Thiago Lemos

The plasma membrane Ca2+ pump from proximal kidney tubules is exclusively localized and active in caveolae

Plasma membrane Ca2+‐ATPase is involved in the fine‐tuned regulation of intracellular Ca2+. In this study, the presence of Ca2+‐ATPase in caveolae from kidney basolateral membranes was investigated. With the use of a discontinuous sucrose gradient, we show that Ca2+‐ATPase is exclusively located and fully active in caveolin‐containing microdomains. Treatment with methyl‐β‐cyclodextrin – a cholesterol chelator – leads to a spreading of both caveolin and completely inactive Ca2+‐ATPase toward high‐density fractions. These data support the view that Ca2+ fluxes mediated by Ca2+‐ATPase in kidney epithelial cells occur only in caveolae, being strictly dependent on the integrity of these microdomains.

Ca2+/Calmodulin-dependent Protein Kinase II Is an Essential Mediator in the Coordinated Regulation of Electrocyte Ca2+-ATPase by Calmodulin and Protein Kinase A

The aim of this study was to investigate (a) whether Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) participates in the regulation of plasma membrane Ca2+-ATPase and (b) its possible cross-talk with other kinase-mediated modulatory pathways of the pump. Using isolated innervated membranes of the electrocytes from Electrophorus electricus L., we found that stimulation of endogenous protein kinase A (PKA) strongly phosphorylated membrane-bound CaM kinase II with simultaneous substantial activation of the Ca2+ pump (≈2-fold). The addition of cAMP (5-50 pm), forskolin (10 nm), or cholera toxin (10 or 100 nm) stimulated both CaM kinase II phosphorylation and Ca2+-ATPase activity, whereas these activation processes were cancelled by an inhibitor of the PKA α-catalytic subunit. When CaM kinase II was blocked by its specific inhibitor KN-93, the Ca2+-ATPase activity decreased to the levels measured in the absence of calmodulin; the unusually high Ca2+ affinity dropped 2-fold; and the PKA-mediated stimulation of Ca2+-ATPase was no longer seen. Hydroxylamine-resistant phosphorylation of the Ca2+-ATPase strongly increased when the PKA pathway was activated, and this phosphorylation was suppressed by inhibition of CaM kinase II. We conclude that CaM kinase II is an intermediate in a complex regulatory network of the electrocyte Ca2+ pump, which also involves calmodulin and PKA.

Motor imagery modulation of postural sway is accompanied by changes in the EMG–COP association

Motor imagery (MI) performed in an upright stance promotes increases in postural sway without changes in usual amplitude measures of calf muscle EMG. However, postural muscle activity can also be determined from the temporal association between EMG and center of pressure (COP) displacements. In this study we investigated whether the MI modulation of postural sway is accompanied by changes in EMG-COP association. Surface EMG from the lateral gastrocnemius (LG) muscle and COP coordinates were collected from 12 subjects while they imagined themselves performing a rising on tiptoes movement via kinesthetic or visual imagery. As a control condition subjects were requested to imagine singing a song. The standard deviation of the forward-backward COP sway and the coefficient of variation of the EMG were calculated and compared across tasks. The degree of association between COP sways and LG activity was evaluated through a cross-correlation function. Kinesthetic imagery promoted a larger COP displacement than both visual and control imagery (p<0.02). No difference in EMG amplitude was observed across imagery tasks (p=0.08). Crucially, we found a stronger EMG-COP association during kinesthetic imagery compared to control imagery (p=0.02), whereas the EMG-COP association in visual imagery was not different from that observed during kinesthetic or control imagery (p>0.19). In conclusion, kinesthetic imagery resulted in a higher EMG-COP temporal association. Subliminal fringe mechanisms may account for the imagery effects on muscle activity and postural sway during upright stance.

Modulation of tibialis anterior muscle activity changes with upright stance width

When individuals stand with their feet apart, activation of tibialis anterior (TA) muscle seems to slightly exceed rest levels. In narrow stances, conversely, the stabilization of body lateral sways may impose marked, active demand on ankle inversors/eversors. In this study we investigate how much the modulation in TA activity, associated to center of pressure (COP) lateral sways, changes when stance width reduces. Surface EMG and COP coordinates were collected from 17 subjects at three different stances: feet apart, feet together and tandem. Pearson correlation analysis was applied to check whether the expected greater modulations in TA activity corresponded to a stronger association between fluctuations in EMG amplitude and COP lateral sways. When standing at progressively narrower stances participants showed larger fluctuations in COP lateral sways (p<0.01) and higher EMG-COP association (p<0.01); marked increases in TA activity were only observed in tandem stance (p<0.001). Interestingly, more pronounced modulations in TA activity were observed for subjects showing greater association between EMG amplitude and COP sways in feet together and tandem stance (Pearson R>0.56, p<0.02), though not when standing with feet apart (R=-0.22, p=0.40). These results indicate that the contribution of TA activity to lateral sway control increases for narrower stances.

An alternative approach in muscle fatigue evaluation from the surface EMG signal

The aim of this study was to compare the intervals of time between adjacent zero crossings (ZCI), an alternative frequency-temporal parameter, with the root-mean-square (RMS) value and the median frequency (Fmed) from the surface EMG (sEMG) signal in muscle fatigue analysis. Twenty right-handed volunteers performed isometric contractions of right biceps brachii muscle while sEMG signals were collected from it at three different and arbitrary load levels until fatigue. The mean ZCI presented a significant correlation with Fmed but not with RMS value and it also presented lower coefficients of variation than others. The results pointed that mean ZCI properties can contribute more than Fmed and RMS value on the interpretation of the muscle function under fatigue conditions.

Is heart rate variability affected by distinct motor imagery strategies

Although some studies have reported significant changes in autonomic responses according to the perspective-taking during motor imagery [first person perspective (1P) and third person perspective (3P)], investigations on how the strategies adopted to mentally simulate a given movement affect the heart rate variability (HRV) seem so far unexplored. Twenty healthy subjects mentally simulated the movement of middle-finger extension in 1P and 3P, while electrocardiogram was recorded. After each task, the level of easiness was self-reported. Motor imagery ability was also assessed through the revised version of Movement Imagery Questionnaire (MIQ-R) and a mental chronometry index. The traditional measures of HRV in the time- and frequency-domain were compared between 1P and 3P tasks by using Students t-test for dependent samples. The MIQ-R results showed that subjects had the same facility to imagine movements in 1P or 3P. The mental chronometry index revealed a similar temporal course only between 1P and execution, while the 3P strategy had a shorter duration. Additionally, the subjective report was similar between the experimental tasks. Regarding the HRV measures, the low frequency component, in log-transformed unit, was significantly higher (p=0.017) in 1P than 3P, suggesting a higher activity of the sympathetic system during 1P. This log-transformed HRV parameter seems to be more sensitive than normalized values for the assessment of the motor imagery ability, together with questionnaires, scales and mental chronometry.

Cerebral Dynamics during the Observation of Point-Light Displays Depicting Postural Adjustments

Objective: As highly social creatures, human beings rely part of their skills of identifying, interpreting, and predicting the actions of others on the ability of perceiving biological motion. In the present study, we aim to investigate the electroencephalographic (EEG) cerebral dynamics involved in the coding of postural control and examine whether upright stance would be codified through the activation of the temporal-parietal cortical network classically enrolled in the coding of biological motion. Design: We registered the EEG activity of 12 volunteers while they passively watched point light displays (PLD) depicting quiet stable (QB) and an unstable (UB) postural situations and their respective scrambled controls (QS and US). In a pretest, 13 volunteers evaluated the level of stability of our two biological stimuli through a stability scale. Results: Contrasting QB vs. QS revealed a typical ERP difference in the right temporal-parietal region at an early 200–300 ms time window. Furthermore, when contrasting the two biological postural conditions, UB vs. QB, we found a higher positivity in the 400–600 ms time window for the UB condition in central-parietal electrodes, lateralized to the right hemisphere. Conclusions: These results suggest that PLDs depicting postural adjustments are coded in the brain as biological motion, and that their viewing recruit similar networks with those engaged in postural stability control. Additionally, higher order cognitive processes appear to be engaged in the identification of the postural instability level. Disentangling the EEG dynamics during the observation of postural adjustments could be very useful for further understanding the neural mechanisms underlying postural control.

Biphasic regulation of type II phosphatidylinositol-4 kinase by sphingosine: Cross talk between glycero- and sphingolipids in the kidney

Phosphatidylinositol-4 kinase (PI-4K) is responsible for the generation of phosphatidylinositol-4 phosphate (PtdIns(4)P), a bioactive signaling molecule involved in several biological functions. In this study, we show that sphingosine modulates the activity of the PI-4K isoform associated with the basolateral membranes (BLM) from kidney proximal tubules. Immunoblotting with an anti-α subunit PI-4K polyclonal antibody revealed the presence of two bands of 57 and 62kDa in the BLM. BLM-PI-4K activity retains noteworthy biochemical properties; it is adenosine-sensitive, not altered by wortmanin, and significantly inhibited by Ca(2+) at the μM range. Together, these observations indicate the presence of a type II PI-4K. Endogenous phosphatidylinositol (PI) alone reaches PI-4K half-maximal activity, revealing that even slight modifications in PI levels at the membrane environment promote significant variations in BLM-associated-PI-4K activity. ATP-dependence assays suggested that the Mg.ATP(2-) complex is the true substrate of the enzyme and that free Mg(2+) is an essential cofactor. Another observation indicated that higher concentrations of free ATP are inhibitory. BLM-associated-PI-4K activity was ~3-fold stimulated in the presence of increasing concentration of sphingosine, while in concentrations higher than 0.4mM, in which S1P is pronouncedly formed, there was an inhibitory effect on PtdIns(4)P formation. We propose that a tightly coupled regulatory network involving phosphoinositides and sphingolipids participate in the regulation of key physiological processes in renal BLM carried out by PI-4K.

Reference Values for Human Posture Measurements Based on Computerized Photogrammetry: A Systematic Review

Objective: The main objective of this study was to review the literature to identify reference values for angles and distances of body segments related to upright posture in healthy adult women with the Postural Assessment Software (PAS/SAPO). Methods: Electronic databases (BVS, PubMed, SciELO and Scopus) were assessed using the following descriptors: evaluation, posture, photogrammetry, physical therapy, postural alignment, postural assessment, and physiotherapy. Studies that performed postural evaluation in healthy adult women with PAS/SAPO and were published in English, Portuguese and Spanish, between the years 2005 and 2014 were included. Results: Four studies met the inclusion criteria. Data from the included studies were grouped to establish the statistical descriptors (mean, variance, and standard deviation) of the body angles and distances. A total of 29 variables were assessed (10 in the anterior views, 16 in the lateral right and left views, and 3 in the posterior views), and its respective mean and standard deviation were calculated. Reference values for the anterior and posterior views showed no symmetry between the right and left sides of the body in the frontal plane. There were also small differences in the calculated reference values for the lateral view. Conclusion: The proposed reference values for quantitative evaluation of the upright posture in healthy adult women estimated in the present study using PAS/SAPO could guide future studies and help clinical practice.

Balance Impairments after Brachial Plexus Injury as Assessed through Clinical and Posturographic Evaluation

Objective: To investigate whether a sensorimotor deficit of the upper limb following a brachial plexus injury (BPI) affects the upright balance. Design: Eleven patients with a unilateral BPI and 11 healthy subjects were recruited. The balance assessment included the Berg Balance Scale (BBS), the number of feet touches on the ground while performing a 60 s single-leg stance and posturographic assessment (eyes open and feet placed hip-width apart during a single 60 s trial). The body weight distribution (BWD) between the legs was estimated from the center of pressure (COP) lateral position. The COP variability was quantified in the anterior-posterior and lateral directions. Results: BPI patients presented lower BBS scores (p = 0.048) and a higher frequency of feet touches during the single-leg stance (p = 0.042) compared with those of the healthy subjects. An asymmetric BWD toward the side opposite the affected arm was shown by 73% of BPI patients. Finally, higher COP variability was observed in BPI patients compared with healthy subjects for anterior-posterior (p = 0.020), but not for lateral direction (p = 0.818). Conclusions: This study demonstrates that upper limb sensorimotor deficits following BPI affect body balance, serving as a warning for the clinical community about the need to prevent and treat the secondary outcomes of this condition.