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JP SIMONS & COMPANY

Manufacturing Components and Supplies-Pneumatic and hydraulic and electric control systems

Dance Theatre of Harlem, Inc.

Replace outdated and inefficient HVAC boiler systems which will include design, technology and plumbing/electrical upgrades to support a high technical system.

Added Value

Construction of a rain water capture system for a community garden to reduce the garden's ecological footprint, along with community education to promote rainwater capture and use.

Marrakech, Inc.

I/T - Electronic Document System

NIAID - National Institute of Allergy and Infectious Diseases

Abstract Hepatitis B virus (HBV) chronically infects 254 million people worldwide and accounts for 1.1 million deaths per year due to cirrhosis and liver cancer. Finding a cure for this disease is urgently needed, which requires a more thorough understanding of the key molecular events driving establishment and persistence of HBV infections. Thus far, identification of HBV-hepatocyte interactions is mostly achieved through conventional focused screens or one-gene-at-a-time approaches. Due to the overlapping feature of the HBV genome, unbiased genetic screens by reporter HBV viruses allowing phenotypic selection have not been possible. To overcome this challenge, our lab recently developed an RNA sensing and editing based HBV reporter without modifying the viral genome, but instead introduces reporter genes containing stop codon (UAG) that becomes contingently translatable upon precise RNA editing (UIG, translate as UGG) guided by the complementary HBV RNA sequences. Our preliminary data demonstrated that it functions in an HBV RNA-sensing and ADAR RNA-editing dependent manner, and an HBV puromycin reporter allows for HBV infection-dependent survival of cells following puromycin treatment. Therefore, this novel cell culture-based HBV infection reporter system permits unbiased genome- wide screens using sgRNA, cDNA and chemical libraries, etc. The purpose of this R21 grant application entitled “Identification of cellular functions involved in Hepatitis B Virus infection via a novel RNA sensing and editing dependent reporter system” is to further validate the newly developed system and to identify still elusive host factors required for early steps in HBV infection and for amplification of the viral genome. We anticipate that our novel HBV infection reporter assay will allow us to perform the most comprehensive loss-of-function and gain-of-function screens in tissue culture cells to date, and most likely reveal new knowledge on virus-host interactions critical for the HBV life cycle, which will in a short-term enhance our understanding of HBV replication and advance current HBV model systems, and in a long-term, provide potential novel targets for antiviral therapies to combat chronic HBV infections.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary/Abstract Protective humoral immunity is mediated by both long-lived memory B cells (MBC) and antibody secreting plasma cells (ASC). Recently it has become increasingly clear that MBC and ASC are in fact composed of functionally diverse subpopulations that can be distinguished based on unique surface marker expression, tissue localization, and the B-cell receptor (BCR) isotype expressed. During a humoral immune response, naïve B cells (nB) give rise to differentiated subsets, with the ultimate composition of the MBC and ASC pool primarily influenced by the antigen, the involvement of T cells, and the cytokine environment. Given the evidence for increased heterogeneity, it is surprising that we currently lack the genetic tools to further interrogate the complexity, molecular properties, and immunological importance of the known B cell fates. A precise phenotypic analysis of MBC and ASC subsets cannot currently be performed with current Cre-lox mouse models. Cis- regulatory elements (CEs), or enhancers, are DNA sequences that act to promote cell-type and context-specific gene expression programs. These sequences are regulated by epigenetic mechanisms, which act to control the accessibility of CEs to DNA-binding transcription factors. Over the past several years, we have characterized the epigenetic architecture that controls primary humoral immune responses to T cell dependent and independent antigens, integrated newly published datasets defining MBC subsets, and generated new preliminary data defining the CEs of ASC expressing distinct BCR isotypes. These data have revealed specific CEs that are active in defined stages of B cell differentiation, including IgA ASC and extrafollicular (EF)-MBC that arise independently of a germinal center reaction. Therefore, we hypothesize that cell-type specific CEs can be co-opted to provide precise genetic manipulation that enables functional exploration of B-cell subsets. To address this, we propose two aims designed to 1) develop new Cre recombinase tools for genetic manipulation of IgA ASC and 2) map the CEs for EF-MBC that arise during influenza infection and integrate CEs specific for EF-MBC to allow precise genetic editing of this MBC subset. These aims will utilize novel hematopoietic stem cell engineering to rapidly generate chimeric mice expressing genes of interest; therefore, bypassing the need to generate transgenic animals. Completion of these aims will provide a novel set of DNA sequences that allow for MBC and ASC specific genetic manipulation. These tools are critically important begin to derive the biology, molecular properties, and importance of the entire spectrum of B-cell differentiation to humoral immunity. If successful, it also has the potential to redefine how Cre recombinase vectors are engineered and further our understanding of CE biology in B cells and the immune system.

NIAID - National Institute of Allergy and Infectious Diseases

Identification of immune activation DNA inside cells Cyclic GMP-AMP synthase (cGAS) is a key sensor of cytosolic double-stranded DNA (dsDNA) that activates the STimulator of INterferon Genes (STING) pathway, triggering immune responses against infections and cellular damage. While cGAS plays a protective role in pathogen defense and tumor suppression, its aberrant activation by self-DNA is implicated in autoimmune diseases (e.g., Aicardi- Goutières Syndrome, systemic lupus erythematosus) and chronic inflammation associated with aging. Despite extensive studies on cGAS-STING signaling, the physiological dsDNA ligands that activate cGAS under various cellular conditions remain poorly characterized due to the transient and weak nature of cGAS-DNA interactions, which conventional crosslinking methods fail to capture. To address this challenge, we propose to develop and apply a novel photochemical crosslinking approach to covalently capture cGAS-DNA complexes in living cells. Aim 1 focuses on adapting and optimizing this crosslinking technology for cGAS-DNA interactions, ensuring high efficiency and specificity. Aim 2 applies this method to establish a first-in-class cGAS BFPX ChIP-seq protocol, enabling the genome-wide identification of endogenous DNA ligands that activate cGAS in Trex1 KO MEF cells—a cellular model of Aicardi- Goutières Syndrome. This innovative approach will provide a powerful tool for studying cGAS-STING pathway activation across diverse physiological and pathological contexts. By uncovering the sources and mechanisms of immune-activating DNA, our work will pave the way for novel diagnostic and therapeutic strategies targeting cGAS in autoimmune, inflammatory, and age-related diseases.

NIEHS - National Institute of Environmental Health Sciences

ABSTRACT Plastic additives are widely used in consumer products, yet thousands of plastic additives remain uncharacterized for their potential to disrupt endocrine function - posing significant public health risks. This project aims to develop an integrated computational (Aim 1) and experimental (Aim 2) workflow to systematically predict and validate the endocrine-disrupting potential of plastic additives. In Aim 1, we will design novel machine learning models trained on publicly available datasets to predict AR and ERα modulating activity of plastic additives and then used to predict the potential effects of all plastic additives to select the most promising based on novelty and predictive uncertainty for further in vitro and in vivo testing. In Aim 2, we will validate our predictions through a multi-step experimental characterization approach using our in-house AR and ERα assays, followed by dose-response studies in AR- and ERα-responsive cell lines to measure target gene activation and cell proliferation. The top three plastic additives with the strongest in vitro effects will be further evaluated in vivo using mice to assess systemic hormonal changes caused by the plastic additives. This work will have a substantial positive societal impact by establishing a first-in-kind machine learning-assisted predictive toxicological model to pinpoint plastic additives of highest concern to induce adverse health effects as well as generate a large dataset of plastic additive effects on endocrine function. Taken together, this work can serve to provide policy guidance on plastic additives to ban or remove from products, with potentially beneficial health outcomes for billions of consumers.

NIA - National Institute on Aging

PROJECT SUMMARY/ABSTRACT The preservation of organ functions, such as eyesight and mobility, is a significant health concern in the elderly population. Organ frailty and disease progression are associated with the dysregulation of tissue homeostasis, which is typically regulated by regenerative units, consisting of adult stem cells (ASCs) and neighboring niche cells that regulate ASC function. Therefore, to better understand why regenerative functions decrease in aged individuals, uncovering how aging alters niche cell types and expression (mRNA) is critical for discovering regenerative therapeutical approaches. The potential to uncover molecular mechanisms to reverse age-related disorders prompted me to spatially profile microenvironmental niches in young, old, and regenerated tissues. I have recently developed Ex-Scope, which integrates Expansion Microscopy and Seq-Scope, a submicrometer-resolution ST (spatial transcriptomic) technology, to obtain a high-resolution multi-Omic method that represents an order of magnitude improvements over Seq-Scope. With the assistance of Dr. Guo, who has extensively worked on planarian tissues, we optimized Ex-Scope to spatially profile planarian tissue. Planarians are capable of regenerating any lost body part, but most importantly, regenerated tissues have a youthful tissue architecture; thus, making them ideal to study tissue homeostasis and rejuvenation. Using mRNA single-cell data on young, old, and regenerated planarians, as a reference dataset (obtained by Dr. Guo), we will provide spatial insight into rejuvenating mechanisms between microenvironmental niches and stem cells. Concurrent, we will demonstrate the advantageous resolution of Ex-Scope by profiling RNA granules in planarian stem cells and oocytes (young, old, and regenerated), which are compartmentalized biomolecules that regulate transcription in stem cells and the establishment of pluripotency. In aim 1) we propose to characterize RNA granules and soluble transcriptomes in planarian stem cells and oocytes, with a hypothesis that the granular structures in oocytes and ASCs would have transcriptome contents distinct from soluble cytoplasm, and 2) we propose to profile microenvironmental niches and their changes during aging and rejuvenation, with a hypothesis that aging and rejuvenation will affect cellular (single cell), tissue-level (microenvironment) and subcellular level (RNA granule) transcriptome, each of which is important for tissue function and homeostasis. We expect that the current work will give us a systematic understanding of how aging deteriorates tissue function by altering transcriptomic structure at both microscopic and macroscopic levels, and how regeneration can reverse it and rejuvenate tissue homeostasis.

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY T follicular helper (Tfh) cells are a subset of CD4+ T cells that specialize in communicating with germinal center (GC) B cells to facilitate B cell differentiation into either Memory B cells or Plasma cells that secrete high-affinity class-switched antibodies. As a key regulator of both antibody quantity and quality, coupled to the crucial role antibodies play in mediating pathogen control following immunization or infection, or in driving autoimmune or allergic diseases respectively, Tfh cells directly contribute to either protective or pathological B cell responses depending on the context of the disease. Notably, Tfh cells play an especially important role during chronic viral infections, such as during HIV-1 and hepatitis C virus infection, or during chronic lymphocytic choriomeningitis (LCMV) infection in mice, all of which critically depend on Tfh-mediated antibody responses for viral control. Moreover, recent evidence indicates that Tfh cell-derived IL-21 functions to sustain the effector function of CD8+ T cells during chronic viral infection and cancer. However, despite their essential role, the precise molecular circuits underpinning Tfh cell differentiation remain unclear. Moreover, in settings of persistent infection, the development of neutralizing antibodies is often delayed for several months, and the initial wave of antibodies that do develop are often of low-affinity and poor quality. Furthermore, hypergammaglobulinemia, or the overproduction of non-specific antibodies, and impaired FCγ receptor effector functions are hallmarks of chronic viral infections in both mice and humans, indicating that dysregulated humoral immunity is a generalizable feature of persistent viral infection. We hypothesize that persistent exposure to antigen and inflammation perturbs functional Tfh cell responses, thereby resulting in insufficient “help” signals to both B cells and CD8+ T cells, suboptimal antibody responses and impaired viral control during chronic infection. Herein, I will employ cutting-edge technologies, including single cell profiling of T cell transcriptomes and epigenomes, innovative bioinformatic pipelines, and CRISPR/Cas9 screening and genetic mouse models to identify novel and conserved regulators of Tfh cell differentiation across different infections and species, as well as determine how persistent exposure to antigen and inflammation perturbs the transcriptional and epigenetic program of Tfh cells and their functional capacity to provide “help” to B cells. Additionally, using computational approaches to integrate publicly available single-cell RNA-sequencing datasets containing human CD4+ T cell populations, accompanied by a creative in vitro Tfh:B cell co-culture system, we will extend our findings from experimental models to further delineate critical regulators of human Tfh cells. Successful completion of this project will provide mechanistic insight into the molecular circuitry underpinning Tfh cell development and aid in the identification of novel strategies aimed at targeting Tfh cells to modulate their function for therapeutic benefit during chronic viral infection and/or other diseases.

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY/ABSTRACT Dense microbial communities known as the microbiota are intimately associated with human health. Invading pathogens must overcome the microbiota to establish and successfully cause infections. In settings like the gastrointestinal tract, antagonistic interactions between microbes can strongly influence colonization outcomes. One highly prevalent pathway known to mediate these interactions the type VI secretion system (T6SS) which mediates contact-dependent killing via the translocation of toxic effector proteins into recipient cells. How the T6SS affects pathogen fitness and microbiota composition in other body sites, like the respiratory tract, is not well understood. A model disease for studying microbial ecology and interspecies interactions is Cystic Fibrosis, where polymicrobial communities of microorganisms colonize the respiratory tract due to defects in ion secretion and pathological mucus accumulation. Pseudomonas aeruginosa (Pa) causes chronic infections, dominates the airways of people with cystic fibrosis (pwCF) and is a major cause of morbidity and mortality. Pa harbors three T6SS with a variety of effectors delivered to host and bacterial competitors. We have recently reported that a T6SS effector, TseT, is delivered by the H2-T6SS and regulates microbial diversity in the upper respiratory tract of pwCF. In addition to TseT, the tseT operon encodes an immunity protein, TsiT, which our preliminary studies show also regulates Pa biofilm, in addition to its role as a bona fide immunity protein. TsiT has sequential and structural homology to general LysR-type transcriptional regulators (LTTR), which are known to regulate diverse genes including those involved in biofilm, metabolism, and quorum sensing. We have also identified diverse homologs for tsiT in a variety of Gram-negative organisms, including Burkholderiales. These results have led us to the overarching hypothesis that TsiT is a dual-function immunity protein that both regulates biofilm and neutralizes the TseT effector. In this proposal, we will define the mechanism by which TsiT regulates biofilm, investigate the evolution of tsiT homologs, and determine their impact on microbial ecology in polymicrobial communities in the respiratory tract. To this end, we propose three aims: 1) Test the hypothesis that TsiT is a transcriptional regulator that mediates Pa biofilm; 2) Test the hypothesis that tsiT diversity outside of P. aeruginosa confers protection against Pa TseT intoxication; 3) Test the hypothesis that tsiT genes modulate P. aeruginosa fitness during biofilm growth in a model CF respiratory microbiota.

NINDS - National Institute of Neurological Disorders and Stroke

Project Summary/Abstract Progressive supranuclear palsy (PSP) is a rare neurodegenerative motor disorder affecting approximately 20,000 people in the U.S. Clinical hallmarks of PSP are early loss of balance and falls, together with impairment in saccadic eye movements. Additional features often mimic Parkinson’s disease (PD), but PD treatments provide little to no therapeutic benefits in PSP. Histopathologically, PSP is characterized by the presence of insoluble aggregates of the microtubule-associated protein, Tau. While disease-causing mutations to the Tau- encoding MAPT gene have been described, the majority of PSP cases are idiopathic. Postmortem analysis of PSP patient brains show Tau pathology in several motor control regions, such as the basal ganglia (e.g. striatum, subthalamic nucleus, substantia nigra) and midbrain eye movement nuclei. Recent findings indicate basal ganglia output nuclei, such as the substantia nigra, are among the first sites of neuronal pathology. Tau pathology in these regions may lead to physiological dysfunction of motor circuits, in turn driving motor symptoms seen in PSP. To test this hypothesis, we recently repurposed a previously described Tau transgenic mouse and determined that Tau pathology in the mice was sufficient to mimic two key clinical aspects of PSP: gait and eye movement abnormalities. Using custom-built gait and eye movement recording systems, we observed that Tau hP301S mice had impaired coordination and vertical eye movements. Furthermore, we find these phenotypes correlate with pathology in many PSP-associated motor control regions, such as the substantia nigra and mesencephalic locomotor region. We also find that expressing mutant Tau specifically in the substantia nigra is sufficient to mimic these same abnormal phenotypes. In this proposal, I will use these two new mouse models to test my central hypothesis that Tau pathology leads to behavioral deficits via physiological abnormalities in midbrain motor regions, the substantia nigra pars reticulata and the mesencephalic locomotor region. I will use a combination of in vivo electrophysiology, ex vivo patch-clamp recordings, and quantitative measures of eye movement and gait to identify these links between pathology, physiology, and behavior. Understanding these mechanisms may help focus future work on both the underlying mechanism of PSP and identify new treatment strategies. The experiments proposed in this study expands upon my prior ex vivo slice electrophysiology training and adds training in in vivo electrophysiology and complex behaviors. In addition, I have assembled an expert mentoring team to provide guidance in my career development with the goal of launching a career as a successful independent investigator at a biomedical research institution.

NIAID - National Institute of Allergy and Infectious Diseases

In the United States and globally, respiratory syncytial virus (RSV) is a leading cause of morbidity and mortality in infants under 6 months, leading to ~50,000 deaths per year. Preterm infants are at particularly high risk of severe disease. To boost protection, a bivalent RSV vaccine formulated as recombinant pre-Fusion glycoprotein (preF) was recently licensed for use in pregnancy and provides ~70% protection from severe RSV disease in infants under 6 months of age. To date, the efficacy of this vaccine has been attributed solely to the transplacental transfer of maternal IgG to the fetus. However, maternal vaccination may also impact breastmilk immunity, where breastmilk exposure and breastmilk RSV-specific IgG are both associated with protection from RSV infection and reduced disease severity. We and others have also demonstrated that breastmilk also contains RSV-specific IgA and mucosal memory T cells, which increase during RSV-infection in the maternal-infant dyad. Breastmilk IgG and IgA may be systemically absorbed in the first few days of life and could contribute to viral neutralization. In addition, animal data demonstrate that breastmilk T cells can integrate into offspring organs including the lung, where they may facilitate viral clearance. Together, these observations suggest that adaptive immunity derived from breastmilk is functionally distinct from transplacental IgG and, importantly, can provide mucosally active IgA, IgG, and durable T cell protection to the infant. However, the impact of RSV-VX on breastmilk immunity is currently unknown. To address this knowledge gap, we have recently demonstrated that there is increased preF- specific IgA and IgG in the breastmilk of women who received RSV-VX compared to those who did not. Thus, our central premise is that breastmilk provides a distinct form of mucosal immunity to the infant which can be boosted with vaccination. Further, prior data demonstrate that transplacental transfer of IgG is reduced in preterm dyads, but early breastmilk IgA and T cell concentrations are increased. Thus, we hypothesize that the relative importance of RSV-VX on breastmilk immunity is greater in preterm versus term infants, where preterm infants receive less transplacental IgG but more breastmilk IgA and cellular immunity. To test our hypothesis, we will enroll a cohort of pregnant women at delivery who did or did not receive RSV-VX, including preterm and term deliveries. We will collect maternal and cord blood at delivery, maternal breastmilk and blood at 2-, 6-, and 12- weeks postpartum, and infant blood at 12-weeks. In Aim 1, we will determine the impact of RSV-VX and preterm delivery on breastmilk and peripheral RSV-specific antibody levels and function. In Aim 2, we will determine the impact of RSV-VX and preterm delivery on frequency and function of peripheral blood and breastmilk RSV- specific CD4 and CD8 T cells. The proposed studies will address a critical knowledge gap regarding the ability of RSV-VX to boost mucosal breastmilk and peripheral immune responses in term and preterm dyads. These data will provide rationale for the development of next generation maternal vaccines designed to induce mucosal immunity as a targeted strategy to improve infant protection.

NIAID - National Institute of Allergy and Infectious Diseases

Accidental or deliberate radiation exposure of humans remains a major health concern, due to the paucity of medical countermeasures (MCMs) to ameliorate radiation-induced damage. While high dose radiation exposure is generally fatal, even low dose whole body (WBI) or partial radiation exposure can have acute- and/or delayed- negative impacts that appear to act through disruption of the immune system. The cytoreductive effects of WBI have long been exploited in conjunction with chemotherapy as a preparative regimen prior to hematopoietic stem cell transplant in patients with blood cancer to deplete malignant cells and suppress the immune system. While there is strong evidence that radiation kills rapidly dividing cells, a hallmark of the immune system, and induces inflammation that can mediate tissue destruction, the precise nature of radiation induced immune-dysfunction is not well understood. This knowledge gap is a key impediment to development of MCMs to treat radiation exposure. For one example, memory CD8 T cells provide enhanced resistance to re-infection and malignancies. However, most studies in the literature examine the impact of radiation exposure on the capacity of the host’s naïve CD8 T cells to mount a new (primary) immune response and just a few reports have looked at how radiation exposure influences the longevity and protective capacity of pre-existing pathogen or vaccine-induced CD8 T cell memory. Memory CD8 T cell populations have the job of surveying the entire body for signs of re-infection. They accomplish this task using two complimentary and interactive strategies. This first strategy involves populations of memory CD8 T cells that survey the body by using the circulatory system (circulating memory CD8 T cells - Tcircm). The second strategy involves the generation of a population of non-circulating memory CD8 T cells (called T resident memory, Trm), generally in the tissue of pathogen entry. These cells, which persist long-term in tissues, provide rapid detection of re-invading pathogens and then send out mediators to recruit other cells of the immune system to the site of infection. Importantly, our recent data obtained after WBI or partial (targeted) thorax radiation suggest that sublethal ionizing radiation inflicted numerical and functional damage to Tcircm and Trm cells that diminished their ability to provide protection to pathogen-re-encounter. Our long-term goal is to precisely identify mechanisms that govern maintenance, differentiation and function of infection and/or vaccine- induced memory CD8 T cell subsets and explore modalities to recover memory CD8 T cell responses in radiation survivors. We will address our long-term goal through the following specific aims: SA1 - Delineate the tissue-specific impact of WBI on pathogen-specific Trm and evaluate targeted vaccine strategies to restore memory CD8 T cell numbers and function after irradiation. SA 2 - Define mechanisms underlying WBI-induced numerical and functional diminishment of Tcircm and exhausted (Tex) CD8 T cells generated after acute or chronic viral infections.

Watervliet

Implementation of ERP and Financial System

Substance Abuse and Mental Health Services Adminis

Implementing Zero Suicide in Health Systems

Star, Inc., Lighting the Way…

Improvement - Sprinkler System

Meadowbrook Community Gardens and Orchards

Meadowbrook Community Gardens and Orchards, (MCGO), will improve our irrigation system, (currently irrigation is all done by hand by volunteers), replace old and worn tools, repair raised beds and add a few more, and buy bulk soil amendments and compost to improve soil quality. All of the ese improvements will allow us to better achieve our purpose of producing more food to meet the growing needs of our community.

NIA - National Institute on Aging

PROJECT SUMMARY Up to 3.1% of women in the United States experience premature menopause (menopause before age 40) and another 6.2% experience early menopause (menopause between age 40-45) due to a spectrum of surgical and non-surgical conditions. Premature/early menopause results in estrogen deprivation which has significant health consequences and adversely affects quality of life. Compared to natural menopause, women who undergo premature/early menopause have an increased risk for cardiovascular disease, osteoporosis, and neurological/psychological conditions, leading to a higher mortality risk. Systemic hormone therapy (HT) is highly effective in mitigating the short- and long-term consequences of estrogen deprivation and is recommended by guidelines from national and international professional organizations. However, research on HT use in women with premature/early menopause is sparse and the limited data available suggest severe underuse. Because women at younger ages are less prepared of menopausal transition and may face greater challenges in work, life, and health care, there is a critical need for research on premature/early menopause. The overarching goal of this project is to improve the long-term health of women with premature/early menopause by understanding factors influencing their HT use and developing a preliminary tool for identifying women at high risk for underuse. We will focus on women without malignancy or hereditary predisposition for cancer – a homogeneous population to better isolate influencing factors. Within this population, we propose the following specific aims: 1) to characterize the overall trajectory and heterogeneity in HT use and identify patient and physician factors associated with HT initiation and continuation; 2) to ascertain patients' and physicians' perceived barriers and facilitators to HT use; and 3) to develop a dynamic risk prediction model for HT underuse and explore the feasibility of a patient prioritization tool for HT counseling. We will achieve these aims by analyzing a unique dataset that integrates insurance claims data with electronic health record data and applying novel analytical approaches to characterize patients’ longitudinal trajectory of HT use (both the overall pattern and heterogeneity among patients), examine physician peer influence in HT practice, and develop artificial intelligence/machine learning-based risk prediction models. This will be supplemented by primary data collection using a mixed methods approach – combining qualitative interviews and focus groups with quantitative surveys of patients and physicians – to uncover individual, structural, and system-level factors influencing HT initiation and continuation and inform proactive interventions for improving HT use. By understanding care patterns related to HT use in women with premature/early menopause, its influencing factors, and characteristics of patients at particularly high risk for HT underuse, this project addresses an important but understudied clinical question. The findings can facilitate targeted interventions and better equip us with the knowledge and tools to improve care for this large population of women.

U.S. National Science Foundation

Improving Undergraduate STEM Education: Directorate for STEM Education

NIGMS - National Institute of General Medical Sciences

Abstract: Cell signaling regulates information transfer across tissues through graded ligands that interact with receptors. Across evolution, ligand and receptor proteins co-evolve, with changes in one protein being counterbalanced by changes in its partner, thus preserving successful interactions that promote development rather than pathology. Despite the importance of co-evolution in speciation and disease, our understanding of these reciprocal processes primarily relies on bioinformatics predictive tools and in vitro systems like yeast two-hybrid assays. Currently, there is no robust in vivo system, at moderate cost, to validate the co-evolution of multiple cell-signaling components. Given the differences in physiological conditions across tissues that impact protein interactions, we will develop in vivo genetic and molecular platforms for investigating co-evolving proteins. Research using Drosophila melanogaster has greatly contributed to both basic and medical research at a low cost. Building on our extensive experience with Drosophila species, we plan to generate two new in vivo systems to study the co- evolution of Gurken (GRK), a TGF-alpha ligand that activates the epidermal growth factor receptor (EGFR), endogenously. Drosophila oogenesis is a powerful model system for studying EGFR-guided axis formation and eggshell morphology, providing simple phenotypic readouts. Although the pathway and the underlying mechanism, a localized source of GRK, are conserved among Drosophila species, we have observed dramatic variations in the ability of GRK orthologs to signal interchangeably across species, ranging from no signal to a striking overactivation of EGFR signaling. The new in vivo platforms will allow us to thoroughly explore the effects of a histidine-rich domain loss in GRK on EGFR activation across different subgenera. Additionally, we will investigate the causes of EGFR overactivation when both GRK and EGFR are substituted with their homologs. S2 culture cells will be engineered with biosensors for EGFR signaling, providing a robust system to prioritize computationally predicted co-evolving sequences in GRK and EGFR for in vivo studies. Furthermore, we will examine the evolution of EGFR signaling targets, including the Tbx-20 homologs, Midline (MID) and H15, in relation to changes in signaling intensity and duration. Finally, we will study the evolution of cis-regulatory modules (CRMs) that control the expression of Mid/H15 tandem paralogs in oogenesis and other fly tissues. Our comprehensive and multidisciplinary approach-- combining new in vivo systems, computational, biophysical, analytical, and CRISPR/Cas9 genome engineering tools-- provides an exciting new path for identifying the co- evolving domains in ligands and receptors that control signaling levels. By investigating these domains in vivo, under natural physiological conditions, we will advance our understanding of cell signaling mechanisms, which could lead to new strategies for addressing tissue pathologies. The research field will benefit from new in vivo platforms to explore co-evolving processes during development and homeostasis. The EGFR signaling field will gain high-resolution analyses of co-evolving sequences that control signaling levels and duration in vivo.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

In 2023, about 4750 Americans died while awaiting a kidney transplant (KT), but over 8,500 kidneys were recovered and not transplanted. The National Academies of Sciences, Engineering, and Medicine (NASEM) declared kidney non-utilization a critical problem that mandates immediate attention and remedy. Highly prioritized candidates receive many offers for superior kidneys, but also receive offers for many non-ideal kidneys that they refuse sequentially while the clock counts down to kidney non-use. Many candidates with low priority would benefit from receiving a non-ideal kidney quickly but rarely get these offers. Stratified kidney allocation is a proposed system under which patients would make an obligatory but modifiable choice between receiving a superior kidney after a longer wait or a non-ideal kidney sooner. Stratified kidney allocation might increase organ utilization while shortening wait times and improving transplant outcomes for selected patients. Prior work has shown that patients have misperceptions about the dimensions of kidney quality, underestimate the survival benefit of transplant with non-ideal kidneys, and alarmingly, are given a very limited role in decisions about offer quality. While stratified allocation might expedite and increase the use of non-ideal kidneys, impacts are unknown because myriad and sundry details such as which deceased donor kidneys would be categorized as non-ideal, which candidates would select which quality category, etc., have never been specified, nor have patient perceptions about acceptability been investigated. We will build expert consensus on the design of stratified kidney allocation, estimate the impact of stratified allocation to assist policymakers, and investigate attitudes and perceptions of patients, care partners, and providers on shared decision making about kidney offer acceptance, and on making a choice to receive either superior or non-ideal kidneys. Our aims are (1) To understand patient and care partner attitudes, perceptions, and preferences about stratified allocation, shared decision-making, and needs for transparency around organ offers and declines, (2) to assess transplant provider attitudes, perceptions, and preferences about a stratified allocation system, and (3) to estimate the impact of stratified allocation on kidney utilization, and to estimate waiting time for KT candidates choosing either to receive either superior or non-ideal offers. This research will take stratified allocation from a speculative proposal to a concrete policy design that supports deliberation and possible adoption, informed by the judgements of transplant experts, by patient and caregiver perceptions, and by numerical estimates of its impacts. Whether or not stratified allocation succeeds, we will, as advocated in NASEM’s 2022 report, investigate approaches for increasing transparency around offer declines, and involving patients in shared decision making about the kidney quality vs. wait time tradeoff.

NIAID - National Institute of Allergy and Infectious Diseases

It has long been evasive why strictly hepatotropic viruses, such as hepatitis C virus (HCV), have been associated with the generation of delayed antibody responses that are of relatively poor quality to other, more systemic infections. Studies to date investigating antiviral adaptive immune priming within the liver have been hindered by lack of a natural, biologically relevant small animal model of viral infection exclusively restricted to hepatocytes. In this proposal, we will accordingly use the HGV-related rodent hepacivirus (RHV) recently discovered by our team, which possesses several similarities to its genetic relative, including strict liver tropism, conserved replication dependence on miR-122 interaction with the viral 5' UTR, identical genomic organizational structure and polyprotein cleavage pattern, and the propensity to cause chronic infection with fibrosis and hepatocellular carcinoma tumorigenesis. Our preliminary identification of robust antibody-secreting cell (ASC) responses, both total lgG and those targeting the envelope glycoprotein E2, almost exclusively arising within the liver during both human HCV and murine RHV infection, served as the rational impetus for further investigation of how, when, and where such responses are generated. The secondary lymphoid organ (SLO) dormancy accompanying such strong ASC responses within the liver alongside their durable maintenance therein by putative cognate anchoring pairs, many of which are also critical for tethering of long-lived plasma cells (LLPCs) to stromal niches in bone marrow, collectively suggest that the liver may serve as a site conducive to both the local generation and perpetual maintenance thereafter of LLPCs during strictly hepatotropic viral infection. As our recently published findings demonstrate that viral-specific lgG is critical for viral resolution, the development of chronic infection and HCC tumors in its absence suggests that generation of plausibly locally generated humoral responses within the liver at sites of virus-induced tertiary lymphoid structures, hereafter denoted as inducible Hepatic-Associated Lymphoid Tissue (iHALT), likely serve an important functional role. The survival of LLPCs residing in the liver 1.5 years post-clearance in these mice represents a unique scenario in which the liver may also provide a suitable niche for these cells, contrary to the canonical paradigm in which the bone marrow or select SLO circumstances are exclusively capable of fostering such an environment. We propose to mechanistically determine the signaling prerequisites responsible for driving this local, successful antiviral antibody response within the liver (Aim 1) as well as the cues instructing putatively locally derived progenies to be retained directly adjacent to their generative origins (Aim 2). Understanding these processes would significantly enhance our understanding of the relationship between hepatotropic viruses capable of suppressing SLOs in a state of functional dormancy with humoral responses plausibly being housed ectopically at extra lymphoid sites. Further, these studies would likely yield important insights capable of influencing the current paradigm of durable antibody responses in antigen free settings, specifically exploring a potentially novel niche site conducive to LLPC survival within the liver.

Metropolitan Museum of Art

Hot water heating piping system will be tested, chemically flushed and replaced as required throughout the building.