Adducin is an in vivo substrate for protein kinase C: phosphorylation in the MARCKS-related domain inhibits activity in promoting spectrin-actin complexes and occurs in many cells, including dendritic spines of neurons

Adducin is a heteromeric protein with subunits containing a COOH-terminal myristoylated alanine-rich C kinase substrate (MARCKS)-related domain that caps and preferentially recruits spectrin to the fast-growing ends of actin filaments. The basic MARCKS-related domain, present in alpha, beta, and gamma adducin subunits, binds calmodulin and contains the major phosphorylation site for protein kinase C (PKC). This report presents the first evidence that phosphorylation of the MARCKS-related domain modifies in vitro and in vivo activities of adducin involving actin and spectrin, and we demonstrate that adducin is a prominent in vivo substrate for PKC or other phorbol 12-myristate 13-acetate (PMA)-activated kinases in multiple cell types, including neurons. PKC phosphorylation of native and recombinant adducin inhibited actin capping measured using pyrene-actin polymerization and abolished activity of adducin in recruiting spectrin to ends and sides of actin filaments. A polyclonal antibody specific to the phosphorylated state of the RTPS-serine, which is the major PKC phosphorylation site in the MARCKS-related domain, was used to evaluate phosphorylation of adducin in cells. Reactivity with phosphoadducin antibody in immunoblots increased twofold in rat hippocampal slices, eight- to ninefold in human embryonal kidney (HEK 293) cells, threefold in MDCK cells, and greater than 10-fold in human erythrocytes after treatments with PMA, but not with forskolin. Thus, the RTPS-serine of adducin is an in vivo phosphorylation site for PKC or other PMA-activated kinases but not for cAMP-dependent protein kinase in a variety of cell types. Physiological consequences of the two PKC phosphorylation sites in the MARCKS-related domain were investigated by stably transfecting MDCK cells with either wild-type or PKC-unphosphorylatable S716A/S726A mutant alpha adducin. The mutant alpha adducin was no longer concentrated at the cell membrane at sites of cell-cell contact, and instead it was distributed as a cytoplasmic punctate pattern. Moreover, the cells expressing the mutant alpha adducin exhibited increased levels of cytoplasmic spectrin, which was colocalized with the mutant alpha adducin in a punctate pattern. Immunofluorescence with the phosphoadducin-specific antibody revealed the RTPS-serine phosphorylation of adducin in postsynaptic areas in the developing rat hippocampus. High levels of the phosphoadducin were detected in the dendritic spines of cultured hippocampal neurons. Spectrin also was a component of dendritic spines, although at distinct sites from the ones containing phosphoadducin. These data demonstrate that adducin is a significant in vivo substrate for PKC or other PMA-activated kinases in a variety of cells, and that phosphorylation of adducin occurs in dendritic spines that are believed to respond to external signals by changes in morphology and reorganization of cytoskeletal structures.     

Induced magnetic fields as evidence for subsurface oceans in Europa and Callisto

The Galileo spacecraft has been orbiting Jupiter since 7 December 1995, and encounters one of the four galilean satellites-Io, Europa, Ganymede and Callisto-on each orbit. Initial results from the spacecraft's magnetometer have indicated that neither Europa nor Callisto have an appreciable internal magnetic field, in contrast to Ganymede and possibly Io. Here we report perturbations of the external magnetic fields (associated with Jupiter's inner magnetosphere) in the vicinity of both Europa and Callisto. We interpret these perturbations as arising from induced magnetic fields, generated by the moons in response to the periodically varying plasma environment. Electromagnetic induction requires eddy currents to flow within the moons, and our calculations show that the most probable explanation is that there are layers of significant electrical conductivity just beneath the surfaces of both moons. We argue that these conducting layers may best be explained by the presence of salty liquid-water oceans, for which there is already indirect geological evidence in the case of Europa.     

Prevalence of voice problems in teachers

Teachers are frequently cited as experiencing a high rate of vocal dysfunction (1-7). Despite considerable research in the area of voice problems in teachers, the prevalence of voice disorders in this group is unknown. This study investigated the prevalence of self-reported voice problems in teachers using a mail survey of a simple random sample of 1168 state school teachers (preschool-Grade 12) in South Australia. As part of the survey, teachers were asked to report voice problems for the day of the survey, during the current teaching year, and during their careers. The response rate was 75%, with 16% of teachers reporting voice problems on the day of the survey, 20% reporting problems during the current teaching year, and 19% reporting problems at some time during their career. Females were twice as likely as males to report voice problems. These findings clearly indicate a need for further investigation of the causes of vocal dysfunction in teachers and for the development of educational programs aimed at preventing voice problems in this group of professional voice users.     

Analyzing subjectivity in therapeutic discourse: Rogers, Perls, Ellis, and Gloria revisited.

Analyzed the film Three Approaches to Psychotherapy (E. L. Shostrom, 1966), which depicts C. Rogers (client-centered therapy), A. Ellis (rational-emotive therapy), and F. S. Perls (Gestalt therapy) interviewing the same client, Gloria. Verbal exchanges between participants were analyzed, using T. S. Essig and R. L. Russell's (1988) categories for analyzing psychological relations between subjects and events in discourse. Predictable patterns in how therapists of different theoretical orientations represent subjectivity in discourse can be screened with these categories. Classification categories are appended. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Maximizing learning without sacrificing the fun: Stealth assessment,adaptivity and learning supports in educational games

In this study, we investigated the validity of a stealth assessment of physics understanding in an educational game, as well as the effectiveness of different game-level delivery methods and various in-game supports on learning. Using a game called Physics Playground, we randomly assigned 263 ninth- to eleventh-grade students into four groups: adaptive, linear, free choice and no-treatment control. Each condition had access to the same in-game learning supports during gameplay. Results showed that: (a) the stealth assessment estimates of physics understanding were valid—significantly correlating with the external physics test scores; (b) there was no significant effect of game-level delivery method on students' learning; and (c) physics animations were the most effective (among eight supports tested) in predicting both learning outcome and in-game performance (e.g. number of game levels solved). We included student enjoyment, gender and ethnicity in our analyses as moderators to further investigate the research questions.     

Infants' sensitivity to optical flow for controlling posture.

Infants' sensitivity to optical flow for controlling sitting and standing was tested using a "moving room" in which all of the walls moved together, or only the side walls or front wall moved. Two questions motivated this research. (a) is the optical flow necessary for inducing postural compensations spatially distributed in the optic array? (b) Do visually induced compensations follow a developmental progression, or alternatively, emerge all at once? Experiment 1 was designed to test postural compensations by 14-month-old infants capable of standing in the moving room. Experiment 2 was designed to test postural compensations by 5- to 9-month-old infants who were passively supported while sitting in the moving room. The results revealed that partial flow is generally sufficient for inducing postural compensations, but that the amplitude and consistency of the response depend on the location of the flow in the optic array. In addition, there was evidence suggesting that compensatory responses become increasingly systematic during the second half of the first year. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Rapid software integration using structured methods

In this paper we discuss an approach to organizing the integration of 110,000 lines of C, SQL, Assembler, and microcode distributed over a network of 36 processors of four types so that it could be accomplished in six months. The software runs on a test system architecture consisting of a LAN-based workstation group and a set of VME-based embedded processors. By using structured methodology, parallelism in the integration process was achieved. The necessary stub tools were identified and developed before integration began. The principles followed and experiences of integrating the system are discussed.     

Regulation of lactate production in Streptococcus bovis: A spiraling effect that contributes to rumen acidosis

When Streptococcus bovis was grown in batch culture with 6 g/L glucose at pH 6.7, maximum specific growth rate was 1.47 h(-1), and lactate was the primary fermentation product. In continuous culture at pH 6.7 and growth rate equal to .10 h(-1), little lactate was formed, and formate, acetate, and ethanol accounted for most of the product. When extra-cellular pH decreased to 4.7, intra-cellular pH declined to 5.4, and organisms switched back to lactate production. Intracellular concentration of fructose 1,6-diphosphate of batch culture cells was greater than 12 mM, a concentration that allowed maximal lactate dehydrogenase activity. When Streptococcus bovis was grown in continuous culture at pH 6.7, intracellular fructose-l,6-diphosphate declined to .4 mM, a concentration which gave little lactate dehydrogenase activity at pH 6.5 or greater. Decreasing pH of continuous culture to 4.7 increased intracellular fructose-1,6-diphosphate concentration to .8 mM. This concentration was still limiting if lactate dehydrogenase was assayed at pH 6.5, but nearly maximal activity was obtained when enzyme was assayed at pH 5.5. The small increase in fructose-l,6-diphosphate and decreased requirement of lactate dehydrogenase for fructose-l,6-diphosphate under acidic assay conditions, accounted for increased lactate production during low pH (4.7) continuous culture. These and other aspects of lactate regulation by Streptococcus bovis are discussed as factors leading to rumen acidosis. This pattern of regulation also helps to explain why rumen acidosis is difficult to reverse.