Elucidation of Binding Determinants and Functional Consequences of Ras/Raf-Cysteine-rich Domain Interactions

Raf-1 is a critical downstream target of Ras and contains two distinct domains that bind Ras. The first Ras-binding site (RBS1) in Raf-1 has been shown to be essential for Ras-mediated translocation of Raf-1 to the plasma membrane, whereas the second site, in the Raf-1 cysteine-rich domain (Raf-CRD), has been implicated in regulating Raf kinase activity. While recognition elements that promote Ras.RBS1 complex formation have been characterized, relatively little is known about Ras/Raf-CRD interactions. In this study, we have characterized interactions important for Ras binding to the Raf-CRD. Reconciling conflicting reports, we found that these interactions are essentially independent of the guanine nucleotide bound state, but instead, are enhanced by post-translational modification of Ras. Specifically, our findings indicate that Ras farnesylation is sufficient for stable association of Ras with the Raf-CRD. Furthermore, we have also identified a Raf-CRD variant that is impaired specifically in its interactions with Ras. NMR data also suggests that residues proximal to this mutation site on the Raf-CRD form contacts with Ras. This Raf-CRD mutant impairs the ability of Ras to activate Raf kinase, thereby providing additional support that Ras interactions with the Raf-CRD are important for Ras-mediated activation of Raf-1

Interplanetary CubeSats: Opening the Solar System to a Broad Community at Lower Cost

Interplanetary CubeSats could enable small, low-cost missions beyond low Earth orbit. This class is defined by mass < ~ 10 kg, cost < $30 M, and durations up to 5 years. Over the coming decade, a stretch of each of six distinct technology areas, creating one overarching architecture, could enable comparatively low-cost Solar System exploration missions with capabilities far beyond those demonstrated in small satellites to date. The six technology areas are: (1) CubeSat electronics and subsystems extended to operate in the interplanetary environment, especially radiation and duration of operation; (2) Optical telecommunications to enable very small, low-power uplink/downlink over interplanetary distances; (3) Solar sail propulsion to enable high !V maneuvering using no propellant; (4) Navigation of the Interplanetary Superhighway to enable multiple destinations over reasonable mission durations using achievable !V; (5) Small, highly capable instrumentation enabling acquisition of high-quality scientific and exploration information; and (6) Onboard storage and processing of raw instrument data and navigation information to enable maximum utility of uplink and downlink telecom capacity, and minimal operations staffing. The NASA Innovative Advanced Concepts (NIAC) program in 2011 selected Interplanetary CubeSats for further investigation, some results of which are reported here for Phase 1

Extracellular Signal-regulated Kinase (ERK) Phosphorylates Histone Deacetylase 6 (HDAC6) at Serine 1035 to Stimulate Cell Migration

Histone deacetylase 6 (HDAC6) is well known for its ability to promote cell migration through deacetylation of its cytoplasmic substrates such as α-tubulin. However, how HDAC6 itself is regulated to control cell motility remains elusive. Previous studies have shown that one third of extracellular signal-regulated kinase (ERK) is associated with the microtubule cytoskeleton in cells. Yet, no connection between HDAC6 and ERK has been discovered. Here, for the first time, we reveal that ERK binds to and phosphorylates HDAC6 to promote cell migration via deacetylation of α-tubulin. We have identified two novel ERK-mediated phosphorylation sites: threonine 1031 and serine 1035 in HDAC6. Both sites were phosphorylated by ERK1 in vitro, whereas Ser-1035 was phosphorylated in response to the activation of EGFR-Ras-Raf-MEK-ERK signaling pathway in vivo. HDAC6-null mouse embryonic fibroblasts rescued by the nonphosphorylation mimicking mutant displayed significantly reduced cell migration compared with those rescued by the wild type. Consistently, the nonphosphorylation mimicking mutant exerted lower tubulin deacetylase activity in vivo compared with the wild type. These data indicate that ERK/HDAC6-mediated cell motility is through deacetylation of α-tubulin. Overall, our results suggest that HDAC6-mediated cell migration could be governed by EGFR-Ras-Raf-MEK-ERK signaling

Osteoarthritis-Like Changes in Bardet–Biedl Syndrome Mutant Ciliopathy Mice (Bbs1M390R/M390R): Evidence for a Role of Primary Cilia in Cartilage Homeostasis and Regulation of Inflammation

Osteoarthritis (OA) is a debilitating inflammation related disease characterized by joint pain and effusion, loss of mobility, and deformity that may result in functional joint failure and significant impact on quality of life. Once thought of as a simple “wear and tear” disease, it is now widely recognized that OA has a considerable metabolic component and is related to chronic inflammation. Defects associated with primary cilia have been shown to be cause OA-like changes in Bardet–Biedl mice. We examined the role of dysfunctional primary cilia in OA in mice through the regulation of the previously identified degradative and pro-inflammatory molecular pathways common to OA. We observed an increase in the presence of pro-inflammatory markers TGFβ-1 and HTRA1 as well as cartilage destructive protease MMP-13 but a decrease in DDR-2. We observed a morphological difference in cartilage thickness in Bbs1M390R/M390R mice compared to wild type (WT). We did not observe any difference in OARSI or Mankin scores between WT and Bbs1M390R/M390R mice. Primary cilia appear to be involved in the upregulation of biomarkers, including pro-inflammatory markers common to OA

The bii4africa dataset of faunal and floral population intactness estimates across Africa’s major land uses

Sub-Saharan Africa is under-represented in global biodiversity datasets, particularly regarding the impact of land use on species’ population abundances. Drawing on recent advances in expert elicitation to ensure data consistency, 200 experts were convened using a modified-Delphi process to estimate ‘intactness scores’: the remaining proportion of an ‘intact’ reference population of a species group in a particular land use, on a scale from 0 (no remaining individuals) to 1 (same abundance as the reference) and, in rare cases, to 2 (populations that thrive in human-modified landscapes). The resulting bii4africa dataset contains intactness scores representing terrestrial vertebrates (tetrapods: ±5,400 amphibians, reptiles, birds, mammals) and vascular plants (±45,000 forbs, graminoids, trees, shrubs) in sub-Saharan Africa across the region’s major land uses (urban, cropland, rangeland, plantation, protected, etc.) and intensities (e.g., large-scale vs smallholder cropland). This dataset was co-produced as part of the Biodiversity Intactness Index for Africa Project. Additional uses include assessing ecosystem condition; rectifying geographic/ taxonomic biases in global biodiversity indicators and maps; and informing the Red List of Ecosystems

Reducing Inpatient Falls and Fall Related Injuries in Acute Care Settings

Falls are a continually rising issue in today\u27s healthcare. In acute care settings, patient falls make up 38% of all adverse events in which include physical injury, undesirable emotional and financial outcomes for the client (Angn, Mordiffi, Wong, Devi, & Evans, 2007). Falls in the hospitals lead to fear, pain, decreased healing, longer in-patient stays, further health-related complications. Falls may also cause patient discomfort and affect quality of life. Prevention of falls is an important goal of hospitals world-wide. Research has been conducted to determine the clinical effectiveness and implementation of a fall prevention. Although falls in hospitals cannot always be prevented, using the most accurate measures to decrease the incidence of falls is necessary

Surrogate assessment of coronary artery disease patients' functional capacity

An investigation of the surrogate assessment of coronary artery disease (CAD) patients' functional capacity was conducted using 193 patient and surrogate rater dyads. Mean age of patients and surrogate raters were 60.4 and 54.4 years, respectively. Patients and surrogates independently completed a brief questionnaire that assessed health and psychosocial factors. The Duke Activity Status Index (DASI) was contained in the patients' questionnaire, while a similar form modified to assess patients' functional capacity was imbedded in the surrogates' questionnaire. Results indicated similar psychometric characteristics and clinical validity for patients' self-report and surrogates' ratings, suggesting that the Surrogate Rating Form of the Duke Activity Status Index (DASI-SRF) is a reliable and valid proxy method of assessing patient's functional capacity when this information may not be obtained directly from the patient. Further, while there were no effects of surrogates' health and psychological characteristics on their ratings of patients' functional capacity, in comparison with other surrogates, spouses were more likely to rate patients higher in functional capacity. Exploration of the patient/care provider relationship via concurrent use of the DASI and DASI-SRF is discussed.functional capacity coronary artery disease surrogate assessment