Worldwide, over 2.5 million people live with spinal cord injury, with over 100,000 new cases occurring annually. Spinal cord injury often causes motor dysfunction below the level of the injury. For example, thoracic and lumbar spinal cord injury can cause paraplegia and cervical spinal cord injury can cause quadriplegia. Such injury is permanent and often severe and there is no effective treatment. Various neurologic diseases also involve damaged or dysfunctional spinal cord neurons. Neural stem cell grafts have potential for treating such conditions. However, it has not been possible to obtain sufficient numbers of appropriately patterned neural stem cells, having a spinal cord positional identity, for implanted cells to survive and functionally engraft.

The implementation of ortho-quinone methide (o-QM) intermediates in complex molecule assembly represents a remarkably efficient strategy designed by Nature and utilized by synthetic chemists. o-QMs have been taken advantage of in biomimetic syntheses for decades, yet relatively few examples of o-QM-generating enzymes in natural product biosynthetic pathways have been reported. The biosynthetic enzymes that have been discovered thus far exhibit tremendous potential for biocatalytic applications, enabling the selective production of desirable compounds that are otherwise intractable or inherently difficult to achieve by traditional synthetic methods. Characterization of this biosynthetic machinery has the potential to shine a light on new enzymes capable of similar chemistry on diverse substrates, thus expanding our knowledge of Nature's catalytic repertoire.

The primary therapeutic goal in female pelvic medicine is to restore normal pelvic floor function. Despite this, the current standard treatments are 5 compensatory, as they do not directly target sphincteric and supportive muscle dysfunction and do not reverse the existing injury or halt functional deterioration. Surgical treatments, such as muscle transplantation and transposition techniques, have had some success; however, there still exists a need for alternative therapies. Tissue engineering approaches offer potential new solutions; however, current options offer incomplete regeneration. Many naturally derived as well as synthetic materials have been explored as scaffolds for skeletal tissue engineering, but none offer a complex mimic of the native skeletal extracellular matrix, which possesses important cues for cell survival, differentiation, and migration. The extracellular matrix consists of a complex tissue-specific network of proteins and polysaccharides, which help regulate cell growth, survival and differentiation.Despite the complex nature of native ECM, in vitro cell studies traditionally assess cell behavior on single ECM component coatings, thus posing limitations on translating findings from in vitro cell studies to the in vivo setting. Overcoming this limitation is important for cell-mediated therapies, which rely on cultured and expanded cells retaining native cell behavior over time.Skeletal muscles are composed of bundles of highly oriented and dense muscle fibers, each a multinucleated cell derived from myoblasts. The muscle fibers in native skeletal muscle are closely packed together in an extracellular three dimensional matrix to form an organized tissue with high cell density and cellular orientation to generate longitudinal contraction. Skeletal muscle can become dysfunctional due to a variety of different factors including trauma, atrophy or degeneration.The reconstruction of skeletal muscle, which is lost by injury, tumor resection, or various myopathies, is limited by the lack of functional substitutes.  

Although a wide variety of hydrogels have been developed for a multitude of uses, various functional characteristics have been hard to capture in a controllable manner. A significant feature is the ability to ‘tune’ the gel so its gelling time can be controlled in a manner suitable to its application. In this disclosure, because the gel is both tunable and its composition allows it to bond to tissue, the inventors believe it can be used to address an unmet medical need – the formation of adhesions after cardiac surgery. Current methods used are either drug therapy or various physical barriers but their success is limited.

Efficient super-Mendelian inheritance of transgenic insertional elements has been demonstrated in flies, mosquitoes, yeast, and mice. While numerous potentially impactful applications of such so-called gene-drive systems have been proposed they are currently limited to copying relatively large DNA cargo sequences (~1-10 Kb). Many desired genetic traits (e.g., drought tolerance in plants, crop yield, pest-resistance, or insecticide sensitivity), however, result from allelic variants altering only one or a few base pairs. An efficient system for super-Mendelian inheritance of such subtle genetic variants would accelerate a wide array of efforts to disseminate favorable traits throughout populations, or to assemble complex genotypes consisting of point-mutant alleles in combination with insertional transgenes for a multitude of research and applied purposes.

Fluorescent proteins (FP) have been widely used as research tools in both academia and pharma for many years.  Naturally occurring FP have been mutated to either be brighter, be monomers, and/or for easier folding and expression in cells.  The most common FP to date has been the green fluorescent protein (GFP) of the jelly fish Aequorea victoria which can be expressed in cells and fused with proteins of interest, and has proven to be an excellent tool to study protein localization, expression, signaling, etc. in real time via microscopy and other techniques. 

These transgenic mice express an inducible version of cre recombinase mice under the direction of a Sox9 promoter. They are suitable for performing cre-recombination in pancreatic ductal cells and their progenitors.

Cardiovascular disease (CVD) is the leading cause of death and disability worldwide. The primary prevention of CVD is dependent upon the ability to identify high-risk individuals long before the development of overt events. This highlights the need for accurate risk stratification. An increasing number of novel biomarkers have been identified to predict cardiovascular events. Biomarkers play a critical role in the definition, prognostication, and decision-making regarding the management of cardiovascular events. There are several promising biomarkers that might provide diagnostic and prognostic information. The myocardial tissue-specific biomarker cardiac troponin, high-sensitivity assays for cardiac troponin, and heart-type fatty acid binding potential help diagnose myocardial infarction (MI) in the early hours following symptoms. Inflammatory markers such as growth differentiation factor-15, high-sensitivity C-reactive protein, fibrinogen, and uric acid predict MI and death and many others. However, there is a high unmet medical need for the more specific biomarkers that reflect different aspects of the development of atherosclerosis. 

Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. Its mode of action is to inhibit protein synthesis by disrupting peptide transfer on ribosomes, leading to premature chain termination during protein translation. Puromycin blocks protein synthesis in both eukaryotes and prokaryotes and is routinely used as a research tool in cell culture. The native Puromycin is also used assays such as mRNA display. As such, derivatives have been synthesized in which the amino acid of the 3' end of adenosine based antibiotics is altered to change the compound's antibiotic activity. Other compounds have been synthesized with differing amino acids and functionalities to examine the effect it has on bacterial viability. The majority do not show useful absorption or emission profiles. What is needed is a method to track the compounds in biological systems.

Kainic acid is a chemical first derived from seaweed. Neuroscientists routinely use Kainic acid to simulate brain degeneration in lab experiments. Certain inotropic receptors in the brain--known as kainate receptors--are selectively activated only by kainic acid. Research into kainate receptors helps researchers to understand Alzheimer's disease, epilepsy, and other brain disorders. Some scientists use kainic acid to find answers to more fundamental questions such as the function of glutamate receptors. Currently, there are two procedures for generating kainic acid commercially. The first involves the farming and collection of kainic acid-containing seaweed and that method is impacted by seasonal fluctuations of seaweed growth and kainic acid production. The second involves synthetic processes, but the current procedures generally require at least 6 synthetic steps with yields less than 40% and generate environmentally toxic byproducts including heavy metals, cyanides, or halogenated organics.

Advances in science are driven by new discoveries which can pave the way to new create new research directions. For example, crystals by the nature of their order in three-dimensional space, cannot flex or expand, but with the integration of macromolecular ferritin crystals with hydrogel polymers can change their dimensions.

Cyanide is a highly toxic agent that inhibits mitochondrial cytochrome-c oxidase, thereby depleting cellular ATP. Cyanide exposure contributes to smoke inhalation deaths in fires and could be used as a weapon of mass destruction. Cobalamin (vitamin B12) binds cyanide with a relatively high affinity and is used to treat smoke inhalation victims. Cobinamide, the penultimate compound in cobalamin biosynthesis, binds cyanide with about 1010 greater affinity than cobalamin and is 5-10 times more potent than cobalamin in rescuing animals from cyanide poisoning. Cobinamide is also an effective intra- and extracellular nitric oxide scavenger. Currently, three cyanide antidotes are currently available in the United States: nitrites, thiosulfate, and hydroxocobalamin. All three drugs are approved only for intravenous (IV) administration, and thus are not suitable for treating mass casualties as could occur after a major industrial accident or a terrorist attack. Thus, new formulations for cyanide exposure treatment that are faster and easier to administer are needed.

Microbial ecologists are increasingly turning to small, synthesized ecosystems as a reductionist tool to probe the complexity of native microbiomes. Concurrently, synthetic biologists have gone from single-cell gene circuits to controlling whole populations using intercellular signaling. 

Cardiovascular disease manifested as a myocardial infarction (MI) usually results in the irreversible death of heart muscle cells. While medical treatments can mitigate some symptoms, they often fail to prevent heart failure after a MI. The current standard of care for MI relies on surgical intervention via a coronary artery bypass. An alternative therapeutic approach has been taken in the last few years with the introduction of biomaterials designed to promote neovascularization after an MI and help prevent negative left ventricle remodeling by increasing infarct wall thickness and decreasing volume, fibrosis, and infarct size. 

Synthetic polymer constructs are an important tool in modern medical practice, but the lack of control over their activity limits their utility. The ability to combine structural function with localized interaction has proven extremely successful in stents, but polymer technology has not advanced sufficiently to serve a wider range of needs. PLGA polyesters can be degraded by hydrolysis facilitating their widespread use in medicine and biomedical research. Their dependence on slow hydrolysis makes for long degradation times (half-life of one year in vivo) limiting their applicability. While degradation can be sped up by copolymerization with more hydrophilic monomers; degradation is still too slow for triggered release or degradation.

2D crystalline materials possess high surface area-to-volume ratios, light and can be very porous. These properties have rendered synthetic 2D materials immensely attractive in applications including electronics, sensing, coating, filtration and catalysis. The rational design of self-assembling 2D crystals remains a considerable challenge and a very active area of development. The existing methods for the bottom-up fabrication of biological or non-biological 2-D crystalline materials are not generalizable and scalable. 2D protein design strategies, in particular, require extensive computational work and costly protein engineering. In addition, these strategies have low success rates, the resulting materials contain large defects, and are multi-layered and therefore not appropriate for scaling or materials-applications. Moreover, these strategies often require the presence of lipids for supported assembly.

Complex 3D interfacial arrangements of cells are found in biosystems such as blood vasculature, renal glomeruli, and intestinal villi. Tissue engineering techniques have been used to fabricate 3D microenvironments that mimic such biosystems but most methods fail to reproduce the concurrent effects of complex topography and cell encapsulation. There is a need to develop new approaches that control cell density and distribution within complex 3D features, and for biological scaffolds that reflect the true native physiology.

Strategies for the polymerization of (graft-through) and polymerization from (graft-from) proteins and peptides have been used to build macromolecules through sequential addition of monomers to a growing chain, taking advantage of polymerization catalyst proficiency and avoiding kinetically unfavorable conjugations (graft-to) between multiple large macromolecules. However, unlike for other bio-molecules (saccharides, peptides, and proteins), there are no examples of graft-through polymerization and few examples of graft-from polymerization of nucleic acids. Therefore, despite their promise, polymer bioconjugates of true nucleic acid sequences have been mostly limited to those prepared via post-polymerization modification and hence are difficult to reproduce and suffer from incomplete incorporation of the nucleic acid at each position of the polymer.

Nucleic acids have exceptional potential in the preparation of complex nanostructured materials for use as therapeutic agents and as powerful investigative tools. However, unmodified nucleic acids are inherently susceptible to enzymatic degradation in biological milieu, limiting their practical utility in detection and as therapeutics in real world applications. Specifically, new strategies are needed for the preparation of well-defined, stable and competent nucleic acid-based materials.

Some of nature’s most complex molecules are made by cellular factories that rely on an acyl carrier protein (ACP) to shuttle growing molecules along biological assembly lines. Post-translational protein modification is important for adding functions to proteins that can be exploited for therapeutics, protein engineering, affinity design and enzyme immobilization, among other applications. Commercial techniques for attaching labels to acyl carrier protein (ACP) and other carrier proteins are currently in use.

Nitrogen-rich tetrazines, have broad applications in biochemistry including small-molecule imaging, genetically targeted protein tagging, post-synthetic DNA labeling, nanoparticle-based clinical diagnostics, in-vivo imaging, as well as significant use in materials science, coordination chemistry, and the production of high energy materials such as those used in specialty explosives research. Among other uses, tetrazines can serve as coupling agents for molecular imaging compounds such as fluorophores or magnetic contrast agents, or even as ligands for metal catalysts or inorganic materials such as metal-organic frameworks. Tetrazines are also valuable synthetic intermediates, and have been elegantly deployed on route to several natural product syntheses. Despite the promise of tetrazines, the lack of convenient synthetic methods is a significant roadblock to their broader use and study.

The elastic properties of a biomaterial tissue scaffold reflect its ability to handle external loading conditions and must be tailored to match the attributes of the native tissue that it aims to repair. A scaffold’s elastic modulus and Poisson’s ratio describe how it supports and transmits external stresses to the host tissue site. (The Poisson ratio is positive/negative when the material contracts/expands transversally with axial expansion; “auxetic” materials are materials that exhibit negative Poisson ratio.) While the elastic modulus is tunable in scaffolds, the Poisson’s ratio of virtually every porous tissue construct is positive. There have been no reports of solid-phase micro-cellular biomaterials synthesized with a precisely-tuned negative Poisson’s ratio. Others have formed auxetic polyurethane foams by compressing the foams and annealing them while compressed; however, the annealing process renders little practical control over the cellular microstructure comprising the foams, making it very difficult to tune the strain-dependent behavior of Poisson’s ratio. Additionally, the foams have little to no use in biological applications involving the interactions between biomaterials and living tissue (e.g., tissue engineering applications) and other biological applications.

Nanoparticles capable of reversible changes in morphology in response to specific stimuli are expected to have broad utility in designing targeted drug-delivery, detection strategies, self-healing materials, and templates for hierarchical directed assembly. While there are several elegant examples of stimuli-responsive soft nanoparticles, programmable materials with the requisite shape-change properties remain elusive.

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