and anatomical cues for fibrogenesis in TB infection. Successfully completing our three aims will create the foundational knowledge and novel tools necessary to ultimately build a mechanistic model for the path to TB-associated fibrosis and for preclinical testing of candidate interventions. We anticipate this work will directly contribute to new strategies for treating TB.

Forests, Parks & Recreation

A Trail for All at the Pratt Refuge

Forests, Parks & Recreation

A Trail for All at the Pratt Refuge

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY. Precise tissue- and temporally- specific manipulation of gene expression has driven mechanistic studies across many different model organisms. Bimodal expression systems such as the Q-system from N. crassa and the GAL4-system from S. cerevisiae are key genetic tools for elucidating gene function and cellular properties. These systems were introduced and optimized in Drosophila melanogaster, contributing to groundbreaking discoveries in embryonic development, animal growth, physiology, metabolism, and neuroscience, and have had a significant impact in the understanding of human biology, in both health and disease. Using CRISPR/Cas-9 technology, the Q-system (the QF2 driver and QUAS reporters) has been successfully introduced to other insects with major impact on human health, such as Aedes and Anopheles mosquitoes. Additionally, it has been applied to vertebrate model systems including the zebrafish Danio rerio. In contrast, while the GAL4 system, has been used in zebrafish, its transfer to mosquitoes has been unsuccessful due to cell toxicity. However, even the Q-system poses a number of limitations, including toxicity and the lack of an effective suppressor that functions independently of chemical compounds. In his application, the Q-system will be re-engineered to remedy these drawbacks by incorporating temperature-sensitive self-splicing intein modules (INTts), tested for efficacy in D. melanogaster and transferred to the zebrafish Danio rerio. Inteins are self-splicing endopeptidases embedded within proteins found in bacteria and unicellular eucaryotes (e.g., yeast), with the enigmatic property of excising themselves from pre-proteins, leading to the generation of a functional protein. Remarkably, inteins can be inserted into foreign proteins, where they self-splice as in their normal host protein. The experimental strategy leverages temperature-sensitive self-splicing inteins that have been validated in S. cerevisiae to self-splice at temperatures up to 270 C in foreign proteins. Specifically, inteins with permissive and restrictive setpoints between 15 and 300 C, compatible with growth and development of insects and fish, will be integrated into the DNA binding domain of QF2 (QF2_ INTts) and the protein interaction domain of QS (QS_ INTts), to disrupt their function when retained at restrictive temperature (24 to 300 C). When kept at permissive set points (17 to 230 C), intein removal via self-splicing activates QF and QS. Transgenic Drosophila will be tested for functionality of QF2_ INTts and QS_ INTts using QUAS-GFP reporter genes at different temperature. Genes encoding validated QF2_ INTts will be conferred to plasmids for D. rerio transgenic fish, tested for functionality using QUAS-GFP reporters. These new tools provide a much-needed temporal control element for the Q-system, making it invaluable for the research community to analyze gene and cell function relevant to human health.

First Hill Improvement Association

In Phase 2, the First Hill Improvement Association will continue working with Site Workshop (consultant), Parks, and DoN to lead the community through final design and construction documentation for improvements to First Hill Park. This phase of work will build off of the approved Phase 1 concept plan.

Friends of Small Business Saturday in Magnolia

The Magnolia Chamber of Commerce will engage the neighborhood to define a collective “Vision for Magnolia Village” and a set of actionable steps to help shape future development and growth.

Montgomery County MD / State of Maryland

Housing and Community Development

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Zika virus (ZIKV) is a re-emerging mosquito-borne flavivirus of significant public health concern. To date there are no effective licensed antiviral treatments or a vaccine. Therefore, elucidating the molecular biology of these viruses and the interactions with the host cell are foundational to identifying and developing effective treatment options. The single-stranded positive-sense RNA genome of ZIKV mimics a cellular mRNA. Specifically, the viral RNA encodes a single open reading frame, has structured untranslated regions (UTRs) adjacent to the open reading frame, and contains a N5’-methyl guanosine cap. Unlike mRNAs however, flaviviruses lack a poly(A) tail. From a multiple sequence alignment of all mosquito-borne flaviviruses we identified conserved regions containing 3-6 tandem adenosines in the 3’ UTR. These A-rich regions are localized in single-stranded regions within and between the XRN-1 resistant pseudoknots (xrRNA), dumbbell and 3’ stem loop (3’SL) RNA structures. In preliminary experiments we investigated the role of the A-rich region between xrRNA2 and the pseudo- dumbbell RNA structure (pre-pseudo/Y-dumbbell). Specifically, we generated three different mutations in a ZIKV Renilla luciferase reporter replicon and infectious clone. These mutants revealed that the pre-YDB A-rich region had a role in translation in both mammalian and mosquito cell lines. Different mass spectrometry studies have shown that the cellular poly(A) binding protein (PABP) interacts with the ZIKV 3’ UTR. Additionally, PABP was previously shown to interact with a region in the dengue virus 3’ UTR that harbors an A-rich motif. In preliminary studies we find that depletion of PABP1 decreased ZIKV, but not cellular, protein levels and viral titers without affecting cell viability. We therefore hypothesize that A-rich regions in the ZIKV 3’ UTR function to recruit cellular RNA binding proteins such as PABP1 to promote distinct steps in the virus infectious cycle. In Aim 1 we will mutate the A-rich regions in the infectious clone and a subgenomic luciferase reporter replicon to investigate the function of select A-rich regions on translation, replication, and viral fitness in mammalian and mosquito cells. In Aim 2, we will investigate the role of PABP1 on ZIKV gene expression and determine if PABP1 and other cellular RNA binding proteins bind A-rich regions in the ZIKV 3’ UTR. Overall, this study will advance our understanding of how flavivirus 3’ UTRs interact with the host to promote distinct steps in the infectious cycle in two vastly different hosts. Understanding how specific sequences in RNA genome function could lead to the therapeutic advancement namely the development of an attenuated vaccine ZIKV strain. Moreover, defining similar and unique RNA-protein interactions between mammalian and mosquito hosts could inform future vector control strategies.

Young Faces Smiling

Anti-Poverty

West Harlem Group Assistance, Inc.

Local

Montgomery County MD / State of Maryland

Housing and Community Development

HPZS

Management and Business Professionals and Administrative Services

Specialty Consulting, Inc.

Management and Business Professionals and Administrative Services

FGM Architects Inc.

Management and Business Professionals and Administrative Services

FGM Architects Inc.

Management and Business Professionals and Administrative Services

FGM ARCHITECTS INC.

Management and Business Professionals and Administrative Services-Management advisory services

Project U N I Q U E Inc

A/R for V.I.T.A.L.

RADA ARCHITECTS LTD

Management and Business Professionals and Administrative Services-Management advisory services

City of Richmond

A1 Nails

PRODUCTION DISTRIBUTION COMPANIES INC

Electrical Systems and Lighting and Components and Accessories and Supplies

Enfield

Abbe RD Reconstruction

NYC Department of Cultural Affairs

ABC No Rio

City of Richmond

Abdu Saleh