E-cadherin engagement stimulates proliferation via Rac1

E-cadherin has been linked to the suppression of tumor growth and the inhibition of cell proliferation in culture. We observed that progressively decreasing the seeding density of normal rat kidney-52E (NRK- 52E) or MCF-10A epithelial cells from confluence, indeed, released cells from growth arrest. Unexpectedly, a further decrease in seeding density so that cells were isolated from neighboring cells decreased proliferation. Experiments using microengineered substrates showed that E-cadherin engagement stimulated the peak in proliferation at intermediate seeding densities, and that the proliferation arrest at high densities did not involve E-cadherin, but rather resulted from a crowding-dependent decrease in cell spreading against the underlying substrate. Rac1 activity, which was induced by E-cadherin engagement specifically at intermediate seeding densities, was required for the cadherin-stimulated proliferation, and the control of Rac1 activation by E-cadherin was mediated by p120- catenin. Together, these findings demonstrate a stimulatory role for E-cadherin in proliferative regulation, and identify a simple mechanism by which cell–cell contact may trigger or inhibit epithelial cell proliferation in different settings

Attitude Control System Design for the Solar Dynamics Observatory

The Solar Dynamics Observatory mission, part of the Living With a Star program, will place a geosynchronous satellite in orbit to observe the Sun and relay data to a dedicated ground station at all times. SDO remains Sun- pointing throughout most of its mission for the instruments to take measurements of the Sun. The SDO attitude control system is a single-fault tolerant design. Its fully redundant attitude sensor complement includes 16 coarse Sun sensors, a digital Sun sensor, 3 two-axis inertial reference units, 2 star trackers, and 4 guide telescopes. Attitude actuation is performed using 4 reaction wheels and 8 thrusters, and a single main engine nominally provides velocity-change thrust. The attitude control software has five nominal control modes-3 wheel-based modes and 2 thruster-based modes. A wheel-based Safehold running in the attitude control electronics box improves the robustness of the system as a whole. All six modes are designed on the same basic proportional-integral-derivative attitude error structure, with more robust modes setting their integral gains to zero. The paper details the mode designs and their uses

An inhibitory role for FAK in regulating proliferation: a link between limited adhesion and RhoA-ROCK signaling

Focal adhesion kinase (FAK) transduces cell adhesion to the extracellular matrix into proliferative signals. We show that FAK overexpression induced proliferation in endothelial cells, which are normally growth arrested by limited adhesion. Interestingly, displacement of FAK from adhesions by using a FAK−/− cell line or by expressing the C-terminal fragment FRNK also caused an escape of adhesion-regulated growth arrest, suggesting dual positive and negative roles for FAK in growth regulation. Expressing kinase-dead FAK-Y397F in FAK−/− cells prevented uncontrolled growth, demonstrating the antiproliferative function of inactive FAK. Unlike FAK overexpression–induced growth, loss of growth control in FAK−/− or FRNK-expressing cells increased RhoA activity, cytoskeletal tension, and focal adhesion formation. ROCK inhibition rescued adhesion-dependent growth control in these cells, and expression of constitutively active RhoA or ROCK dysregulated growth. These findings demonstrate the ability of FAK to suppress and promote growth, and underscore the importance of multiple mechanisms, even from one molecule, to control cell proliferation

Zwitterionic PEG-PC hydrogels modulate the foreign body response in a modulus-dependent manner

Reducing the foreign body response (FBR) to implanted biomaterials will enhance their performance in tissue engineering. Poly(ethylene glycol) (PEG) hydrogels are increasingly popular for this application due to their low cost, ease of use, and the ability to tune their compliance via molecular weight and crosslinking densities. PEG hydrogels can elicit chronic inflammation in vivo, but recent evidence has suggested that extremely hydrophilic, zwitterionic materials and particles can evade the immune system. To combine the advantages of PEG-based hydrogels with the hydrophilicity of zwitterions, we synthesized hydrogels with co-monomers PEG and the zwitterion phosphorylcholine (PC). Recent evidence suggests that stiff hydrogels elicit increased immune cell adhesion to hydrogels, which we attempted to reduce by increasing hydrogel hydrophilicity. Surprisingly, hydrogels with the highest amount of zwitterionic co-monomer elicited the highest FBR we observed. Lowering the hydrogel modulus (165 kPa to 3 kPa), or PC content (20 wt% to 0 wt%), mitigated this effect. A high density of macrophages was found at the surface of implants associated with a high FBR, and mass spectrometry analysis of the proteins adsorbed to these gels implicated extracellular matrix, immune response, and cell adhesion protein categories as drivers of macrophage recruitment to these hydrogels. Overall, we show that modulus regulates macrophage adhesion to zwitterionic-PEG hydrogels, and demonstrate that chemical modifications to hydrogels should be studied in parallel with their physical properties to optimize implant design

Solar Dynamics Observatory Launch and Commissioning

The Solar Dynamics Observatory (SDO) was launched on February 11, 2010. Over the next three months, the spacecraft was raised from its launch orbit into its final geosynchronous orbit and its systems and instruments were tested and calibrated in preparation for its desired ten year science mission studying the Sun. A great deal of activity during this time involved the spacecraft attitude control system (ACS); testing control modes, calibrating sensors and actuators, and using the ACS to help commission the spacecraft instruments and to control the propulsion system as the spacecraft was maneuvered into its final orbit. This paper will discuss the chronology of the SDO launch and commissioning, showing the ACS analysis work performed to diagnose propellant slosh transient and attitude oscillation anomalies that were seen during commissioning, and to determine how to overcome them. The simulations and tests devised to demonstrate correct operation of all onboard ACS modes and the activities in support of instrument calibration will be discussed and the final maneuver plan performed to bring SDO on station will be shown. In addition to detailing these commissioning and anomaly resolution activities, the unique set of tests performed to characterize SDO's on-orbit jitter performance will be discussed

Hydrogel crosslinking modulates macrophages, fibroblasts, and their communication, during wound healing

Biomaterial wound dressings, such as hydrogels, interact with host cells to regulate tissue repair. This study investigates how crosslinking of gelatin-based hydrogels influences immune and stromal cell behavior and wound healing in female mice. We observe that softer, lightly crosslinked hydrogels promote greater cellular infiltration and result in smaller scars compared to stiffer, heavily crosslinked hydrogels. Using single-cell RNA sequencing, we further show that heavily crosslinked hydrogels increase inflammation and lead to the formation of a distinct macrophage subpopulation exhibiting signs of oxidative activity and cell fusion. Conversely, lightly crosslinked hydrogels are more readily taken up by macrophages and integrated within the tissue. The physical properties differentially affect macrophage and fibroblast interactions, with heavily crosslinked hydrogels promoting pro-fibrotic fibroblast activity that drives macrophage fusion through RANKL signaling. These findings suggest that tuning the physical properties of hydrogels can guide cellular responses and improve healing, offering insights for designing better biomaterials for wound treatment

Solar Dynamics Observatory Guidance, Navigation, and Control System Overview

The Solar Dynamics Observatory (SDO) was designed and built at the Goddard Space Flight Center, launched from Cape Canaveral on February 11, 2010, and reached its final geosynchronous science orbit on March 16, 2010. The purpose of SDO is to observe the Sun and continuously relay data to a dedicated ground station. SDO remains Sun-pointing throughout most of its mission for the instruments to take measurements of the Sun. The SDO attitude control system (ACS) is a single-fault tolerant design. Its fully redundant attitude sensor complement includes sixteen coarse Sun sensors (CSSs), a digital Sun sensor (DSS), three two-axis inertial reference units (IRUs), and two star trackers (STs). The ACS also makes use of the four guide telescopes included as a part of one of the science instruments. Attitude actuation is performed using four reaction wheels assemblies (RWAs) and eight thrusters, with a single main engine used to provide velocity-change thrust for orbit raising. The attitude control software has five nominal control modes, three wheel-based modes and two thruster-based modes. A wheel-based Safehold running in the attitude control electronics box improves the robustness of the system as a whole. All six modes are designed on the same basic proportional-integral-derivative attitude error structure, with more robust modes setting their integral gains to zero. This paper details the final overall design of the SDO guidance, navigation, and control (GN&C) system and how it was used in practice during SDO launch, commissioning, and nominal operations. This overview will include the ACS control modes, attitude determination and sensor calibration, the high gain antenna (HGA) calibration, and jitter mitigation operation. The Solar Dynamics Observatory mission is part of the NASA Living With a Star program, which seeks to understand the changing Sun and its effects on the Solar System, life, and society. To this end, the SDO spacecraft carries three Sun-observing instruments: Helioseismic and Magnetic Imager (HMI), led by Stanford University; Atmospheric Imaging Assembly (AIA), led by Lockheed Martin Space and Astrophysics Laboratory; and Extreme Ultraviolet Variability Experiment (EVE), led by the University of Colorado. The basic mission is to observe the Sun for a very high percentage of the 5-year mission (10-year goal) with long stretches of uninterrupted observations and with constant, high-data-rate transmission to a dedicated ground station to be located in White Sands, New Mexico. These goals guided the design of the spacecraft bus that will carry and service the three-instrument payload. Overarching design goals for the bus are geosynchronous orbit, near-constant Sun observations with the ability to fly through eclipses, and constant HGA contact with the dedicated ground station. A three-axis stabilized ACS is needed both to point at the Sun accurately and to keep the roll about the Sun vector correctly positioned with respect to the solar north pole. This roll control is especially important for the magnetic field imaging of HM I. The mission requirements have several general impacts on the ACS design. Both the AIA and HMI instruments are very sensitive to the blurring caused by jitter. Each has an image stabilization system (ISS) with some ability to filter out high frequency motion, but below the bandwidth of the ISS the control system must compensate for disturbances within the ACS bandwidth or avoid exciting jitter at higher frequencies. Within the ACS bandwidth, the control requirement imposed by AIA is to place the center of the solar disk no more than 2 arc sec, 3 , from a body-defined target based on one of the GTs that accompany the instrument. This body-defined target, called the science reference boresight (SRB), was determined from the postlaunch orientation of the GTs by averaging the bounding telescope boresights for pitch to get a pitch SRB coordinate, and by averaging the bounding boresights for yaw toet the yaw SRB coordinate. The location of this SRB in the 0.5-deg field-of-view for each GT then becomes the central target for each telescope; one GT is selected for use as the ACS controlling guide telescope (CGT) at any given time. Fine Sun-pointing is effected based on this SRB for all three instruments when the Sun is within the linear range of the CGT. In addition to limiting jitter, HMI science requires averaging several observations, making the instrument sensitive to low frequency motion that induces differential motion between each observation. This requires the spacecraft attitude to be stable about the roll axis to approximately 10 arcsec over a ten-minute period. Instrument calibrations require that the spacecraft point the SRB up to 2.5 degrees in pitch and yaw away from the center of the Sun, placing the Sun outside the field-of-view of the guide telescopes. In such instances, when the GTs cannot provide the definitive target for the ACS, on-board attitude determination combined with ephemeris prediction of the Sun direction must provide the definitive target. EVE is capable of observing the Sun with less dependence on attitude control. However, the ground data processing needs for calibrations result in the most strict attitude knowledge requirements for the mission: [35,70,70] arcsec, 3 , of knowledge with respect to the center of the solar disk. In addition to driving the ACS sensor selection, the knowledge requirements, which have their effect primarily during Inertial mode calibrations, drive the accuracy requirements for the solar ephemeris. The need to achieve and maintain geosynchronous orbit (GEO) drove the need for high-efficiency propulsive systems and appropriate attitude control. The main engine provided high specific impulse for the maneuvers to attain GEO, while the smaller ACS thrusters managed the disturbance torques of the larger engine and provided the capability for much smaller adjustment burns on orbit. SDO s large solar profile means that solar radiation pressure is a large torque disturbance, and the momentum buildup from this disturbance and the GEO altitude drives the ACS to use thrusters to manage vehicle momentum. The demanding data capture budget for the mission, however, requires SDO to avoid frequent thruster maneuvers, while concerns about on-orbit jitter restrict the maximum desired wheel speeds desired from the RWAs. The plan for on-orbit wheel speed and momentum management will be discussed as well as what is now being done in operation after the jitter environment was characterized. The SDO ACS hardware complement is single-fault tolerant. Two main processors carry virtually identical copies of the command and data handling and ACS software, and two identical attitude control electronics (ACE) boxes carry Coldfire processors with contingency ACS software and other hardware interface cards; the ACE structure allows reaction wheels to be commanded by the Sun-pointing Safehold independent of the Mil Std 1553 data bus. The sixteen Adcole CSSs are grouped into primary and backup sets of eight sensors, each set providing the ability to calculate a sun vector. Each set of eight eyes provides full 4 -steradian coverage. The Adcole DSS comprises an optics head and a separate electronics box providing a 1553 data interface. The electronics box is mounted inside the Faraday cage created by the spacecraft bus module. The DSS head with its 32- deg square FOV is mounted on the instrument module with its boresight along the spacecraft X axis, nearly aligned with the Sun during observations. Adcole has designed the DSS calibration parameters so that the accuracy is 0.24 arcminutes within 10 deg of the boresight, and diminishes to 3 arcminutes as the Sun moves towards the edges of its FOV . This DSS calibration scheme provides higher accuracy attitude determination over the range of the instrument calibration maneuvers

Adapting Ferritin, a Naturally Occurring Protein Cage, to Modulate Intrinsic Agonism of OX40

Ferritin is a multivalent, self-assembling protein scaffold found in most human cell types, in addition to being present in invertebrates, higher plants, fungi, and bacteria, that offers an attractive alternative to polymer-based drug delivery systems (DDS). In this study, the utility of the ferritin cage as a DDS was demonstrated within the context of T cell agonism for tumor killing. Members of the tumor necrosis factor receptor superfamily (TNFRSF) are attractive targets for the development of anticancer therapeutics. These receptors are endogenously activated by trimeric ligands that occur in transmembrane or soluble forms, and oligomerization and cell-surface anchoring have been shown to be essential aspects of the targeted agonism of this receptor class. Here, we demonstrated that the ferritin cage could be easily tailored for multivalent display of anti-OX40 antibody fragments on its surface and determined that these arrays are capable of pathway activation through cell-surface clustering. Together, these results confirm the utility, versatility, and developability of ferritin as a DDS