Electromyographic instantaneous amplitude and instantaneous mean power frequency patterns across a range of motion during a concentric isokinetic muscle action of the biceps brachii.

The purpose of this study was to examine the electromyographic (EMG) instantaneous amplitude (IA) and instantaneous mean power frequency (IMPF) patterns for the biceps brachii muscle across a range of motion during maximal and submaximal concentric isokinetic muscle actions of the forearm flexors. Ten adults (mean +/- SD age = 22.0 +/- 3.4 years) performed a maximal and a submaximal [20% peak torque (PT)] concentric isokinetic forearm flexion muscle action at a velocity of 30 degrees s(-1). The surface EMG signal was detected from the biceps brachii muscle with a bipolar electrode arrangement, and the EMG IA and IMPF versus time relationships were examined for each subject using first- and second-order polynomial regression models. The results indicated that there were no consistent patterns between subjects for EMG IA or IMPF with increases in torque across the range of motion. Some of the potential nonphysiological factors that could influence the amplitude and/or frequency contents of the surface EMG signal during a dynamic muscle action include movement of the muscle fibers and innervation zone beneath the skin surface, as well as changes in muscle fiber length and the thickness of the tissue layer between the muscle and the recording electrodes. These factors may affect the EMG IA and IMPF patterns differently for each subject, thereby increasing the difficulty of drawing any general conclusions regarding the motor control strategies that increase torque across a range of motion.     

The Gcs1 Arf-GAP mediates Snc1,2 v-SNARE retrieval to the Golgi in yeast

Gcs1 is an Arf GTPase-activating protein (Arf-GAP) that mediates Golgi-ER and post-Golgi vesicle transport in yeast. Here we show that the Snc1,2 v-SNAREs, which mediate endocytosis and exocytosis, interact physically and genetically with Gcs1. Moreover, Gcs1 and the Snc v-SNAREs colocalize to subcellular structures that correspond to the trans-Golgi and endosomal compartments. Studies performed in vitro demonstrate that the Snc-Gcs1 interaction results in the efficient binding of recombinant Arf1Delta17N-Q71L to the v-SNARE and the recruitment of purified coatomer. In contrast, the presence of Snc had no effect on Gcs1 Arf-GAP activity in vitro, suggesting that v-SNARE binding does not attenuate Arf1 function. Disruption of both the SNC and GCS1 genes results in synthetic lethality, whereas overexpression of either SNC gene inhibits the growth of a distinct subset of COPI mutants. We show that GFP-Snc1 recycling to the trans-Golgi is impaired in gcs1Delta cells and these COPI mutants. Together, these results suggest that Gcs1 facilitates the incorporation of the Snc v-SNAREs into COPI recycling vesicles and subsequent endosome-Golgi sorting in yeast.     

Neuromuscular responses to three days of velocity-specific isokinetic training

The purpose of this investigation was to examine the effects of 3 days of velocity-specific isokinetic training on peak torque (PT) and the electromyographic (EMG) signal. Thirty adult women were randomly assigned to a slow-velocity training (SVT), fast-velocity training (FVT), or control (CON) group. All subjects performed maximal, concentric, isokinetic leg extension muscle actions at 30 and 270 degrees .s(-1) for the determination of PT on visits 1 (pretest) and 5 (posttest). Electromyographic signals were recorded from the vastus lateralis, rectus femoris, and vastus medialis muscles during each test. The training groups performed 4 sets of 10 maximal repetitions at 30 degrees .s(-1) (SVT group) or 270 degrees .s(-1) (FVT group) on visits 2, 3, and 4. For the SVT group, PT increased from pretest to posttest at 30 and 270 degrees .s(-1). The increase in PT at 30 degrees .s(-1) was greater than at 270 degrees .s(-1). For the FVT group, PT increased at 270 degrees .s(-1) only. For the CON group, there were no changes in PT at either velocity. There were no pretest to posttest changes in EMG amplitude or mean power frequency (MPF) for any group at any velocity, with the exception of an increase in EMG MPF from the vastus medialis muscle at 270 degrees .s(-1) for the FVT group. The results indicated that 3 sessions of slow velocity (30 degrees .s(-1)) isokinetic training resulted in an increase in PT at slow and fast velocities (30 and 270 degrees .s(-1)), whereas training at the fast velocity (270 degrees .s(-1)) increased PT only at 270 degrees .s(-1). The lack of consistent increases in EMG amplitude or MPF suggested that the training-induced increases in leg extension PT were not caused by increased activation of the superficial muscles of the quadriceps femoris. The important implication for coaches, trainers, and physical therapists is that significant muscular performance gains may be achieved even after very short training periods, but determination of the specific physiological adaptation(s) underlying these performance gains requires further investigation.     

Vesicular trafficking, cytoskeleton and signalling in root hairs and pollen tubes

Root hairs and pollen tubes show strictly polar cell expansion called tip growth. Recent studies of tip growth in root hairs and pollen tubes have revealed that small GTPases of the Rab, Arf and Rho/Rac families, along with their regulatory proteins, are essential for spatio-temporal regulation of vesicular trafficking, cytoskeleton organization and signalling. ROP/RAC GTPases are involved in a multiplicity of functions including the regulation of cytoskeleton organization, calcium signalling and endocytosis in pollen tubes and root hairs. One of the most exciting recent discoveries is the preferential localization of vesicles of the trans-Golgi network (TGN), defined by specific RAB GTPases, in the apical "clear zone" and the definition of TGN as a bona fide organelle involved in both polarized secretion and endocytosis. The TGN is thought to serve the function of an early endosome in plants because it is involved in early endocytosis and rapid vesicular recycling of the plasma membrane in root epidermal cells.     

Direct coupling of expanded bed adsorption with a downstream purification step

In the course of developing a cost-effective, scaleable process for the purification of a recombinant protein from Chinese hamster ovary (CHO) suspension cell culture, we investigated direct capture of this molecule using expanded bed adsorption (EBA). EBA combines clarification, purification, and concentration of the product into a single step. The unclarified bioreactor material was directly applied to a STREAMLINE 25 column containing an affinity STREAMLINE adsorbent. This work focused on simplifying the EBA operations and minimizing the overall processing time by running the EBA column unidirectionally, eluting in the expanded bed mode, and coupling the EBA column directly with ion exchange or hydrophobic interaction chromatography. Unidirectional EBA was clearly a simpler unit operation and did not require the use of specialized equipment. The increase in the elution pool volume was insignificant, especially when the EBA column was eluted directly onto the downstream column. Scale-down was simple and could be automated. Coupling of unidirectional EBA with a downstream purification step reduced processing time, equipment requirements and cost.     

Distinct phosphatidylinositol 3-kinase lipid products accumulate upon oxidative and osmotic stress and lead to different cellular responses

Signaling by phosphatidylinositol (PI) 3-kinases is mediated by 3-phosphoinositides, which bind to Pleckstrin homology (PH) domains that are present in a wide spectrum of proteins. PH domains can be classified into three groups based on their different lipid binding specificities. Distinct 3-phosphoinositides can accumulate upon PI 3-kinase activation in cells in response to different stimuli and mediate specific cellular responses. In Swiss 3T3 mouse fibroblasts, oxidative stress induced by 1 mM H(2)O(2) caused almost exclusive accumulation of phosphatidylinositol 3,4-bisphosphate (PtdIns(3, 4)P(2)), whereas osmotic stress increased both phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) and PtdIns(3,4)P(2) levels. The increase in PtdIns(3,4)P(2) levels, caused by oxidative stress, correlated with the activation of protein kinase B, which has a promiscuous PH domain that binds both PtdIns(3,4,5)P(3) and PtdIns(3, 4)P(2). p70 S6 kinase, another signaling component downstream of PI 3-kinase, however, was not activated by this oxidative stress-induced increase in PtdIns(3,4)P(2) levels. Increased PtdIns(3,4,5)P(3) and PtdIns(3,4)P(2) levels in response to osmotic stress did not correlate with protein kinase B activation, because of concomitant activation of an inhibitory pathway, but p70 S6 kinase was activated by osmotic stress. These results demonstrate that PtdIns(3,4)P(2) can accumulate independently of PtdIns(3,4, 5)P(3) and exerts a pattern of cellular responses that is distinct from that induced by accumulation of PtdIns(3,4,5)P(3).