Integration of linear and dendritic actin nucleation in Nck-induced actin comets

The Nck adaptor protein recruits cytosolic effectors such as N-WASP that induce localized actin polymerization. Experimental aggregation of Nck SH3 domains at the membrane induces actin comet tails—dynamic, elongated filamentous actin structures similar to those that drive the movement of microbial pathogens such as vaccinia virus. Here we show that experimental manipulation of the balance between unbranched/branched nucleation altered the morphology and dynamics of Nck-induced actin comets. Inhibition of linear, formin-based nucleation with the small-molecule inhibitor SMIFH2 or overexpression of the formin FH1 domain resulted in formation of predominantly circular-shaped actin structures with low mobility (actin blobs). These results indicate that formin-based linear actin polymerization is critical for the formation and maintenance of Nck-dependent actin comet tails. Consistent with this, aggregation of an exclusively branched nucleation-promoting factor (the VCA domain of N-WASP), with density and turnover similar to those of N-WASP in Nck comets, did not reconstitute dynamic, elongated actin comets. Furthermore, enhancement of branched Arp2/3-mediated nucleation by N-WASP overexpression caused loss of the typical actin comet tail shape induced by Nck aggregation. Thus the ratio of linear to dendritic nucleation activity may serve to distinguish the properties of actin structures induced by various viral and bacterial pathogens.     

Evaluation of Novel EMG Biofeedback for Postural Correction During Computer Use

Postural correction is an effective rehabilitation technique used to treat chronic neck and shoulder pain, and is aimed toward reducing the load on the surrounding muscles by adopting a neutral posture. The objective of this investigation was to evaluate the effectiveness of real-time high-density surface EMG (HDsEMG) biofeedback for postural correction during typing. Twenty healthy participants performed a typing task with two forms of postural feedback: (1) verbal postural coaching and (2) verbal postural coaching plus HDsEMG biofeedback. The interface used activity from two HDsEMG arrays placed over the trapezius designed to shift trapezius muscle activity inferiorly. The center of gravity across both arrays was used to quantify the spatial distribution of trapezius activity. Planar angles taken from upper extremity reflective markers quantified cervicoscapular posture. During the biofeedback condition, trapezius muscle activity was located 12.74 ± 3.73 mm more inferior, the scapula was 2.58 ± 1.18° more adducted and 0.23 ± 0.24° more depressed in comparison to verbal postural coaching alone. The results demonstrate the short-term effectiveness of a real-time HDsEMG biofeedback intervention to achieve postural correction, and may be more effective at creating an inferior shift in trapezius muscle activity in comparison to verbal postural coaching alone.     

Quantifying Microbial Utilization of Petroleum Hydrocarbons in Salt Marsh Sediments by Using the 13C Content of Bacterial rRNA

Natural remediation of oil spills is catalyzed by complex microbial consortia. Here we took a whole-community approach to investigate bacterial incorporation of petroleum hydrocarbons from a simulated oil spill. We utilized the natural difference in carbon isotopic abundance between a salt marsh ecosystem supported by the ¹³C-enriched C₄ grass Spartina alterniflora and ¹³C-depleted petroleum to monitor changes in the ¹³C content of biomass. Magnetic bead capture methods for selective recovery of bacterial RNA were used to monitor the ¹³C content of bacterial biomass during a 2-week experiment. The data show that by the end of the experiment, up to 26% of bacterial biomass was derived from consumption of the freshly spilled oil. The results contrast with the inertness of a nearby relict spill, which occurred in 1969 in West Falmouth, MA. Sequences of 16S rRNA genes from our experimental samples also were consistent with previous reports suggesting the importance of Gamma- and Deltaproteobacteria and Firmicutes in the remineralization of hydrocarbons. The magnetic bead capture approach makes it possible to quantify uptake of petroleum hydrocarbons by microbes in situ. Although employed here at the domain level, RNA capture procedures can be highly specific. The same strategy could be used with genus-level specificity, something which is not currently possible using the ¹³C content of biomarker lipids.     

The Nature and Extent of Mutational Pleiotropy in Gene Expression of Male Drosophila serrata

The nature and extent of mutational pleiotropy remain largely unknown, despite the central role that pleiotropy plays in many areas of biology, including human disease, agricultural production, and evolution. Here, we investigate the variation in 11,604 gene expression traits among 41 mutation accumulation (MA) lines of Drosophila serrata. We first confirmed that these expression phenotypes were heritable, detecting genetic variation in 96% of them in an outbred, natural population of D. serrata. Among the MA lines, 3385 (29%) of expression traits were variable, with a mean mutational heritability of 0.0005. In most traits, variation was generated by mutations of relatively small phenotypic effect; putative mutations with effects of greater than one phenotypic standard deviation were observed for only 8% of traits. With most (71%) traits unaffected by any mutation, our data provide no support for universal pleiotropy. We further characterized mutational pleiotropy in the 3385 variable traits, using sets of 5, randomly assigned, traits. Covariance among traits chosen at random with respect to their biological function is expected only if pleiotropy is extensive. Taking an analytical approach in which the variance unique to each trait in the random 5-trait sets was partitioned from variance shared among traits, we detected significant (at 5% false discovery rate) mutational covariance in 21% of sets. This frequency of statistically supported covariance implied that at least some mutations must pleiotropically affect a substantial number of traits (>70; 0.6% of all measured traits).