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Pancreatic Differentiation Process

Diabetes is an autoimmune disease in which the immune system attacks and destroys the insulin-producing beta cells in the pancreas. Islet transplantation is a suitable therapy for diabetes, but the scarceness of islet donors makes this approach practically impossible in a higher scale. Promising alternative cell sources for treating diabetes comes from human embryonic stem cells (hESCs), and numerous groups have generated functional insulin-producing cells in vitro using stepwise differentiation protocols that mimic pancreatic development, including from diabetic patients. However, the limited transcriptomic data on these cells indicate that there remain marked differences between the most mature hESC-derived cells and primary human adult beta cells. Therefore, improvements in methodologies for proper maturation of beta-cells in vitro continues to be an intense area of research.

Near Infrared Fluorescent Imaging Used to Assess Tissue Perfusion in Surgery

Near infrared (NIR) fluorescence imaging (FI) utilizing the fluorophore indocyanine green (ICG) has become more popular for use in medical diagnostics. It is useful for assessing tissue perfusion in a number of surgeries, particularly abdominal, heart, plastic, hepatic as well as other areas of medicine. The light needed for the excitation of the fluorescence is generated by a near infrared light source which is attached directly to a camera. A digital video camera allows the absorption of the ICG fluorescence to be recorded in real time, which means that perfusion can be assessed and documented. Currently, ICG provides a visual representation of tissue perfusion as a global view. Although some efforts have been put into density analysis, no device or software currently performs dynamic evaluation of blood flow for a surgeon. Without objective dynamic measurements, practitioners are only limited to snap shot view of the static environment. This is a problem because it is the dynamics of blood flow that determines tissue perfusion, not how much blood present at a stationary point in time. Furthermore, because there are no numerical evaluations out on the market that can capture this dynamic aspect of blood flow, practitioners are forced to use the naked eye to make a clinical decision that is not only subjective, but is difficult to assess between cases.

Methods for Global RNA-Chromatin Interactome Discovery

Recent decades of genomic research reveal that mammalian genomes are more prevalently transcribed than previously anticipated. It is now quite clear that mammalian genomes express not only protein-coding RNAs but also a large repertoire of non-coding RNAs that have regulatory functions in different layers of gene expression. Many of those regulatory RNAs appear to directly act on chromatin, as exemplified by various long noncoding RNAs (IncRNAs). Some of those regulatory RNAs mediate genomic interactions only in cis, while others, such MALAT1 and NEAT1, are capable of acting in trans. These findings suggest an emerging paradigm in regulated gene expression via specific RNA-chromatin interactions. Various techniques have been developed to localize specific RNAs on chromatin. These methods, such as chromatin Isolation by RNA purification or comprehensive identification of RNA binding proteins (ChIRP), capture hybridization analysis of RNA targets (CHART), and RNA affinity purification (RAP), all rely on using complementary sequences to capture a specific RNA followed by deep sequencing to identify targets on chromatin. Importantly, all of these methods only allow analysis of one known RNA at a time, and up to date, a global view is lacking on all RNA-chromatin interactions, which is critical to address a wide range of functional genomics questions.

Diagnostic Gene Signature For Cancer Vascular Mimicry in Solid Tumors

One of the characteristic trademarks of tumorigenesis is the need for an extensive vascular system to supply blood for the tumor to grow and disseminate from the original node to distant sites via the process of metastasis. This involves the growth of new vessels from existing vessels, as well as the migration of tumor cells through the extracellular matrix (ECM) and into the lymphatic or vascular systems. However, some very aggressive solid tumors can form vascular channels by themselves, which is termed vascular mimicry (VM). Moreover, only certain cells in these tumors have the ability to produce blood-transporting channels, contributing to metastasis. There is growing evidence that supports the idea that VM can be a prognostic factor for poor clinical outcomes in various types of cancer. Currently, VM is identified by a pathologist’s evaluation of histological slides, wherein vascular-like structures that do not stain positive for endothelial cells are identified as VM. Thus far, conserved molecular biomarkers that define this phenotype have remained unknown.

Imaging Cells In Flow Cytometer Using Spatial-Temporal Transformation

Flow cytometry analyzes multiple physical characteristics of a large population of single cells as cells flow in a fluid stream through an excitation light beam. Flow cytometers measure fluorescence and light scattering from which information about the biological and physical properties of individual cells are obtained. Although flow cytometers have massive statistical power due to their single cell resolution and high throughput, they produce no information about cell morphology or spatial resolution offered by microscopy, which is a much wanted feature missing in almost all flow cytometers.

Live Cell Detection by Near-Infrared Fluorogenic Tetrazine Uncaging Oligo Probes

There is significant interest in developing methods that visualize and detect RNA in live cells. Bioorthogonal template driven tetrazine ligations are quickly becoming a powerful route to visualizing nucleic acids in native cells, yet past work has been limited with respect to the diversity of fluorogens and existing tetrazine-reactive fluorogenic probes are quenched by through‐bond energy transfer (TBET) or Fӧrster resonance energy transfer (FRET) between the donor fluorophore and acceptor tetrazine.

In Situ Lipid Synthesis for Protein Reconstitution (INSYRT)

While current methods for membrane protein functional reconstitution in biomimetic membranes approaches are powerful and have uncovered fundamental properties of protein function, they are methodologically cumbersome, requiring chromatography steps to remove detergents. Moreover, structural features normally found in cell membranes such as curvature and polarity are mostly absent. In this regard, an efficient reconstitution methodology that better mimics the native chemical environment of a whole-cell embedded protein would be highly useful.

Next Generation PCR

In many critical healthcare situations, including sepsis and septic shock, the identification and diagnosis of infectious agents is burdensome and slow. Timely medical intervention is often delayed while laboratory testing is performed and the results analyzed. A point-of-care rapid diagnostic tool is a well-known unmet need within the clinical community. Some tools do exist, but they typically present limitations and draw-backs. Importantly, none give actionable results in the clinically relevant timeframe of 3-4hrs. Recently, UCSD researchers have developed an improved system for rapid gene profiling and diagnostic identification of infectious disease agents and their resistance profiles, by applying High Resolution Melt (HRM) technology and machine learning to a digital polymerase chain reaction (dPCR) platform.

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