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Improved Growth of Stem Cells in Culture

Prof. Talbot and her colleagues from the University of California, Riverside have developed a research tool to prolong the viability and pluripotency of stem cells in culture. The culture medium is supplemented with an additive that includes a source of acetate ions, a carboxylic acid, or a salt of the carboxylic acid, or a combination of these substances. Results have shown that this substrate medium allows for less stem cell death, faster colony growth, and causes cells to attach to and spread faster on the substrate. This provides tremendous advantages in stem cell colony morphology, growth, survival, maintenance of pluripotency, and dynamic behavior when compared to existing media.  Fig 1: Images of stem cells in culture before and after treatment  

Development of Methods and Assay for Measurement of Total Oxidized Phospholipid (OxPL)

Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease in the United States. It can be broadly sub-classified into nonalcoholic fatty liver (NAFL), which is thought to have minimal risk of progression to cirrhosis, and nonalcoholic steatohepatitis (NASH), which is thought to have an increased risk of progression to cirrhosis. The current diagnostic gold standard for differentiating whether a patient with NAFLD has NAFL versus NASH is liver biopsy. However, liver biopsy is an invasive procedure, which is limited by sampling variability, cost, and may be complicated by morbidity and even death, although rare. Accurate, non-invasive, biomarkers for the detection of liver disease and liver disease progression e.g., progression to NASH, are currently also not available.

Monoclonal Antibodies Specific to Canine PD-1 and PD-L1

Researchers at the University of California, Davis have developed monoclonal antibodies with multiple applications relevant to canine PD-1 and PD-L1.

Kelch Like Family Member 11 (Klhl11) Autoantibodies As Markers Of Seminoma Associated Paraneoplastic Encephalitis In Men

Researchers at UCSF and Chan Zuckerberg Biohub have discovered a novel biomarker for an autoimmune disease that affects patients with testicular cancer.  The disease, known as “testicular cancer-associated paraneoplastic encephalitis,” can cause severe neurological symptoms.  The symptoms include loss of limb control, eye movement, and in some cases, speech.  The disease begins with testicular cancer, which in some cases causes the immune system to attack the brain.  Affected patients are often misdiagnosed and appropriate treatment is delayed. 

New Indications For ENPP1 Inhibitors

UCLA researchers in the Department of Medicine have developed small molecule ENPP1 inhibitors and monoclonal antibodies for treating myocardial infarction and ocular calcification.

Hydrodealkenylative C(Sp3)–C(Sp2) Bond Scission

UCLA researchers in the Department of Chemistry and Biochemistry have developed a new chemical reaction that combines ozone, an iron salt, and a hydrogen atom donor to enable hydrodealkenylative cleavage of C(sp3)–C(sp2) bonds in a widely applicable manner.

Drug Repurposing To Explore Novel Treatment For Cushing Disease

UCLA researchers in the Department of Medicine and the Department of Molecular and Medicinal Pharmacology have identified several small molecule reagents to treat Cushing disease.

Development of Novel Fluorescent Puromycin Derivatives

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.

Methods of Diagnosing Ocular Diseases

Researchers at UCLA have developed a novel technique to detect ocular diseases.

Rapid, Sensitive Detection of Nucleic Acid Sequences in Environmental Samples

UCLA Researchers at the California NanoSystems Institute have developed a methodology that permits PCR-based detection of nucleic acid sequences in soil that does not require the isolation of DNA.

Reagent to Label Proteins via Lysine Isopeptide Bonds

Researchers in the UCLA Department of Chemistry and Biochemistry and the University of Texas-Medical Center, Houston Department of Microbiology and Molecular Genetics have modified the Corynebacterium diphtheriae (C. diphtheriae) sortase enzyme so that it can be used as a bioconjugation reagent in vitro.

Assay for Oligonucleotides in Serum Without Extraction or RT-PCR

Prof. Ameae Walker’s laboratory at the University of California, Riverside (UCR) has developed an assay to quantify oligos in sub-picomole amounts without the need for sample purification and amplification. This new competitive assay is called an ELOHA (Enzyme-Linked Oligonucleotide Hybridization Assay). The method is illustrated in Fig. 1, below.  Capture Oligos that are to hybridize with an oligo to be measured are covalently linked to a plate (1), a Detection Oligo, with the same sequence as the oligo to be measured, has a conjugated label, such as horseradish peroxidase or biotin.  The Detection Oligo then competes with the oligo of interest for binding to the Capture Oligo (2).  Once the hybridization is complete, the unbound oligos are washed away (3).  A colorimetric readout is produced (4) to inversely quantify the oligo of interest.  Fig. 1 Schematic of the ELOHA assay Fig. 2 shows the use of an ELOHA for amounts of Antimaia in mouse serum. Antimaia is a splice modulating oligomer therapy for breast cancer developed in the UCR lab of Prof. Walker.  

Electrochemical Flash Fluorination and Radiofluorination

Researchers led by Saman Sadeghi from the Department of Molecular & Medical Pharmacology at UCLA have developed a new and simple process to make fluorinated organic compounds.

Protein-Coated Microparticles For Protein Standardization In Single-Cell Assays

Single-cell analysis offers powerful capabilities of identification of rare sub-populations of cells, understanding heterogeneity of cancerous tumors, and tracking cell differentiation and reprogramming. Despite great potentials for uncovering new biological systems and targeting diseases with precision medicine, single-cell approaches are composed of complex device processes that can cause bias in measurement.  In deep sequencing, technical variation in single cell expression data occurs during capture and pre-amplification steps. Similarly, in single-cell protein assays, technical variability can obscure functionally relevant variance.    To better control protein measurement quality in single-cell assays, researchers at the University of California, Berkeley developed a novel method to loading and release protein standard. This method utilizes the surface of modified and functionalized microparticles as vehicles to capture target proteins with desired concentrations. Chelation-assisted click chemistry is applied to demonstrate that protein standards with different molecular masses can be loaded and bounded in a single-cell protein assay. Microparticles are introduced into single-cell devices by either passive gravity, magnetic attraction, or other physicochemical forces. These protein standards from microparticles provide a reference to measure protein mass sizes from individual cells and a quality control for any biases in device fabrication, cell lysis, protein solubility, protein capture, and protein readouts (i.e. antibody probing).   

Scar Minimization Treatment: Fibrotic to Fat Cell Conversion

Clinical treatment for scar-less wound healing remains a highly desired, yet unmet need. UCI researchers have developed a method to minimize scarring during wound healing through cellular reprograming that encourages formation of new skin fat cells. This novel therapy is non-surgical and applicable to multiple types of scars and aging skin.

The Brightest, Red-Shifted Luciferase-Luciferin Bioluminescent Pairs

Researchers at the University of California, Riverside, have developed several new luciferase-luciferin pairs that have superior brightness and excellent performance in vitro and in vivo. Through directed evolution of the existing NanoLuc Luciferase and the use of diphenylterazine (DTZ) as a substrate, the emission extensity is more than doubled compared to NanoLuc-furimazine. Moreover, red-shifted emission of teLuc-DTZ makes it an excellent tool for in vivo imaging. teLuc-DTZ streamlines a variety of applications to afford high sensitivity and reproducibility. Furthermore, fusing teLuc to a fluorescent protein creates the Antares2-DTZ pair, with emissions further red-shifted to the > 600 nm range and 65 times more photons emitted above 600 nm than FLuc-D-Luciferin. Fig. 1 shows the relative emission intensity and the range of emitted wavelengths of light  

17S-FD-895: An Improved Synthetic Splice Modulator mirroring FD-895

The spliceosome, the cellular splicing machinery, regulates RNA splicing of messenger RNA precursors (pre-mRNAs) into maturation of protein coding RNAs. Recurrent mutations and copy number changes in genes encoding spliceosomal proteins and splicing regulatory factors have tumor promoting or suppressive functions in hematological malignancies, as well as some other cancers. Over the past 10 years, a list of natural products, including FD-895, pladienolide B, herboxidiene, and spliceostatin A, have been identified as spliceosome modulators. They have been shown to have anti-cancer effect in vitro and in vivo models. However, these compounds demonstrate poor metabolic stability and short half-lives in vivo, excluding them from entering clinical evaluation. This invention, 17S-FD-895, is an analog of FD-895 and was synthesized through the combination of total synthesis and synthetic methods, demonstrating improved stability and on-target effect. This new spliceosome targeting compound was evaluated in different secondary acute myeloid leukemia models and showed potent efficacy in inhibition of acute myeloid leukemia (AML) LSC and disruption of AML maintenance in vitro and in mouse xenograft models (Crews et al. 2016). The study by Crews et al. indicates the pivotal role of spliceosome in secondary acute myeloid leukemia and the therapeutic potential of targeting leukemia stem cells in this subtype of AML often unresponsive to current therapy.

Modulation Of p53 as a Cancer Therapeutic Target

Researchers at the University of California, Davis have designed peptides and oligonucleotide sequences to enhance p53 expression.

Biologically Applicable Water-Soluble Heterogeneous Catalysts For Parahydrogen-Induced Polarization

UCLA researchers in the Department of Chemistry and Biochemistry have developed a novel method of parahydrogen-induced polarization in water using heterogeneous catalysts.

Sieve Container For Contactless Media Exchange For Cell Growth

Media that contains nutrients and growth factors is necessary to grow all types of cells, a process that is widely used in many fields of research. Such media should be routinely changed either to different media or a fresh batch of the same media. This change currently involves either using a pipette to transfer cells from their current dish of media to a new dish, or aspirating the media out of the dish and replacing it with new media. Both methods have inherent risks to stressing and damaging the cells. Researchers at UCI have developed a unique dish for growing cells that allows for safer aspiration of the old media, which reduces stress and damage to the cells.

Gene Delivery Into Mature Plants Using Carbon Nanotubes

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} Current methods of biomolecule delivery to mature plants are limited due to the presence of plant cell wall, and are additionally hampered by low transfection efficiency, high toxicity of the transfection material, and host range limitation. For this reason, transfection is often limited to protoplast cultures where the cell wall is removed, and not to the mature whole plant.  Unfortunately, protoplasts are not able to regenerate into fertile plants, causing these methods to have low practical applicability. Researchers at the University of California have developed a method for delivery of genetic materials into mature plant cells within a fully-developed mature plant leaf, that is species-independent. This method utilizes a nano-sized delivery vehicle for targeted and passive transport of biomolecules into mature plants of any plant species. The delivery method is inexpensive, easy, and robust, and can transfer biomolecules into all phenotypes of any plant species with high efficiency and low toxicity.

Xenobiotic-Free Culture System To Expand Human Limbal Stem Cells

UCLA researchers in the Departments of Opthalmology have developed a xenobiotic-free manufacturing process to produce transplantable human limbal stem cells for use in treating limbal stem cell deficiency.

Au(III) Complexes For [18F] Trifluoromethylation

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} The biological properties of trifluoromethyl compounds (e.g, CF3) have led to their ubiquity in pharmaceuticals, yet their chemical properties have made their preparation a substantial challenge, necessitating innovative chemical solutions.  For example, strong, non-interacting C-F bonds lend metabolic stability while simultaneously limiting the ability of chemical transformations to forge the relevant linkages and install the CF3 unit.  When these same synthetic considerations are extended toward the synthesis of trifluoromethylated positron emission tomography (PET) tracers, the situation becomes more complex.   UC Berkeley researchers discovered an unusual alternative mechanism, in which borane abstracts fluoride from the CF3 group in a gold complex. The activated CF2 fragment can then bond to a wide variety of other carbon substituents added to the same gold center. Return of the fluoride liberates a trifluoromethylated compound from the metal. This mechanism would be useful for the introduction of radioactive fluoride substituents for potential tracers to be used for positron emission tomography applications.

Small Molecule Generation of Multinucleated and Striated Myofibers from Human Pluripotent Stem Cells Equivalent to Adult Skeletal Muscle

Researchers in the UCLA Department of Microbiology, Immunology and Molecular Genetics have developed a novel means of generating adult skeletal muscle-equivalent myofibers from human pluripotent stem cells.

Siderophore-Based Immunization Against Gram-Negative Bacteria

Bacterial pathogens such as E. coli and Salmonella hijack the host’s iron to cause infection. This invention describes an immunization strategy for triggering an immune response against the iron-sequestering agent secreted by the pathogen, thus turning the bacterial virulence mechanism against itself, and thereby resulting in host immunity.

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