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Assay for Chronic Wound Biofilm Disruptors

Dr. Manuela Martins-Green’s UCR laboratory has developed a new assay to identify medicines that disrupt chronic wound biofilms in patients.  A patient’s wound biofilm is collected with a sterile swab and the bacteria collected from the swab are cultured and identified.  The bacteria are then used singly or in combination, to make a biofilm in 96 well plates to then be used for high throughput screening using a multitude of antibiotics, chemical and small molecules that may be tested alone or in combination for their ability to disperse the wound’s biofilm. Fig. 1 shows the biofilm cultures from chronic wound patient isolates. The biofilm is made more easily visible by staining with crystal violet. Fig. 2 Different concentrations of an antimicrobial drug ranging from 3 to 20 mg/ml were applied at 0, 6, 12, and 24 hours after the biofilm cultures were initiated. When applied at time zero the bacteria will not grow or form biofilm. This is akin to application after debridement. When applied after the biofilm has been formed it will dismantle the biofilm when used at the right concentrations. Akin to treating without debridement.  

Stamping-based Method for Microwell Production and Cell Aggregate Formation

Researchers at the University of California, Davis have developed a 3-D printed stamping system (the “Aggrestamp”) with the capability for in-situ production of microwells that facilitate cell aggregate formation.

Systems and Methods for Monodisperse Drop Generation and Use

UCLA researchers in the Department of Bioengineering have developed systems and methods to produce single particle, monodisperse droplets for use in digital assays, targeted drug delivery, and theranostics.

A Microfluidic Single-Cell Pairing Array for Studying Cell-Cell Interaction in Isolated Compartments

Cell interactions are fundamental to biological processes. Microfluidics provides a reliable platform to study these intricate phenomena. The researchers have developed a microfluidic trapping array which efficiently pairs single cells in isolated compartments in an easy to operate manner to study cell-cell interaction, especially at single-cell level.

Antibody and Vaccine Therapy for C. diff. Infection

Clostridium difficile (C. diff.) infection is estimated to cause nearly 0.5 million illnesses in the US. C. diff. can cause severe gastrointestinal effects, including life-threatening inflammation, is contagious, and is an urgent antibiotic-resistant threat, according to the Centers for Disease Control and Prevention. UCI researchers have determined the crystal structure for the virulent C. diff. toxin, TcdB, and characterized sites to target for neutralization along with immunogens that can be used in vaccine strategies to prevent and treat C. diff. infection.

The CryoEM Method MicroED as a Powerful Tool for Small Molecule Structure Determination

UCLA researchers in the Department of Chemistry and Biochemistry have developed a novel use of the cryogenic electron microscopy (CryoEM) method electron micro-diffraction (MicroED) to provide routine and unambiguous structural determination of small organic molecules.

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.

Novel Fret Method

Dr. Jiayu Liao and colleagues at the University of California, Riverside have developed a FRET assay using nitrobenzoxadiazole (NBD) and coumarin (CUM) amino acid analogs as a FRET pair.  These fluorophores are genetically encoded into peptides and proteins surrounding a protease cleavage site or ligand binding site and used for FRET-based high throughput screening for enzymes or small molecule inhibitors involved in pathways such as SUMOylation. Researchers have demonstrated FRET for peptides encoded with NBD and CUM separated by 4 and 6 amino acids and excited at 340 nm (Figure 1). Figure 1.  Fluorescent intensity of peptide I (6 amino acids between CUM and NBD) and II (4 amino acids between CUM and NBD) excited at 340 nm.  

Very-Small-Nuclear Circulating Tumor Cell (vsnCTC) as a Diagnostic Biomarker of Visceral Metastasis in Advanced Prostate Cancer

UCLA researchers in the Department of Molecular and Medical Pharmacology have identified a novel biomarker that can be used to diagnose prostate cancer patients for the presence of visceral metastasis with 54% sensitivity and 100% specificity.

DEVICES AND METHODS FOR GENERATING OLIGODENDROCYTE PROGENITOR CELLS

The emergence of several cell based therapy candidates in the clinic is an encouraging sign for human diseases/disorders that currently have no effective small molecule or biologic based therapy. Stem cells – including adult and pluripotent subtypes – offer tremendous clinical promise for the treatment of a variety of degenerative diseases, as these cells have the capacity to self-renew indefinitely and to mature into functional cell types and thereby serve as a source of cell replacement therapies (CRTs) and pluripotent stem cells (hPSCs) are of increasing interest for the development of CRTs because of their capacity to differentiate into all cell types in an adult, for which adult tissue-specific stem cells may in some cases not even exist. One potential CRT enabled by hPSCs is oligodendrocyte progenitor cells (OPCs) for the treatment of spinal cord injury (SCI). Such hPSC-OPCs have recently advanced to a Phase II clinical trial and are even being considered for additional diseases in the central nervous system (CNS), such as multiple sclerosis (MS), or injury from radiation.   UC researchers have developed a microscale 3D culture screening and analysis methodology that is relevant to the production of several up and coming cell replacement therapy candidates for which derivation from a precursor cell type requires searching through a large in vitro design space of doses, durations, dynamics, and combinations of signaling cues over several weeks of culture, such as oligodendrocyte progenitor cells (OPCs) and midbrain dopaminergic neurons (mDA neurons) derived from human pluripotent stem cells. 

DNA Methylation: A New Method for the Quantitative Predictor Of Age In Dogs

The ability to properly estimate the age of dogs would be quite useful in a variety of ways. For example, proper age estimation is important because age often plays a significant role when making medical decisions for pets. Currently, the accepted method to estimate age in dogs is based on the quality of teeth as well as ocular features. Estimating age based on tooth-wear (the commonly used metric in shelters) is very inaccurate after the teeth have fully erupted, generally by 6-7 months of age in dogs. Unfortunately, these methods have an accuracy of ~50% at best for domesticated pets and is error-prone for dogs between 2-8 years, encompassing a large portion of a dog’s adult life. Thus, shelters commonly underestimate the ages of these dogs to increase the likelihood of dogs being adopted, as people generally have a preference for younger pets. 

Capture And Stimulated Release Of Circulating Tumor Cells On Polymer Grafted Silicon Nanostructures

UCLA researchers in the department of Molecular and Medical Pharmacology have developed a novel capture system of circulating tumor cells for the early detection of metastatic cancer.

Single Circulating Tumor Cell Isolation Using Laser Microdissection And A Polymer Enrichment Assay

UCLA researchers in the department of Molecular and Medical Pharmacology have developed a novel matrix polymer capture system of circulating tumor cells that preserves biomolecular integrity through laser microdissection for the early detection of metastatic cancer.

Method For Indefinite Storage And Preservation Of Membrane Precursors

UCLA researchers in the Department of Bioengineering have developed a novel strategy for the creation of biomimetic lipid bilayer membrane using a high freezing point lipid-containing solvent.  Using this method, the membrane precursor is frozen/immobilized prior to the completion of the spontaneous process of bilayer self-assembly, and the process can be resumed later by simply thawing and allowing membrane formation to resume.

Novel Steroid Hormone Assay

Researchers at the University of California have identified in insects that the membrane transporter, Ecdysone Importer (EcI), is involved in the cellular uptake of the primary steroid hormone ecdysone. Specifically after transport through Ecl, ecdysone’s active form (20-hydroxyecdysone or 20E and related ecdysteroids) enters its target cells and binds to the ecdysone receptor (EcR), which forms a heterodimer with another nuclear receptor and activates transcription of multiple genes involved in molting and metamorphosis. This new discovery of Ecl’s role counters the prevailing consensus that steroid hormones diffuse through cell membranes.  This will enable the screening of new compounds that interact with Ecl.  Such new compounds may be used for insect pest control. Fig. 1 membrane transporters (blue) guide steroid hormones (blue dots) into cells. This new discovery counters the conventionally held scientific consensus that steroid hormones passively diffuse through cell membranes.   Fig. 2 EcI mutants (bottom) were not able to enter into metamorphosis when compared to the control (top).

Mitochondrial Respirometry In Frozen Specimens

UCLA researchers from the Department of Medicine have developed a novel technique for performing mitochondrial respirometry in frozen specimens to accurately assess the cellular energy production capacity.

Phenotypic Profiling Of Hepatocellular Carcinoma Circulating Tumor Cells For Treatment Selection

Researchers in the UCLA Departments of Surgery and Molecular and Medical Pharmacology have developed a novel blood-based assay that can capture and characterize circulating tumor cells indicative of both early- and late-staged hepatocellular carcinoma (HCC).

A High Throughput Biochemical Fluorometric Method For Measuring HDL Redox Activity

UCLA researchers in the Department of Medicine have developed a method of screening for the functional properties of high-density lipoprotein (HDL) in the blood that may serve as a more accurate risk indicator of cardiovascular disease.

Predicting the Placebo Response and Placebo Responders in Medicated and Unmedicated Patients Using Baseline Psychometric and Clinical Assessment Score

UCLA researchers have developed a method and model to predict the placebo effect and placebo responsiveness using the 30-item baseline positive and negative syndrome scales (PANSS) scores, within both the medicated and unmedicated Schizophrenia patients.

Lipid-Modified Oligonucleotides For Sample Barcoding in Droplet Microfluidics-Based Single-Cell RNA Sequencing

A new strategy for barcoding single living cells using lipid-modified oligonucleotides that can vastly enhance sample multiplexing in droplet microfluidics-based RNA sequencing

Optimizing B Cell Epitope-Based Autoantibody Detection In Cancer Patients

UCLA researchers from the Department of Urology have invented a new approach to optimize detection of autoantibody response using Luminex microbead-based multiplex assay. This approach bypasses the difficult process of purifying whole proteins by using select combinations of short B-cell epitopes.

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.  

A Micro-Bubble Plate For Patterning Biological and Non-Biological Materials

A method for creating a 3D micro-bubble plate for patterning biological and non-biological materials. Because each sample is at a known location, large numbers of samples may be studied and allow for significant statistical data sets, which will aid in diagnosing unknown agents or diseases inexpensively.

A Micro-Patterned Plate Composed Of An Array Of Releaseable Elements Surrounded With Solid Or Gel Walls

This technology is a micro-patterned plate made of an array of releasable elements surrounded by a gel or solid wall, and a process for manufacturing the micro-patterned plate. This is an efficient way of studying samples for statistically significant data sets of cells or biological materials for important scientific research and medicines.

A Novel Method and Protocol to Induce Pluripotent Stem Cells Toward Astrocyte Differentiation

Rett syndrome (RTT) is a devastating disease that affects 1 in every 10,000 children born in the United States, primarily females. RTT patients undergo apparently normal development until 6-18 months of age, followed by impaired motor function, stagnation and then regression of developmental skills, hypotonia, seizures and a spectrum of autistic behaviors. Rett syndrome is a rare disease that shares certain pathways with major developmental disorders such as autism and schizophrenia, increasing the potential impact. There is no cure for Rett syndrome and the animal model does not entirely recapitulate the human disease. Thus, having the possibility to screen drugs directly in human neurons is a major milestone.

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