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Solid Phase Peptide Synthesizer

open

OD - NIH Office of the Director

ABSTRACT/SUMMARY This proposal requests funds to purchase a Liberty Blue 2.0 solid-phase peptide synthesizer. At present, Vanderbilt lacks a comparable capacity for customized peptide synthesis, compelling researchers to rely on commercial vendors. While standard peptides can often be sourced at reasonable cost, the synthesis of peptides incorporating non-proteinogenic amino acids, macrocyclizations, or site-specific chemical modifications incurs prohibitive costs and prolonged lead times. These limitations negatively affect numerous NIH-funded research programs and severely lowers the chemical novelty accessible to investigators who make use of peptides in their research. Acquisition of an institutional instrument will directly address this gap, enabling timely and affordable access to high-quality, customized peptides that are increasingly central to modern biomedical research. This instrument will serve a large and scientifically expansive group of investigators across 15 departments in the College of Arts and Science, the School of Medicine Basic Sciences, and the Vanderbilt Institute of Chemical Biology. Investigators from the Vanderbilt University Medical Center will also have access. The user base spans a wide array of NIH-funded projects that rely on synthetic peptides. For example, one group synthesizes fluorophore-labeled peptides to monitor receptor trafficking. Another develops cleavable linkers that release antibiotics from antibody-drug conjugates designed to target methicillin-resistant Staphylococcus aureus. A third focuses on macrocyclic peptides that modulate the activity of CFTR and thus show promise as future therapeutics for cystic fibrosis. Several other groups engage heavily in structure- and AI-guided design and require rapid synthesis of candidate molecules to support downstream biochemical and cellular validation. The Liberty Blue 2.0, manufactured by CEM Corporation, uses microwave-assisted chemistry to accelerate synthesis cycles, improve coupling efficiency, and enhance overall yield and purity. The instrument accommodates a wide range of chemistries and scales, offering flexibility to support exploratory screening, structure-activity relationship campaigns, and early-stage preclinical development. Importantly, it also provides significant cost and time savings compared to commercial synthesis, especially for chemically complex sequences. The instrument will be housed within the Molecular Design and Synthesis Core, which has provided synthetic chemistry expertise and training to the Vanderbilt community since 2006. This core will oversee daily operation and user access, supported by administrative and financial contributions from the School of Medicine Basic Sciences and the College of Arts and Science. Acquisition of the Liberty Blue 2.0 will significantly enhance Vanderbilt’s infrastructure for chemical biology, lower the barrier to peptide-based experimentation, and accelerate discovery across multiple scientific disciplines and therapeutic categories.

Up to $129K
2027-05-14
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Solvent-Driven Assembly of Intrinsically Disordered Peptides: Integrating Protein-Language Models with Atomistic-to-Mesoscopic Simulation

open

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY The project supports ongoing efforts in the Shea group geared at developing new computational methodologies and tools to study the liquid-liquid phase separation (LLPS) of intrinsically disordered proteins (IDPs). Biomolecular condensates formed by LLPS play a range of vital physiological roles in the body, but under aberrant conditions, they can transition into amyloid fibrils, a process linked to disease. The project will meld artificial intelligence protein-language models with a multiscale computational framework to accurately simulate the dilute and dense LLPS phases, characterize the role of water, co-solvents, and high pressure in modulating assembly, and identify new LLPS-prone sequences in the human proteome. The proposal consists of three research projects. Project 1 involves the development of a tightly integrated multiscale computational approach bridging the atomistic to mesoscopic time and length scales. The relative entropy approach will be used to generate chemically accurate protein and water coarse-grained models from atomistic simulations, which will be used as input for efficient field theoretic simulations. The latter will be used to generate phase diagrams for the LLPS of the microtubule-binding Tau protein and Elastin-Like Polypeptides (ELPs), with field theoretic outputs backmapped to generate atomistic, solvated condensate structures that can be directly compared to experiment. Project two involves the development of new high pressure Kirkwood-Buff force fields for the osmolyte trimethylamine N-oxide (TMAO) from experimental Kirkwood-Buff Integrals, and their application to the study of TMAO’s counteraction of high-pressure denaturation of ELP condensates. Project 3 involves developing new artificial intelligence protein language model tools to mine the IDRome – the 28k proteome of intrinsically disordered regions – for new LLPS-prone and co-condensating sequences. The research will lead to state-of- the-art computational tools that will be deposited in Github and made freely available to the broad scientific community, to new physical insights into osmolyte and pressure modulation of LLPS, and to the discovery of new LLPS-prone sequences. The research will inform on conditions that promote functional forms of LLPS as well as lay the foundation for rational therapies for condensate-linked diseases.

Up to $391K
2031-04-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Sound Asleep: Technologies for uninterrupted whole night sleep recording in an MRI scanner

open

NIBIB - National Institute of Biomedical Imaging and Bioengineering

Abstract The project aims to develop a platform that would enable whole-brain functional MRI (fMRI) during full night natural sleep in-sync to polysomnographic recordings (PSG), nearly as easily as in a PSG-only sleep lab. Sleep is vital for maintaining all aspects of human health. Extensive research has confirmed its role in homeostatic processes that are critical for brain health, especially in relation to aging and neurodegeneration. Yet, most of our knowledge of sleep neurophysiology in healthy individuals and patients has been limited by the capability of scalp EEG. EEG recordings, although extremely reliable, do not provide information with spatial specificity in the cortex nor do they provide information on the activity in subcortical areas, such as the medial temporal lobe and hippocampus, key brain areas involved in the study of cognitive decline due to dementia. fMRI provides superior spatial resolution of hemodynamics driven by neural activity, simultaneously in all brain areas, and provides a time resolution (seconds) relevant to sleep transitions (minutes) and their associated second scale events. However, attempts to obtain fMRI data during sleep have mostly been limited to light sleep in healthy young adults due to the difficulty of falling asleep and maintaining sleep in the MRI scanner environment where the acoustic noise is high, and participants are head-fixed in the coil. We will develop a novel platform to address these limitations and to enable simultaneous whole-night fMRI and polysomnographic recordings while the participants sleep naturally with minimal interruption. This platform will permit us to evaluate the coupling of fMRI brain activity and established polysomnographic recordings in a setup relevant for aged and clinical populations. Specifically, we will: (1) develop wearable high-density MRI head-cap arrays that are EEG compatible and allow the participants to adjust sleep posture while sleeping in the scanner environment; (2) adapt, and further develop silent fMRI using the looping-star sequence to eliminate acoustic noise while performing the scans; (3) develop an online real-time motion event classifier and scanner control for automatic sequence prescription for multiple head positions; (4) develop and integrate the analytical tools required to harmonize brain data recorded in multiple head positions; and (5) evaluate the platform in an aged population. Our proposed technology is highly innovative compared to the state of the art and addresses a significant need in the field of sleep studies. A success of the project will provide solution for simultaneous whole-night fMRI and polysomnographic recordings with quality comparable to that of clinical sleep labs outside MRI scanners. Our platform would open new diagnostic and research avenues, not possible today in patients and aged individuals.

Up to $611K
2030-03-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

SourceSpikeNet – A Biophysically Grounded AI Approach to Spike Detection in EEG

open

NINDS - National Institute of Neurological Disorders and Stroke

ABSRACT: SourceSpikeNet – A biophysically grounded AI approach to spike detection in EEG Epilepsy affects approximately 3.4 million Americans, with 40% experiencing seizures despite medication and 50-60% continuing to have seizures after surgical intervention. Electroencephalogram (EEG) detection of interictal epileptiform discharges (IEDs) is crucial for diagnosis, medication selection, and surgical planning. However, current practice relies on subjective, qualitative identification of a small subset of IEDs, with significant inter-reviewer variability. State-of-the-art automated detectors suffer from high false alarm rates, creating an urgent unmet need for precise, objective IED identification. We propose SourceSpikeNet, a biophysically grounded artificial intelligence approach to IED detection that leverages known neurophysiology and electromagnetic physics to improve spatiotemporal accuracy. Our central hypothesis is that a biophysically informed AI can identify IEDs much more accurately than current methods. In Aim 1, we will characterize IED prototype timeseries patterns from 32,433 expert-annotated IEDs using inverse source modeling and self- supervised machine learning. In Aim 2, we will develop SourceSpikeNet by training a deep learning algorithm on an augmented dataset of 3,200,000 simulated spikes from 1,000 uniformly spaced cortical locations. In Aim 3, we will evaluate SourceSpikeNet's ability to detect all IEDs from 2,484 patients' continuous EEG recordings (3-7 days each) and compare comprehensive IED localization to traditional methodology. We expect that SourceSpikeNet will provide superior spatiotemporal localization of IEDs compared to current approaches, with lower false alarm rates and higher spatial resolution. The successful completion of this project will enhance the accuracy of IEDs as a biomarker for identifying seizure foci, potentially enabling higher surgical success rates and improved outcomes for epilepsy patients.

Up to $252K
2028-04-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Spatial Profiling of Pancreatic N-Glycosylation in Type 1 Diabetes

open

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

PROJECT SUMMARY / ABSTRACT Type 1 diabetes (T1D) is caused by T-cell mediated destruction of insulin-producing pancreatic beta cells. While notable progress has been made in predicting and delaying onset of T1D, our limited understanding of the factors that initiate and maintain this autoimmune attack continue to act as a major barrier to further progress. Increasing evidence points to the pancreas itself, including endocrine cells, exocrine cells, and the extracellular matrix, as possible contributors to the pathogenic immune activation. One factor that is known to contribute to immune cell activation, and that is altered in other pancreatic disease including cancer, is protein N-glycosylation, wherein complex carbohydrate chains called glycans are enzymatically attached to specific asparagine (called N-glycans) residues as proteins transit the endoplasmic reticulum and golgi complex. Glycosylation patterns influence protein stability, localization, and receptor binding, which can dramatically alter cell function and intercellular communication. Though the pancreatic glycome has been studied in pancreatic cancer, very little is known about how the glycome changes in diabetes pathogenesis, partially owing to the relative scarcity of appropriate human tissues to study and to the complexity of analysis methods required to measure protein glycosylation. While single cell transcriptomic data shows that expression of many of the enzymes involved in glycosylation are altered in T1D, it remains unknown how the pancreatic glycome changes during T1D pathogenesis, and whether altered glycosylation contributes to changes in pancreatic structure, cell composition, or immune cell infiltration. I hypothesize that N-glycosylation in the pancreas is altered as type 1 diabetes progresses, contributing to changes in immune cell localization and phenotype. I will employ two state-of-the-art imaging technologies to test this hypothesis in pancreas tissues from donors without diabetes, with positive auto-antibodies, or with recent-onset or long-standing T1D: 1) Imaging mass spectrometry will allow comprehensive quantitation of different N-glycans across entire tissue sections and at single-cell resolution, and 2) Multiplex immunofluorescence microscopy (CODEX) will be used to define pancreatic regions of interest and to quantify cell types and subtypes across the same tissue section. In Aim 1, I will test the hypothesis that the pancreatic N-glycome quantitatively changes throughout T1D progression. In Aim 2, I will test the hypothesis that regions of altered N-glycome signature are associated with changes in cellular composition and immune cell phenotypes. Completion of these aims will identify high level changes to post-translational protein processing signatures as T1D progresses. These results will lay the groundwork for future studies into mechanisms responsible for glycomic changes, identification of specific proteins that are affected, and definition of novel glycoprotein signatures that may be promising biomarkers or drug targets.

Up to $175K
2028-04-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Specialized Programs of Research Excellence (SPOREs) in Human Cancers for Years 2024, 2025, and 2026 (P50 Clinical Trial Required)

open

National Institutes of Health

Through this funding opportunity announcement (FOA), the National Cancer Institute (NCI) invites applications for P50 Research Center Grants for Specialized Programs of Research Excellence (SPORE). The program will fund P50 SPORE grants to support state-of-the-art investigator-initiated translational research that will contribute to improved prevention, early detection, diagnosis, and treatment of an organ-specific cancer or a highly related group of cancers. For the purpose of this FOA, a group of highly related cancers are those that are derived from the same organ system, such as gastrointestinal, neuroendocrine, head and neck, and other cancers. Other programmatically appropriate groups of cancers may include those centered around a common biological mechanism critical for promoting tumorigenesis and/or cancer progression in organ sites that belong to different organ systems. For example, a SPORE may focus on cancers caused by the same infectious agent or cancers promoted and sustained by dysregulation of a common signaling pathway. In addition, a SPORE may focus on cross-cutting themes such as pediatric cancers or cancer health disparities. The research supported through this program must be translational and must stem from research on human biology using cellular, molecular, structural, biochemical, and/or genetic experimental approaches. SPORE projects must have the goal of reaching a translational human endpoint within the project period of the grant.

2026-09-25
EducationHealth

Free to search & build · $99 one-time to unlock the application pack · No subscription

Specialized Programs of Research Excellence (SPOREs) in Human Cancers for Years 2024, 2025, and 2026 (P50 Clinical Trial Required)

open

National Institutes of Health

Through this funding opportunity announcement (FOA), the National Cancer Institute (NCI) invites applications for P50 Research Center Grants for Specialized Programs of Research Excellence (SPORE). The program will fund P50 SPORE grants to support state-of-the-art investigator-initiated translational research that will contribute to improved prevention, early detection, diagnosis, and treatment of an organ-specific cancer or a highly related group of cancers. For the purpose of this FOA, a group of highly related cancers are those that are derived from the same organ system, such as gastrointestinal, neuroendocrine, head and neck, and other cancers. Other programmatically appropriate groups of cancers may include those centered around a common biological mechanism critical for promoting tumorigenesis and/or cancer progression in organ sites that belong to different organ systems. For example, a SPORE may focus on cancers caused by the same infectious agent or cancers promoted and sustained by dysregulation of a common signaling pathway. In addition, a SPORE may focus on cross-cutting themes such as pediatric cancers or cancer health disparities. The research supported through this program must be translational and must stem from research on human biology using cellular, molecular, structural, biochemical, and/or genetic experimental approaches. SPORE projects must have the goal of reaching a translational human endpoint within the project period of the grant.

2026-09-25
Education

Free to search & build · $99 one-time to unlock the application pack · No subscription

Specialized Programs of Research Excellence (SPOREs) in Human Cancers for Years 2027, 2028, and 2029 (P50 Clinical Trial Required)

upcoming

National Institutes of Health

Through this Notice of Funding Opportunity (NOFO), the National Cancer Institute (NCI) invites applications for P50 Research Center Grants for Specialized Programs of Research Excellence (SPORE). This is a re-issuance of PAR-23-284. The program will fund P50 SPORE grants to support state-of-the-art investigator-initiated translational research that will contribute to improved prevention, early detection, diagnosis, and treatment of an organ-specific cancer or a highly related group of cancers. For the purpose of this NOFO, a group of highly related cancers are those that are derived from the same organ system, such as gastrointestinal, neuroendocrine, head and neck, and other cancers. Other programmatically appropriate groups of cancers may include those centered around a common biological mechanism critical for promoting tumorigenesis and/or cancer progression in organ sites that belong to different organ systems. For example, a SPORE may focus on cancers caused by the same infectious agent or cancers promoted and sustained by dysregulation of a common signaling pathway. In addition, a SPORE may focus on cross-cutting themes such as pediatric cancers or epigenetics. The research supported through this program must be translational and must stem from research on human biology using cellular, molecular, structural, biochemical, and/or genetic experimental approaches. SPORE projects must have the goal of reaching a translational human endpoint within the project period of the grant.

2027-01-25
Healthhealthcare

Free to search & build · $99 one-time to unlock the application pack · No subscription

Specialized Programs of Research Excellence (SPOREs) in Human Cancers for Years 2027, 2028, and 2029 (P50 Clinical Trial Required)

upcoming

National Institutes of Health

<p>Through this Notice of Funding Opportunity (NOFO), the National Cancer Institute (NCI) invites applications for P50 Research Center Grants for Specialized Programs of Research Excellence (SPORE). This is a re-issuance of <a href="https://grants.nih.gov/grants/guide/pa-files/PAR-23-284.html">PAR-23-284</a>. The program will fund P50 SPORE grants to support state-of-the-art investigator-initiated translational research that will contribute to improved prevention, early detection, diagnosis, and treatment of an organ-specific cancer or a highly related group of cancers. For the purpose of this NOFO, a group of highly related cancers are those that are derived from the same organ system, such as gastrointestinal, neuroendocrine, head and neck, and other cancers. Other programmatically appropriate groups of cancers may include those centered around a common biological mechanism critical for promoting tumorigenesis and/or cancer progression in organ sites that belong to different organ systems. For example, a SPORE may focus on cancers caused by the same infectious agent or cancers promoted and sustained by dysregulation of a common signaling pathway. In addition, a SPORE may focus on cross-cutting themes such as pediatric cancers or epigenetics. The research supported through this program must be translational and must stem from research on human biology using cellular, molecular, structural, biochemical, and/or genetic experimental approaches. SPORE projects must have the goal of reaching a translational human endpoint within the project period of the grant.</p>

2027-01-25
Health

Free to search & build · $99 one-time to unlock the application pack · No subscription

SPECT/CT for Translational Theranostics Research

open

OD - NIH Office of the Director

PROJECT SUMMARY/ABSTRACT: This S10 Shared Instrumentation Grant application from Washington University (WashU) in St. Louis requests funds in partial support of the purchase of a Sybmia Pro.specta X3 scanner (Siemens Medical Solutions USA). This hybrid single photon emission computed tomography and x-ray computed tomography (SPECT/CT) system will be housed in a dedicated Nuclear Medicine research facility for the non-invasive assessment of therapeutic and diagnostic (theranostic) radiopharmaceuticals. This state-of-the-art instrument will be a critical, broadly used resource for the clinical and translational neuroscience, cardiovascular and oncology research programmes at WashU. The requested SPECT/CT will be the only research dedicated SPECT/CT system across the WashU clinical enterprise. At present, WashU through its affiliated Hospitals, has access to 7 SPECT/CT scanners across the medical campus. These are dedicated for standard of care and clinical trial workflows, 1 of them being at the Children’s hospital (out-of-reach for research), and 2 of the SPECT scanners are obsolete and only used for planar imaging. These systems are all >10 yr, and they are heavily utilized, at nearly 8 h of scan time per day average (utilization >85%), which does not include protocol development and maintenance. Access for research is highly restricted and there is no support for the special attention required for clinical research. Additionally, in the greater St. Louis region beyond WashU there is no research SPECT/CT hardware, and the nearest research SPECT/CT scanners are located at University of Missouri Veterinary Health Center (2.5 h drive), dedicated for non-human use. The Symbia Pro.specta incorporates advanced workflows including advanced iterative data driven motion correction features are critical for advanced quantitative imaging-; a redesigned quantitative framework for therapeutic absorbed dose assessment; and best-in-the-field collimators. The requested SPECT/CT scanner will anchor major new research efforts in theranostics for cancer, cardiovascular disease and neuroscience at WashU. Towards this end, Pamela Woodard, Radiology Chair and MIR Director, and Timothy Eberlein, Director, Alvin J. Siteman Cancer Center, have made substantial financial and administrative commitments to ensure the successful utilization of this instrument. These include funds for (1) installation and renovation costs, (2) adjacent radioactive handling and patient-administration space, (3) maintenance for the instrument, (4) pilot funds for protocol development and (5) personnel support. A new Section of Medical Physics is being established to harness the outstanding imaging science and translational radiopharmaceutical expertise at WashU that will be co-located with this centerpiece scanner. The combination of advanced instrumentation and robust support from our institution will enable groundbreaking discoveries and innovations that will benefit both our research community and patients, reflecting our dedication to excellence in scientific inquiry and healthcare.

Up to $750K
2027-04-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Spectral Cell Sorter

open

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY/ABSTRACT This S10 application requests a Cytek Aurora CS Spectral Cell Sorter for Sanford Burnham Prebys (SBP) to replace a 22-year-old FACSAriaIIu sorter that has experienced >45 days of downtime in the past 18 months and lacks service contract support, creating a critical bottleneck for time-sensitive experiments. Moreover, SBP's current cell sorting infrastructure cannot meet the growing demand for complex, high-parameter sorting required for cutting-edge immunological, cancer, aging, neuroscience, and skeletal muscle research by SBP's NIH-funded scientists. The Cytek Aurora CS offers revolutionary advantages including: (1) spectral technology with 64 fluorescence detectors across five lasers, enabling simultaneous analysis of 40+ parameters in a single experiment; (2) superior sensitivity for detecting low-abundance molecules; (3) advanced autofluorescence extraction capabilities to improve signal resolution in samples like brain tissue, primary tumor cells and liver samples. These features substantially improve the precision with which cell populations can be isolated. Further, the Aurora CS sorter offers seamless compatibility with our existing Aurora spectral analyzer for direct transfer of experimental protocols without panel redesign, accelerating research and reducing waste. The instrument will be housed in the SBP Flow Cytometry Shared Resource, which has demonstrated exceptional management of institutional flow cytometry equipment and provides expert technical support to ensure optimal utilization. The acquisition of this instrument is part of SBP's strategic plan to modernize the Flow Cytometry Shared Resource, in operation since 2002, and enhances the research environment by providing graduate students and postdoctoral fellows with access to state-of-the-art technology essential for developing advanced technical skills critical to the nation's future biomedical workforce. The instrument will serve fourteen NIH-funded investigators spanning multiple disciplines, with projects requiring isolation of rare cell populations for use in disease models and molecular studies including single-cell genomic and transcriptomic analyses. The acquisition of this instrument will significantly accelerate discoveries in cancer biology, immunology, neuroscience, aging and regenerative medicine at SBP, while establishing a technological foundation for future collaborative research initiatives.

Up to $632K
2027-06-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Spectral flow cytometry identifies new immune signatures that provide personalized ALS risk and progression biomarkers and therapeutic targets

open

ATSDR - Agency for Toxic Substances and Disease Registry

ABSTRACT Amyotrophic lateral sclerosis (ALS) is a progressive motor neuron disease with no cure, and although inflammation plays a significant role in the disease, gaps remain in leveraging this knowledge for personalized clinical outcome models and personalized therapeutics. Peripheral blood immune profiles—defined as the total numbers and activation states of specific peripheral immune cells—reflect overall inflammation, but methodologic gaps exist to characterize these immune profiles given limitations in conventional flow cytometry, hampering its widespread use for ALS. The long-term goal is to leverage immune profiles to identify dysregulated immune pathways that can be treated to slow or stop ALS progression. The overall objective in this proposal, being submitted in response to RFA-TS-25-036 Funding Option A, is to establish spectral flow cytometry as the state-of-the-art approach to characterize peripheral immune profiles in ALS. The central hypothesis is that spectral flow cytometry will yield rigor and reproducibility with fresh and frozen blood samples and will identify pro-inflammatory immune profiles for ALS clinical outcome prediction. The rationale is that establishing rigorous protocols for the widespread multicenter use of spectral flow cytometry in ALS will unlock the complex, but vast, potential of the immune system for improving diagnosis, prognosis, and drug development for all persons with ALS. The central hypothesis will be tested by pursuing two specific aims: 1) Utilize spectral flow cytometry to quantify inflammation in ALS peripheral blood biosamples and determine the consistency of immune markers between samples processed fresh versus frozen to inform multisite ALS studies; and 2) Determine the natural history, diagnostic, and prognostic significance of comprehensive longitudinal spectral flow cytometry immune profiles as an ALS inflammatory signature. Under the first Aim, spectral flow cytometry protocols will be optimized to characterize ALS inflammation in fresh and frozen samples, paving the way for its use in multisite ALS studies. Under the second Aim, immune profiles will be associated with important ALS clinical outcomes, such as case status and disease progression. The research proposed in this application is innovative, in the applicant’s opinion, because it moves the field in a new direction—bridging both mechanistic and knowledge gaps—by bringing the transformational potential of spectral flow cytometry to ALS, establishing the rigor needed to make the technology widely available to the ALS community, leveraging the resulting data to better understand the role of comprehensive immune profiles for ALS, and providing the foundation for future multisite studies. The proposed research is significant because peripheral blood immunophenotyping will enable improved ALS clinical outcome associations, and eventually therapeutic target identification, testing, and responder analysis.

Up to $500K
2028-09-29
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

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