Merlin Phosphorylation by p21-activated Kinase 2 and Effects of Phosphorylation on Merlin Localization

The Nf2 tumor suppressor gene product merlin is related to the membrane-cytoskeleton linker proteins of the band 4.1 superfamily, including ezrin, radixin, and moesin (ERMs). Merlin is regulated by phosphorylation in a Rac/cdc42-dependent fashion. We report that the phosphorylation of merlin at serine 518 is induced by the p21-activated kinase PAK2. This is demonstrated by biochemical fractionation, use of active and dominant-negative mutants of PAK2, and immunodepletion. By using wild-type and mutated forms of merlin and phospho-directed antibodies, we show that phosphorylation of merlin at serine 518 leads to dramatic protein relocalization. Neurofibromatosis type 2 (NF2)1 is an inherited disorder characterized by the development of Schwann cell tumors of the eighth cranial nerve. Mutations and loss of heterozygosity of theNF2 gene have been detected in NF2 patients and in various sporadic tumors, including schwannomas, meningiomas, and ependymomas (1). In further support of a role for NF2 in tumor suppression, mice heterozygous for an Nf2 mutation are predisposed to a wide variety of tumors with high metastatic potential (2). In a separate model in which Nf2 is inactivated specifically in Schwann cells, mice develop schwannomas and Schwann cell hyperplasia (3). The longest and predominant splice form of the Nf2gene codes for a 595-amino acid protein highly similar to the band 4.1 family of proteins. It is most closely related to the ERM proteins,moesin, ezrin, and radixin. The ERM proteins are thought to function as cell membrane-cytoskeleton linkers and are localized to cortical actin structures near the plasma membrane such as microvilli, membrane ruffles, and lamellipodia (4, 5). Likewise, merlin is localized to cortical actin structures, in patterns that partially overlap with the ERMs (1). It has been proposed that intramolecular binding of the N-terminal and C-terminal domains conformationally regulates the ERM proteins by masking binding sites for interacting proteins. The ERMs can also form homodimers and heterodimers, among themselves and with merlin, adding an additional level of complexity to the regulation of these proteins (6). The recently solved crystal structure of the moesin N/C-terminal complex strengthens this model of conformational regulation (7). Given the sequence and, most likely, structural similarities of merlin to the ERM proteins, it is possible that merlin itself could be regulated in a similar fashion. Recent studies (8, 9) have implicated additional factors in the regulation of the ERMs, including phospholipids and phosphorylation. Previous work from our group and others (10, 11) has shown that merlin is differentially phosphorylated as well and that merlin protein levels are affected by growth conditions such as cell confluency, loss of adhesion, or serum deprivation. Merlin is found in an hypophosphorylated form when the combination of cellular and environmental conditions are growth-inhibitory (10). ERMs can be phosphorylated by Rho kinase, and this phosphorylation can affect intramolecular association and cellular localization. Phosphorylation and/or phospholipids may promote the transition of the proteins to an active form by “opening” intra- and intermolecular associations. These active monomers can then bind to other interacting proteins and the actin cytoskeleton and induce actin-rich membrane projections (5,8, 12, 13). The induction of merlin phosphorylation by activated alleles of the Rho family GTPases has also been examined. Interestingly, although activated Rho did not induce noticeable phosphorylation of merlin, activated forms of Rac and cdc42 did. The site of Rac-induced phosphorylation was determined to be a serine at position 518; mutation of serine 518 results in reduced basal phosphorylation and eliminated Rac-induced phosphorylation (11). Although Rac and cdc42 are implicated in the regulation of many pathways, they are most associated with regulation of cytoskeleton reorganization and gene expression (for recent reviews see Refs.14-16). In light of the data demonstrating that activated Rac/cdc42 leads to phosphorylation and possible inactivation of merlin, the elucidation of the responsible effector pathways and their effects on merlin function are of major importance. Understanding this regulation of merlin could lead to a more complete appreciation of the effects of merlin loss in tumors

The Effects of Participation in Marching Band on Physical Activity and Physical Fitness in College Aged Men and Women

Purpose: Regular physical activity is essential in preventing many chronic diseases and conditions including heart disease, obesity, type two diabetes and some forms of cancer. The majority of the population does not meet recommended guidelines for daily physical activity. Barriers to participation in regular physical activity include boredom and a lack of enjoyment for traditional exercises such as running. Novel forms of exercise that improve cardiorespiratory fitness and increase the number people meeting recommended guidelines has the potential to promote good health and prevent chronic disease. Therefore, the purpose of this study was to measure the effects of participating in marching band on cardiorespiratory fitness and body composition and to quantify the amount of physical activity accomplished during a typical marching band practice session. A secondary purpose was to compare these variables between different sections of the band (woodwinds/brass and drumline). Methods: Twenty-one members (14 males, 7 females) of the University of Rhode Island Marching Band (age 20.2 ± 2.97 yr) were recruited for this study. Body composition and VO2max were measured before and after participation in the marching band season. Each subject wore a pedometer and a Suunto heart rate monitor belt during three to five practices to measure the number of steps taken and to determine the amount of time spent participating light-, moderate- and vigorous-intensity training during routine practices. Results: There was a significant change from pre- to post-season in VO2max (38.5 ± 9.23 vs 40.8 ± 8.5 ml∙kg-1∙min-1; p\u3c0.05) but not in body composition (24.8 ± 12.1 % vs 25.8 ± 10.1%). The average number of steps taken during practices was 2930.1 ± 1075.8 steps. During practice, subjects were engaged in moderate-intensity physical activity for 12.4 ± 6.4 min and vigorous-intensity for 6.37 ± 6.8 min. In total, subjects were active 29.7 ± 14.9 min (light- plus moderate- plus vigorous-intensity). Although the drumline took more steps during practice compared to the woodwinds/brass (3485.1 ± 766.7 vs 2513.9 ± 1111.6 steps, p\u3c0.05), the woodwinds/brass section had a greater change (p\u3c0.05) in VO2max (3.6 ± 2.4 ml∙kg-1∙min-1) compared to the drumline (0.39 ± 2.6 ml∙kg-1∙min-1). Conclusions: Participation in marching band resulted in a significant improvement in the cardiorespiratory fitness of the marching band members. Although marching band practice did not meet national exercise guidelines, the amount of time spent in moderate- and vigorous- intensity exercise and/or the number of steps taken during practice contributed to the overall volume of daily physical activity

Locating Place and Landscape in Early Insular Literature

No abstract

Shrouded small wind turbines

The goal of this project is to improve the energy production of small wind turbines (rated less than 100 kW) by increasing wind velocity at the turbine blades through the design of a shroud attachment. The design process involves the analysis of various computer aided design (CAD) nozzle/diffuser shroud geometries. Computational fluid dynamic (CFD) modeling is used to analyze the effect of shroud features on velocity and pressure fields. A 3D printed scale model is tested in a wind tunnel with strain gauges and pressure transducers to validate the CFD data. The resulting design locally increases velocity by a factor of 1.47, and subsequent energy yield by a factor of 3.18 when compared to the performance of an unshrouded turbine. Additionally, the CFD modeling of the shroud was validated through pressure measurements along the shroud

Modified Mediterranean Diet for Enrichment of Short Chain Fatty Acids: Potential Adjunctive Therapeutic to Target Immune and Metabolic Dysfunction in Schizophrenia?

Growing interest in gut and digestive processes and their potential link to brain and peripheral based inflammation or biobehavioral phenotypes has led to an increasing number of basic and translational scientific reports focused on the role of gut microbiota within the context of neuropsychiatric disorders. However, the effect of dietary modification on specific gut metabolites, in association with immune, metabolic, and psychopathological functioning in schizophrenia spectrum disorders has not been well characterized. The short chain fatty acids (SCFA) acetate, butyrate, and propionate, major metabolites derived from fermentation of dietary fibers by gut microbes, interact with multiple immune and metabolic pathways. The specific pathways that SCFA are thought to target, are dysregulated in cardiovascular disease, type II diabetes, and systemic inflammation. Most notably, these disorders are consistently linked to an attenuated lifespan in schizophrenia. Although, unhealthy dietary intake patterns and increased prevalence of immune and metabolic dysfunction has been observed in people with schizophrenia; dietary interventions have not been well utilized to target immune or metabolic illness. Prior schizophrenia patient trials primarily focused on the effects of gluten free diets. Findings from these studies indicate that a diet avoiding gluten benefits a limited subset of patients, individuals with celiac disease or non-celiac gluten sensitivity. Therefore, alternative dietary and nutritional modifications such as high-fiber, Mediterranean style, diets that enrich the production of SCFA, while being associated with a minimal likelihood of adverse events, may improve immune and cardiovascular outcomes linked to premature mortality in schizophrenia. With a growing literature demonstrating that SCFA can cross the blood brain barrier and target key inflammatory and metabolic pathways, this article highlights enriching dietary intake for SCFA as a potential adjunctive therapy for people with schizophrenia

Antioxidant Effectiveness of Natural Rosemary Extract in Pork Sausage

A natural rosemary extract, FORTIUMTM R20, provides a significant means of protecting color and freshness for pork sausage products.The rosemary extract, used at 2500 ppm, was as effective as commercial antioxidants (BHA/BHT) in fresh pork sausage and was superior to the commercial antioxidants in frozen pork sausage

Connectivity-enhanced diffusion analysis reveals white matter density disruptions in first episode and chronic schizophrenia.

Reduced fractional anisotropy (FA) is a well-established correlate of schizophrenia, but it remains unclear whether these tensor-based differences are the result of axon damage and/or organizational changes and whether the changes are progressive in the adult course of illness. Diffusion MRI data were collected in 81 schizophrenia patients (54 first episode and 27 chronic) and 64 controls. Analysis of FA was combined with "fixel-based" analysis, the latter of which leverages connectivity and crossing-fiber information to assess both fiber bundle density and organizational complexity (i.e., presence and magnitude of off-axis diffusion signal). Compared with controls, patients with schizophrenia displayed clusters of significantly lower FA in the bilateral frontal lobes, right dorsal centrum semiovale, and the left anterior limb of the internal capsule. All FA-based group differences overlapped substantially with regions containing complex fiber architecture. FA within these clusters was positively correlated with principal axis fiber density, but inversely correlated with both secondary/tertiary axis fiber density and voxel-wise fiber complexity. Crossing fiber complexity had the strongest (inverse) association with FA (r = -0.82). When crossing fiber structure was modeled in the MRtrix fixel-based analysis pipeline, patients exhibited significantly lower fiber density compared to controls in the dorsal and posterior corpus callosum (central, postcentral, and forceps major). Findings of lower FA in patients with schizophrenia likely reflect two inversely related signals: reduced density of principal axis fiber tracts and increased off-axis diffusion sources. Whereas the former confirms at least some regions where myelin and or/axon count are lower in schizophrenia, the latter indicates that the FA signal from principal axis fiber coherence is broadly contaminated by macrostructural complexity, and therefore does not necessarily reflect microstructural group differences. These results underline the need to move beyond tensor-based models in favor of acquisition and analysis techniques that can help disambiguate different sources of white matter disruptions associated with schizophrenia

KINEMATIC AND KINETIC COMPARISON BETWEEN PRE-PROFESSIONAL DOMINICAN REPUBLIC AND AMERICAN BASEBALL PITCHERS

The purpose of this study was to compare elbow valgus torque and shoulder distraction force in pre-professional American and Dominican Republic (DR) pitchers. Kinematics that are known to influence elbow valgus torque and shoulder distraction force were also compared. Three dimensional biomechanical analyses were performed on Dominican Republic (n = 37) and American (n = 37) baseball pitchers. Potential difference between Dominican Republic and American pitchers were assessed through analysis of covariance with 95% confidence intervals. Age, hand dominance, and pitch velocity are known to influence elbow torque and shoulder force, therefore these confounding variables were controlled for within the analyses. Pre-professional Dominican Republic pitchers were found to throw fastballs with slower ball velocity but experienced increased elbow valgus torque compared to their American counterparts. Increased elbow valgus torque and inefficient pitching mechanics among Dominican Republic pitchers should be considered when developing training programs and pitching plans for professional pitchers from the Dominican Republic

Applying a Team Performance Framework to better Understand the Handoff Process: Part 2

In healthcare, handoffs are crucial when it comes to transitioning care of patients from one provider to another. Handoffs are communication events between providers that include the background and treatment plan, as well as all pertinent information involving the patient (Paterson, 2010). There are many factors that influence the reliability and transfer of the information being passed during a handoff. In order to understand the way these factors influence and change the handoff process, they were organized according to the input - mediator - output – input (IMOI) model. The IMOI model claims that the productivity and effectiveness of interactions between team members can be influenced by cognitive, affective, and external factors (Weaver, Feitosa, Salas, Seddon, & Vozenilek, 2013). The IMOI also clarifies that the output affects the performance of the team in the future through a feedback loop. This paper focuses specifically on the Output – Input stages, as well as future implications that come from understanding handoffs within the IMOI model