#5 - Identifying cellular mechano-biological responses to PEG-based hydrogels

Cells sense and respond to mechanical stimuli from their external environment through a process called mechanotransduction. Focal adhesions are integrin-containing, multiprotein structures through which mechanical force is transmitted between the extracellular matrix and the interacting cell. Cells convert the transmitted force into biological responses including migration, proliferation and differentiation. The Garcia lab has previously engineered an integrin-specific hydrogel system resulting in changes in mesenchymal stem cell (MSC) gene expression, secretome, and ultimately regenerative capacity in a murine bone repair model. However, the mechano-biological mechanism driving this cell response to varying hydrogel biophysical and biochemical properties has yet to be studied. Here we have developed relationships between various hydrogel properties and cellular responses (cell adhesion, YAP localization, cell area, and cell shape). We engineered PEG-based hydrogel systems with two different polymerization chemistries, malemide and norbornene, to explore the effect of hydrogel chemistry on MSC cell adhesion and spreading. Using rheology, we demonstrated that hydrogel mechanical properties can be tuned by altering the weight percent of PEG macromer, while adhesion ligand type and density had no effect on hydrogel mechanical properties. PEG-4MAL gels were used for the remaining studies because that chemistry resulted in improved spreading and cell adhesion over norbornene hydrogels. By varying the density of RGD, the adhesive ligand for αvβ3 integrin, on the hydrogels, we showed that higher RGD densities resulted in greater YAP nuclear localization. We sought to understand the mechano-biological signaling pathway involved in YAP nuclear localization by inhibiting ROCK and FAK, proteins critical in mechanosensing via focal adhesion complexes. The inhibition of ROCK, and FAK decreased cell spread area, increased cell circularity and decreased YAP nuclear localization. Taken together this data demonstrates that external signals from PEG-based hydrogels as well as the intracellular signaling cascades involving ROCK and FAK can modulate YAP mechanosensing in MSCs

Dissolved oxygen dynamics during a phytoplankton bloom in the Ross Sea polynya

The Ross Sea polynya is one of the most productive regions in the Southern Ocean. However, limited access and high spatio-temporal variability of physical and biological processes limit the use of conventional oceanographic methods to measure early season primary productivity. High-resolution observations from two Seagliders provide insights into the timing of a bloom in the southern Ross Sea polynya in December 2010. Changes in chlorophyll and oxygen concentrations are used to assess bloom dynamics. Using a ratio of dissolved oxygen to carbon, net primary production is estimated over the duration of the bloom showing a sensitive balance between net autotrophy and heterotrophy. The two gliders, observing spatially distinct regions during the same period, found net community production rates of -0.9±0.7 and 0.7±0.4 g C m-2 d-1. The difference highlights the spatial variability of biological processes and is probably caused by observing different stages of the bloom. The challenge of obtaining accurate primary productivity estimates highlights the need for increased observational efforts, particularly focusing on subsurface processes not resolved using surface or remote observations. Without an increased observational effort and the involvement of emerging technologies, it will not be possible to determine the seasonal trophic balance of the Ross Sea polynya and quantify the shelf's importance in carbon export

Penguin decays of B mesons

Penguin, or loop, decays of B mesons induce effective flavor-changing neutral currents, which are forbidden at tree level in the Standard Model. These decays give special insight into the CKM matrix and are sensitive to non-standard model effects. In this review, we give a historical and theoretical introduction to penguins and a description of the various types of penguin processes: electromagnetic, electroweak, and gluonic. We review the experimental searches for penguin decays, including the measurements of the electromagnetic penguins b -> s gamma and B -> K* gamma and gluonic penguins B -> K pi, B+ -> omega K+ and B -> eta' K, and their implications for the Standard Model and New Physics. We conclude by exploring the future prospects for penguin physics.Comment: 49 pages, LATEX, 30 embedded figures, submitted to Annual Reviews of Nuclear and Particle Scienc

Nature of Viscoelasticity in Lamellar Block Copolymers: Contraction Correlated to Strain Localization

We determined the local strain profile in sheared lamellar block copolymers. A trilayer model, based on elastic brushes separated by viscous interpenetration zones, captures the rheological response of these materials and provides a measure of the relative contributions of elastic versus viscous strain. The elastic chain distortions were evident from a reversible lamellar contraction, as measured from in situ small-angle x-ray scattering. The contraction was directly correlated to the elastic strain from our trilayer model; three distinct diblock copolymers conform to the predicted relationship, thus suggesting universal behavior

Harmful Algae Bloom Identification Laboratory for Virginia Shellfish Hatcheries and Nurseries

HAB Identification Laboratory Training Module This laboratory is one part of a collaborative effort funded by NOAA Sea Grant to deliver timely and practical shellfish culture information to the commercial industr

The Application of the Emergency Medical Treatment and Labor Act (EMTALA) to Hospital Inpatients

This issue brief provides a brief overview of the Emergency Medical Treatment and Labor Act (EMTALA) and focuses on its application to hospital inpatients. EMTALA applies differently to patients than non-patients, and also applies differently to patients admitted through the emergency department than patients admitted as regular inpatients. In addition, courts and the Centers for Medicare and Medicaid Services (CMS) have differed in their interpretation of the statute. Depending on the specific facts of any particular case, EMTALA may or may not have implications for specialty-related transfers and discharges

Exploring the origins of EEG motion artefacts during simultaneous fMRI acquisition: implications for motion artefact correction

Motion artefacts (MAs) are induced within EEG data collected simultaneously with fMRI when the subject’s head rotates relative to the magnetic field. The effects of these artefacts have generally been ameliorated by removing periods of data during which large artefact voltages appear in the EEG traces. However, even when combined with other standard post-processing methods, this strategy does not remove smaller MAs which can dominate the neuronal signals of interest. A number of methods are therefore being developed to characterise the MA by measuring reference signals and then using these in artefact correction. These methods generally assume that the head and EEG cap, plus any attached sensors, form a rigid body which can be characterised by a standard set of six motion parameters. Here we investigate the motion of the head/EEG cap system to provide a better understanding of MAs. We focus on the reference layer artefact subtraction (RLAS) approach, as this allows measurement of a separate reference signal for each electrode that is being used to measure brain activity. Through a series of experiments on phantoms and subjects, we find that movement of the EEG cap relative to the phantom and skin on the forehead is relatively small and that this non-rigid body movement does not appear to cause considerable discrepancy in artefacts between the scalp and reference signals. However, differences in the amplitude of these signals is observed which may be due to differences in geometry of the system from which the reference signals are measured compared with the brain signals. In addition, we find that there is non-rigid body movement of the skull and skin which produces an additional MA component for a head shake, which is not present for a head nod. This results in a large discrepancy in the amplitude and temporal profile of the MA measured on the scalp and reference layer, reducing the efficacy of MA correction based on the reference signals. Together our data suggest that the efficacy of the correction of MA using any reference-based system is likely to differ for different types of head movement with head shake being the hardest to correct. This provides new information to inform the development of hardware and post-processing methods for removing MAs from EEG data acquired simultaneously with fMRI data

The Family Check-Up in a Pediatric Clinic: An Integrated Care Delivery Model to Improve Behaviors in the Home Environment

This study examines the feasibility of adapting the Family Check Up (FCU), an evidence-based program to identify and manage behavioral concerns in children ages 4 and 5 years, within a pediatric primary care clinic with an integrated mental health professional. Methods: Caregivers attending their child’s 4 and 5 year-old well child visit were asked to complete a screening tool (Pediatric Symptom Checklist-17; PSC-17) measuring behavioral concerns as part of routine care. Families who screened positively, were referred to the FCU and asked to participate in a study evaluating the intervention. The FCU is a 2-session intervention during which information on home environment and parenting style was collected through tailored questionnaires, videotaped interactions, and a clinical interview. Feasibility was examined using portions of the Reach, Effectiveness, Adoption, Implementation, and Maintenance (REAIM) framework from the Dissemination and Implementation Science field. This study presents preliminary data on the domains of Reach and Adoption over the first 5 months of the FCU. Results: The number of families referred who attended at least one session (Reach) was 77.2%. Current data shows that use of the PSC-17 screening instrument (Adoption) is 91.4% for well child checks and 25% for acute visits. Adoption of those referred to the FCU is 84%, indicating most families screening positively for behavioral concerns were successfully referred to the FCU. Conclusion: Initial results suggest Reach and Adoption rates support the feasibility of adapting a behavioral intervention for delivery in the pediatric clinic. Notably, having an existing integrated care delivery model is a critical piece to this early success. Future directions will continue to explore feasibility of the remaining REAIM domains